Volume 3, Number 1A, 2013 PDF

Journal of
Agricultural Science
and Technology A
Volume 3, Number 1, January 2013 (Serial Number 21)
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Journal of
Agricultural Science
and Technology A
Volume 3, Number 1, January 2013 (Serial Number 21)
Contents
Review
1
Biosystems Engineering Curricula in Europe

Pierluigi Febo and Antonio Comparetti
Research Papers
10
Effect of Biofertilizer on Biomass Productivity, Nutrient Balance and Soil Fertility in Rainfed
Organic Ginger Production System
Nongmaithem Jyotsna, Mainak Ghosh, Dulal Chandra Ghosh, Wahengbam Ingo Meitei and Jagadish
Timsina
20
Organic Agriculture: Socioeconomic Sustainability of Brazilian Coffee

Rubia Wegner, Patrcia Helena Nogueira Turco and Flvia Maria de Mello Bliska
33
Enhancing Maize Grain Yield in Acid Soils of Western Kenya Using Aluminium Tolerant
Germplasm
Ouma Evans, Ligeyo Dickson, Matonyei Thomas, Agalo Joyce, Were Beatrice, Too Emily, Onkware
Augustino, Gudu Samuel, Kisinyo Peter and Philip Nyangweso
47
Response of Peach (Prunus persica) cv. to Foliar Application of Potassium and Copper
Shawkat Mustafa Mohammed Al-Atrushy and Sarfaraz Fatah Ali Al-bamarny
53
Carboxylesterase and Glutathione-S-Transferase (GSTs) Induced Resistance to Bacillus

thuringiensis Toxin Cry1Ab in Rice Leaf Folder, Cnaphalocrocis medinalis (Guenee) Populations
Veegala Ramesh Babu, Vemuri Shashi Bhushan, Chintalapati Padmavathy, Muthugonder Mohan, Sena
Mahendran. Balachandran and Bellamkonda Ramesh
60
Environmental Impacts of Feeding High-fiber Diet to Pigs

Abraham Woldeghebriel, Shanequa Smith, Teo Barios, Brad Pope and Sebhatu Gebrelul
66
Effects of Feeding Siamese Neem Leaves and Zanthoxylum Pods, on Dry Matter Intake, Dry
Matter Digestibility, Milk Production and Composition in Thai Holstein Dairy Cows, Fed Rice
Straw as Fiber Source
Penjor, Virote Pattarajinda, Suporn Katawatin, Chaiyapas Thamrongyoswittayakul and Wandee
Gritsanapan
72
Feeding Effect of Triticale Fodder as Replacement of Straw on Production Performance of Dairy

Cows
Nathu Ram Sarker, Mohammad Asaduzzaman, Khan Shahidul Huque, Mohammad Toyebur Rahman,
Nazrul Islam, Mohammad Enamul Haque and Stephen R. Waadington
Journal of Agricultural Science and Technology A 3 (2013) 1-9

Earlier title: Journal of Agricultural Science and Technology, ISSN 1939-1250
DAVID
PUBLISHING

Pierluigi Febo and Antonio Comparetti
Dipartimento dei Sistemi Agro-Ambientali, Universit di Palermo, Palermo 90128, Italy
Received: September 26, 2012 / Published: January 20, 2013.

Abstract: This paper describes the history of the harmonisation of agricultural/biosystems engineering degree study programs in
Europe from 1989, when the need for this process was widely felt, until now, when this need was partly satisfied through the
implementation of the projects of two EU funded thematic networks, i.e., USAEE-TN and ERABEE-TN. The objective of this paper
is to contribute to promote, in each EU country and elsewhere, the process of harmonisation of agricultural/biosystems engineering
degree study programs, and student and graduate mobility within the EU, as well as between the EU and the USA. At present, in
Europe, this harmonisation process is aided by the key results of the projects of USAEE-TN, ERABEE-TN and POMSEBES.
USAEE developed some core curricula, to be used as benchmarks for European agricultural/biosystems engineering degree study
programs, and a web-based database of these study programs. ERABEE promoted the transition from agricultural engineering to
biosystems engineering and established the recognition procedures of new European study programs in biosystems engineering. The
EU-US POMSEBES consortium built up a platform for exchange of experiences and ideas between the USA and the EU, aimed at:
enhancing the quality and linkage of research and education; establishing appropriate policy oriented measures; promoting
compatible degree study programs in biosystems engineering, within the EU as well as between the EU and the USA.
Key words: Harmonisation, degree study programs, agricultural/biosystems engineering, student and graduate mobility.
1. Agricultural Engineering Curricula: from

1989 to USAEE-TN
1.1 The Need for the Harmonisation of Agricultural
Engineering Curricula
At the end of 1989, the International Commission
of Agricultural Engineering (CIGR), under the
chairmanship of Prof. Giuseppe Pellizzi, with the
cooperation of the former Italian Association of
Agricultural Engineering (AIGR, now AIIA) and the
University of Milan, sponsored a project designed to
compare university curricula in Agricultural
Engineering in the 12 countries of the former
European Community (EC). The aim of this project
was to facilitate the creation of academic
harmonisation, in view of the Unique Market,
beginning in 1993, and to facilitate the free exchange
of university graduates in agriculture and agricultural
Corresponding author: Antonio Comparetti, researcher,

research
field:
agricultural
machines.
antonio.comparetti@unipa.it.
engineering throughout the EC.

As a consequence, two study seminars, attended by
representatives of EC countries, were held at
Gargnano (Italy), in May 1991, and Silsoe (UK), in
May 1992; several papers were presented at
conferences or published in Italian or international
journals [1-6]; two working groups were created,
called WG 1, within CIGR, and SIG 12 Education
and Communication, within the European
Association of Agricultural Engineers (EurAgEng).
From 1990 to 1994, surveys were carried out in
several countries, in order to investigate the
organisation of the university degree study programs
or specialisations, with special regard to agricultural
engineering [7-10].
From 1996 to 1999, the surveys were updated and
extended to other countries and universities [11, 12].
In June 2000, the results of the last survey of the
study programs in agricultural engineering were
presented in the report The University Structure and
Curricula on Agricultural Engineering. An overview

of 36 Countries, published by the Food and
Agriculture Organization of the United Nations (FAO);
this work was presented in July 2000, during the
EurAgEng Conference, held in Warwick (UK) [13].
1.2 Towards a European Standard for Agricultural
Engineering Curricula
In 2001, the working package 3 of the EU Socrates
thematic network for agriculture, forestry, aquaculture
and the environment (AFANet) issued the report
Towards a European Standard for Agricultural
Engineering Curricula, which outlines the basis for
four European core curricula:
1) 5-year diploma degree, more scientifically
oriented;
2)
3-year
Bachelor
degree,
more
application-oriented;
3) Masters degree, corresponding to the long term
5-year diploma degree;
4) Masters degree, corresponding to the short term
3-year Bachelor degree.
In this report, an accreditation system is also
proposed and the European university faculties and
departments offering diplomas in agricultural
engineering, agriculture with specialisations in
agricultural engineering and engineering (agricultural
engineering courses) are listed [14].
In Europe, the beginning of the third millennium
brought a crisis in the agricultural engineering sector.
Several institutes, departments and research centres of
agricultural engineering were closed or had their staff
significantly reduced.
On the other side of Atlantic Ocean, a process of
transition from agricultural engineering into
biosystems engineering, started in the 1960s (when it
was recognised that both biomedical engineering and
agricultural engineering could perhaps be included as
disciplines of a broader biological engineering), was
in progress. Since then, several agricultural
engineering departments had modified their degree
study programs towards biological engineering

discipline and included the term Biological
Engineering (or similar) in the department and/or
curriculum names. This process finished in 2005,
when the American Society of Agricultural Engineers
(ASAE), after changing its name in 1993 to the
Society for the Engineering of Agriculture, Food and
Biological Systems, changed it again to the American
Society of Agricultural and Biological Engineers
(ASABE).
Meanwhile in Europe, following the above process,
with the advent of new topics (e.g., precision
agriculture, robotics, information systems for
agriculture), several departments of agricultural
engineering changed their name, course contents and
research topics towards applied biology. This trend
also led to the progressive substitution of the name
Agricultural Engineering with Bio-Engineering or
Biosystems Engineering. In order to take into
account this trend, in 2002, EurAgEng changed the
name of its official journal from Journal of
Agricultural Engineering Research to Biosystems
Engineering.
In agricultural engineering study programs, the
learning outcomes (knowledge, competencies and
skills) of many courses are integrated with each other,
so that graduates can:
develop new technologies and materials, in order
to improve the quality and reliability of agricultural
products;
plan field operations with high energy efficiency;
control electronically agricultural production;
design environmental structures and systems;
develop efficient technologies for processing
agricultural products.
In several European countries (e.g., Italy), the
university study programs in agricultural engineering
show a high variability of curriculum structure and
course contents. Often, they are a specialisation of a
degree in agricultural sciences rather than a specific
study program in agricultural engineering, so that the
engineering learning outcomes are limited and,

therefore, the graduates are agronomists rather than
agricultural engineers.
The third millennium also brought the new structure
of the study programs (3 + 2) (1st cycle or 3-year
Bachelor, 2nd cycle or 2-year Master), according to
the Bologna Declaration of 1999. Several EU
countries felt the need to update all the university
study programs, including those in agricultural
engineering, in order to satisfy the general economic
conditions, the scientific and technological
development and the need to develop competitive
study programs.
1.3 A European Thematic Network on Agricultural
Engineering
From the above background, the need to establish a
European thematic network on agricultural
engineering was felt, as it is described in the report of
AFANet-Working Package 3.
Therefore, the idea of carrying out, at European
level, a project aimed at developing basic core
curricula, to be used as benchmarks for local
development of agricultural engineering and for
training future agricultural engineers, was born.
In 2002 the thematic network University Studies of
Agricultural Engineering in Europe (USAEE),
comprising 31 institutions from 27 European countries,
was established with the aim of developing this
project, approved and supported by EurAgEng
through the SIG RD12, and funded by the
Socrates-Erasmus EU programme.
The main objectives of USAEE-TN were to:
define and develop core curricula of 1st and 2nd
cycles, to be used as benchmarks for agricultural
engineering studies in Europe;
determine a set of minimum criteria/requirements,
against which any curriculum can be tested, in order to
decide whether it meets these criteria/requisites and,
therefore, can be recognised as a program in
agricultural engineering;
define common accreditation procedures, also in

terms of European Credit Transfer System (ECTS)
credits, and establish the bodies/committees for
carrying out these procedures.
In order to be recognised, a core curriculum must
meet both the criteria of the European Federation of
the National Associations of Engineers (FEANI) for
an engineering study program, concerning the basic
engineering course contents and the related ECTS, and
the criteria of EurAgEng, concerning the agricultural
and biological course contents and the related ECTS
[15].
In 2005, the USAEE-TN produced the draft report
Core
Curricula
of
Agricultural/Biosystems
Engineering for the First Cycle Pivot Point Degrees of
the Integrated M.Sc. or Long Cycle Academic
Orientation Programs of Studies.
In this report, it is recognised that the weak area of
agricultural engineering studies in Europe is the
inadequate
engineering
foundation
of
the
corresponding curricula. Therefore, the main
challenges are to:
enhance the engineering part of the European
core curricula, so that they meet the FEANI criteria
for engineering study programs;
significantly reduce the agricultural and/or
biological sciences part of the core curricula.
In several European countries, intermediate 3-year
degree study programs, named pivot point, were
established, in order to facilitate the exchange of
students between universities and countries. These
study programs are different from those relevant for
the job market, defined by Bologna Declaration.
The degree study programs should be adapted to the
Bologna Declaration Scheme and coexist with the new
intermediate 3-year pivot point degree study
programs (Bachelors Science), according to the
history, industrial and social conditions, and the
traditions of each country.
The main challenge is to agree on a set of minimum
standards for core curricula and to clearly describe the
criteria to be used for professional and academic

applications. In this respect the work carried out by
the Thematic Network Enhancing Engineering
Education in Europe (E4), run by the University of
Florence in cooperation with the European Society for
Engineering Education (SEFI) and other organisations,
is relevant. Therefore, from the outset the USAEE-TN
established strong collaboration with both SEFI and
E4 TN towards common objectives.
In the first step of the development of agricultural
engineering core curricula, the 1st study cycle was
examined and two different schemes were defined and
focused in a draft report.
Scheme A, with academic orientation, consists of:
core curricula of integrated 5-year degree study
programs (M.Sc.);
core curricula of pivot-point 1st cycle 3-year
degree study programs (B.Sc.).
Scheme B, with application-technological orientation,
is represented by the core curricula of professional 1st
cycle (mostly 3-year) degree study programs.
This report contains not only the core curricula (Fig.
1), but also seven modules or specialisations in
agricultural engineering:
Water Resources Engineering;
Mechanical Systems and Mechanisms used in
Agricultural and Bioprocess Engineering (Tables 1

and 2);
Structural Systems and Materials in Agricultural
and Bioprocess Engineering;
Waste Management in Agricultural and
Bioprocess Engineering;
Bioprocessing;
Energy Supply and Management in Agricultural
and Bioprocess Engineering;
Information Technology and Automation in
Agricultural and Bioprocess Engineering.
In September 2004, this draft report was distributed
to the Executive Committees of FEANI and
EurAgEng for evaluation and comment. At the end of
2005, the European Monitoring Committee (EMC) of
FEANI assessed this draft and, then, requested some
modifications:
to explain the contents of general within basic
sciences;
to specify the number of ECTS of mathematics
(which must be at least 24);
to specify the percentage of engineering basic
sciences (which must be at least 20% and 36 ECTS)
of the study program;
to provide all the seven specialisations with at
least 60% of engineering subjects;
Courses not included

in the core curricula:
needed to achieve the
University strategic objectives
13-17 %
Specialisation courses
Engineering
Basic courses
22-28%
Engineering
31-44 %
Agricultural/Biological Sciences 9-11 %
Agricultural/Biological
Sciences
11-14%
General courses (adopted from E4-TN)

+ optional courses
Fig. 1 Agricultural engineering core curricula of 1st cycle pivot point integrated degree study programs (3 + 2) and
5-year degree study programs with academic orientation.
Table 1 Proposed module or specialisation in Mechanical Systems and Mechanisms used in Agricultural and Bioprocess
Engineering within the 1st cycle pivot point degree study programs.
Engineering part of the core curricula :
optional courses
24-30 ECTS credits: equal to 13-17% of total 180 ECTS credits
Assuming 6 course units with 4 or 5 ECTS credits per unit,
respectively, or equivalent, the learning outcomes that follow
may be delivered through the following structured coursework.
1. Kinematics of Mechanisms
2. Power Generation Engines
3. Mechatronics
4. Soil Mechanics
5. Electrotechnics
6. Electronic Circuits
7. Instrumentation and Measurements
8. Engineering Surveying - GIS
Learning outcomes and contents follow this table;
Source: USAEE-TN.
Table 2 Indicative list of agricultural engineering courses
included in the proposed module or specialisation in
Mechanical Systems and Mechanisms used in Agricultural
and Bioprocess Engineering.
1. Agricultural Machinery Design
2. Farm Power Units
3. Farmstead Equipment
4. Analysis and Design of Biomachinery
5. Techniques in Precision Agriculture
6. Automatic Controls
7. Computer Control of Machines and Processes
8. Ergonomics, Health and Safety
9. Design Methods for Machines for Biosystems
10. Remote Sensing
11. Soil Erosion
12. Landscape Planning
13. Free Technical or Agricultural/Biological Electives
Agricultural/Biological Sciences part of the core curricula :

optional courses
16-20 ECTS credits: equal to 9-11% of total 180 ECTS credits
Assuming 4 course units with 4 or 5 ECTS credits per unit,
respectively, or equivalent, the learning outcomes that follow
may be delivered through the following structured coursework.
1. Crop Science and Management
2. Crop Protection
3. Agro-chemicals
4. Animal Science and Management
5. Environmental Impact Assessment
promoted, in August 2005, the dissemination proposal

submitted to the DG for Education and Culture of the
EU was selected, so that on the October 1, 2005 the
4th and last year of the USAEE-TN project, mainly
aimed at the dissemination of its results, started.
Therefore, the duration of USAEE-TN project was
four years (01/10/2002-30/09/2006).
The main outputs achieved during the USAEE-TN
dissemination year were to:
develop a web-based database, containing the
courses or modules (set of courses) of the study

programs, including the course ECTS, in order to
facilitate the recognition of the core curricula and,
therefore, promote student mobility throughout the EU
Source: USAEE-TN.
(http://sunfire.aua.gr:8080/ects/Welcome.do);
to specify the percentage of non-technical subjects

(which must be at least 10% of the study program).
The EMC of FEANI also implicitly required an
accreditation process for agricultural/biosystems
engineering curricula, as well as an overview of
academic and professional qualification of the
teaching staff and laboratory facilities.
EurAgEng agreed to undertake the task of
establishing the recognition process of the core
curricula.
Since it was necessary for the results of the
USAEE-TN project to be widely disseminated and
database will be continuously updated and made
this
available using a specific authorisation;
enhance the USAEE web-site, by creating links
with organisations, other related thematic networks

and
projects
(Interuniversity
Conference
of
Agriculture and Related Sciences-ICA, Training &

Resources in Early Education-TREE, Archipelagos
and TUNING, etc.);
disseminate and promote the USAEE-TN results
to the wider area of higher engineering education in
Europe,
through
synergic
activities
with
TECHNO/Archipelagos on issues concerning ECTS,
quality assessment and employability;

disseminate and promote the USAEE-TN results
to the wider international agricultural/biosystems
engineering community outside Europe, through the
TUNING III web-site and synergic activities with
TUNING III;
disseminate the USAEE-TN results, through
contacts with student associations (in order to promote
student mobility and participation), alumni
associations (in order to promote alumni involvement),
deans, rectors, Erasmus officials, policy makers and
other academic bodies, representatives of enterprises,
companies and industries involved in agricultural
engineering;
cooperate with FEANI and, through FEANI itself,
with the major professional stakeholders in Europe,
aimed at the final approval of the core curricula
developed by USAEE-TN;
cooperate with the Accreditation of European
Engineering Programmes and Graduates (EUR-ACE)
towards a common accreditation system for the higher
engineering education in Europe, according to the
current developments of Bologna process;
organise dissemination events, at national level, to
which the national professional societies of agricultural
engineers, also representing strong national contact and
dissemination points towards the students and the
industrial and the broader non-academic sector of
agricultural engineering, will participate;
develop synergic activities (together with ICA,
SEFI, TREE, etc.), for promoting the USAEE-TN
results to academic and non-academic communities
and targeted groups, including industries and
professional societies;
support short-term student mobility, through the
participation
of
student
associations
(e.g.,
International Association of Students in Agricultural
and related sciences-IAAS, Board of European
Students of Technology-BEST) to workshops, with
contributions to presentations and proceedings, and to
the dissemination activities addressed to them, the
European
market
and
the
students
of
agricultural/biosystems engineering university studies
in Europe (in synergy with the International Relations
Officers Network of the Association of European
Life Science Universities-IROICA) [16].
2. From Agricultural Engineering to

Biosystems
Engineering
Curricula:
ERABEE-TN
In November 2005, the same partners of
USAEE-TN, and others, proposed a new thematic
network, aimed at using and developing the results
achieved through the previous project.
Thus, in 2007 the thematic network Education and
Research in Biosystems Engineering in Europe
(ERABEE), comprising 35 institutions from 27
Erasmus countries, of which 33 were higher education
area institutions and two were student associations,
was established with the aim of developing this
project, co-funded by EU, under the umbrella of the
Lifelong
Learning
Programme-LLP
(http://www.erabee.aua.gr).
The objectives of ERABEE-TN project were to:
promote the transition from agricultural
engineering to biosystems engineering;
establish the recognition procedures of the new
European study programs in biosystems engineering
by FEANI and EurAgEng, based on the core curricula
of the first two cycles developed by USAEE-TN;
enhance the compatibility between the new
European study programs in biosystems engineering,
in order to promote their recognition and accreditation,
in synergy with EUR-ACE and in support of the
establishment of European Quality Labels in
bio-engineering;
organise case studies of the implementation of
new European study programs in biosystems
engineering, based on the core curricula of the first
two cycles developed by USAEE-TN, aimed at
recognition by FEANI and EurAgEng;
map and promote the 3rd cycle University
study programs and the European doctorate in

biosystems engineering, following the recognition by
FEANI and EurAgEng, through the implementation of
the core curricula of the first two cycles developed by
USAEE-TN;
develop synergies for strengthening the link
between research and education in all three cycles of
the university studies (above all in the 3rd cycle) in
biosystems engineering in Europe;
promote the adoption of European Standards on
quality assessment and assurance of European study
programs in biosystems engineering, in accordance
with the emerging European Qualifications
Framework (EQF);
enhance the attractiveness of European study
programs in biosystems engineering, both within and
outside Europe;
promote the mobility of researchers and students
within the EU;
implement the main lines of TUNING, based on
the outcomes of USAEE-TN.
The beneficiary target groups of ERABEE-TN
project are:
universities offering biosystems engineering
graduate and postgraduate studies;
graduates in biosystems engineering, professional
societies, companies and enterprises involved in
agricultural production and processing, the industry
and market in the field of the technical support of
agriculture, etc.;
European and national accreditation bodies,
policy-makers, ministries of education and the
academic society of higher education in Europe;
students, scholars and researchers from regions
outside Europe;
other related disciplines, associations and
thematic networks in synergy with ERABEE-TN
itself.
The main achievements of the ERABEE project, in
each partner country, were to:
define the emerging biosystems engineering

discipline in Europe by describing the situation;
describe the current situation and perspectives of
the development of biosystems engineering study
programs towards the areas of bio-fuels, bio-materials
and quality of products;
describe the current schemes and the possible
structured study programs of the 3rd cycle university
studies in agricultural engineering and in the emerging
discipline of biosystems engineering;
describe the research activities in the first two
cycles of biosystems engineering university studies;
describe the infrastructures for the quality
assessment and accreditation of biosystems
engineering university studies;
describe the tools for enhancing the attractiveness
of European study programs in biosystems
engineering [17-22].
The duration of ERABEE-TN project was three
years (01/10/2007-30/09/2010).
3. A Case of EU-US Cooperation in

Biosystems
Engineering
Curricula:
POMSEBES
During the ERABEE-TN project, from 01/11/2006
to 31/10/2008, the consortium Policy Oriented
Measures in Support of the Evolving Biosystems
Engineering Studies in USA-EU (POMSEBES) was
established (http://www.pomsebes.aua.gr).
The project of this consortium, comprising 12
higher education area institutions, of which eight are
from the EU and four from the USA, was funded by
the European Commission, jointly with the US
Department of Education, Fund for the Improvement
of Post Secondary Education (FIPSE), under the
programme Actions for Transatlantic Links and
Academic Networks for Training and Integrated
Studies (ATLANTIS), in the framework of the
2006-2013 EU-US Agreement in higher education and
vocational training.
The objectives of the POMSEBES project, which

were mostly achieved, were to:
provide a platform for a systematic exchange of
experiences and ideas between the USA and the EU,
in order to contribute to the enhancement of the
quality and linkage of research and education and to
establish appropriate policy oriented measures, i.e.,
the development of biosystems engineering study
programs
including
strong
basic
engineering
courses/topics and disseminating these courses into

other study programs in (applied) biological sciences,
in order to open engineering concepts to the
appropriate students;
develop appropriate degree study programs in
biosystems engineering, whereas the relationship
between the quality of these curricula and the learning
outcomes and core abilities of students can be
established and encouraged by EUR-ACE in the EU
and the Accreditation Board for Engineering and
Technology (ABET) in the USA, respectively;
encourage compatible study programs, within the
EU as well as between the EU and the USA, through a
systematic comparison of curricula, aimed at a
standard definition of basic courses, clarification of
The highest achievements of ERABEE-TN, which

developed the results of USAEE-TN, were to:
promote the transition from agricultural engineering to
biosystems engineering, including the areas of
bio-fuels, bio-materials and quality of products;
establish the recognition procedures of new European
study programs in biosystems engineering by FEANI
and EurAgEng, based on the core curricula developed
by USAEE-TN; promote the mobility of researchers
and students within the EU, as a consequence of the
development of compatible study programs in
biosystems engineering and the enhancement of their
attractiveness.
Another important contribution towards the
harmonisation of the European curricula in
agricultural/biosystems engineering was achieved
through the cooperation between EU and US higher
education area institutions, during the project of
POMSEBES consortium.
However, the above process is still in progress and
will also be performed through the dissemination
activities of ERABEE-TN and future projects, which
will be submitted to the EU by the partners in this
network.
areas of application and a common definition of
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DAVID
PUBLISHING
Effect of Biofertilizer on Biomass Productivity, Nutrient

Balance and Soil Fertility in Rainfed Organic Ginger
Production System
Nongmaithem Jyotsna1, Mainak Ghosh2, Dulal Chandra Ghosh3, Wahengbam Ingo Meitei1 and Jagadish Timsina4
1. College of Agriculture, Central Agricultural University Imphal, Manipur 795001, India
2. Department of Agronomy, Bihar Agricultural University, Sabour 813210, Bihar, India
3. Institute of Agriculture, Visva-Bharati, Sriniketan 731236, West Bengal, India
4. IRRI-Bangladesh Office, Banani DOHS, Dhaka 1206, Bangladesh
Received: November 13, 2012 / Published: January 20, 2013.
Abstract: Farmers of North-Eastern India grow ginger organically and obtain low yield. Biofertilizer may help in increasing yield
and maintaining soil fertility. An investigation made with different biofertilizers showed that seed treatment with biofertilizer
increased biomass by 18.3%, enhanced N, P and K removal and improved short-term soil fertility status by increasing N and P
balance and reducing negative K balance over control plots. Use of high dose (5.0 kg ha-1) of Azotobacter (a3) and medium dose
(3.75 kg ha-1) of both Azospirillum (b2) and Phosphotica (c2) increased rhizome biomass by 6.8%-12.5% and shoot biomass by
5.6%-14.3% over other levels. They enhanced N, P and K removal by both rhizome and shoot when compared with other levels. The
above biofertilizer treatments improved organic carbon and available N and P status of the soil by increasing N and P balance. The
result showed overall strong negative K balance; but biofertilizer treatments greatly reduced the negative K balance in soil as
compared to the control plots. Seed treatment with high level of Azotobacter along with medium level of Phosphotica (a3c2)
produced the highest biomass yield (7.4 t ha-1), increased N and P balance and fertility status in spite of high N, P and K removal.
Key words: Biofertilizer, ginger productivity, nutrient balance, soil fertility.
1. Introduction
Ginger (Zingiber officinale Rosc.) is an aromatic
spice crop grown in almost all the states of the
North-Eastern region of India. The climatic condition
of the region is highly suitable for cultivation of
ginger [1]. It is the main cash crop supporting the
livelihood and improving the economic level of many
ginger growers in the region. However, it is a heavy
feeder and an exhaustive crop, and requires large
quantities of manures and fertilizers for its proper
growth and development. Inorganic fertilizers besides
being costly are causing problems to the ground water
Corresponding author: Jagadish Timsina, Ph.D., research

field: agronomy. E-mail: timsinaj@hotmail.com.
and environment as well as quality of the produce.

Majority of the farmers in the region involved in
ginger production can not afford to purchase the
fertilizers. The ginger production in the region is
organic by default because the farmers only apply the
locally available farmyard manures and do not apply
any chemical fertilizers or pesticides to ginger crop [2].
In this way, the ignorance of the farmers about the
technological advances is turning out to be a key to
prosperity because of increasing demand for organic
produce all over the world [3].
Organic farming has attracted increasing attention
among our environmental protectionists, improving
quality and reorientation of agriculture towards areas
of market demands [4]. The beneficial effects of
Effect of Biofertilizer on Biomass Productivity, Nutrient Balance and

Soil Fertility in Rainfed Organic Ginger Production System
11
organic manures are manifested through increase in

soil organic matter and humus over the period. Soil
organic matter and humus act in several ways. They
serve as slow release source of plant nutrients to the
crops, increase water holding capacity of the soil and
act as a buffer against change in soil pH [5]. However,
ginger cultivated with only organic manures, though
improved organic carbon and available N, P and K

contents in soil through greater microbial activities
[13, 14]. However, information in detail about the
effect of different biofertilizers on biomass
productivity, nutrient removal, nutrient balance and
soil health are limited particularly in ginger
production in the North-Eastern region of India.
has the potential to improve rhizome quality, is unable

to obtain high yield. In recent years, biofertilizers have
emerged as a promising source of nutrient supply in
organic agriculture [6]. Biofertilizers, being essential
components of organic farming, play vital role in
improving the soil nutrient supplying capacity and
Keeping this idea in view and considering the

importance of the problem, an effort has been made to
study the effect of different biofertilizers (Azotobacter,
Azospirillum and Phosphotica) on biomass
productivity, nutrient removal, nutrient balance and
soil fertility status in rainfed organic ginger
maintain soil fertility and sustainability by fixing

atmospheric dinitrogen (N2) and mobilizing fixed
macro- and micro-nutrients [7]. They are also
responsible for converting insoluble P in the soil into
forms available to plants and producing plant growth
hormones and vitamins by microorganisms thereby
production system in the North-Eastern region of

India.
increasing their efficiency and availability [8, 9].

Azotobacter, Azospirillum and Phosphotica are found
very important in soil nutrient supply system and
enhance atmospheric N2 fixation, mineralization and
mobilization of plant nutrients due to greater
microbial activities and thereby increasing the nutrient
removal by the crop [10]. Son et al. [11] also noticed
that application of organic and biofertilizers together
in rice-soybean-rice cropping system increased N, P,
and K uptake by the crop and maintained higher level
of organic carbon and available N, P and K of the soil
as compared to only inorganic fertilizers. The soils of
the North-Eastern region are acidic to strongly acidic
due to leaching of bases owing to high rainfall (>
2,000 mm). The low availability of P due to fixation
as Fe/Al- complex is the major problem of crop
production in the region [1]. Biofertilizers like P
solubilizing bacteria (PSB), Azotobacter and
Azospirillum may be useful for improving P and N
nutrition to the crops [11, 12]. Organic manures and
biofertilizers not only improved crop nutrition, but
also increases nutrient removal by the crop and
2. Materials and Methods

2.1 Experimental Site
The field experiment was conducted during 2007
and 2008 at the Horticulture Experimental Farm,
College of Agriculture, Central Agricultural
University, Imphal, Manipur. The place is located at
2445N latitude, 93o56E longitude with an altitude of
790 m above mean sea level. The experimental soil
was acidic in reaction (pH 5.5), clayey in texture
(15.5% sand, 21.2% silt and 61.1% clay), medium in
fertility (230, 13.3 and 267 kg ha-1 available N, P and
K, respectively), well-drained with gentle slope. The
experimental site comes under warm humid moist
region where monsoon normally starts from April and
extends up to September. The weather conditions
during the two years of study are provided in detail in
a companion paper [15]. The crop was grown on
rainfed condition. Briefly, stated, the crop received
1,341 and 1,207 mm rainfall during its growing period
in 2007 and 2008, respectively. The maximum
temperature ranged from 24.1 to 29.6 C, while the
minimum temperature ranged from 9.6 to 22.5 C
during the cropping seasons. The relative humidity of
the cropping season varied from 58.5% to 84.7% in
2007 and 58.9% to 88.5% in 2008. Both temperature
12

and relative humidity remained very conducive for

growth and rhizome productivity of ginger.
2.2 Experimental Details
The experiment was laid out in complete
randomized block design with three biofertilizers each
at three levels along with a common control (no use of
biofertilizer) in three replications in 3.6 m 3.0 m
plots. The three levels of three biofertilizers are (1)
Azotobacter at 3 levels (2.5, 3.75 and 5.0 kg ha-1); (2)
Azospirillum at 3 levels (2.5, 3.75 and 5.0 kg ha-1) and
(3) Phosphotica at 3 levels (2.5, 3.75 and 5.0 kg ha-1).
Altogether, there were 28 treatment combinations. The
treatment details for this study are presented in a
companion paper [15]. Briefly stated, culture solutions
of different biofertilizers were prepared by dissolving
10 g, 15 g and 20 g of each biofertilizer and their
combinations in 500 mL of water separately for each
biofertilizer treatment. Thus, a total of 27 biofertilizer
culture solutions were prepared. Each biofertilizer
treatment (culture solution) was mixed thoroughly
with 8 kg ginger setts (required for each treatment) of
variety Bhaisey and dried in shade before planting.
The rhizomes (20 g sett) were planted on March 16,
2007 and March 18, 2008 with a spacing of 30 cm
30 cm in 3.6 3.0 m plots. A general dose of 20 t
FYM ha-1 containing 0.50% N, 0.133% P and 0.55%
K was applied during land preparation. The crop
received hand weeding twice along with light earthing
up on May 25 and July 14 in 2007 and May 27 and
July 16 in 2008. No chemical fertilizer, pesticide and
irrigation water was applied under this investigation.
2.3 Shoot and Rhizome Biomass
Five clumps from each plot were collected at 210
days after planting (DAP) for determination of dry
matter yield (DMY) and its partitioning. The plant
samples were cleaned and washed in water to remove
surface contamination and separated into shoot (leaves
+ stem) and rhizome. Fresh weight of rhizome of each
plot was recorded and converted into g m-2. A piece of
rhizome of each plot was taken; its fresh weight was

recorded before chopping. Thereafter, shoot and
chopped rhizome were kept in separate paper packets
which in turn were placed in an oven for drying at
65-70 C till constant weight was obtained. The dry
weight of shoot (shoot biomass) and rhizome (rhizome
biomass) were recorded. The sum of shoot biomass
(SB) and rhizome biomass (RB) was the total biomass
(TB). Biomass partitioning (BP) was estimated as: BP
= RB (kg ha-1)/TB (kg ha-1) 100, and expressed in
percentage. The crop was harvested on November 12
in 2007 and November 14 in 2008 when the leaves
turned yellow and started drying up. Fifty clumps
from each plot were lifted carefully with the help of a
spade and the rhizomes were separated and kept in
shade for two days. The fresh weight of rhizome was
recorded in t ha-1.
2.4 Nutrient Removal and Soil Fertility
Plant samples (shoot and rhizome) collected at
maturity were oven dried and ground properly for N,
P and K determination in the laboratory following
micro-kjeldahl,
spectrophotometer
and
flame
photometer methods respectively as described by
Jackson [16]. N, P and K removal by shoot and
rhizome were then estimated by multiplying shoot and
rhizome dry weights with their respective N, P and K
contents. Soil samples collected from 0-15 cm depth
before start of the experiment and after harvest of 2nd
year of ginger were used for determining organic
carbon (OC) following Walkley and Black method
[17], available N by alkaline potassium permanganate
method [18], available P by Brays method [19] and
available K by ammonium acetate extraction method
[16]. The data were used for determining N, P and K
balances and fertility status of the experimental soil.
2.5 Data Analysis
All data were analyzed statistically following the
standard procedure as described by Gomez and
Gomez [20]. The data were tested for analysis of
13

variance and least significant difference (P = 0.05) to

compare the effect of biofertilizer treatments on
biomass productivity, nutrient content and nutrient
removal by ginger crop, nutrient balance and soil
fertility status were calculated for various treatments.
The interaction effects were presented wherever they
were significant.
biomass partitioning (BP) did not differ among the

different treatments in either of the two years. The RB,
SB and TB increased gradually with increase in each
level of Azotobacter up to 5.0 kg ha-1 (a3) during both
the years (Table 1). It increased RB, SB and TB by
12.5%, 14.3% and 12.9% of over the low level and by
8.6%, 10.3% and 9.3% of RB, SB and TB over the
medium level. The biomass productivity of ginger
between medium and low levels of Azotobacter
application did not vary significantly in either of the
two years. The seed treatment with Azospirillum and
Phosphotica also markedly influenced the biomass
productivity of ginger. The RB, SB and TB increased
3. Results and Discussion

3.1 Biomass Productivity and Partitioning
Biomass productivity of ginger is a function of
growth and development of the crop during its
growing period. Seed treatment with biofertilizers
exerted significant effect on rhizome biomass (RB),
shoot biomass (SB) and total biomass (TB)
productivity of ginger during both years. Biofertilizers
increased RB by 17.6% to 19.5%, SB by 17.8% to
significantly due to application of medium level (3.75

kg ha-1) of Azospirillum (b2) and Phosphotica (c2)
over their lower and higher levels during both years
(Table 1). Application of medium level of
Azospirillum increased RB, SB and TB by 8.6%, 10.6%
18.3% and TB by 17.7% to 18.9% over control but

Table 1
Effect of biofertilizer on biomass yield and its partitioning in ginger.

Rhizome biomass
2007
2008
3,095
3,122
3,697
3,673
164
154
465
435
Biomass yield (kg ha-1)

Shoot biomass
2007
2008
2,653
2,612
3,138
3,078
152
164
431
465
Total biomass
2007
2008
5,748
5,734
6,835
6,751
245
231
739
696
3,515
3,631
3,944
56
158
3,482
3,612
3,924
51
145
2,966
3,068
3,380
51
144
2,897
3,013
3,324
55
155
6,481
6,699
7,324
82
231
3,567
3,878
3,645
56
158
3,546
3,852
3,620
51
145
3,005
3,332
3,077
51
144
2,954
3,258
3,021
55
155
3,604
3,846
3,639
56
158
3,582
3,824
3,612
51
145
3,051
3,277
3,086
51
144
2,988
3,224
3,021
55
155
Particulars
Control
Treatment
S. Em ()
LSD 0.05
Azotobacter
a1
a2
a3
S. Em ()
LSD 0.05
Azospirillum
b1
b2
b3
S. Em ()
LSD 0.05
Phosphotica
c1
c2
c3
S. Em ()
LSD 0.05
Biomass partitioning (%)

2007
53.8
54.1
0.98
NS
2008
54.3
54.4
0.92
NS
6,379
6,625
7,248
75
214
54.2
54.2
53.9
0.33
NS
54.6
54.5
54.1
0.31
NS
6,572
7,210
6,722
82
231
6,500
7,110
6,642
75
214
54.3
53.8
54.2
0.33
NS
54.6
54.2
54.5
0.31
NS
6,655
7,123
6,726
82
231
6,570
7,048
6,634
75
214
54.2
54.0
54.1
0.33
NS
54.5
54.3
54.5
0.31
NS
a = Azotobacter; b = Azospirillum; c = Phosphotica; doses of biofertilizer 1, 2 and 3are 2.5, 3.75 and 5.0 kg ha-1, respectively.
14

and 9.6% over its low level and by 6.4%, 8.0% and
7.2% over its high level. Similarly seed treatment with
medium level of Phosphotica increased RB, SB and
TB by 6.8%, 7.6% and 7.2% over its low level and by
5.8%, 6.5% and 6.1% over its high level. Seed
treatment with Azotobacter and Phosphotica exerted
significant interaction effect on biomass productivity
of ginger. The highest RB, SB and TB were recorded
with the use of high level (5.0 kg ha-1) of Azotobacter
along with medium level (3.75 kg ha-1) Phosphotica
(a3c2) and was significantly higher over other
combinations except for the combinations of high
level of Azotobacter with other levels of Phosphotica
(Table 2). Application of low level of both
biofertilizers was found less efficient resulting in the
lowest biomass productivity of ginger. The increased
shoot and rhizome biomass by biofertilizer use might
be due to higher availability and efficient use of plant
nutrients throughout the growing period as a result of
greater microbial activities [10]. Increased biomass
productivity could be attributed to increase in shoot
and rhizome biomass resulting from higher fixation of
atmospheric N, dissolution of insoluble phosphates in
soil to soluble forms and production of plant growth
hormones and vitamins by microorganisms [21-23].
3.2 Nutrient Removal
The nutrient removal varied widely between
rhizome and shoot of ginger. While N removal shared
equally by rhizome and shoot, the P and K removal
were markedly higher in rhizomes when compared
with shoot. Seed treatment with biofertilizer exerted
significant effect on N, P and K removal by both

rhizome and shoot, and thus, recorded higher N, P and
K removal by 26.7%, 43.3% and 33.0%, respectively
over those of the control plots. The N, P and K
removal increased steadily and significantly as the
level of Azotobacter seed treatment increased and the
highest N, P and K removal by rhizome and shoot was
obtained at high dose of Azotobacter treatment (a3),
and was significantly higher than those obtained at its
medium (a2) and low (a1) levels (Table 3).
Accordingly, seed treatment with high level of
Azotobacter removed N, P and K by 16.3%, 10.1%
and 12.4% higher over its low level and 9.8%, 7.7%
and 9.2% higher over its medium level. The medium
dose of Azotobacter also showed higher N and K
removal by the crop than those obtained at its lower
level. Seed treatment with Azospirillum and
Phosphotica also exerted marked effect on N, P and K
removal by ginger. Use of medium level of both the
biofertilizers removed significantly higher N, P and K
by both rhizome and shoot over their other levels
(Table 3). Medium level of Azospirillum (b2) removed
higher N, P and K by 10.2%, 12.2% and 9.2% over its
low level (b1) and 6.9%, 9.2% and 6.6% over its high
level (b3), respectively. Similarly medium dose of
Phosphotica treatment (c2) removed 8.4%, 13.8% and
8.5% higher N, P and K, respectively over its low
level (c1) and 6.0%, 10.5% and 8.2% more than those
of its high level (c3). Seed treatment with Azotobacter
and Phosphotica exerted significant interaction effect
on N, P and K removal by rhizome, shoot and total
removal by the crop. The highest N, P and K removal
Table 2 Interaction effect of Azotobacter and Phosphotica (axc) on rhizome, shoot, and total biomass productivity of ginger
(average of 2 years data).
Phosphotica
Azotobacter
a1
a2
a3
S Em (+)
LSD 0.05
c1
c2
c3
Rhizome biomass (RB) (kg ha-1)
3,373
3,715
3,408
3,522
3,798
3,545
3,884
3,991
3,925
76.1
215.8
c1
c2
c3
Shoot biomass (SB) (kg ha-1)
2,818
3,123
2,854
2,937
3,220
2,964
3,303
3,409
3,343
83.1
235.6
c1
c2
c3
Total biomass (TB) (kg ha-1)
6,191
6,839
6,262
6,459
7,018
6,509
7,187
7,400
7,268
142.3
403.4
a = Azotobacter; c = Phosphotica; doses of biofertilizer 1, 2 and 3 are 2.5, 3.75 and 5.0 kg ha-1, respectively.
15

Table 3
Effect of biofertilizer on NPK removal by rhizome, shoot and total plant (average of 2 years data).
Particulars
Control
Treatment
S. Em ()
LSD 0.05
Azotobacter
a1
a2
a3
S. Em ()
LSD 0.05
Azospirillum
b1
b2
b3
S. Em ()
LSD 0.05
Phosphotica
c1
c2
c3
S. Em ()
LSD 0.05
N removal (kg ha-1)

Rhizome
Shoot
Total
39.2
39.1
78.3
49.9
49.5
99.4
1.17
1.60
3.15
3.29
4.51
8.89
P removal (kg ha-1)

Rhizome
Shoot
Total
9.4
4.2
13.5
13.5
6.0
19.4
0.50
0.26
0.89
1.41
0.74
2.51
K removal (kg ha-1)

Rhizome
Shoot
Total
69.0
46.2
115.1
91.9
61.3
153.2
2.44
2.44
3.76
6.90
6.90
10.64
46.6
49.3
53.8
0.67
1.90
46.0
48.7
53.9
0.54
1.50
92.5
98.1
107.6
1.05
2.96
12.9
13.2
14.2
0.17
0.47
5.7
5.9
6.4
0.09
0.25
18.6
19.1
20.5
0.30
0.84
87.7
90.2
97.9
0.82
2.30
58.1
59.9
66.0
0.62
1.70
145.7
150.1
163.8
1.26
3.55
47.9
52.4
49.4
0.67
1.90
47.3
52.5
48.7
0.54
1.50
95.2
104.9
98.0
1.05
2.96
12.9
14.4
13.2
0.17
0.47
5.7
6.5
5.9
0.09
0.25
18.5
20.7
19.0
0.30
0.84
88.8
96.2
90.8
0.82
2.30
58.8
64.8
60.3
0.62
1.70
147.5
161.0
151.1
1.26
3.55
48.3
52.1
49.3
0.67
1.90
47.7
52.0
48.9
0.54
1.50
96.0
104.0
98.1
1.05
2.96
12.8
14.5
13.1
0.17
0.47
5.7
6.5
5.8
0.09
0.25
18.4
20.9
18.9
0.30
0.84
89.4
96.7
89.6
0.82
2.30
59.4
64.8
59.7
0.62
1.70
148.8
161.5
149.3
1.26
3.55
a = Azotobacter; b = Azospirillum; c = Phosphotica; doses of biofertilizer 1, 2 and 3 are 2.5, 3.75 and 5.0 kg ha-1, respectively.
by rhizome and shoot were recorded with the

combined use of high dose of Azotobacter with
medium dose of Phosphotica (a3c2). This treatment
combination removed the highest quantity of N
(109.5 ha-1), P (21.5 ha-1) and K (167.8 kg ha-1) and
were significantly higher than those of other
combinations except high level of Azotobacter with
high and low levels of Phosphotica (Table 4). Use of
lower doses of both Azotobacter and Phosphotica
recorded the lowest N, P and K removal (88.5, 17.5
and 140.2 kg ha-1, respectively) by the crop. The
increase in nutrient removal due to biofertilizer
inoculation could be attributed to increase in biomass
yield. The higher removal of nutrients due to
biofertilizer application could be on account of high
nutrient availability in the root zone under high
microbial activities [24]. This showed very vital role
of biofertilizer in the nutrient supply system of
organic farming [25, 26].
3.3 Nutrient Balance

Seed treatment with biofertilizer recorded
significantly greater N (20.4 kg ha-1) and P (3.3 kg
ha-1) balances than that of the control plots. The study
showed overall negative K balance; but biofertilizer
treated plots reduced the negative K balance to a great
extent (-28.5 ha-1) as compared to that of the control
plots (-45.3 ha-1). As same amount of nutrient (200 kg
N, 53.2 kg P and 220 kg K ha-1 through 20 + 20 t
FYM ha-1 in two crop cycles) was added in all the
treatments, the variation in nutrient balance was
primarily due to the variation in soil microbial
activities induced by biofertilizer treatments and
nutrient removal by the crop. N and P balance in soil
increased steadily up to the highest level of
Azotobacter treatment (a3). Accordingly the high level
of Azotobacter treatment (a3) recorded the highest N
(22.4 kg ha-1) and P (3.5 kg ha-1) balance in soil and
was markedly higher than that of its medium (a2) and
16
Table 4

Interaction effect of Azotobacter and Phosphotica (axc) on N, P and K removal (average for 2 years).
Phosphotica
Azotobacter
a1
a2
a3
S Em ()
LSD 0.05
Phosphotica
Azotobacter
a1
a2
a3
S Em ()
LSD 0.05
Phosphotica
Azotobacter
a1
a2
a3
S Em ()
LSD 0.05
c1
c2
c3
N removal by Rhizome (kg ha-1)
44.7
49.6
45.6
47.6
51.9
48.4
52.5
54.7
53.9
1.17
3.29
c1
c2
c3
P removal by Rhizome (kg ha-1)
12.3
13.5
13.0
12.4
13.8
12.8
14.1
14.9
14.0
0.31
0.87
c1
c2
c3
K removal by Rhizome (kg ha-1)
84.4
94.0
84.5
87.4
95.6
87.1
96.5
100.1
97.2
1.41
3.98
c1
c2
c3
N removal by shoot (kg ha-1)
43.8
49.1
44.8
46.7
51.7
47.7
52.6
54.9
54.0
0.92
2.60
c1
c2
c3
P removal by shoot (kg ha-1)
5.3
6.1
5.7
5.4
6.2
5.6
6.3
6.7
6.3
0.17
0.49
c1
c2
c3
K removal by shoot (kg ha-1)
55.8
62.4
56.0
57.6
64.0
57.6
64.9
67.7
65.4
1.22
3.45
c1
c2
c3
Total N removal (kg ha-1)
88.5
98.8
90.4
94.4
103.6
96.2
105.2
109.5
107.8
1.82
5.14
c1
c2
c3
Total P removal (kg ha-1)
17.5
19.6
18.6
17.7
20.0
18.3
21.0
21.5
20.3
0.51
1.44
c1
c2
c3
Total K removal (kg ha-1)
140.2
156.4
140.5
145.0
159.7
144.7
161.5
167.8
162.7
2.18
6.18
*a = Azotobacter; c = Phosphotica; doses of biofertilizer 1, 2 and 3 are 2.5, 3.75 and 5.0 kg ha-1, respectively.
low level (a1). Seed treatment with low level of

Azotobacter (a1) was found less efficient in
maintaining N and P balance in soil. Use of
Azospirillum and Phosphotica in seed treatment also
showed positive and significant effect on N and P
balance in soil. Medium dose of both Azospirillum (b2)
and Phosphotica (c2) recorded the highest N and P
balance which were significantly greater than that
obtained at their high and low levels (Table 5). Seed
treatment with medium level of both Azospirillum (b2)
and Phosphotica (c2) greatly reduced the negative K
balance in soil. Our results showed that biofertilizer
treatments increased the N and P balance because of
high microbial activities of biofertilizer treated plots.
In spite of high N and P removal, biofertilizer showed
markedly higher N and P balance at high dose of
Azotobacter (a3) and medium dose of both
Azospirillum (b2) and Phosphotica (c2). Ginger is a
heavy K feeder crop and K supply through organic
manure (20.0 t FYM ha-1 year-1) did not match the K
demand of the crop. Biofertilizers though enhanced K
removal, yet they improved K balance over control
plots through greater microbial activities in soil. The

study clearly showed that current level of organic
manuring (FYM 20 t ha-1) was not sufficient to meet
the K requirement of ginger in the North-Eastern
region of India.
3.4 Soil Fertility
The organic carbon and available N and P of the
soil improved considerably over control due to use of
biofertilizer. The available K decreased to a great
extent in all cases as compared to its initial status; but
biofertilizer exerted favourable effect on minimizing
the depletion of soil available K as compared to that of
the control plots (Table 5). Different biofertilizer
treatments responded differently on short-term fertility
of the ginger field. The organic carbon and available
N content of the soil increased gradually due to
increasing the level of Azotobacter and application of
high dose of Azotobacter (a3) increased markedly both
organic carbon and available N over its lower levels;
but it did not influence the available P and K content
in the ginger field. Use of medium dose of Azospirillum
17

Table 5
Effect of biofertilizer on nutrient balance and fertility status of the experimental soil after 2 ginger crop cycles
Nutrient
management
Nutrient added in 2 crop

Organic
cycle
C (%)
(kg ha-1)
N
P
K
2.41
200.0 53.2
220.0
2.66
200.0 53.2
220.0
0.07
0.21
-
Nutrient removed by 2
crop cycle
(kg ha-1)
N
P
K
156.6 27.1
230.2
198.7 38.8
306.3
6.54 1.39
9.55
19.75 4.19
28.85
Soil available nutrient

after 2 crop cycle
(kg ha-1)
N
P
K
235.8 14.32 221.7
250.4 16.61 238.5
4.48 0.59 4.41
13.50 1.80 13.30
Actual nutrient gain/loss

over initial status
(kg ha-1)
N
P
K
5.8
1.0
-45.3
20.4
3.3
-28.5
1.14 0.21
1.73
3.46 0.65
5.22
Control
Treatment
S. Em ()
LSD 0.05
Azotobactor level
a1
2.62
200.0 53.2
220.0 185.0 37.9
291.4 248.5 16.41 237.5 18.5
3.1
-29.5
a2
2.65
200.0 53.2
220.0 196.1 37.5
300.0 250.4 16.58 238.5 20.4
3.3
-28.5
a3
2.71
200.0 53.2
220.0 215.1 41.0
327.6 252.4 16.84 239.6 22.4
3.5
-27.4
0.02
2.20 0.46
3.20
1.35 0.20
1.47 0.37 0.07
0.58
S. Em ()
LSD 0.05
0.07
6.20 1.31
9.00
3.80 NS
NS
1.06 0.20
1.63
Azospirillum level
b1
2.64
200.0 53.2
220.0 190.1 36.9
294.9 249.0 16.48 237.7 19.0
3.2
-29.3
b2
2.69
200.0 53.2
220.0 209.7 41.5
321.9 252.9 16.76 239.5 22.9
3.5
-27.5
b3
2.66
200.0 53.2
220.0 196.3 38.0
302.1 249.4 16.59 238.2 19.4
3.3
-28.8
0.02
2.20 0.46
3.20
1.35 0.20
1.47 0.37 0.07
0.58
S. Em ()
LSD 0.05
NS
6.20 1.31
9.00
3.80 NS
NS
1.06 0.20
1.63
Phosphotica level
c1
2.62
200.0 53.2
220.0 191.9 36.8
297.6 249.9 16.26 238.2 19.9
3.0
-28.8
c2
2.72
200.0 53.2
220.0 208.0 41.8
323.0 251.6 17.25 239.3 21.6
4.0
-27.7
c3
2.64
200.0 53.2
220.0 196.2 37.8
298.5 249.8 16.32 238.0 19.8
3.0
-29.0
0.02
2.20
0.46
3.20
1.35
0.20
1.47
0.37
0.07
0.58
S Em ()
C D at 5%
0.07
6.20 1.31
9.00
3.80 NS
NS
1.06 0.20
1.63
a = Azotobacter; b = Azospirillum; c = Phosphotica; doses of biofertilizer 1, 2 and 3 are 2.5, 3.75 and 5.0 kg ha-1, respectively; Initial
fertility status =230, 13.3 and 267 kg N, P and K ha-1, respectively.
(b2) considerably improved the soil available N over

control and its higher and lower levels; but it had very
little effect on organic carbon and available P and K
contents of the experimental soil (Table 5). Similarly,
seed treatment with medium dose of Phosphotica (c2)
improved organic carbon and available P level of the
soil over its other levels and control plots; but it had
no effect on available N and K levels. The results
showed a sharp decrease in available K content of the
experimental soil after two ginger crop cycles in spite
of addition of 20.0 t FYM ha-1 year-1. The high K
removal by the crop was mainly responsible for the
decrease of available K level of the experimental soil
[27]. High dose of Azotobacter (a3) and medium dose
of both Azospirillum (b2) and Phosphotica (c2) were
found the most effective in improving the fertility
status of the ginger field.
4. Conclusions
Use of biofertilizer increased biomass productivity
of both rhizome and shoot, enhanced N, P and K
removal by the crop and helped in improving soil
fertility by increasing N and P balance and reducing
negative K balance over those of the control plots.
Seed treatment with Azotobacter 5.0 kg ha-1 (a3),
Azospirillum 3.75 kg ha-1 (b2) and Phosphotica 3.75
kg ha-1 (c2) increased biomass productivity of rhizome
and shoot, enhanced N, P and K removal through
rhizome and shoot as well as total removal by the crop
as compared to those of the other levels. In spite of
high N, P and K removal, biofertilizers showed
markedly higher N and P balance at high dose of
Azotobacter
(a3)
and
medium
dose
of
both
Azospirillum (b2) and Phosphotica (c2). The study
18

showed overall strong negative K balance; but

biofertilizer treatments greatly reduced the negative K
balance in soil as compared to the control plots. They
improved organic carbon and available N and P levels
of the soil over other treatments. Seed treatment with
high level of Azotobacter along with medium level of
Phosphotica (a3c2) produced high biomass yield (7.4 t
ha-1), removed large quantity of N, P and K from the
soil, increased soil nutrient balance and maintained
high
soil
fertility
responsible
for
high
crop
productivity and sustainability.
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DAVID
PUBLISHING
Organic Agriculture: Socioeconomic Sustainability of

Brazilian Coffee
Rubia Wegner1, Patrcia Helena Nogueira Turco2 and Flvia Maria de Mello Bliska3
1. Institute of Humanities and Social Sciences, Federal Rural University of Rio de Janeiro, Seropdica 23890-000, Brazil
2. Department of Decentralization of Development, So Paulo Agency of Agribusiness Technology, Agriculture Secretary of So
Paulo State, Campinas, P.O. Box 28, So Paulo 13012-970, Brazil
3. Coffee Centre, Agronomic Institute, So Paulo Agency of Agribusiness Technology, Agriculture Secretary of So Paulo State,
Campinas, P.O. Box 28, So Paulo 13012-970, Brazil
Received: October 8, 2012 / Published: January 20, 2013.

Abstract: Worldwide, there is a growing demand for products made by technologies that contribute to environmental preservation
and to sustainable rural development. In Brazil, organic farming is one of the most established initiatives in this area. Therefore, it is
important to subsidize the decision-making regarding policies for organic coffee production. Thereby, this study analyzed the
socio-economic sustainability of this production system in relation to conventional. We applied a semi-structured questionnaire on
coffee farms in the Brazilian coffee producers states and analyzed the coffee production cost structure. We identified the reasons that
led the producers to organic management, implications of certification on the management, and prices received by organic coffee and
the market mechanism. Results showed that there is no standard for organic coffee production and marketing in Brazil. Among the
producers, there are different levels of access to technical information and the main limitation of the organic coffee chain is not the
technology of production, but the difficulty of coffee marketing. Another important limitation, for medium and large farms, is the
cost of manpower, mainly in areas dominated by mountain, where machines operations are not viable. Moreover, the yield of organic
coffee system is generally lower than the conventional. Furthermore, soil and climatic differences lead to different regional behavior.
Finally, we could conclude that the production of organic coffee can be sustainable in Brazil mainly in two structural conditions: 1)
family farmers; and 2) small producers, who employ only one or two workers, preferably only in the harvest time.
Key words: Sustainable development, coffee crop, production costs, organic agriculture.
1. Introduction
1.1 Theoretical Fundamentals of Organic Farm
There are many controversies about the roots of
Agroecologyplans, projects or policies for
economic development. However, there is consensus
that it seeks to understand the complex functioning of
agroecosystems (the units of study), as well as
different interactions present in these systems, with
the principle of biodiversity conservation and
expansion of agricultural systems as a basis for
Corresponding author: Flvia Maria de Mello Bliska,
Ph.D., research field: agricultural sustainability. E-mail:
bliska@iac.sp.gov.br.
engendering self-regulation and hence sustainability

[1]. In addition, it provides a methodological
framework for this and relies on the incorporation of
the farmer to agricultural practice [2]. However, for
some authors [3] to keep up with good levels of
productivity and profitability in an organic system are
needed technical knowledge about the combination of
fertilizers and biofertilizers. Moreover, the
agroecological strategy could also be defined as the
ecological management of natural resources, which,
by incorporating a social action group with
characteristic participatory, allows designing methods
for sustainable development [4].
Then, formed the following streams: biodynamic
farming, organic, organic-biological and natural

agriculture. Such currents promoted from the 1970s,
movements such as ecological agriculture, regenerative,
biological and permaculture, which were agglutinated
in alternative agriculture, then agroecology [5] and,
finally, sustainable agriculture. All these agricultural
practices are covered by the principles of agroecology
and pointed the search for a sustainable production
system in time and space, by managing and protecting
natural resources, without using harsh chemicals to
human health and the environment [6].
Since the 1990s, it has established itself the term
organic farming as a reference to all those forms,
mainly as a synonym of agroecology [7]. But it is also
characterized as a system in which soluble chemical
are removed from inputs, and which seeks to preserve
the biodiversity of agricultural ecosystems involved
[8]. Or else, as a productive system that aims to
prevent and even totally exclude synthetic pesticides
and fertilizers of agricultural production, which
replace external sources, such as chemicals and fuels
purchased commercially, by resources obtained within
the same farm or nearby. But, after all, organic
farmers are characterized by use of certified seed,
modern equipment and practices of soil and water
conservation [5].
Also since the late 1990s, studies that analyze the
organic farming emphasized the importance of
certification to increase its profitability and therefore
their economic, social and environmental benefits.
Organic agriculture is important for marketing,
because it guarantees to the farmer certificate to enter
and remain in the market. For the consumer, the label
organic is assured of a product originating from
properties that meet social and environmental values.
Even if the certification homogenizes the farmers in
view of the criteria it establishes, organic systems
production has different characteristics from one
region to another within the same country. Product
differentiation is the core profitability of the organic
chain, i.e., his competitiveness comes from the range
21
of specific markets and thus guaranteed [7-9]. Organic

agriculture offers opportunities for farmers in
developing countries to increase their income by
exporting agricultural products, as well as reduce its
dependence on other parts of the production chain,
such as agribusiness [10].
1.2 Organic Coffee in Brazil: A Brief Characterization
In Brazil, coffee is one of the most important crops,
particularly regarding job creation and foreign
exchange. Its cultivation is spread over much of the
Brazilian territory, but its production is concentrated
in some regions of Minas Gerais, Espirito Santo, Sao
Paulo, Paran, Bahia and Rondnia States. Each of
these regions presents competitiveness and production
costs differentiated by the use of different
technological packages.
Most of the Brazilian coffee producers made the
conversion from conventional to organic in the early
2000s, when conventional coffee prices were very low.
These years were marked by optimism among coffee
producers and Brazilian researchers, as to the
profitability of organic coffee, because from 2000 to
2006, its production grew by 5% [11]. Rondnia,
Mato Grosso do Sul, Sao Paulo, Bahia, Paran and
Esprito Santo are states where we can find coffee
production in agroforestry or organic systems.
In recent years, organic products, especially grocery
are closer to the Brazilian consumer demand. The
main reasons are the real1 strong, the boom phase
of the Brazilian economy and the financial crisis,
which spared the producing countries but has reached
the mature economies of Europe and the US, major
buyers of organic [12].
Research conducted by the Brazilian Coffee
Industry [13] referring to 2010 showed that among the
Brazilians, is consolidated into the habit of drinking
coffee, especially for consumption outside the home.
The specialty coffee group where organic coffee is
included grown in preference of consumption: the
1
Brazilian currency.
22
participation of this group in the consumption at home

was 1.2%, and outside the home was 3.3%.
However, the concept of organic agriculture was
understood by the Brazilian coffee growers, especially
as non-use of agrochemicals. A controversial point in
the characterization of organic farming in Brazil is the
maintenance of a monoculture, which also affect the
competitive insertion in the market for these products
[14].
The pressure of NGOs (Non-Governmental
Organizations), cooperatives of farmers and certifiers
led in the 1990s, the Federal Government to prepare a
draft law to regulate organic production in Brazil
(Law 10831, 2003). In addition, were created the
Sectorial Camera of Organic Agriculture and the
National Collegiate of Organic Products, to accredit,
monitor and control certification. In the 2000s, the
federal government created the Department of
Agroecology (COAGRE), linked to the Department of
Production Systems and Sustainable Development
Secretariat of Agriculture and Cooperatives, and it is
responsible for the actions of the promotion,
development of standards and implementation of
quality control mechanisms, which must develop
agriculture organic in Brazil.
Based on Article 1 of Law 10,831, Brazil considers
organic agricultural production system, that when
specific techniques are adopted, by optimizing the use of
natural and socioeconomic resources available, and
respect the cultural integrity of rural communities,
aiming at economic and ecological sustainability,
maximizing benefits, minimizing the energy
dependence is not renewable, using, where possible,
cultural methods, biological and mechanical, as opposed
to using synthetic materials, eliminating the use of
genetically modified organisms and ionizing radiation at
any stage of production, processing, storage, distribution
and marketing, and environmental protection.
We can scale the Brazilian public policies with
respect to encouraging conversion to organic
production, and with respect to the support and
infrastructure offered to those who already act as

organic growers. These dimensions are due to the way
the state understands the promotion of sustainable
agriculture, i.e., in that it links the environmental
protection and incentive compensation payments in
order to promote the necessary changes [14]. It
highlights how tenuous a specific policy for organic
farming can be compared to the market. In this area,
the main Brazilian public policy is restricted to the
scope of certification and marketing.
The main certifiers operating in Brazil are
ECOCERT (Organic Certification Organization), the
Biodynamic Institute and BCS Brazil. The
relationship between farmers and certifying that it
comes down to the support and control of matters
related to certification, and not to technical assistance,
that would set off the conflict of interests between the
domestic and international regulatory authorities.

To survey the cost structures and technical
coefficients of production, we use semi-structured
interviews in loco, that is, applied to coffee producers
in their respective farms in the states of Minas Gerais
and So Paulo, where we visited the following regions:
South of Minas Gerais, Mogiana and Pontal do
Paranapanema, in So Paulo, between February and
June 2011. Interviews with producers of Rondnia,
Espirito Santo, Bahia, Paran and Mato Grosso do Sul
were conducted by telephone. The interviews
consisted of two parts. At first, it was found the
reasons that led the producers to organic management,
the implications of certification on the management
and the prices received by producers, marketing
method, and the main difficulties with this
management. The second part was to search for
information on the structure of production costs, per
hectare of coffee grown and per bag produced. We
evaluated the operations, inputs and materials
consumed and machinery and implements used in the
2009-2010 crops.
The description of costs followed the concept of

total operating cost (COT), which consists of variable
costs, expenses for manpower, operations of
machinery and equipment, raw materials used
throughout the year and includes depreciation of
durable goods used in production process. Also
followed the concept of operational cost (COE), which
adds to the total operating cost: 1) social security
contributions; 2) depreciation of machinery, other
fixed costs of machinery related to shelter, insurance
and interest on capital invested in this item; 3) when
applicable, the rate of the National Program for
Strengthening Family Farming (PRONAF). This
method therefore serves to estimate costs and analyze
the economic feasibility [15].
We analyzed the areas of coffee production to
organic management and conventional management,
in terms of hours of hand labor, tractor and equipment,
materials and quantity consumed for each operation
activities, from fertilization through the processing, in
view of the sequence used by the manufacturer
respondent. This systematization is called the matrix
of technical coefficients [15]. Also we attempted to
determine the profitability of both of these
management systems through indicators such as:
(1) Gross Revenue (GR)
GR = R*Pu
(1)
Where:
R = yield of activity per unit area;
Pu = unit price of the product of the activity.
(2) Leveling point (LP) determines the minimum
production necessary to cover production costs, given
the unit-selling product price:
LP = TOC/Pu
(2)
Where:
TOC = total operating cost
Pu = unit price
(3) Operating profits (OP): measures the
profitability in the short term, is the difference
between gross revenue and total operating cost (COT)
per hectare.
23
(4) Profitability index shows the relationship

between the operating profit (OP) and the gross
revenue (GR) in percentage. This index shows the rate
of revenue available from activity after paying all
operating costs including depreciation. So:
PI = (OP/GR) 100
(3)
2.1 Measurement of Coffee Production Costs

In a country with the size of Brazil, where each of
the producing regions has specific soil and climatic
characteristics and the producers have totally different
characteristics and objectives, it is expected that
production costs are very different. After all, the cost
of production depends mainly on the following factors:
1) system of production; 2) level of technology; 3)
size of farm; 4) philosophy of life and the producer
objective; 5) national and international economic
conditions (e.g., via changes in relative prices,
exchange rates, international prices, yields in countries
competitors or changes in demand); 6) characteristics
of climate and soil of the original area where the
property is located; 7) climate variations that may
occur in specific years (frosts, prolonged droughts)
and other aspects.
2.2 Factors that Can Interfere with the Yield of Coffee
Plantations
(1) Biennial: it is the physiology of the coffee itself,
that is, the coffee plantations have a high production
year and another year of low and this is established
fact to the producers. The biennial has variations
within the following conditions:
In plantings in full sun, the biennial is very
significant;
In rotation with other crops and forested crops (but
not shaded), the biennial is apparent, though not as
significant as in the unshaded crop;
In shaded plantations, the biennial is not significant.
That is, the variation in productivity from year to year
is so small that it can be considered that, in practice, it
does not exist;
24
In scientific studies, especially in plantings in full

sun, when you want to evaluate the productivity of a
coffee stand, it is important to calculate the average
productivity of this stand for the last four years (two
cycles of high and low yield).
When the plants are young, or are undergoing
strong growth, the Biennale is not significant. For
systems in full sun, this occurs in the first two seasons,
when the plants reach 3.5 years. From the third season
of the biennial becomes significant.
The intensity of the Biennial of coffee depends on
many factors. The main ones are climatic conditions
and the packet technology used (such as variety, pest
and disease management, fertilization, spacing, and
pruning).
The renovation and expansion of the coffee
plantations usually occur at a higher intensity when
coffee prices are considered good. Consequently,
much of the crop is planted or renewed in the same
time, which leads to the existence of years of low and
high productivity.
However, in one year high, there are producers
with low yields, as well as in years of low, there are
producers with high yields. Sometimes, in the same
farm, there are stands of coffee in year of low
crop and other in high.
Frosts, prolonged drought or excessive rainfall can
change the volumes produced annually and therefore
the sequence of high and low yields.
Therefore, despite the implications of the Biennale,
to calculate production costs, we need to do this every
year. The relative prices in the economy vary from
season to season, especially when using imported
inputs, especially fertilizers and pesticides, as in
conventional cultivation of coffee. And for that we
must use the yield of the year. We can not use the
average of four years.
(2) Yield: according to information provided by
farmers and rural extension workers, the productivity
of organic coffee is generally 30% lower than the
productivity of conventional coffee, even though, it is
possible to obtain high yields in the organic system.

In Brazil, today, there are conventional arabica
coffee plantations that produce 70 to 100 bags per
hectare, average four years (for instance, in South and
Cerrado of Minas Gerais State; in Gara and Mogiana
regions, in So Paulo State; or in the Cerrado of Bahia
State). So you can get yields of 30 to 60 bags per
hectare in organic crops.
One situation where we can get regular high yields
in organic crops in normal years (that is, free of frost,
drought or too much rain) is that found in the region
of Serra da Mantiqueira in the State of Minas Gerais,
1,200 m altitude, using variety of coffee rust resistant,
and in soil without nematodes infestation. In that
region, the soils are not sandy, the weather conditions
and temperature inhibit the attacks of leaf miner and
cicadas and natural fertility is high. Thus, the
probability of obtaining high yields and excellent
quality of the beverage is high, since it makes a good
crop management and good work of post-harvest. The
drop in productivity compared to conventional coffee
can occur, but a careful producer can get high yields
and competitive costs.
Many farmers still do not understand that organic
system requires specific management, with investment
in nutrients and control pests and diseases. The most
of organic coffee producers began using this system of
cultivation due to high costs for the conventional and
the initial idea was to reduce production costs. Many
farms still had reserves of available nutrients in the
soil and also in the plants themselves. So even after
three years without chemical fertilizers, necessary for
the conversion to organic system to be recognized, the
crop still had reasonable productivity. However, prices
for organic inputs are competitive with conventional
chemical inputs in years of high prices of conventional,
while in other years the prices of conventional inputs
are lower than those of organic.
Without using organic inputs and without
management of pests and diseases, these coffee
producers observed over the subsequent years, the
sharp drop in their productivity. Some of these

producers have abandoned the organic system. Other
producers began to use organic inputs, improved crop
management and persisted in the organic system. In
these situations, the highest prices paid for the bag of
organic coffee represented an incentive for farmers to
remain in this production system.
In general organic crops with low productivity were
not implemented with varieties resistant to rust or leaf
miner did not receive nutritional treatment and
management of pests and diseases adequate or are not
located in regions with favorable climatic conditions
for coffee production or in soil naturally fertile.
3. Results
The results are based on information obtained
through interviews with 21 organic coffee producers,
three organic agroforestry, five conventional, a large
coffee-growing company and exporting of organic and
conventional coffee, cooperatives, and researchers,
institutions of technical assistance and rural extension,
and export companies.
All producers of this sample were clearly
committed to organic practices, although some of
them were dissatisfied with the return of the last
harvest (low season), and conventional coffee resulted
in better pay per benefit bag. All those producers
follow the recommendations of their certification as
fertilizers and pesticides. The machinery used was not
especially developed for organic system. Producers
who are not certified seek to follow the guidelines of
the IBD or the BCS. The main inputs used in organic
coffee in Brazil are:
Fertilizers: fish fertile, castor seeds pie, coffee straw,
chicken manure, castor meal, natural phosphate and
urea.
Pesticides: Bordeaux mixture, cooper hydroxide,
supera, borax, hydrated lime, cooper sulphate,
magnesium sulphate, zinc sulphate, fungi Beauveria
bassiana, caustic, Bordeaux and Viosa mixture.
Machinery: mowing, sprayer, harvesters, trator and
25
dryer.
3.1 Characterization of Organic Coffee Producers:
Brazilian States
It was observed that organic growers interviewed
chose this production system due to: 1) its social and
environmental awareness; 2) specific programs
captained by rural technical assistance; 3) the search
for reducing costs; 4) or even, given the expectation of
obtaining higher price compared to the conventional.
Some growers, especially those who remain in the
market with modest production, face this culture
system as a way to contribute to the preservation of
the environment and health of their families.
In general, organic farmers use family labor and,
during the harvest, they hire day laborers. However,
some producers have hired salaried employees. Part of
the organic coffee growers consort with products such
as cassava, banana and honey (which are important
sources of income). Despite the existence of growing
demand for organic products, maintaining its
production scale has been for some producers, a
problem to ensure their profitability and also
continuity in management.
One of the interviewed cooperatives, located in
Minas Gerais State, did not export organic coffee in
the last 10 years because their members were unable
to match the scale of production with good quality
coffee. An exporter of Sao Paulo State, informs that,
even certified, it is difficult to get good prices for
organic coffee, so the producers income. Furthermore,
there is great difficulty in obtaining uniform batches,
especially when, to export, it is necessary a container.
As this exporter marketing only high quality coffees,
lots of heterogeneity prevents their commercial
relationship with cooperatives of small producers.
Since, for organic coffee, sieve mixture is allowed
[14-17], this mixture, in roasting results a lower
quality of drink, therefore, lower prices. So it has
exported coffee regularly only from two organic
farmers, which produce high quality coffees.
26
In Bahia State, two respondents have created a

coffee exporting company to sell their production of
organic coffee. Each one has an average area of 20
hectares of organic coffee, and both invest heavily in
the quality of coffee. In 2009-2010, they exported 640
bags of coffee, priced 100% higher than conventional,
and have sold about 50 bags for national companies
from So Paulo and Bahia States. They are certified
by IBD and claim that organic coffee production is
feasible for them, because they make use of its own
channel of marketing and because their coffee is high
quality standard, i.e., the premium is linked more to
the quality of coffee than the organic seal.
Many growers declined to produce organic coffee
due of the low pay they got. For some, the high cost of
manpower was the biggest obstacle (mainly medium
and large farmers). Others add to that the insufficient
technical assistance (especially in the case of family
farmers) and a significant drop in productivity of the

plantation, after conversion to organic. Table 1 shows
us the main characteristics of organic farmers
surveyed.
In the Southern region of Minas Gerais State, the
organic coffee production is better established in
Brazil. In this region there are predominantly small
farmers, the coffee has great socio-economic
relevance and producers has better knowledge about
the requirements concerning the practice of organic
coffee than in other producing areas, considering
technical or agronomic dimension and marketing. The
cooperative is the element that stimulates this
structure. All of them are connected to the Family
Farmers Cooperative and export all their bags of
organic coffee. These farmers started using the
principles of organic practices in the 2000s. They have
relative understanding that the certification, as well as
Table 1 Characterization of organic coffee production: producer, municipality, Brazilian States, certifying, cultivate, coffee
area (ha), number of coffee trees, selling price (*R$/bag of 60 kg).
Producer
Municipality/State
Andradas/MG
2
3
4
5
6
7
8
Andradas/MG
Andradas/MG
Poo Fundo/MG
Poo Fundo/MG
Ouro Fino/MG
Ouro Fino/MG **
Paraispolis/MG**
Santo Antnio do
Pinhal/SP
Irupi/ES
Santa Maria de Jetib/ES
Santa Maria de Jetib/ES
Teodoro Sampaio/SP
Teodoro Sampaio/SP
Teodoro Sampaio/SP
Lunardelli/PR
Ibicoara/BA**
Ibicoara/BA**
Ji-Paran/RO
Certifier
Certifica Minas;
ECOCERT; BCS
BCS; ECOCERT
BCS; ECOCERT
BCS; ECOCERT
BCS; ECOCERT
BCS
ECOCERT
IBD
Cultivate
Area (ha)
Coffee
(trees/ha)
Price
(R$/bag)
Catua, Mundo Novo
13200
430
Catua, Mundo Novo

6
18000
480
Catua, Mundo Novo
2
5300
480
Catua, Mundo Novo
2.4
2500
430
Catua, Mundo Novo
06
3500
435
Mundo Novo
03
1500
410
Catua, Mundo Novo
02
-***
Catua, Mundo Novo
25
725
Mundo Novo, Tupi,
9
IBD
75
3333
650
Obat
10
BCS
Arabica
10
2500
380
11
Cho Vivo
Arabica
04
4000
250
12
Cho Vivo
Arabica
03
2000
380
13
Not certified
Arabica
01
2000
250
14
Not certified
Arabica
02
2000
250
15
Not certified
Arabica
01
2500
250
****
16
IBD
Obat, IAPAR-59 Tupi 40
5500
17
IBD
Arabica
25
7000
630
18
IBD
Arabica
20
7500
630
19
ECOCERT
Conilon
2.5
2100
185
Tupi, Mundo Novo,
20
Glria dos Dourados/MS Not certified
10
8.000
120
Catua
* Reais: currency in Brazil; ** Not cooperate with information on production costs; ***Not cooperate with information; ****Three
times the conventional price; Source: Data from the study.
the cooperatives, does not guarantee a stable return, at

high levels, since these factors depend on the
international market. They believe that to keep
producing organic coffee, farmers should not
prioritize getting high prices per bag produced. In this
region, the production dynamics is linked to
cooperatives, starting with the certification, BCS and
Ecocert, with direction for the use of inputs and other
components of management and, particularly, with
regard to marketing. In the Cerrado of Minas, there is
only one producer of organic coffee, with 20 hectares
devoted to this production system. It is connected to a
regional cooperative, and he sells his coffee to
national companies.
One of the biggest supporters of organic coffee
production in Brazil also was a major producer of
organic coffee and managed several family farms, all
in the Southern region of Minas Gerais. Their farms
were relatively large, some with 70 to 100 ha of coffee.
By 2006, all the organic coffee produced was sold to
Japan; grains above 14 screens were sold in bulk. The
rest of the coffee was dried and exported to Japan
already packed. About two years the farms managed
by him broke. According to him, the reason was the
extremely high cost of hired labor which prevents the
organic system, on farms of medium to large. Today,
he recommends the organic system only for small
farms, mainly in the family production system.
Interview with the technical manager of a big farm,
in Southern of Minas Gerais, also indicated that the
large production with organic system is not
economically viable. A few years ago, it had converted
50 ha of conventional coffee to organic. The yield, that
was around 30 bags/ha, fall from 15 to 20 bags/ha. In
addition to this low yield, only 50% of the coffee was
sold at the organic price, due to many problems with
its quality. For nearly two years, that farm began to
managing another farm, also in Minas Gerais State, a
coffee exporter, with all the possible certifications for
coffee, including organic. Today, the new
administration excluded the area of organic coffee.
27
Two years ago, the average yield of the conventional

area was between 30 and 40 bags, while in the area of
organic coffee the yield was 20 bags/ha. Those farms
used inputs as recommended for organic system, but
had great difficulty in applying the products in
mountain area with no possibility of management and
harvest mechanization. All operations needed to be
done manually, which became unviable due to the high
cost of manpower. For him, the organic system is
feasible only for small producers.
In Western of Sao Paulo State, producers of
agroforestry organic coffee sell their coffee in regional
markets, or companies of Paran State, with the price
of conventional coffee. In the region, known as the
Pontal of Paranapanema, Ipe Foundation is developing
a program called Coffee with Forest in settlements
of farmers landless. This project has brought great
socioeconomic outcomes. The coffee production in
organic agroforestry system (SAF) has shown a
significant supplemental income source, with
extremely low cost. The coffee is not certified and the
producers are not sensitized to ascertain or to seek
foreign markets. The coffee is sold regionally and in
the State which borders that region. Using a very low
volume of external inputs, farmers have obtained
yields of around 15 benefit bags/ha, which are sold at
the price of conventional coffee.
In Esprito Santo State, the conversion of
conventional coffee to organic was due to the
discomfort that the use of agrochemicals caused to
farmers and their families, and due to the damage,
they cause to the environment. Moreover,
conventional inputs were expensive and the possibility
of the bag of organic coffee to get better pay was an
important persuasive factor. Growers did not receive
technical assistance and they used fertilizers and
biofertilizers on their own, to analyze those were more
efficient, for added profitability and sustainability.
In Paran State, four farmers were contacted. Two
of them left the organic system. Moreover, they had
not recorded their spending with the crop. The third
28
producer did not provide information about their

production costs. The conversion of this crop for the
organic system followed a severe frost in 1994.
Moreover, prices of inputs required for cultivation of
conventional coffee were high.
In the Bahia State, organic coffee production is also
declining. The main reason is the difficulty face to fit
within the market. The Biodynamic Association of
Farmers (certified by IBD), in the beginning of its
activities consisted of a group of 34 organic growers.
In 2009-2010, this group was reduced to seven
members. Two of them created its own exporting
company, which allowed them to continue to produce
organic coffee. These producers have access to an
important organic raw material, the castor bean and
therefore use manure, which represents a high cost
due to the cleaning process. Their farms have an
average area of 20 hectares of coffee under organic
management. They pointed out that the domestic
market is still inactive and paying lower prices. They
have achieved good export prices, because they invest
heavily in quality and created dedicated channel
marketing, i.e., their export business.
In Rondnia State, much of the marketing of coffee
is made through the Mutual Aid Associations
(ACARAM: Joint Central Rural Associations for
Mutual Aid), present in more than a dozen
municipalities. These associations receive and store
coffee, and generally sell it for a large company in the
Paran State.
In Mato Grosso do Sul State, there were 200
families associated with APOMS (Association of
Organic Farmers in Mato Grosso do Sul), but only 20
were dedicated to organic coffee and the volume
produced by all of them is between 1,500 and 3,000
bags. They do not export their coffee and they passed
over IBD, due to the high cost to them and because
they do not provide any support or guidance. They sell
their coffee to the price of conventional product,
basically in the local market in trade fairs and coffee
houses. The harvest is manual and selective, so they
hire day laborers during this the harvest time.

3.2 Production Costs of Organic and Conventional
Coffee
The analysis of production costs is a key
management tools available to farmers. It allows
growers to consolidate the monitoring of farming,
streamlining production, and represents a tool to
specific policies for the coffee.
In Table 2, we present the results of two ways of
calculating the cost of organic coffee production. First,
we do not consider the remuneration of family work,
and the gross profit represents the remuneration of
family labor and remuneration of agricultural activity.
Second, we consider the remuneration of family labor
as a value similar to that of daily workers hired in
each region, and net income represents only the
remuneration of agricultural activity.
A first aspect to note is that farmers in agroforestry
systems have very low cost for supplies, which are
restricted mainly to spending on fuel and bags.
Producers needed to spend a little more with
inputsespecially in Minas Gerais in order to achieve
certain productivity with profitability showed higher
production costs. In Esprito Santo, Rondnia and
Mato Grosso do Sul, taking into account differences in
the cultivated area, the farmers faced higher costs in
farming operations. Also, the results indicate that net
income per ha generally decreases sharply when the
remuneration of family labor is added to the cost of
production. In one case examined, the result indicates
a loss.
4. Conclusions
There is not a standard for organic coffee
production in Brazil. The literature available on coffee
production and informations obtained from the
farmers interviewed in this study indicate different
conditions of production and marketing. The
producers have different levels of access to technical
information, especially on the combinations of inputs
29
Table 2 Cost of production of organic and conventional coffee, Brazilian regions, with remuneration of family labor or not,
2009-2010 crop, yield (bags/ha), total cost, total income (R$/ha), gross profit (R$/ha).
Brazilian
States
Production
systems
Organic
Minas
Gerais
Conventional
Organic
So Paulo
Conventional
Rondnia
Organic
Conventional
Organic
Esprito
Santo
Conventional
Mato Grosso
Organic
do Sul
Yield bags/hectare Remuneration of family labor

(ha)
Yes
No
X
28
X
X
41
X
X
15
X
X
10
X
39
X
05
X
20
X
20
X
22
X
20
X
20
X
49
X
X
15
X
X
15
X
X
15
X
25
X
15
X
X
35
X
30
X
14
X
15
X
30
X
20
X
15
X
16
X
15
X
14
X
X
12
X
Total cost Real (R$)*

R$/ha
R$/bag
3,976
142
4,592
164
4,018
98
17,384
424
3,630
242
3,705
247
1,560
156
1,760
176
7,137
183
2,340
468
4,940
247
5,980
299
7,634
347
6,440
322
6,420
321
8,036
164
1,020
68
2,340
156
1,095
73
1,890
126
1,200
80
2,175
145
6,450
258
4,305
287
3,045
87
3,325
95
3,570
119
2,604
186
2,610
174
3,600
120
2,800
140
3,645
243
4,336
271
2,610
174
3,808
272
1,020
85
3,348
279
Total income
R$/ha
12,183
12,183
17,853
17,853
6,150
6,150
4,720
4,720
16,684
2,400
6,000
5,600
6,006
5,600
5,600
17,333
3,750
3,750
1,875
1,875
3,750
3,750
7,625
4,200
6,300
6,300
5,100
5,320
4,650
3,800
3,360
4,050
3,840
3,300
4,130
8,640
8,640
Gross profit
R$/ha
8,207
7,591
13,835
469
2,520
2,445
3,160
2,960
9,547
60
1,060
-380
-1,628
-840
-820
9,297
2,730
1,410
785
-15
2,550
1,575
1,175
-105
3,255
2,975
1,530
2,716
2,040
200
560
405
-496
690
322
7,620
5,292
Real: Brazilian currency; Source: Data from the study.
to be used. In addition, soil and climatic differences,

intrinsic to a country of continental dimensions, like
Brazil, lead to different micro-regional or regional
results.
We can summarize the types of organic coffee
producers in Brazil as follows:
Farmers without capital to invest and do not depend

on coffee culture. Coffee results in additional revenue,
but the producer depends, for instance, on honey, milk
or banana production. In many cases, the coffee
culture does not receive any treatment, it is just
harvested, an extractive system. These producers
30
generally sell its product in local or regional fairs and

the coffee is not always of good quality. They may be
certified or not.
Family farmers without capital to invest, but that
depend on coffee plantations. They can not invest in
buying the ideal volume of inputs for farming, but still
use some compost or organic products to control pests
and diseases allowed in the organic system. They are
not expensive in manpower, but hire workers
especially in harvest time. Some can do a good job of
post-harvest and get good quality coffee. They may be
certified or not.
Small producers, which use higher levels of
technology, increasing volumes of inputs, greater care
in post-harvest, which has one or two contract
employees, and sometimes hire more workers at
harvest time, or even who does not contract
employees but hire day laborers for the harvest. Most
of these producers is certified and is associated with a
cooperative and can export the coffee regularly.
Producers of medium size, in general, they have
high production costs due to the high cost of
harvesting. Many of those who used the manual
harvesting system broke in recent years. They need to
invest in mechanized harvesting, but in general they
do not produce volumes of coffee to justify this
investment.
Large producers, farms export various different
kinds of coffee. In general, this type of producer has
organic coffee fields only to have this type of coffee
among the products offered by the company, as
propaganda for social and environmental concern and
not because they are economically viable.
Large producers, who produce coffee with a profit,
but do not worry about the size of that profit; they do
not rely on coffee for survival and have other sources
of income. Organic production is a philosophy of life.
Generally, coffee quality is good, they are not
associated with cooperatives and they export
individually or selling for large companies roasters.
One aspect to be highlighted in the organic system
is the issue of inputs: although the use of natural

fertilizer from other agricultural products of the farm,
constitutes one of the pillars Organic Agriculture
definition, much of the Brazilian coffee growers are
monoculture. Some of them produce coffee in
association with fruit and even in association with
other agricultural activities, but they have no
knowledge about the most appropriate combinations
of fertilizers and biofertilizers allowed by certifiers.
Although in some regions, the average productivity of
organic coffee is around 15 bags per hectare, the
regions where we observe increased use of fertilizers
(biofertilizers), recommended by certification, but not
natural, as the Minas Gerais State, are observed high
levels of productivity, sometimes, around 38 or 40
bags per hectare.
The main conclusions of the study can be
summarized as follows:
The production of organic coffee can be
economically viable in Brazil, mainly in the following
structural conditions;
Small producers who do not rely on hand
labor-hired (family farms);
Small producers who hire fewer workers (one or
two employees), and preferably only at harvest time;
Organic producers, with rare exceptions, do not
receive technical assistance and technological
information in general. Most of them are self-taught.
The differences in yield obtained by farmers are
significant. The objectives of each producer are also
different. Nevertheless, it is considered that the main
problem for organic production is not technological.
The primary limitation to the sustainability of
organic production in Brazil is the difficulty of
marketing:
Producers who are cooperative membership have
better marketing opportunities, because the
cooperatives strengthen the market access, especially
for the international market;
The coffee produced must have excellent standard
quality, so crop management and post-harvest
processing should be extremely careful: the quality of

coffee is as important as its own certification seal;
The difficulty to get homogeneous coffee shipments
and to maintain the quality of each crop year after year,
make it difficult for international sales;
The coffee that is not exported could get higher
price than conventional coffee, in the domestic market,
through the development of this niche of market,
currently limited;
The dual certifiedFair-trade and organicresults
in better prices, greater integration into the
international market and increases stability for the
coffee producer.
The main limitation to the sustainability of organic
systems, for medium and large farms, is the cost of
manpower. In areas with better soil and climatic
conditions for coffee production, the South of Minas
Gerais and Mogiana, in So Paulo State, dominated by
mountain coffee production, where machines
operations are not viable and the cost of manpower is
very high.
The yield of organic coffee system is generally
lower than the conventional. The difference between
them depends on the crop management, post-harvest,
characteristics of soil, temperature and rainfall,
especially the natural fertility of soil and altitude.
The organic system can be enhanced through
integrated management with other crops (like bananas)
or other agricultural activities (such as beekeeping and
dairy cattle), provided with manpower family or fewer
employees.
The agroforestry systems have great potential in
relation to the productive capacity and income
generation. However, lack support to producers and,
especially regarding the dissemination of information
and credit for its implementation. Such a system could
assist in the sustainable development of areas that are
highly disturbed by man.
As recommendation, programs to support
sustainable coffee production in Brazil should
prioritize:
31
Sustainable production systems, whether organic or

not, because the definition of sustainable agriculture
does not imply exclusively organic production;
Family and small farmers located in regions with
soil and climatic characteristics suitable for coffee
production, such as the southern region of Minas
Gerais State and Mogiana region, in So Paulo State;
Family farming, agro-forestry system, in highly
degraded areas by human activities, with marginal soil
and climatic conditions for growing Arabica coffee, as
the Western region of the So Paulo State (Alta
Paulista);
Access to technological and market information, for
achieving higher productivity and, especially, higher
quality (organic management, harvesting and post
harvest), to meet the requirements of the international
market (minimum volumes, homogeneous coffee
shipments, excellent quality of coffee grain and drink);
The approximation between Brazilian organic
coffee producers and buyers, especially international
buyers;
The promotion of organic coffee in the domestic
market, seeking alternative market and higher prices
for coffee that is not exported, once today, much of
this coffee is sold on the domestic market at the price
of conventional coffee.
The access to sources of credit: dissemination of
information on available sources, support to prepare
the documentation required by funding agencies or
implementation of alternative sources.
Incentive certifications, which allow simultaneous
access to other niche markets, beyond organic, as well
as Fairtrade and the geographical indication.
In summary, the promotion of organic coffee
production needs to be done very carefully, because
organic agriculture is sustainable for some situations
and for very specific Brazilian regions. We can not
encourage the cultivation of organic coffee without
analyzing each producer (or producers association) in
particular, because there is strong risk of the producer
to be financially distressed.
32
Acknowledgments
Tierra Nova Fonds, Netherlands, the funding for the
study.www.tierranova.nl.
[8]
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p. 14.

DAVID
PUBLISHING
Enhancing Maize Grain Yield in Acid Soils of Western

Kenya Using Aluminium Tolerant Germplasm
Ouma Evans1, Ligeyo Dickson2, Matonyei Thomas3, Agalo Joyce1, Were Beatrice3, Too Emily3, Onkware
Augustino3, Gudu Samuel4, Kisinyo Peter4 and Philip Nyangweso1
1. Department of Biological Sciences, Moi University, P.O. Box 3900-30100, Eldoret, Kenya.
2. Kenya Agricultural Research Institute (KARI) Kitale Centre, P.O. Box 460-30200, Kenya
3. Department of Biological Sciences Chepkoilel University College, P.O. Box 1125-30100, Eldoret, Kenya
4. Department of Agronomy, Rongo University College, P.O. Box 103-40404, Rongo-Kenya
Abstract: Maize (Zea mays L.) is one of the worlds most important cereals and is a staple food for many people in developing
countries. However, in acid soils (pH < 5.5), its productivity is limited by aluminium (Al) toxicity, besides other factors. The
objectives of this study were to: develop Al tolerant maize inbred lines for a maize breeding program in Kenya, develop single cross
hybrids (SCHs) from some of the tolerant inbred lines and determine Al tolerance levels of the SCHs. One hundred and seventy five
inbreds and 49 SCHs were developed and screened in nutrient culture containing 0 or 222 M using Relative Net Root Growth
(RNRG), hematoxylin staining (HS) and under Al saturated field conditions (44%-45.6%) at Sega and Chepkoilel. Seedling root
growth was inhibited in 95% of the inbreds. F1 hybrids obtained from inbreds varying in Al tolerance, exhibited tolerance equal to or
greater than that of the more tolerant parent indicating a positive transgressive inheritance to Al toxicity. Fifty eight percent of the F1
SCHs were heterotic for tolerance to Al toxicity. Al tolerance estimated by RNRG was well correlated to that of HS (r2 = 0.88, P <
0.005) but minimally correlated with the field estimates (r2 = 0.24-0.35), implying that RNRG can predict field selection under Al
toxic soils by between 24% and 35%. Plant breeders should therefore employ both approaches in selecting cultivars under Al stress.
This study has developed and identified Al tolerant inbreds and SCHs for use in the acid soils of Kenya and similar regions.
Key words: Maize, inbred lines, hybrids, heterosis, aluminium toxicity, acid soils.
1. Introduction
Aluminium (Al) toxicity and low available P are
some of the most limiting plant growth factors on
most acid soils worldwide [1]. Highly weathered acid
soils occupy 40% of the worlds arable soils [2]. They
are found mainly in South America (26.7%), North
America (19.4%), Africa (19.1%) and Asia (15.1%).
The rest occur in Australia and New Zealand, Europe
and Central America [3]. On highly acidic soils, (pH <
5.5), the rhizotoxic aluminum species, Al3+ is
solubilized, inhibiting root growth and function in the
majority of crops [4]. Al toxicity limits plant growth
Corresponding Author: Ouma Evans, Ph.D., research fields:
plant breeding and genetics. E-mail: ochivao@yahoo.com.
mainly through its adverse effects on root growth and

development [5]. In addition, it increases drought
susceptibility and limits plant access to subsoil
nutrients, which restricts the full expression of the
genetic potential of the plant [6]. According to Giller
et al. [7], Al toxicity reduces the agronomic and
recovery efficiencies of nutrients such as P by plants.
As a result, crops grown in tropical acid soils with
high Al toxicity can only recover and utilize between
10% and 25% of the P fertilizer applied due to its high
fixations by Al and Fe oxides [8]. The level of Al
saturation in Kenyan acid soils ranges between 20%
and 45% which is too high for most crop species to
tolerate [9]. According to these authors, most
34
Enhancing Maize Grain Yield in Acid Soils of Western Kenya Using Aluminium Tolerant Germplasm
improved maize varieties and landraces grown by

farmers are sensitive to high Al saturation (> 20%)
commonly found in most maize growing areas in the
region. This implies that such germplasm are unable
to efficiently utilize the native soil phosphorus (P) or
added P fertilizer as a result of reduction in root
growth due to Al toxicity [10]. Moreover, these
farmers incur up to 16.8% grain yield loss due to Al
toxicity [11]. Acid soils cover over 13% of maize
growing areas in Kenya [12]. In these areas
(especially the marginal rainfall medium altitude
areas), maize yields are very low, with averages of
1.0-1.5 t ha-1 compared to the research potential of
over 5.0 t ha-1 in the same regions [13]. Al toxicity is
partly responsible for the declining yields.
Conventionally, acid soils are mainly managed by
liming the top soil layer to neutralize the exchangeable
Al [14]. Besides, the use of lime is highly
recommended for the management of acid soils in
Kenya [12]. Lime reduces the levels of exchangeable
Al3+, Fe3+ and Mn4+ in acid soils and thus reduces P
sorption. This makes both the native soil P and
applied P fertilizers available for plant uptake [15].
Besides, lime is known to have longer residual effects
on acid soils compared to other soil amendments such
as organic and inorganic materials [16]. However, the
adoption of such input technologies has largely been
restricted to large scale farmers who can afford them
despite the fact that such technologies would be best
suitable for low input agriculture practiced by small
scale farmers in the maize ecosystems of Kenya. For
example, most resource-poor small holder farmers,
who are also the majority in the acid soil areas of
Kenya where maize is grown, have hardly adopted
such technologies due to lack of credit and the relative
high cost [17]. The two main sources of lime in Kenya
(Homa and Athi lime) are located approximately 250
km away from the major maize growing regions in the
country, where Al toxicity is a problem. This makes it
expensive to transport the large tonnage of lime
needed to mitigate Al toxicity in these regions.
Furthermore, the few farmers who apply lime do not

apply the recommended rates; hence this approach has
been ineffective in managing Al toxicity in these
regions [13].
There is therefore a challenge and need for
alternative, affordable and integrated approaches in
the management of the problem of Al toxicity in order
to increase maize productivity among the small holder
farmers in the marginalised areas of Western Kenya.
Selection, development and utilization of Al-tolerant
maize genotypes, together with minimal inputs, are
proposed as potentially sustainable and viable options
for managing Al toxicity in such regions.
Screening of maize genotypes in nutrient solution
using Relative Net Root Growth (RNRG) and
hematoxylin staining (HS) has been successful over
the past decade in selecting Al tolerant and sensitive
genotypes [18-20]. Root staining with hematoxylin
solution is a quick, rapid, efficient and reliable method
of discerning among Al-tolerant and Al-sensitive
maize genotypes since it is highly specific to Al
accumulation [20]. The method allows for rapid
evaluation of a large number of genotypes without
destroying the root apical meristem [21]. Besides,
field screening is one of the most direct screening
methods for tolerance to Al toxicity in cereals as it
allows a direct measurement of tolerance [22].
Accordingly, this study adopted these approaches in
assessing various maize germplasm for tolerance to Al
toxicity.
Genetic variation for aluminum (Al) tolerance in
crop species can allow the development of cultivars
that can give high yields when grown on acidic soils
with high Al toxicity problems. In fact, such traits
have been used to develop high-yielding, Al-tolerant
maize hybrids for use in acid soils [23]. Kenyan
farmers who grow maize on Al toxic soils do not yet
have access to such cultivars. Earlier screening of
Kenyan maize germplasm for Al toxicity showed that
some of the Kenyan landraces are tolerant [24]. This
study focussed on: developing maize inbred lines from
various sources including landraces and Brazilian

introductions which contained CATETO (Al-tolerant
Brazilian inbred line); selecting some of the inbred
lines for tolerance to Al toxicity; using them to
develop single crosses and testing the Al-tolerance in
the single crosses.
Caroline II mating design as described by Comstock

and Robinson [25]. A total of 49 single crosses were
developed. One of the single crosses, however, did not
yield enough seeds and was therefore not included in
soils of Western Kenya (HD614) were therefore tested
Maize germplasm used in this study were developed

from various sources: Kenya Agricultural Research
KARI-Kakamega
and
KARI-Muguga. Others were Brazilian introductions to

Kenya (single crosses) and derivatives of CATETO
(Brazilian most Al tolerant inbred line) while the rest
were local collections including Al tolerant 203B
landrace, collected from Al toxic soils of Muranga
county in central Kenya. All the sources were obtained
in the year 2002 and were used to develop 175 inbred
lines between the year 2003 and 2007 (Table 1). The
inbred lines were either developed from single cross
hybrids from the various sources or from topcrosses of
these single cross hybrids crossed with the Kenyan
testers for medium and high altitude. All the sources
were individually selfed to F6 to obtain the respective
inbred lines which were screened for tolerance to Al
toxicity in nutrient culture solution according to
Magnavaca et al. [18] and also under field conditions
(0 t ha-1 and 4 t ha-1 of lime).
Fourteen inbred lines were selected for tolerance to
Al toxicity based on relative net root growth (RNRG),
hematoxylin staining (HS) and grain yield at high Al
saturation (43.1%-45%) (data not shown). The single
Table 1
the selected Al tolerant inbred lines using North
and one commercial variety grown under Al toxic
2.1 Genetic Materials Used
(KARI)-Kitale,
cross hybrids were then generated in 2009 by crossing
the screening work. Forty-eight single cross hybrids
Institute
35
for tolerance to Al toxicity in nutrient solution culture.

CON 5, 203B and K4 were used as Al tolerant checks
while SCH 3 and REGNUR 0114 were used as
susceptible checks [11].
2.2 Description of Experimental Sites
Chepkoilel site is located at 0o3437.24N;
35o1510.04E, 2,143 m above sea level (a.s.l), and
has between 900 and 1,100 mm rainfall with a 10-26
o
C temperature range. The soils are chromic ferralsols
characterized by low pH 4.8, and Al saturation of
45.6% with P levels of 4.4 mg P kg-1 of soil [13]. Sega
site is located at 0o15N and 34o20E. It has an
elevation of between 1,140 and 1,400 m (a.s.l) with a
bimodal annual average rainfall pattern of between
800 and 1,200 mm. The mean minimum temperature
ranges between 15 and 17 oC, while the mean
maximum range is 27-30 oC. The soils are
OrthicAcrisols characterized by low pH 4.5 and a
mean Al saturation of 43.1% and 2.2 mg P kg-1 of soil
[13].
2.3 Experimental Design and Procedures
Seeds of each line were surface sterilized in 1%
sodium hypochlorite and rinsed thoroughly with sterile
Description of maize inbred lines used as parents of the single cross hybrids.
Original source of germplasm

Brazilian single crosses
Landrace (203B)
KARI-Muguga lines
KARI-Kakamega lines
KARI-Kitale lines
Al standards from Kenya and Brazil
No. of inbred lines developed from various sources

95
34
18
14
14
5
36
distilled water to remove all traces of the hypochlorite.

The seeds were set to germinate inside paper rolls
moistened with aerated distilled water. These were
placed vertically on plastic trays covered with
aluminium foil, which were incubated in darkness for
three days in a growth chamber set at 26 3 C. The
experiment was conducted at the Botany laboratory in
Chepkoilel University College. The setup was a
completely randomized design (CRD) replicated three
times. Treatments consisted of single cross maize
hybrids (49) or inbred lines (175) and two levels of Al
(0 M or 222 M Al). Eight litre trays were used to
hold nutrient solution under continuous aeration.
The nutrient solution was prepared according to
Magnavaca et al. [18]. Three days old uniform-sized
seedlings with no visible injury or damage on their
roots were transferred to the cups on a perforated
styrofoam sheet and stabilized for 24 h in nutrient
solution without added Al at pH 4.0 after which the
Initial root length (IRL) was measured. The seedlings
were then transferred to fresh nutrient solution where
Al was added to the trays as Al K (SO4)2 12H2O to
attain the stated concentration which corresponds to
free Al3+ M activities of (0) and (39) respectively
[26]. The seedlings were then grown in a growth
chamber at a photoperiod of 14 h of light and 10 h of
darkness. The day length growth room conditions
were approximately 340 mol photons m-2 s-1 of light
intensity, 30 2 C and 70% relative humidity; the
dark conditions were 22 2 C and 90% relative air
humidity.
Seventy two hours after transplanting, final seminal
root length (FSRL) was measured and the net seminal
root length (NSRL) calculated from the difference
between FSRL and initial seminal root length ISRL
[18]. The tolerance level was assessed using relative
net root growth (RNRG), where,
RNRG = NSRL under Al treatment/NSRL under
control 100
(1)
The heterosis for the F1 single crosses was
calculated using both mid-parent heterosis (MPh) and
high parent heterosis (HPh) for comparison [27]. The

two indices were expressed in percentages as:
F1 - M
(2)
MP% =
100
MP
F1- HP
HP =
100
HP
(3)
Where, F1 = performance of hybrid, MP = average

performance of both parents and HP = performance of
high parent.
Hematoxylin staining was used as a confirmatory
test for tolerance to Al toxicity in selecting the Al
tolerant inbred lines. The seedlings of 20 selected
(tolerant, moderately tolerant and sensitive) maize
inbred lines were subjected to hematoxylin staining as
described by Cancado et al. [20]. Visual scores for
root staining intensity were made on a scale of 1-5, as
follows: non-stained roots were classified as very
tolerant (Scale 1), faintly stained roots as tolerant
(Scale 2), moderately stained roots as moderately
tolerant (Scale 3), well stained roots as sensitive
(Scale 4) and those with deeply stained roots as very
sensitive (Scale 5) [20].
The experiment for screening inbred lines for
tolerance to Al toxicity under field conditions was set
up in a randomized complete block design (RCBD)
with 4 treatments in 3 replications at 2 sites. Some
plots received phosphorus (P) and lime (L) (P + L);
while others received either P (+P) or L (+L). The
control plot received neither P nor L. Phosphorus was
applied as triple super phosphate (TSP) at the rate of
26 kg P ha-1. Agricultural lime from Koru liming
company in Kisumu containing approximately 21%
CaO was applied 2 months before planting at the rate
of 4 t ha-1. CaO in the plots was to receive lime at each
site as recommended by Kisinyo et al. [13]. Planting
was done in March 2010 at Chepkoilel and Sega sites
at a spacing of 0.75 m between the rows and 0.3 m
within the row in a 3 m long plot comprising 2 rows
each. Nitrogen was used in top dressing six weeks
after planting on all the plots in the form of calcium
ammonium nitrate (CAN) at the rate of 75 kg N ha-1.
Weeding was done manually thrice and the crop
protected from stalk borer (Buseola fusca L.) damage

using 2-3 granules of Beta-cyhalothrin (Bulldock GR
0.05) at a rate of 6 kg ha-1 applied in the whorl of each
plant after thinning. Data was recorded on grain yield
(t ha-1) plant height (cm), ear height (cm), days to 50%
tasseling and days to 50% silking.
2.4 Statistical Analysis
The RNRG and hematoxylin staining data was
subjected to 1-way analysis of variance using the
General Linear Models procedure of Genstat and
means compared using Tukeys range test using the
following model:
Xijk = +i +ij
(4)
Where, Xijk: plot observation, : overall mean; i:

treatment effect; i: experimental error due to
treatments [28, 29]. Grain yield and yield component
data were subjected to 2-way analysis of variance by
fitting the following model:
Xijk = +i +j +ij
(29)
Where, Xijk: plot observation, : overall mean; i:
treatment effect; j: block effect; ij: experimental
error due to treatments and blocks [30].
Phenotypic correlation between RNRG and
hematoxylin staining and between RNRG and grain
yield were computed by regression and correlation
analysis, using Genstat software (Payne et al., 2009).
The regression and correlation were analyzed based
on the model:
Yi = o +iXi +i
(6)
where, Yi: the ith observation of the response Y; o:
population parameter giving the intercept; 1:
population parameter giving the slope; i: error term.
Correlation coefficient r was calculated using the
equation:
r=
COV X,Y
S X SY
(7)
where, COV (X, Y): Covariance X (predictor) and

Y = predicted parameter, Sx: standard deviation of the
predictor parameter; Sy: standard deviation of the
predicted parameter [31].
37

3.1 Phenotypic Variation for Tolerance to Aluminium
Toxicity among the Inbred Lines
Significant phenotypic variation (P > 0.05) in
tolerance to Al toxicity was observed among the
inbred lines based on an Al tolerance threshold of
50% RNRG (Figs. 1 and 2). Root growth inhibition
occurred in 95% of the inbred lines. However, root
growth in nine tolerant inbred lines (203B, 203B-14,
CATAL 237/67X63-5, CON 5, HASR, 203B-30, HS
53x280-16, HS 26x294-6 and 203B-15) remained
unaffected after exposure to 39 M Al3+ (Fig. 2).
Similar observations were reported in Sesbania
(Sesbania sesban (L.) Merr, sorghum (Sorghum
bicolor (L.) Moench and in maize but at lower
concentrations of between 148 and 200 M [32, 33].
Such resistance is partly a result of maintaining cell
wall and plasma membrane integrity [34]. Landrace
203B which was used as one of the tolerant standards
(Fig. 3a) had the highest root growth followed by
some of its derivatives, such as 203B-14 and 203B-39.
However, other inbred lines derived from the same
landrace (203B-25 and 203B-28) were among the
most Al sensitive lines. These results imply that these
lines could have initially received pollen from other
Al sensitive lines owing to the out crossing nature of
maize since the starting material was an open
pollinated variety (OPV) and hence such segregants
could have emerged. The 203B landrace and its inbred
lines remains an invaluable source of Al tolerance
which can be exploited in production of acid tolerant
maize varieties.
CON 5, which was used as another Al tolerant
standard, expressed a RNRG of 105% under similar
conditions compared to 203B, 203B-14 and others.
CON 5 is an elite homogenous population from KARI
which has been classified as Al tolerant [23]. A study
by this author indicated that 55% of tolerance to Al
toxicity in CON 5 is attributed to exclusion of Al from
the root tips owing to the activity of ZmMATE1 gene.
The highly tolerant CATAL 237/67XL3-5 is a
38
Number of individuals
35
30
25
20
15
10
5
110-119
100-109
90-99
80-89
70-79
60-69
50-59
40-49
30-39
20-29
9-19.
RNRG (%)
Fig. 1 Percent of relative net root growth (RNRG) frequency distribution for maize inbred lines. The double arrowed line
depicts the threshold for Al sensitivity (RNRG < 50%) and tolerance (RNRG > 50%). 175 maize inbred lines were grown in
nutrient solution containing M Al3+ for three days.
Fig. 2 Relative net root growth of selected 20 inbred lines after 3 days of exposure to Al treatment. Percent of relative net
root growth (RNRG) values are the means of three replications (seven plants per replication). The error bars are standard
error bars (SE). Selection was based on clustering of the means of 175 inbred lines into three homogenous categories; the
inbreds therefore represented each of the categories.
(b) SCH3 inbred line
(a) 203B inbred line
O M Al
Fig. 3
222 M Al
O M Al
222 M Al
a, b: Root growth response to Al stress by inbred line 203B and sensitive inbred line SCH3.
39
derivative of CATETO, the Brazilian Al tolerant

standard. Studies have shown that CATETO has high
expression of ZmMATE1, the Al tolerance gene [23].
This suggests that CATAL 237/67XL3-5 may be using
a similar Al tolerance mechanism as CATETO. Studies
on CATETO have indicated that two genes
(ZmMATE1 and ZmMATE2) co-localize to major Al
tolerance Quantitative Trait Loci (QTLs) in maize [23].
As to whether the Al tolerance in 203B, CON5 and K4
is as a result of ZmMATE allele or a separate gene is
yet to be determined. Interestingly, studies by Matonyei
[23] showed that CON 5, 203B and some of its
derivatives were apparently more tolerant than
CATETO, even though, they expressed lower
ZmMATE1 activity than the latter. These findings
clearly point to the possibility that the Kenyan sources
could have a different gene in play. The least root
growth response (17%) was observed in inbred line
SCH3 (Al sensitive line from Brazil) (Fig. 3b) as
expected.
The correlation between RNRG and hematoxylin

staining showed a negative trend (Fig. 5) probably
because sensitive seedlings have low RNRG as a
result of high quantities of accumulated aluminium in
the root cap and, therefore, they normally show high
hematoxylin adsorption rate. The tolerant genotypes
have some mechanisms to avoid aluminium toxicity,
therefore, they express higher RNRG and lower
hematoxylin absorption rate. These findings are in
agreement with those of Cancado et al. [20] who
reported a strong negative correlation (r = -0.693 and
-0.816) between hematoxylin absorption rate (HS),
NSRL and RNRG, respectively.
A regression analysis of RNRG on the hematoxylin
adsorption rate indicated that 88% of all the observed
variance in tolerance could be explained by
hematoxylin adsorption rate. Therefore, the
colouration of the root apices with hematoxylin can be
employed, without restriction as an informative index
of Al tolerance.
3.2 Variations in Staining Rate of Hematoxylin in

Maize Inbred Lines
3.3 Performance of Inbred Lines under Field

Condition and Correlations with Al Screening Data
The inbred lines differed significantly with regard

to hematoxylin staining adsorption when subjected to
Al stress. Al tolerant lines had lower adsorption rate
(< 3) compared to the sensitive ones ( 4). The very
sensitive line, A089, showed an intense dark-blue
coloration indicating deeply stained roots, the
sensitive line REGNUR 00114 showed blue
coloration in the roots indicating well stained roots,
while the tolerant line CATAL 237/67XL3-5 showed
clear root apices, i.e., non-stained roots (Figs. 4a and
b). These findings compare well with previous
observations in pea roots [36], maize roots [20, 21]
and in rice [37]. According to these authors, the
sensitive lines tend to accumulate more Al in their root
tips, hence adsorbing more hematoxylin stain. These
results into the blue coloration compared to the tolerant
lines which do not bind the hematoxylin stain and
exclude Al from the cells.
At Sega, under control (No P, No L), the inbred

lines produced grain yields of between 0 and 2.4 t ha-1.
However, with the addition of lime (4 t ha-1), the grain
yield increased to between 0.4 and 3.9 t ha-1. Under
control (no P, no L), majority of the inbred lines (70%)
expressed grain yields of between 0.0 and 0.9 t ha-1
while the rest yielded between 1.0 and 2.4 t ha-1 (Figs.
6 and 7).
Regression of grain yield under additional
phosphorus in Al toxic soils on percent RNRG
showed positive, but non-significant trend P 0.05
with coefficient of determination (R2 = 0.24 and 0.35)
for Sega and Chepkoilel sites, respectively (Fig. 8).
However, regression of grain yields under control on
percent RNRG also showed positive trend with lower
R2 values (R2 = 0.11 and 0.30) for Sega and Chepkoilel
sites respectively (data not shown). This showed the
extent of amelioration effects of additional P on
40
Fig. 4a
Mean hematoxylin staining (Hs) values of selected 20 maize inbred lines.
Fig. 4b Maize seedling root apices stained with hematoxylin stain after a 72 h exposure to 222 M Al in nutrient solution:
CATAL 237/67XL3-5tolerant; REG NUR 00114Sensitive; A089Very sensitive.
Fig. 5 Relationship between RNRG and hematoxylin staining of selected inbred lines after exposure to Al containing 222
M concentration for 3 days.
26 kgP/ha+ 4 t/ha lime
4 t/ha lime
26 kg P/ha
41
Control
Number of inbred lines
40
35
30
25
20
15
10
5
0
Inbred line grain yield ranges (t/ha)
Fig. 6
Trends in grain yield of maize inbred lines screened in Aluminium toxic soils at Sega.
Fig. 7
Effects of various treatments on maize growth at Sega site during the long rains of 2010.
Control
+Lime
(a) Sega site
+P
+L+P
(b) Chepkoilel site
Fig. 8 Relationship between grain yield with additional P in the field (26 kg P ha-1) and RNRG of maize inbred lines grown
with P in nutrient solution under Al stress (222 M Al).
tolerance to Al toxicity under field conditions. It also

showed that solution culture screening could predict
the response of maize cultivars when tested under Al
toxic soils culture by up to 35%, although this would
depend on available P and percent Al saturation in the
soil. These findings imply that plant breeders should
employ an integrated approach of using both solution
culture and field screening conditions when selecting
cultivars for tolerance to Al toxicity. The low
correlation between solution culture screening and

field screening could be due to higher interaction of
Al and P in nutrient solution since Al imposed in
nutrient solution, was higher than that found naturally
under field conditions.
These findings compareed well with those of Liao
et al. [36] who reported that P-efficient genotypes
were more Al tolerant than P-inefficient genotypes.
These authors suggested that P could help ameliorate
42
Al toxicity through Al complexation and possible

precipitation of Al in the rhizosphere, in addition to
the Al-P interactions in the root apoplast.
The coefficient of determination (R2 = 24%)
observed in Sega was much lower than the one
observed at Chepkoilel (R2 = 35%) probably because
of the lower available soil P levels at Sega (2.2 mg P
kg-1 of soil) compared to Chepkoilel (4.4 mg P kg-1 of
soil).
3.4 Phenotypic Variation for Tolerance to Aluminium
Toxicity among Single Crosses
The phenotypic expression of NSRL and RNRG
showed transgressive inheritance. The F1s showed
positive, negative and no heterosis (Table 2). Most of
the F1s (58%) were more tolerant to Al toxicity than
either of their parents (Table 3). This can be attributed
to heterosis for RNRG in which the hybrid F1
exhibited a RNRG that is superior to the means of the
two parents (mid-parent heterosis), or the better of the
two parents (better/high-parent heterosis) [39]. The

genetic basis of heterosis includes dominance, over
dominance or epistatic gene effects [40].
The remaining 42% of the single crosses were not
heterotic for RNRG. This observation could have
been due to negative transgressive inheritance
where the offspring performed worse than both
parents. HD614, a Kenyan commercial variety bred
for high altitude areas, was found to be among the
moderately tolerant accessions; however, 32% of
the single crosses developed were more tolerant
than this variety. The great genetic potential for Al
tolerance expressed in the F1 single crosses could be
exploited further to develop varieties (Double
crosses, 3-way crosses and synthetics) with
tolerance to Al toxicity. These may be more
attractive to farmers growing maize in the acid soil
regions of Kenya. Fig. 9 shows root growth
response of selected single cross maize hybrids in
Al stress.
Table 2 Mid-parent and high-parent heterosis of selected F1 single cross maize hybrids tested for tolerance to Al toxicity in
nutrient solution.
F1 Single crosses
NRL 0 M Al
MPh (%)
HPh (%)
13.85
-15
-3.8
10.6
-40
-43.2
72
32.8
-47.6
-64.5
134
87
51
15.8
4.5
-27.2
19.5
-13.4
116.4
108
44.9
-18.4
219
183.2
23.4
-18.3
110.8
100
-12.6
-14.2
79.6
65.5
65.3
35.8
108.3
95.2
88.6
69.9
NRL 222 M Al
MPh (%)
HPh (%)
134
92.7
57.5
29.9
-2.1
-20.6
115.4
67.7
36
-56.6
100
83.4
101.27
80.5
8.8
-27.3
-1.7
-4.4
146.9
134.1
35
14.2
237
209.5
7.5
-31
102.2
93.5
-4.3
-13.9
90.7
73.5
27.2
12.9
67.2
54.5
0
-9.3
MPh (%)
110.8
63.2
55.9
50
39.6
34
33.3
23.3
23
18.75
14
2.8
-0.8
-7
7.6
-10.7
-22
-22.5
-41
RNRG
HPh (%)
90.1
37.9
19.4
35.4
24.6
26.7
14.2
13.8
14.2
5.5
-8
-15.2
-12.3
-8.6
5.6
-19.4
-25
-22.5
-47
KML 036 MUL 863

S596-41-2-2 REG 007-361
KML 036 S396-15-1
MUL 863 MUL 1007
MUL 125 POOLB 26-1
MUL 817 MUL 863
MUL 817 MUL 216
MUL 817 MULX125
MUL 822 S558-2-2-3-7
CML 181 MUL 817
MUL 216 CML 202
KML 026 MUL817
MUL 125 MUL 863
REG N007-361 MUL 817
MUL 116 MUL 104
CML 181 REG N007-361
POOL B26-1 MUL 817
POOL A6-1 CML 202
MUL 817 S558-2-2-3-7
RNRG: Relative net root growth, NSRL 222 M Al Net seminal root length in Al at 222 M concentration; NRL0M A Net
seminal root length at no Al; MP%Percent mid-parent heterosis; HP%Percent high parent heterosis.
43
Table 3 Means for net root lengths, relative net root growth, root reduction and Al tolerance status of selected maize single
crosses and their parents.
Net root
Net root
Relative
Single crosses and Parents
Length
Length
Net root
0 M
222 M
Growth
37.8a-h
34.7j-o
0.97g
KML 036 MUL 863
25.0a-c
23b-m
0.92fg
KML 036 S396-15-1
KML O26
20.2a
16.7a-h
0.85e-g
53.1e-j
34.9k-o
0.84d-g
MUL 863 MUL 1007
23.8a-c
17.8a-i
0.81c-g
MUL 125 POOLB 26-1
48.8a-i
30.1g-n
0.74b-g
MUL 817 MUL 125
S558-27-2-1
29.4a-f
17.1a-i
0.68a-g
MUL 125
67.1h-j
41.4n-p
0.65a-g
57.2e-i
31.6i-o
0.64a-g
MUL 817 MUL 216
MUL 1007
39.9a-h
20.8a-k
0.62a-g
CML 202
23.7a-c
12.1a-d
0.62a-g
MUL 822
33.4a-g
18a-i
0.6a-g
POOL A6-1
27.1a-d
14.3a-f
0.6a-g
REG 007-361
23.7a-c
12.7a-e
0.58a-g
47.6a-i
26.9e-n
0.58a-g
CML 181 REG N007-361
52.6c-i
28.7f-n
0.58a-g
MUL 817 REG 007-361
40.3a-h
20a-j
0.57a-g
MUL 216 CML 202
54.8d-i
28.5f-n
0.57a-g
MUL 125 MUL 863
MUL 817
26.3a-d
13.9a-f
0.56a-g
44a-h
22.8b-l
0.56a-g
MUL 116 MUL 104
57.3e-j
28.2f-n
0.55a-g
MUL 125 MUL 1007
64.3h-j
33.2j-o
0.53a-f
MUL 228 MUL 216
MUL 116
49.6a-i
21.2a-k
0.53a-f
52.7c-i
26.9e-n
0.53a-f
REG N007-361 MUL 817
POOL B26-1
37.4a-h
14.4a-f
0.52a-f
52.9c-i
22.1b-l
0.48a-e
POOL A6-1 CML 202
S558-2-2-1-4
87.1j
37.8m-p
0.43a-d
50.9b-i
20.1a-k
0.42a-d
POOL B26 - 1 MUL 817
KML 036
44.3a-h
18.3a-k
0.42a-c
S596-41-2-2
25.3a-d
8.5ab
0.41a-c
MUL 216
49.4a-i
17.5a-i
0.41a-c
49.6a-i
17.9a-i
0.39a-c
POOL A6-1 S558-2-2-1-4
48.1a-i
15.5a-g
0.37ab
MUL 817 S558-2-2-3-7
REG NUR-00114
23.5a-c
7a
0.32a
Grand mean
42.6
23.3
0.62
SE
0.8
0.4
0.01
Means in the same column followed by the same letter are not significantly different at P 0.05
Ttolerant to Al toxicity; MTmedium tolerant to Al toxicity; Ssensitive to Al toxicity
4. Conclusions
There is a wide variation for tolerance to Al toxicity
among the inbreds and the single crosses. Using this
variation, this study has developed both Al tolerant
inbred lines and single crosses from diverse sources.
Percent
Al
Root
Status
Reduction
3.2a
T
7.6 ab
T
14.6a-c
T
15.7a-d
T
19.3a-e
T
25.9a-f
T
32.2 a-g
MT
34.5 a-g
MT
35.8 a-g
MT
37.8 a-g
MT
38.4 a-g
MT
39.5 a-g
MT
39.7 a-g
MT
41.6 a-g
MT
41.9 a-g
MT
42.3 a-g
MT
43.1 a-g
MT
43.1 a-g
MT
43.6 a-g
MT
44.4 a-g
MT
44.8 a-g
MT
47.1 b-g
MT
47.1 b-g
MT
47.4 b-g
MT
47.9 b-g
MT
52.2 b-g
S
57.1 c-g
S
57.9 d-g
S
58.4 e-g
S
59.2 e-g
S
59.2 e-g
S
61.1 e-g
S
63.3fg
S
68.3g
S
38
S
1.1
according to Tukeys range test.
Nutrient culture screening for Al toxicity can predict

field selection under Al toxic soils by between
24%-35% depending on the Al saturation of the
particular soil and the levels of available phosphorus.
This implies that plant breeders should employ an
integrated approach of using both solution culture and
44
KML 036 MUL 863
REG NUR-00114
O M Al
222 M Al
O M Al
222 M Al
Fig. 9 Root growth response to Al stress by the sensitive Al standard (REG NUR-00114) and the most tolerant Single cross
(KML 036 MUL 863).
field screening conditions when selecting cultivars for

tolerance to Al toxicity. Some of the Kenyan inbreds
identified in this study were more tolerant than the
inbreds derived from CATETO. These include 203B
and some of its derivatives which remain the most Al
tolerant genotype among Kenyan maize germplasm.
Additionally, some of the single cross hybrids
identified in this study showed superior tolerance to
Al toxicity and could be used directly or as parental
material for future hybrids for acid soils. They include:
KML 036 MUL 863, KML 036 S396-15-1, MUL
863 MUL 1007, MUL 125 POOLB 26-1, MUL 817
MUL 125.
Acknowledgments
The authors acknowledge financial support from
Generation Challenge (GCP) project, KARI and
EMBRAPA for providing the initial germplasm for
the study and Moi University for the Research
facilities used to conduct the research.
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2236-2239.

DAVID
PUBLISHING
Response of Peach (Prunus persica) cv. to Foliar

Application of Potassium and Copper
Shawkat Mustafa Mohammed Al-Atrushy and Sarfaraz Fatah Ali Al-bamarny
Department of Horticulture, Faculty of Agriculture and Forestry, University of Duhok, Iraq

Abstract: The experiment was carried out in a private orchard situated at Atrush town, Duhok Governorate Kurdistan Region-Iraq
during 2011 season to study the effect of foliar application of three concentrations of potassium (0.0%, 0.5% and 1%) and three
concentrations of copper (0.0%, 0.02% and 0.04%) on an eight years old peach cultivar, planted in clay soil, spaced at 4.0 m 4.0 m.
The foliar application was done twice, on April 24, 2011 and May 25, 2011. The results showed that spraying of potassium at both
concentration (0.5% and 1%) or copper at high concentration (0.04%) had a positive effect on leaf area, leaf fresh weight, leaf dry
weight, total chlorophyll, fruit weight, fruit number and yield per tree as well as fruit diameter, pith thick, pulp weight, seed weight and
total soluble solid. The interaction between potassium and copper significantly enhanced all detected traits, since trees receiving 1% and
sprayed with 0.04% of copper was characterized by the highest values of all growth and yield characteristics compared to the lowest values at
untreated trees (control).
Key words: Potassium, copper, peach, Atrush.
1. Introduction
The peach tree (Prunus persica) is a species of
Prunus native to China that bears an edible juicy fruit
called a peach [1]. It is a deciduous tree growing to
4-10 m tall, belonging to Rosaceae family.
The scientific name Persica, along with the word
peach itself and its cognates in many European
languages, derives from an early European belief that
peaches were native to Persia [2]. The modern
botanical consensus is that they originate in China,
and were introduced to Persia and the Mediterranean
region along the Silk Road before Christian times [3].
All commercial cultivars belong to P. persica L.
Batsch, are primarily grown in temperate zones
between latitudes 30 and 45 N and S and in the
tropics and subtropics at higher elevation [4].
Potassium has many general functions in plants, it
is involved in the synthesis of proteins, where it plays
Corresponding author: Sarfaraz Fatah Al-Bamarny,
assistant professpr, research fields: pomlogy, fruit production
and improving fruit quality. E-mail: Sarfarazfatah@yahoo.com.
several roles, including the assisting in the transport of

amino acids to the sites of protein synthesis,
Potassium activates a number of enzymes and the
promotion of normal cell division and growth,
potassium improves health and resistance to disease
and adverse environmental conditions, like drought,
cold and flooding. Potassium also helps to regulate
both the electrical balance and the water balance
within the plant [5].
Copper, affects on plant physiology at wide ranging,
including interference with fatty acid and protein
metabolism and inhibition of respiration and nitrogen
fixation processes. At the whole plant level, Cu is an
effective inhibitor of vegetative growth and induces
general symptoms of senescence. Latter appears to
constitute the most widespread response of plants to
stresses provoked by metals, including Cu [6].
Numerous researchers studied the effect of various
concentrations of potassium and copper on peach in
aim to improve its yield and quality. Leece [7] studied
the effects of fertilizer nitrogen, phosphorus and
48
potassium on the leaf composition on Hale-haven

peach trees, on a sandy loam soil. Fertilizer nitrogen
increased the leaf concentrations of nitrogen, iron,
copper, manganese and zinc, and decreased the
concentrations of potassium, calcium, magnesium and
boron. Effects of fertilizer phosphorus and potassium
on leaf composition were very slight and would not
have been of practical importance in diagnosis by leaf
analysis. Awasthi [8] found that application of 500,
600 and 700 g K2O tree-1 progressively increased
significantly the fruit yield and weight; Potassium
application affected fruit quality by significantly
increasing total soluble solids and fruit sweetness and
decreased titratable acidity. Ahmed et al. [5] studied
the effect of adding potassium via foliage at
0.25%-1.0% and/or via fertigation at 0.25-1 g vine-1
on growth and fruiting of flame seedless grapevine.
Results showed that adding potassium via foliage or
fertigation was very effective in improving growth
characters and yield compared with control treatment.
Chatzitheodorou1 et al. [9] studied the response of the
peach cultivars Spring Time and Red Haven
grown in clay loam soil, to nitrogen, phosphorus and
potassium fertilizers, manure, as well some
combinations of these. They noted that significant
increase on fruit yield as well as on fruit quality and
fruit size of both cultivars. However, total soluble
solids content (%) of the cultivars Spring Time and
Red Haven did not alter significantly in comparison
to the control for all the fertilizer combinations used.
Mimoun et al. [10] studied the effects of potassium
foliar spray on peach cultivar Royal Glory grafted on
GF677 Rootstocks and the plum cultivar Black Star
grafted on Mariana rootstocks. At the beginning of the
season, result showed that the use of potassium foliar
fertilization increased fruit weight, total soluble solid
and improved fruit quality of Black Star plum and
Royal Glory peach, at harvest indicates that the fruit
maturity was earlier with the foliar application.
Al-Dulaimi [11] showed that foliar application of
copper at 10-20 mg L-1 to pear cv. Le-Conte caused
significant increases in growth characters as well as

yield and its physical and chemical characteristics
compared with control treatment. Al-Atrushy [12]
studied the effect of copper sulphate at 0.02 and 0.04 g
Cu L-1 on growth and yield of grapevine cv. Zark. His
results indicated that spraying with copper at 0.04 g
Cu L-1 was superior in all vegetative growth
characteristics, yield as well as physical and chemical
characteristics of fruits.
The objectives of this are to study the effect of
foliar application of potassium and copper on the
vegetative and yield of peach cv. Dexired under the
conditions of Atrush region conditions.

This experiment was conducted during 2011 season
on peach trees (Prunus persica L.) cv. Dexired. Trees
were selected to be as uniform as possible in vigor and
grown in a private orchard situated at Atrush town,
Duhok Governorate-Iraq. The trees were 8 years old
planted in clay soil spaced at 4.0 m 4.0 m, the trees
were trained on open central vase form system with
four primary scaffold branches. This experiment
included two factors; the first factor included the
following three concentrations of potassium (0%,
0.5% and 1%). The second factor was represented by
three concentrations of copper (0%, 0.2% and 0.4%).
So, the experiment involved nine various treatments.
A completely randomized blocks design was followed
in the experiment arrangement and every treatment
was done on the same trees. Every treatment consisted
of one tree per replicate with three replications, so the
number of trees used was 27 trees.
A detergent powder as wetting agent at 1-2 g L-1
was added to all the spraying solution including 0.0%
control to reduce surface tension of solution. The
sprays were done to drip point at two times on April
24, 2011 and May 25, 2011, using 16 L hand sprayer.
Horticultural practices were used as usual. Potential
effects of potassium and copper were evaluated in
terms of the change in growth tree; leaf area was
49
calculated by Leaf area meter [13], leaf dry weight,

chlorophyll content according to ASPD, yield and
fruits quality. All results were analyzed statistically by
using SAS programs [14]. Duncans multiple tests at
5% level of portability was to compare the treatment
according to Al-Rawi and Khalafalla [15].
due to the main role of potassium in the synthesis of
chlorophyll for trees sprayed with copper are superior
proteins and activates a number of enzymes and the

promotion of normal cell division and growth, which
are important components in the synthesis of
chlorophyll [2]. Data in the Table 1 also show that the
leaf area, leaf fresh and dry weights and total
significantly on that untreated. Highest values (14.72
3.1 Vegetative Growth Characteristics
cm2, 0.880 g, 0.488 g and 50.4%) respectively, were
Data presented in Table 1 shows that leaf area, leaf
obtained in trees sprayed with copper at 0.04%
fresh and dry weight and total chlorophyll for trees
compared with the lowest values (13.33 cm, 0.813 g,
sprayed with potassium are superior significantly on
0.424 g and 41.8%) respectively at untreated trees.
that untreated trees. The highest area leaf, fresh and
Various
levels
of
copper
were
also
differed
dry weight and total chlorophyll (15.17 cm , 0.881 g,
significantly among each others, except these of leaf
0.497 g and 52.4%) respectively, were given by
area do not show significant differences between the
spraying trees with potassium at level (1%) compared
two levels of copper. The reason behind the increase
with the lowest values (13.05 cm , 0.815 g, 0.426 g
of leaf area, leaf fresh and dry weight and total
and 40.9%) respectively at untreated trees. Various
chlorophyll with increasing copper levels could be due
levels of potassium were also differed significantly
to do the positive role of copper in the process of
among each others. These results are in agreement
photosynthesis, through the entering in the structure of
with what has been concluded by the researchers [7, 8].
proteins private chloroplast [12, 16] which considered
The significant effect of spraying potassium may be
a part of the cycle of electron transition that linking
Table 1 Effect of foliar application with potassium and copper on vegetative growth characteristics of peach (Prunus persica L.) cv.
Treatment
Potassium
(%)
0.0
0.5
1.0
Copper (%)
0.0
0.02
0.04
Potassium Copper
K 0 Cu 0
K 0.0 Cu 0.02
K 0.0 Cu 0.04
K 0.5 Cu 0.00
K 0.5 Cu 0.02
K 0.5 Cu 0.04
K 1.0 Cu 0.00
K 1.0 Cu 0.02
K 1.0 Cu 0.04
Parameters
Leaf dry
Weight (g)
0.426 c
0.450 b
0.497 a
Leaf area
(cm2)
13.05 c
13.95 b
15.17 a
Leaf fresh
Weight (g)
0.815 c
0.846 b
0.881 a
Total
Chlorophyll
40.9 c
46.0 b
52.4 a
13.33 b
14.12 a
14.72 a
0.813 c
0.850 b
0.880 a
0.424 c
0.462 b
0.488 a
41.8 c
46.9 b
50.7 a
12.30 e
13.10 de
13.75 cd
13.21 de
14.07 d-d
14.57 bc
14.49 bc
15.20 ab
15.83 a
0.759 d
0.833 bc
0.854 bc
0.825 c
0.851 bc
0.862 bc
0.855 bc
0.866 b
0.923 a
0.400 e
0.427 de
0.451 cd
0.425 de
0.456 cd
0.469 cd
0.445 cd
0.504 b
0.542 a
35.1 d
43.0 c
44.7 bc
40.1 cd
45.0 bc
52.9 a
50.3 ab
52.6 a
54.3 a
Means with the same letter are not significantly different according to Duncan multiple ranges test at 5% level.
50
both systems of photosynthetic reaction to the process

of photosynthesis [17] and the exploitation of these
materials in increase growth characteristics.
For the interaction, data in Table 1 show that the
highest value of leaf area, leaf fresh and dry weight
and total chlorophyll (15.83 cm2, 0.923 g, 0.542 g and
54.3%) were obtained in trees spraying with
potassium at 1% and copper at 0.04% compared with
the lowest value (12.30 cm2, 0.759 g, 0.400 g and
35.1%) in the untreated trees.
3.2 Yield and Fruit Characteristics
Data in Table 2 indicated that spraying of 0.5% and
1% of potassium, 0.02 and 0.04 of copper
progressively increased fruit weight and yield per tree
over amounts obtained with control. Fruit weight and
yield per tree increased significantly as potassium
application was raised from 0.5% to 1% and copper
from 0.02 to 0.04. The highest fruit weight and yield
were 99.71, 98.94 g and 20.10, 19.94 kg tree-1 with 1%
potassium and 0.04% copper, respectively. Negligible
increase was occurred on number of fruit per tree with
potassium and copper application. Whereas,

potassium application beyond 1% and copper beyond
0.04% only increased fruit diameter. Yield, fruit
weight and diameter and number of fruit per tree
increased with increasing the rate of application of
potassium may be due to the role of potassium in
activating meristematic growth, photosynthesis and
activates a number of enzymes, including those
involved in the synthesis of carbohydrates, and is also
involved in the neutralization of organic acids and the
promotion of normal cell division and growth [8, 10,
18]. The reason beyond the effect of copper on
increasing yield, fruit weight and diameter and
number of fruit per tree may be due to the role of
copper in influencing photosynthesis [19] and provide
important materials that have helped to improve the
qualities of fruits [20].
For the interaction, data presented in Table 2 clearly
show that the highest value of fruit weight, fruit
number, yield per trees and fruit diameter (105.26 g,
203.5, 21.41 kg tree-1 and 57.87 cm) were obtained in
trees sprayed with potassium at 1% and copper at
Table 2 Effect of foliar application with potassium and copper on yield and fruit characteristics of peach (Prunus persica L.) cv.
Treatment
Potassium
(%)
0.0
0.5
1.0
Copper (%)
0.00
0.02
0.04
Potassium Copper
K 0 Cu 0
K 0.0 Cu 0.02
K 0.0 Cu 0.04
K 0.5 Cu 0.00
K 0.5 Cu 0.02
K 0.5 Cu 0.04
K 1.0 Cu 0.00
K 1.0 Cu 0.02
K 1.0 Cu 0.04
Parameters
Yield
(kg/Tree)
16.65 c
18.76 b
20.10 a
Fruit weight
(g)
85.66 c
93.42 b
99.71 a
Fruit number
(Fruit/Tree)
194.4 a
200.8 a
201.7 a
Fruit diameter
(cm)
52.17 b
53.22 b
56.68 a
86.96 c
92.89 b
98.94 a
196.1 a
199.2 a
201.6 a
17.05 c
18.52 b
19.94 a
51.78 c
54.27 b
56.01 a
77.09 e
84.89 d
95.01 bc
89.19 cd
94.52 bc
96.55 b
94.59 bc
99.26 ab
105.26 a
194.4 a
194.8 a
193.9 a
194.4 a
200.6 a
207.4 a
199.3 a
202.1 a
203.5 a
14.99 e
16.55 de
18.40 bc
17.31 cd
18.96 bc
20.02 ab
18.85 bc
20.04 ab
21.41 a
49.70 e
52.44 ef
54.37 ce
50.58 fg
53.29 de
55.81 ac
55.07 bd
57.09 ab
57.87 a
(0.04%) level as compared with the lowest values

(77.09 g, 194.4, 14.99 kg tree-1 and 49.70 cm) in the
untreated trees.
3.3 Fruit Quality Characteristics
Data in Table 3 clearly shows that foliar application
of potassium at 0.5% and 1% was accompanied with
improving quality of the peach fruits in terms of
increasing pith thick, pulp weight, seed weight and
total soluble solids percentage. The best results were
obtained by the addition of potassium via leaves at 1%.
Copper sprays also was of measurable influence on
fruits quality in terms of increasing pith thick, pulp
weight, seed weight and total soluble solids percentage
and the promotion on quality of the fruits was
correlated with increasing concentration of copper
application, the highest values of pith thick, pulp
weight, seed weight and total soluble solids percentage
resulted in fruits of trees received copper at 0.04%.
For the interaction, the best results were regarded
when potassium was sprayed at 1% and copper at
0.04%, the highest values of pith thick, pulp weight,
51
seed weight and total soluble solids percentage (19.87

g, 94.00 g, 1.26 g and 14.84%) respectively, were
detected on trees received potassium at 1% and copper
at 0.04%, compared with the lowest values (14.39 g,
69.43 g, 7.66 g and 12.00%) respectively, at the
control treatment. The increases in fruit quality traits
may be due to the role of potassium influencing
meristematic growth, photosynthesis and activates a
number of enzymes, including those involved in the
synthesis of carbohydrates, and is also involved in the
neutralization of organic acids and the promotion of
normal cell division and growth [8, 10, 21].
The positive effects of copper on fruit quality of
peach could be due to increase the process of
photosynthesis and other enzymatic processes, which
reflected in the processing of fruits materials
necessary to improve fruit quality characteristics [11].
4. Conclusions
According to the experimental results of this study,
the most important conclusions can be expressed as
follows:
Table 3 Effect of foliar application with potassium and copper on fruit quality characteristics of peach (Prunus persica L.) cv.
Treatment
Potassium
(%)
0.0
0.5
1
Copper (%)
0
0.02
0.04
Potassium Copper
K 0 Cu 0
K 0.0 Cu 0.02
K 0.0 Cu 0.04
K 0.5 Cu 0.00
K 0.5 Cu 0.02
K 0.5 Cu 0.04
K 1.0 Cu 0.00
K 1.0 Cu 0.02
K 1.0 Cu 0.04
Parameters
Seed weight
(g)
8.75 b
9.32 b
10.44 a
Pith thick
(mm)
15.32 b
17.48 a
18.90 a
Pulp weight
(g)
76.92 c
84.10 b
89.26 a
16.13 b
17.41 ab
18.16 a
78.20 c
83.29 b
88.79 a
8.75 b
9.60 ab
10.16 a
12.77 c
13.64 b
14.09 a
14.39 d
15.63 cd
15.95 bd
16.12 bd
17.65 ac
18.66 ac
17.87 ac
18.95 ab
19.87 a
69.43 e
76.08 d
85.24 bc
79.94 cd
85.25 bc
87.11 b
85.25 bc
88.53 b
94.00 a
7.66 d
8.81 cd
9.77 ac
9.25 bd
9.27 bd
9.44 bc
9.35 ac
10.73 ab
11.26 a
12.00 e
13.14 d
13.38 cd
12.53 e
13.63 bd
14.03 b
13.76 bc
14.16 b
14.84 a
TSS
(%)
12.84 c
13.40 b
14.25 a
52
Potassium and copper markedly increased leaf area,

leaf fresh weight, leaf dry weight, total chlorophyll,
fruit weight, fruit number and yield per tree as well as
fruit diameter, pith thick, pulp weight, seed weight and
total soluble solid.
Potassium at 1% allowed maintenance of yield
without important loss in fruit quality.
Potassium when applied at 1% and Cu at 0.4%,
markedly increased yield and hence fruit quality such
as pith thick, pulp weight, seed weight and TSS
percent.
References
[1]
[2]
[3]
[4]
[5]
[6]
[7]
[8]
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[12] S.M. Al-Atrushi, Effect of eyes number and foliar spray
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DAVID
PUBLISHING
Carboxylesterase and Glutathione-S-Transferase (GSTs)

Induced Resistance to Bacillus thuringiensis Toxin
Cry1Ab in Rice Leaf Folder, Cnaphalocrocis
medinalis (Guenee) Populations
Veegala Ramesh Babu1, Vemuri Shashi Bhushan1, Chintalapati Padmavathy2, Muthugonder Mohan2, Sena
Mahendran. Balachandran3 and Bellamkonda Ramesh1
1. AINP on Pesticide Residues, Acharya N.G. Ranga Agricultural University, Andhra Pradesh, Rajendranagar, Hyderabad 500030,
India
2. Division of Crop Protection, Directorate of Rice Research, Rajendranagar, Andhra Pradesh, Hyderabad 500030, India
3. Division of Crop Improvement, Directorate of Rice Research, Rajendranagar, Andhra Pradesh, Hyderabad 500030, India
Received: July 9, 2012 / Published: January 20, 2013.
Abstract: The rice leaf folder (RLF), Cnaphalocrocis medinalis (Guenee) (Insecta: Lepidoptera: Pyralidae), is an important pest,
widely distributed in many rice growing areas of Asia. The over-use of broad-spectrum chemical insecticides has been cited as a
major cause of outbreaks of C. medinalis as excessive spraying of insecticide disrupts natural biological control insecticides still
remain the major control tactics against leaf folder. Carbofuran and fenthion, bendiocarb, acephate, carbosulfan, quinolphos,
monocrotophos, phosphamidon and fenvalerate are the common ones used against rice leaf folder. Genetically, modified rice lines
expressing B. thuringiensis insecticidal crystal proteins produced are highly tolerant to leidopteran pests. Though economic and
environmental benefits of GM crops is well established, the matter of concern is the possibility of target insect pest developing
resistance to this B. thuringiensis insecticidal toxins, evident from many laboratory and field experiments against many insect pests.
The involvement of GSH S-transferase, carboxylesterase, and microsomal monooxygenase in insecticide resistance has been reported
in insecticide-resistant strains of many insect species. Hence, the present study was taken up to monitor for cross resistance between
B. thuringiensis cry toxins and synthetic insecticides in larvae of leaf folder as it is mediated by carboxylesterase titre and other
enzymes by bioassay for two selected rice leaf folder field populations at the Entomology division of Directorate of Rice Research
which showed 2-fold resistance ratio. Qualitative and quantitative changes of carboxylesterase (CarE) and glutathione-s-transferase
(GSTs) were worked out with midguts extracts of the two C. medinalis populations in the presence of -napthyl acetate and
chlorodi-nitro benzene substrates.
Key words: Cnaphalocrocis medinalis, carboxylesterase and glutathione-s-transferase, isozymes, B type esterases.
1. Introduction
Carboxylesterases (CES, EC 3.1.1.1) are members
of a superfamily of serine hydrolases that hydrolyze
ester, amide and carbamate bonds. Several different
CarE genes exist with evidence of multiple gene
Corresponding author: Vemuri Shashi Bhushan, Ph.D.,

research fields: entomology, insect toxicology and pesticide
residues. E-mail: Sash_3156@yahoo.co.in.
duplication in insects. Esterases hydrolyse ester bonds

from various substrates with a carboxylic ester.
Esterases are frequently implicated in the resistance of
insects to organophosphorus, carbamates, pyrethroids,
neonicotinoids and many other new classes of
insecticides through gene amplification, up regulation,
coding sequence mutations or a combination of these
mechanisms [1].
54
Carboxylesterase and Glutathione-S-Transferase (GSTs) Induced Resistance to Bacillus Thuringiensis

Toxin Cry1Ab in Rice Leaf Folder, Cnaphalocrocis medinalis (Guenee) Populations
The
rice
leaf
folder
(RLF),
Cnaphalocrocis
medinalis (Guenee) (Insecta: Lepidoptera: Pyralidae),

is an important rice pest, widely distributed in many
rice growing areas of Asia [2]. There have been
frequent and serious outbreaks of this pest in many
countries including India, Korea, Japan, China,
Malaysia, Sri Lanka and Vietnam [3]. The over-use of
broad-spectrum chemical insecticides, such as methyl
parathion, monocrotophos and endosulfan has been
cited as a major cause of outbreaks of C. medinalis
because excessive spraying of insecticide disrupts
activity of midgut carboxylesterase activity [10]. The

involvement of GSH S-transferase, carboxylesterase,
and microsomal monooxygenase in insecticide
resistance has been reported in insecticide-resistant
strains of many insect species. Hence, the present
study was taken up to monitor if any cross resistance
between B. thuringiensis cry toxins and synthetic
insecticides occur in larvae of leaf folder as is
mediated by carboxylesterase titre and other enzymes.
natural biological control [3]. Insecticides still remain
2.1 Insecticide Bioassay
the major control tactics against leaf folder. Carbofuran
Two field populations of C. medinalis were

collected from Directorate of Rice Research,
Rajendranagar and ICRISAT, Patancheru. C.
medinalis adults were collected from rice fields during
the boot leaf stage in Rabi 2011. The collected adults
were released into the pots containing TN-1 plants for
egg laying and were covered with a muslin cloth for
aeration, 20% honey solution was also provided for
feeding. About ten pairs of C. medinalis adults were
released into each TN-1 pot. C. medinalis populations
from different locations were reared separately and
after larval hatching, the 3rd instar larvae were used
for bioassay.
Leaf-dip bioassay method was used. 3-4 cm long
tender leaves from TN-1 rice variety were used in the
bioassay. The leaves were first washed with distilled
water and then were dipped in monocrotophos
solution and thoroughly air dried for about 10 min
different concentrations of insecticide were prepared,
bioassays were carried out first at 10 fold variation.
Based on 20%-80% mortality, concentrations were
prepared at narrow range of five-fold for further
bioassays. Six concentrations were tested with 10
third instar larvae per treatment and replicated thrice.
Larvae were allowed to feed on insecticide treated
leaves for 24 h, and mortality was recorded for 24 h
after treatment. Control treatments with larval
mortality more than 20% were discarded and bioassay
was repeated. Statistical analysis for calculating the
and fenthion [4], bendiocarb, acephate and carbosulfan,

quinolphos, monocrotophos and phosphamidon [5] and
fenvalerate [6] were the common insecticides used
against the control of rice leaf folder.
Genetically, modified rice lines expressing B.
thuringiensis insecticidal crystal proteins have been
produced that are highly tolerant to leidopteran pests.
In China, three GM rice lines transformed with
cry1Ac/cry1Ab
genes
(GM
Minghui
63),
cry1Ac/CpTI genes (GM Minghui 86) and cry1Ab
genes (GM Kemingdao) effective against S. incertulus,
C. medinalis and C. suppressalis are tested both at
field and laboratory levels and are on verge of
commercialization [7-9]. Though economic and
environmental benefits of GM crops is well
established, the matter of concern is the possibility of
target insect pest developing resistance to this B.
thuringiensis insecticidal toxins as it is evident from
many laboratory and field selection experiments
against many insect pest. Though a couple of
resistance mechanisms have been reported for
conferring resistance to B. thuringiensis viz. reduced
binding of the crystal toxins to the brush border
membrane vesicles (BBMVs) of midgut epithelium
and alteration in the midgut proteases that cleave the
protoxin to active toxin. Recent report indicates that a
new resistance mechanism to B. thuringiensis cry
toxins is identified and associated with increased

LC50 values for the bioassay was estimated using

maximum likelihood programme MLP 3.01 [11]. The
corrected percent mortality was calculated by using
Abbotts formula [12].
2.2 Preparation of Enzyme Homogenate
Fifth instar larvae of C. medinalis were used for
enzyme preparation. Larval midguts were excised
with replicate samples and were homogenized in 500
L homogenization buffer (50 mM sodium phosphate
buffer, pH 7.4). After centrifugation at 10,000 rpm for
20 min, the clear supernatant was collected and used
as enzyme sources for analysis of the activity of
carboxylesterase. All the operations were carried out
on ice and centrifugation at 4 C to minimize losses of
enzyme activity. The protein content of enzyme
extract was estimated by Coomassie Brilliant Blue
G-250 dye binding method using bovine serum
albumin as the standard [13].
2.3 Carboxylesterase Assay
Carboxylesterase activity was determined by the
method of Van Asperen [14] with necessary
modifications and -naphthyl acetate as a substrate. A
0.3 mM substrate solution of 1-naphthyl acetate was
prepared in acetone. The assay mixture contained 15
L of enzyme preparation, 0.5 mL of 50 mM sodium
phosphate buffer pH 7.4, and 800 L of 0.3 mM
substrate solution. The mixture was incubated at 30 C
for 30 min. Finally, 200 L of 0.1% tetrazotized
o-dianisidine (fast blue B) in 3.5% SDS was added
and incubated for 20 min at room temperature in the
dark. The -naphthol formation was measured at 590
nm. The enzyme activity was calculated from
-naphthol standard curve.
2.4 Esterase Isozyme Studies
Native PAGE with 10% resolving gel was
performed to separate esterase isozymes. Qualitative
changes in esterase banding pattern was performed
using the F2 generation larvae that were reared after
55
surviving
the
insecticidal
bioassay
with
monocrotophos. 5 g protein concentrations of the
mid-gut homogenate per well were loaded onto the
native PAGE and run at a constant voltage of 90 for
45 min. Gels were stained briefly for esterase activity
with freshly prepared 0.05% (w/v) -napthyl acetate
and 0.1% (w/v) fast blue B in 50 mM phosphate
buffer pH 7.4. For inhibition studies, gels were cut
into strips and incubated in 10-4 M and 10-6 M eserine
sulphate and 10-4 M DDVP individually in 50 mM
phosphate buffer pH 7.4 for 30 min at 28 C with
occasional shaking. Control gels were incubated for
30 min in buffer alone. All the gel strips were stained
incubated for 30 min in -napthyl acetate substrate
diazonium mixture for confirming the esterase
activity.
2.5 Glutathione-S-Transferase Activity Estimation for
Insecticide Resistance
Glutathione-S-Transferase assay was performed
by using reduced glutathione (50 mM), mid-gut
homogenate supernate (10,000 g) from the F2
generation larvae that were reared after surviving the
insecticidal bioassay with monocrotophos, chloro
dinitro benzene (CDNB) 50 mM, sodium phosphate
buffer (pH 6.5, 100 mM) and EDTA (1 mM). The
assay mixture contained 50 L of 50 mM CDNB,
150 L of reduced glutathione and 2.77 mL of 100
mM, pH 6.5 phosphate buffer containing 1 mM of
EDTA. To the above assay 30 L of enzyme
(mid-gut homogenate) was added and the contents
were shaked gently and incubated for 2-3 min at
25 C, then the contents were transferred to 4 mL
cuvettes and absorbance was recorded for 6-7 min at
340 nm. Based on the increase in absorbance over
five minutes the enzyme activity was calculated in
mol min-1 mg-1 protein.

Bioassays results for the two selected C. medinalis
populations, DRR, Rajendranagar and ICRISAT,
56

Patencheru revealed LC50 of value 60 ppm for

ICRISAT C. medinalis population and showed 2-fold
resistance ratio over the DRR population which
showed LC50 30 ppm against monocrotophos (Table 1).
3.1 Qualitative and Quantitative Changes of
Carboxylesterase and Glutathione-S-Transferase in C.
Medinalis Populations
Qualitative and quantitative changes of CarE and
GSTs for DRR population in the presence of -napthyl
acetate and CDNB revealed a titre of 114.3 mol min-1
mg-1 protein and 5.66 mol min-1 mg-1 protein, while
that ICRISAT population showed 155.2 mol min-1
mg-1 protein and GST titre of 12.598 mol min-1 mg-1
protein, respectively. The results revealed that
ICRISAT C. medinalis population had 1.35 folds
greater carboxylesterase and 2.245 folds more GSTs in
its midgut homogenates over DRR population (Table 2).
Similar findings were reported by Mohan and Gujar [15]
where 1.2 to 1.8 fold increased CarE activity was
observed in P. xylostella for monocrotophos, cartap and
fipronil resistant populations, over the laboratory strain
IARI 17-65. Yamamoto et al. [16] reported that GST
from RLF, C. medinalis was inhibited by fenitrothion,
permethrin, and deltamethrin, suggesting GST may be
involved in metabolizing organophosphorus and
pyrethroid insecticides.
3.2 Esterase Isozyme Studies
Esterase activity visualized in native PAGE
following incubation in substrate solution (0.05%
Table 1
-naphthyl acetate in 0.1% fast blue (R) revealed two

isozyme bands. The major band with diffused esterase
activity was relatively of more molecular mass than
the other esterase activity band which was faint in
nature (Fig. 1). Difference in esterase banding pattern
in ICRISAT, C. medinalis population was observed
when using midgut and whole body extracts. The
midgut produced all three types of esterase bands
while whole body homogenates produced only two
bands (Fig. 1). Inhibitor studies with the esterase
isozymes separated under native PAGE when
subjected to inhibition by incubating with class
specific esterase inhibitors in buffers containing
dichlorvos (DDVP) 10-4 M, eserine sulphate, 10-6 and
10-4 M concentrations indicated that these two esterase
isozymes are B type esterases as eserine sulphate, a
specific inhibitor of cholinesterase did not inhibit the
esterase activites at 10-6 and 10-4 M concentrations and
esterases were characterized to have carboxylesterase
activity (Fig. 2).
In insects, the esterase bands separated
electrophoretically under native condition are classified
into three types by the substance which inhibits their
activity [14, 17]. The results of the present study are in
conformity with that of cypermethrin resistant P.
xylostella strain, where in the carboxylesterase levels
from 1st to 4th instar, pupa and adult showed 2.64,
3.16, 2.61, 3.04, 2.93 and 2.75 folds higher
carboxylesterase activity in comparison to P.
xylostella susceptible strain [18] and also reported
monocrotophos, profenofos, quinalphos and phenthoate,
Toxicity of monocrotophos to 3rd instar larvae of C. medinalis 24 HAT.
Location
LC50 (ppm)
RR
Slope SE
DRR population
ICRISAT population
30
60
1.00
2.00
1.29 0.21
1.14 0.21
Lower
0.0018
0.002
F Limits
Upper
0.0057
0.011
2 (Degrees of freedom)
4.0 (4)
2.31 (4)
R R: Resistance ratio over one generation, F Limits: fiducial limits.

Table 2
Carboxylesterase and GST activity of 3rd instar larvae of C. medinalis.
Location
DRR population
ICRISAT population
CarE (mol min-1 mg-1 protein)

114.39
155.2
CarE folds
1.00
1.35
GST (mole min-1 mg-1 protein)

5.660
12.598
GST folds
1.00
2.245

57
Fig. 1 Carboxylesterase, profiles from whole body and midgut homogenates of ICRISAT, C. medinalis population. Lanes 1-5 are
whole body homogenates showing two bands of esterase isozymes, Lanes 6-10 are midgut homogenates showing three bands of
esterase isozymes.
Fig. 2 Characterization of C. medinalis carboxylesterase with specific substrates. Lane: 1-DDVP (10-4 M), Lanes: 2, 3 and
4-Eserine(10-4M), Lanes: 5, 6-Eserine(10-6M), Lanes: 7, 8-Control.
as it showed 54.34%, 72.54%, 78.71%, 80.82% and

82.94% inhibition of carboxylesterase titres associated
with cypermethrin resistance at 0.01, 0.1, 1, 5 and 10
mg mL-1 concentration, and suggested that DDVP is
the best synergist to mitigate cypermethrin resistance
in P. xylostella whereas Rashad [19] reported high
titres of esterase activity in brain, fore-gut, mid-gut
and ovary of 2-day old adults of Schistocerca
gregaria, while in 13 day old adults hind gut exhibited
high esterase levels. Inhibitory studies with EDTA
and profenofos depicted high levels of both
carboxylesterase and phosphotriesterases in the brain

tissues of two ages that attributed to play a role in
insecticide resistance.
In the present study correlation between LC50 value
of monocrotphos of third instar larvae and
carboxylesterase activity was similar to that obtained
by Kranthi et al. [20] who studied the seasonal
dynamics of metabolic mechanisms responsible for
pyrethroid resistance in H. armigera and assigned it
due to involvement of microsomal oxidase and
esterases. Young et al. [21] reported pyrethroid
58

resistance in H. armigera (Hubner) and attributed to

overproduction of esterase isoenzymes that metabolise
and sequester pyrethroid insecticides and found out
that pyrethroid-resistance-associated esterases were
inhibited by piperonyl butoxide (PBO) and maximum
inhibition achieved 3-4 h after dosage and again
restored by 24 h.
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[3]
Esterase zymogram in the present study showed an

additional fast moving band by midgut extracts in
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[4]
conformity with the findings on DBM [15] where no

major difference in banding pattern was identified
except for three additional slow moving faint bands
for all field populations and were associated with
[5]
resistance based on intensity of banding, and

characterized to be as B-esterase similar to P.
xylostella.
4. Conclusions
In the present study, the carboxylesterase present in
ICRISAT population was calibrated in vitro to be
155.2 mol min-1 mg-1 protein and the esterase
zymograms showed intensely stained bands depicting
resistance
association.
The
median
lethal
concentration, LC50 for this strain was 60 ppm, though
more than the discriminating dose for monocrotophos,
0.35 g per larvae [22]. The base-line LC50 estimate
for the ICRISAT population with cry1Ab toxin is 0.50
g/mL, the matter of concern in this regard is that
indiscriminate usage of insecticides for control of C.
medinalis may futher bring an elevation in the esterase
titre which may bind to cry1Ab toxin receptors and
sequester the toxin before it reaches the target site as
exemplified in the case of silver strain H. armigera
towards cry1Ac expressed by transgenic cotton
Ingard in Australia, where sequestration by esterases
was recognized as a potential resistance mechanism
apart from previous resistance mechanisms viz.
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that cleave protoxin to active toxin [10].
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DAVID
PUBLISHING
Environmental Impacts of Feeding High-fiber Diet to

Pigs
Abraham Woldeghebriel1, Shanequa Smith2, Teo Barios1, Brad Pope1 and Sebhatu Gebrelul3
1. North Carolina A&T State University, Greensboro, NC 27411, USA
2. North Carolina Central University, Durham, NC 27707, USA
3. Southern University, Baton Rouge, LA 70809, USA
Abstract: The concentration of volatile fatty acids (VFA) determined from a previous study was used to determine the concentration
of carbon dioxide (CO2) and methane (CH4) in pig digesta using the model developed for ruminant animals. Digesta from the
stomach, cecum and colon of pigs (n = 3 diet-1) were used for the determination of VFA. The pigs were fed either a low fiber diet
(LFD; 8.3% aNDF) as control, or one of the high-fiber diets (HFD, 22.4% aNDF; D1, D2 and D3) containing 1:2, 1:1 and 2:1, oats to
barley ratios, respectively. Results indicated that the concentrations of CO2 and CH4 in pigs fed HFD were, on the average, 25.8 and
24.2%, respectively lower (P < 0.05) than pigs fed LFD. Pigs fed the highest oat to barley ratio also showed higher (P < 0.05) levels
of CO2, (8.3%) and CH4 (5.1%), compared to the average of the two lower ratios (CO2, 5.3% and CH4, 3.3%). Molar proportions of
VFA, CO2 and CH4 in the gut were in the order of VFA > CO2 > CH4, at 53.0%, 28.6% and 18.4%, respectively, and CO2 and CH4
combined represented 47% of total gas.
Key words: Environment, fiber, methane, pig, volatile fatty acids.
1. Introduction
Ruminant nutrition research has for so long been
focused on reduction of enteric CH4 emission not
because of its effect on global warming but because of
its inefficiency in animals [1]. Even though
production of CH4 in the rumen and lower digestive
tract of animals is influenced by a number of factors
[2], the average yearly production estimates of a
typical beef and dairy cow is in the range of 60-70 kg
and 109-126 kg, respectively [3]. As a consequence,
the combined economic loss in the range of 2%-12%
of energy intake in ruminant animals and its
significant contribution to the greenhouse gas (GHG)
effect and global warming [4, 5] are glaring evidences.
A sizable number of approaches including vaccination,
enzyme
inhibitors
phages,
homoacetogens,
Corresponding author: Abraham Woldeghebriel, Ph.D.,

research fields: animal nutrition, digestive physiology. E-mail:
awoldegh@ncat.edu.
defaunation and animal selection pressure have been

investigated and yet, it seems like more than one
strategy may have to be used to have any significant
impact on reducing enteric CH4 emission [6].
Traditionally, diets high in fiber were exclusively
used for ruminant animals while pigs on the other hand
were fed mainly cereal-based concentrate diets. While
exclusive uses of low-fiber diet (LFD) improve
productivity and pollution from excreted nutrients
feeding LFD could be detrimental to the health of the
animal [7]. Also in recent years, higher feed prices and
availability of relatively cheaper co-products on the
market has provoked interest in alternative feed
ingredients including those that are high in fiber. It was
also noted that over the last few years due to public
demand for less intensive farming systems and rising
capital costs on indoor pig housing, outdoor pig farms
started to grow in large numbers. However, given our
current level of production systems and mitigation
strategies we have in place, raising pigs on pasture

could impact environmental protection goals due to
increases in CH4 emission from pigs with free access
to forages and other vegetative parts of plant materials
that could inherently be high in fiber. When high-fiber
diets (HFD) are fed to pigs, more substrates enter the
large intestine. According to Varel and Yen [8],
microbial activity in the digestive tract of pigs fed
HFD was 5.5 times higher and resulted in a 5-9 fold
increase in CO2 and CH4 production than in pigs fed
LFDs. Similar observation was also reported by Jensen
and Jorgensen [9] where CH4 excretion rate of a
7-month old barrow fed HFD was 12.5 L d-1 compared
to 1.4 L d-1 when the same animal was fed LFD
indicating that the concentration of the fiber was the
main dietary contributor to enteric CH4 emission in
pigs [7]. In terms of actual production, while a mature
cow produces up to 250 L of CH4 d-1 (60 kg year-1),
depending on the feeding levels and types of fiber, a
dry gestating sow fed at maintenance level produces
approximately 10.2 L day-1 which is approximately
61
respiratory chambers are expensive, restrictive to

movement of animals and the system relies on indirect
measurements but other less expensive indirect
measurements of fermentation indices exist. For
example, France and Siddon [14] used CH4 production
to obtain an estimate of VFA production. Also, Wolin
[15], Van Nevel and Demeyer [16] each developed
similar model by which emission of CH4 from
ruminants was calculated using VFA concentration
data from ruminant animals. Therefore, we
hypothesized that the model developed by Wolin for
ruminant animals can be used to indirectly estimate
CO2 and CH4 concentration in the digestive tract of
pigs. The aim of the study was to test the hypothesis
that the model developed by Wolin for ruminant
animals can be used to determine the amount of CO2
and CH4 concentration in digesta using VFA
concentration data from pigs.
2. Methods
2.1 The Mathematical Model
0.6%-2.7% of gross energy intake of the animal [7]. A
The equation developed by Wolin [15] and later
recent study conducted in Denmark highlighted the
validated by Blummel et al. [17] to estimate CH4
relative contribution of enteric CH4 emission from
production in ruminant animals was used to determine
livestock and manure management at 86%, 10% and
the concentration of CO2 and CH4 in the digestive
3% from cattle, pig and horses, respectively [4].

Carbon dioxide and CH4 are two of several
anthropogenic and naturally occurring greenhouse
gases (GHG) in the atmosphere. The IPCC [10]
developed a global warming potential index (GWPI)
to estimate and compare the ability of GHG to trap
heat in the atmosphere using CO2 as a reference with
GWPI of 1 and others expressed as CO2 equivalents.
Methane the second most significant contributor to the
GHG effect traps outgoing terrestrial infrared
radiation 21 times more effectively than CO2 [11, 12],
increases surface temperatures and indirectly affects
the atmospheric oxidation reactions that produces CO2
[13]. Measurements of gaseous exchanges such as
CO2 and CH4 production in animals have traditionally
been carried out in respiratory chambers. However,
tract of growing pigs. The reactions shown below

summarize the basic methanogenic processes on
which the model was conceptualized [15]:
Hexose 2 pyruvate + 4H
Pyruvate acetate + CO2 + 2H+
2 pyruvate butyrate + 2CO2
Pyruvate + 4H2 propionate + H2O (propionate is

a potent H sink)
CO2 + 8H+ CH4 + 2H2O
Molar concentrations of the three most abundant
VFAs (acetate, propionate and butyrate) in the
digestive tract of the pigs obtained from one of our
previous unpublished data were used to calculate the
62
fermentative concentration of CO2 (Eq. 1) and CH4

(Eq. 2) as follows.
2.2 Equations
(1)
CO2 (mol) = A/2 + P/4 +1.5B
Where A, P and B are moles of acetate, propionate
and butyrate, respectively and,
CH4 (mol) = (A + 2B) CO2
(2)
Where CO2 is moles of carbon dioxide estimate in
Eq. 1.
2.3 Data Analysis
Data collected were analyzed using the SASs
Glimmix procedure [18] with diet as the main fixed
effect and digestive organs from animals as random
effects. The statistical model investigated the main
effects: diet, digestive organs and associated
interactions. Data presented in the table are least
square means, means were compared using paired
T-test, and statistical significance was accepted at P <
0.05. Probability values between 0.06 and 0.10 were
considered trends.

The individual VFA used to determine the
concentration of CO2 and CH4 in the digestive tract of
the pigs were acetate (C2), propionate (C3) and
butyrate (n-C4). Bacteria in the hindgut metabolize
carbohydrates to obtain energy and proteins to N
through anaerobic fermentation for their own growth
and maintenance requirements. Results of such
anaerobic fermentation were summarized by Ewing
and Cole [19], and the molar ratios of the major VFA
were expressed as 1:0.31:0.23 for C2, C3 and n-C4,
respectively. The average molar ratio of the VFAs in
pigs in the current study was in complete agreement
(1:0.32:0.17, respectively) with the general equation
reported earlier except for n-C4 which was 26% lower
than the general equation indicating that n-C4 might
have been depleted at a relatively faster rate than the
other VFAs when availability of fermentable fiber in
the hindgut of the pigs might probably be on the

decline. According to Fitch and Fleming [20]
transport of n-C4 from the lumen into the blood
increases linearly with increasing concentration of C4,
while transport and metabolism of C2 are significantly
lower than C4. Butyrate has also been shown to be
oxidized to CO2 more readily than C2 or C3, and,
while C4 oxidation is not suppressed by the presence
of other substances, presence of C4 reportedly
suppressed the oxidation of other VFA including C2
[21]. It is also important to note that C4 is
preferentially metabolized by colonocytes for energy
[22], is more effective than C2 and C3 in enhancing Na
absorption [23], and plays an important role in the
prevention of diarrhea in young pigs [24]. Therefore,
given its physiological and multifunctional roles in the
body of the animal, it seems reasonable to assume that
C4 would be depleted at a much faster rate than the
other VFAs, which are relatively less important than
C4 to the pig.
The two most abundant VFAs, i.e., C2 and C3, were
present in 1:0.32 ratio. Combined, they represented
89% of the total VFA. The fact that there was 3.13
times more C2 than C3 may indicate a much higher
probability for methanogenesis through one or both of
the following reactions:
Pyruvate acetate + CO2 + 2H, and
CO2 + 4H2 CH4 + 2H2O, or
CH3COOH CH4 + CO2
where CO2 and acetic acid are terminal electron
acceptors.
As in the rumen, methanogenes in colonic
fermentation use H2 to reduce CO2 to CH4 [25].
However, when non-methanogenic fermentation
occurs, H2 is used to reduce CO2 to acetate according
to the following equation [26]. Acetate can then be
used as a source of carbon and energy.
2CO2+ 4H2 CH3COOH + 2H2
Generally, VFAs are not used as substrates for
methanogenesis, as their conversion to CO2 and H2,
especially in ruminants is lengthy and is inhibited by
63
rumen turnover [27], but also due to the fact that

methanogenesis often uses CO2 and H2 from
carbohydrate fermentation during VFA synthesis [28].
The estimated molar concentration of CO2 and CH4
in the stomach, cecum and colon of the pigs is shown
in Table 1. The concentration of CO2 in the stomach
of pigs fed LFD was 2.6 times higher (P < 0.05) than
the average of the HFD fed pigs. There were no
significant differences among the HFD fed pigs. In the
cecum, the highest (P < 0.05) CO2 concentration was
found in CON and D3 fed pigs, while the lowest
concentration was in pigs fed D2 with no significant
difference between CON and D1 and between D1 and
D2 fed pigs. However, in the colon, the lowest (P <
0.05) CO2 concentration was found in pigs fed D1,
with no significant differences among the remaining
groups of pigs.
The highest (P < 0.05) total concentration of CO2
was observed in pigs fed CON and D3, while the
lower and lowest (P < 0.05) concentrations were
detected in pigs fed D2 and D1, respectively. It was
noted that the concentration of CO2 in pigs fed D2 and
D3 was higher (P < 0.05) than D1 fed pigs, with no
significant differences between D2 and D3, or
between D1 and D2 fed pigs. It was also noted that the
concentration of CO2 in LFD fed pigs was 25.6%
higher than the average of the HFD fed pigs (84.39%
vs. 62.79%). Also, as the amount oats in the diet
increased (D3 > D2 > D1), concentration CO2 also
increased (83.13%, 59.27% and 45.96%, respectively),
Table 1
Concentrations of CO2 and CH4 in visceral contents of pigs.
Visceral organs
% CO2
Stomach
Cecum
Colon
Total (mol L-1)
CH4
Stomach
Cecum
Colon
Total (mol L-1)
a, b, c
indicating that the DF in oats may have been more

readily fermentable by the microorganisms than the
DF in barley. However, differences between D2 and
D3 or between D1 and D2 were not statistically
significant, confirming many of the earlier reports
[29-31] that the amount of CO2 produced by animals
depends mainly on the live body weight, physiological
state, feed intake and physical activity, whereas the
composition of the feed supplied to the animal plays a
minor role.
The CH4 concentration in the stomach of pigs fed
LFD was 40% higher (P < 0.05) than the average
(2.22% vs. 1.33%) of the HFD fed pigs with no
significant differences between CON and D2, between
D1 and D3 and between D2 and D3 fed pigs. Likewise,
in the cecum, the highest (P < 0.05) molar
concentration of CH4 was observed in CON and D3
fed pigs with no significant differences between CON
and D1, or between D1 and D2 fed pigs. On the other
hand, in the colon, the lowest (P < 0.05) concentration
of CH4 was recorded in D1 fed pigs, but there were no
significant differences among the remaining groups of
pigs.
The highest (P < 0.05) total molar concentration of
CH4 was found in pigs fed CON and D3, while the
lower and the lowest (P < 0.05) concentrations were
recorded in pigs fed diets 1 and 2, respectively.
However, there were no significant differences between
CON and D3, between D2 and D3, or between D1 and
D2 fed pigs. It was noted that the concentration of CH4
CON
D1
D2
D3
SEM1
3.88a
12.00ab
9.43a
84.39a
1.23b
10.61bc
1.95b
45.96b
1.33b
8.27c
6.17a
59.27bc
1.95b
14.53a
8.46a
83.13ac
1.60
2.69
4.19
23.83
2.22a
6.98ac
6.21a
51.12a
0.92b
6.37bc
1.55b
29.53b
1.92ac
4.57b
4.41a
36.40bc
1.15bc
8.61a
5.31a
50.32ac
0.97
1.94
2.34
14.58
Least square means in the same row with different superscripts differ (P < 0.05); 1Standard error of means.
64
in LFD fed pigs was 24.2% higher than the average of

the HFD fed pigs (51.12% vs. 38.75%). It was also
observed that as the amount oats in the diet increased
(D3 > D2 > D1), CH4 concentration increased (50.32%,
36.40% and 29.53%, respectively), which suggests that
the DF in oats was more readily fermentable to the
microorganisms than the DF in barley. However, no
significant differences were observed between D3 and
D2 and between D2 and D1 fed pigs.
Acknowledgments
The authors are grateful for the financial support of
this
the
Evans-Allen
Project,
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4.1 Implication
[9]
It is important to remember that mathematical

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In summary, we were able to determine the
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the pigs using the model developed for ruminant
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usefulness of the model to estimate the molar
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DAVID PUBLISHING
Effects of Feeding Siamese neem leaves and

Zanthoxylum Pods, on Dry Matter Intake, Dry Matter
Digestibility, Milk Production and Composition in Thai
Holstein Dairy Cows, Fed Rice Straw as Fiber Source
Penjor1, Virote Pattarajinda1, Suporn Katawatin1, Chaiyapas Thamrongyoswittayakul2 and Wandee Gritsanapan3
1. Department of Animal Science, Faculty of Agriculture, Khon Kaen University, 40002, Thailand
2. Faculty of Veterinary Medicine, Khon Kaen University, 40002, Thailand
3. Department of Pharmacognosy, Faculty of Pharmacy, Mahidol University, 10800, Thailand

Abstract: The plant secondary metabolites (PSM) are highly sought compounds for use as an alternative to conventionally used feed
additives in animal production these days; Siamese neem leaf (Azadirachta indica A. Juss. var. siamenses Valeton) and Zanthoxylum
pods (Zanthoxylum piperatum) are known to contain numerous such compounds.The objectives of this study were to determine
effects of feeding Siamese neem leaf and Zanthoxylum pods as feed additives on dry matter intake (DMI), dry matter digestibility
(DMD) and milk production and milk composition. Lactating Thai Holstein cows (n = 8) were arranged in two replicates of 4 4
Latin square designs, housed in individual stall, treatments consisted of Siamese neem 0.5 and 1.5 g kg-1 dry matter (DM),
Zanthoxylum0.1 and 0.5 g kg-1 DM added to the total mixed ration (TMR). TMRand drinking water were provided ad lib. There were
no significant differences (P > 0.05) in DMI and crude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF)
digestibility, milk production and composition among the treatments.However, significant difference (P < 0.05) was observed in dry
matter (DM) and ether extract (EE) digestibility. Though statistically non-significant, Zanthoxylum at higher dose level tended to
show low DMI (14.85 kg) in cows, while promotingmarginally higher milk (14.18 kg) production as compared with lower dose
levelwhere DMI and milk production were 16.14 kg and 13.83 kg.This indicated that Zanthoxylum has a potential to improve feed
digestion in dairy cows when used as additives.
Key words: Plant additive, TMR, DMD, milk production, dairy cows.
1. Introduction
In an attempt to increase milk production, while
lowering the product cost at the same time, dairy
producers have made use of feed additives like
Monensin, Lasolasid and others antibiotics in dairy
production system for the last few decades. However,
the social acceptances for their use have declined in
the recent years on the grounds of the quality and
Corresponding author: Virote Pattarajinda, professor,

research fields: animal nutrition and feed science. E-mail:
Virote@kku.ac.th,ViroteKKU@hotmail.com.
safety of meat and milk products [1]. The consumer

organizations in European Union have criticized
routine use of antibiotics in livestock nutrition and the
ban remained restricted to EU nations [2].Increased
access to information through internet, television and
other media have enhanced the awareness of the
consumers andchanged their outlook [3]. There is
increasing public concern being raised on the use of
antibiotics in animal production systems [4]. The
consumers notion that anything natural is good for
the human health has created a desire to look for
plants containing PSM, possessing good capacity to
Effects of Feeding Siamese Neem Leaves and Zanthoxylum Pods, on Dry Matter Intake, Dry Matter
Digestibility, Milk Production and Composition in Thai Holstein Dairy Cows
alter nutrient utilization in the rumen as an alternative

to conventionally used antibiotics, where antibiotics as
a source of additives are no longer allowed in animal
production systems [5]. The plants are considered as
potential source of compounds and when fed to dairy
cattle will favorably alter the ruminal fermentation
without causing overall inhibition of fermentation,
thereby enhancing feedstuff degradability and
utilization,promoting microbial growth in rumen,
thereby enhances animal performance. In the current
study, two plants, Siamese neem leaves and
Zanthoxylum pods were selected as additives based on
the results of an in vitro experiment [3]. The doses
were calculated on the basis that DMI of an individual
animal is approximately 3% of body weight. Dosing
of plants on DM basis also reduced the chances of
over dosing animals as may be seen when calculated
on g kg-1 body weight basis. Zanthoxylum was dosed
at lower levels because it is known to cause tingling
and numbing sensation [6] in the oral cavity in man,
which may result in avoidance of feed by cows due to
this effect.This study evaluated the effects of feeding
Siamese neem leaves and Zanthoxylum pods as plant
based feed additive on DMI, DMD, milk production
and milk composition in lactating Thai Holstein cows
fed rice straw as the source of fiber. Eight lactating
cows were arranged in 4 4 Latin square designs,
housed in individual stall, and fed TMR mixed with
Siamese neem and Zanthoxylum. The TMR and
drinking water were provided ad lib. There were four
periods consisting of 21 days, the data were collected
during the last seven days of each period.

2.1 Plants Preparation and Dose Levels
The Siamese neem leaves were collected at Roi-Et
Agricultural Research and Training Center campus,
Zanthoxylum pods were purchased from a commercial
herbal dealer.The plant materials were dried in forced
air oven at 60 C for 24 h and ground through 2 mm
screen in Wiley mill. Siamese neem was dosed at 0.5
67
and 1.5 g kg-1 DM and Zanthoxylum dosed at 0.1 and

0.5 g kg-1 DM.
2.2 Animal and Experimental Design
Eight lactating Thai Holstein cows (420 50 kg)
were housed in individual stall, arranged in two
replicates of 4 4 Latin square design and the
treatments were allotted randomly. The experiment
consisted of four periods of 21 days each, during
which first fourteen days were allotted for
acclimatization of animals to the treatments allotted
and the data were collected during the last seven days
of each period.
2.3 Experimental Feed Formulation and Feeding
The basal diet fed as TMR consisting of rice straw,
premix concentrate and fresh brewers grain was
formulated using the computer program KCF 2006 [7].
The composition of the formulated TMR is given in
Table 1. The premix concentrate was prepared on
weekly basis. The premix concentrate consisted of
ricestraw chopped to 4-5 cm length to facilitate easy
mixing and intake by animals, and fresh Brewers
grain which were weighed separately and mixed in large
Table 1 TMRfeed ingredients and estimated chemical
composition.
Feed ingredients
Premix concentrate
Soya bean meal
Cassava meal
Sugar (FSOW)
Urea
Mineral
Salt
Calcium oxide
Fresh Brewers grain
Rice straw
Chemical composition
Total digestible nutrient
Crude protein
Crude fiber
Acid detergent fiber
Neutral detergent fiber
Ether extract
FSOW = Factory spill over waste.
% Composition
11.09
18.14
9.00
1.20
0.40
1.00
0.50
28.67
30.00
% Composition of DM
66.00
16.00
17.80
20.79
38.02
2.38
68
individual plastic containers, to which plant additives

were added and mixed thoroughly just prior to feeding
individual animal after milking in the morning and
evening, in individual feeding troughs. The TMR and
drinking water were provided ad lib. On 16, 17, 18
and 19 days of each period, 10 g chromic oxide,
divided into two doses of 5 g each was fed in the
morning and evening mixed with TMR diet for
determination of total tract digestibility.
on day 21 at 6 AM after morning milking; as Davis

[10] reported fecal grabs taken at these times
represented nearly mean chromic oxide content in 24
h period. The fecal samples were composited, dried
in the forced air oven and the chromic oxide was
measured by atomic absorption spectrometry, using
potassium dichromate as a standard. The total tract
digestibility was calculated using the concentrations
of the nutrients and chromic oxide in the diet and
feces [11].
2.4 Data Collection

The daily feed intake was determined by recording
orts weight each morning and deducting it from total
TMR fed. Feed samples were taken once a week
dried in forced air oven at 60 C until constant
weight was obtained on repeated weighing. Samples
were composited at the end of the study and
subjected to proximate analysis, crude protein (CP),
crude fiber (CF), EE, DM and ash were determined
according to AOAC [8] protocol while ADF and NDF
were analyzed according to Van Soest [9] method. On
day 17, 18 and 19, milk sample (60 mL from
morning and 40 mL from evening milking) were
collected, composited and analyzed by automatic
milk analyzer. On the 19 day blood sample was
collected from coccygeal vein or artery, centrifuged
at 1,500 rpm for ten minutes and paired serum were
prepared and stored at -20 C for blood glucose (BG)
and blood urea nitrogen (BUN) analysis. Body
weights were measured on day 19 and 20 using
portable weighing balance. Fecal grabs were taken
on day 20 at 3 pm in the evening after milking and
Table2

Data were analyzed by analysis of variance using the
general linear model (GLM) procedure of statistical
analysis system [12]. Treatment means were compared
by Duncans New Multiple Range Test and the level of
significance was determined at P < 0.05.

3.1 Dry Matter Intake and Digestibility
The result of the TMR feed analysis is shown in
Table 2, the CP and ADF were higher than the
estimated values while NDF content was below
estimated value.DMI was not significantly (P > 0.05)
different among the treatments. However DMI tended
to be higher in low levels of both Siamese neem and
Zanthoxylum, while higher dose levels showed low
DMI (Table 3). The reduced DMI at higher dose
levels of Zanthoxylum may be attributed to tingling
and numbing sensation caused by sanshool
compounds of Zanthoxylum. However, DMI in Siamese
Chemical composition of TMR Feed.
Feed samples
NM 0.5
NM 1.5
Zantho 0.1
Zantho 0.5
DM
53.66
53.89
57.11
54.24
GE, kcal
4.32
4.60
4.48
4.32
CP
19.08
18.25
19.12
19.47
EE
3.25
3.61
3.25
3.57
NDF
31.38
33.76
32.48
33.19
ADF
25.55
24.00
22.71
22.32
TA
6.85
6.33
6.65
6.37
NM = Siamese neem, Zantho = Zanthoxylum, GE = Gross energy, CP = Crude protein, EE = Ether extract, NDF = neutral detergent
fiber, ADF = Acid detergent fiber, TA = Total ash.
Items (%)
neem was not different between two doses, and bitter

taste cannot be responsible for low DMI at higher
dose, because cattle possess fewer genes coding for
bitter taste and therefore are tolerant [13] unlike in
human being where bitter taste cause strong aversion
to that particular food [14]. Therefore, this small
difference in DMI in Saimese neem may be attributed
to individual animal differences.
There were no significant difference in CP, ADF
and NDF digestibility among the treatments.
Significant differences (P < 0.05) were observed for
EE and DM digestibility. Zanthoxylum at 0.5 g kg-1
DM had the highest DMD (84.58%), followed by
Siamese neem at 1.5 g kg-1 DM (84.15%), and the
lowest DMD was found in Siamese neem at 0.5 g
kg-1 DM (82.26%). According to Hutjens [15], the
PSM favorably alter the ruminal fermentation
without causing overall inhibition of fermentation in
rumen or cause a desired animal response in a
non-nutrient role like shift in rumen pH. In the
current study, the high value of DMD in
Zanthoxylum is attributed to salivation induced by
sanshool compounds in Zanthoxylum, which is
reported to cause tingling and numbing
sensation in the oral cavity through stimulation of
sensory nerves [16]. The saliva of ruminants
contains phosphates and bicarbonates making it
appropriate medium for buffering rumen pH [17],
Table 3
thereby
enhancing
microbial
fermentation.
Furthermore, it is reported that PSM added to
ruminant feed can enhance or inhibit specific
microbial population in rumen, thereby increasing
efficiency in energy and protein utilization [18].
Also maintaining a ratio of soluble carbohydrate and
soluble protein may improve both palatability and
rumen ecology [19] therefore in current study both
the plants added have probably influenced the DMD
in the rumen in a similar way.
3.2 Milk Yield and Chemical Composition
There were no significant difference (P > 0.05) in
milk yield and chemical composition of milk (Table
3). However, in Zanthoxylum, milk yield tended to be
higher at higher dose levels albeit lower DMI. It can
be deduced that in Zanthoxylum, in spite of lower
DMI, higher dry matter digestibility have resulted in
similar milk yield or slightly higher yield in cows
receiving higher doses of Zanthoxylum than those
receiving lower doses. The percent composition of
milk constituents were unaffected and within normal
range. However, total solid content was below 12%,
this was attributed to early lactation of the animals,
and was expected to improve as lactation approached
peak. Bhosale [20] reported that in a comparative
studyinvolving goats at different lactation, goats
at thirdand fourth lactation produced hightotal solids
Effects of Siamese neem and Zanthoxylum on milk yield and composition.
Siamese neem
Zanthoxylum
SEM
P-value
0.5 g/kg DM 1.5 g/kg DM
0.1 g/kg DM
0.5 g/kg DM
DMI (kg)
15.28
14.95
16.14
14.85
0.25
0.29
DMD
82.26c
84.15ab
82.34bc
84.58a
0.24
0.03
Milk (kg)
13.68
13.07
13.83
14.18
0.22
0.36
Fat
3.71
3.63
3.78
3.60
0.04
0.58
Protein
2.95
2.96
2.98
2.99
0.01
0.59
Lactose
4.18
4.20
4.24
4.23
0.01
0.41
SNF
7.93
7.94
8.01
8.01
0.02
0.34
TS
11.64
11.56
11.59
11.63
0.05
0.93
Mineral
0.75
0.75
0.76
0.76
0.02
0.57
ADG (g)
320
710
330
490
0.06
0.08
DMI = Dry matter intake, DMD = Dry matter digestibility, SNF = Solid not fat, TS = Total solid, ADG = Average daily gain.
a, b, c
Means within a row without a common superscript letter differ (P < 0.05).
Items (%)
69
70
13.19%and 13.67% than those at first and second

lactation 12.33% and 12.59%, respectively, in the
current study, all animals were in first lactation and
therefore, low total solid content. Mech [21] reported
that the total solid and other milk constituent content
increased towards the end of lactation in Mithun
cows (Bos forontlis). There were positive average
daily gain in all treatments, especially in
Zanthoxylum at higher dose level, higher DM
digestibility has promoted higher weight gain in
animals receiving Zanthoxylum.
3.3 Blood Metabolites
The blood metabolites such as blood glucose were
unaffected in all treatments, but blood urea nitrogen
levels were elevated at 0 (before feeding), 1, 2 and 3 h
after feeding in all treatments. The elevated blood urea
nitrogen in animals were probably due to presence of
high levels of crude protein in the TMR diet; estimated
crude protein in TMR diet was 16%, while the crude
protein content in TMR diet was 18% to 19% (Tables
1 and 2). Hammond [22] reported that increased
protein in feed with constant energy supply can cause
increased BUN as energy is required for utilization of
nitrogen by microbes in the rumen. In the current study,
sugar (Factory spill over waste) was supplied as the
energy source; either sugar added was inadequate or
was rapidly fermented by microbes rendering it
unavailable as energy source. While dehydration is
another factor that could lead to elevated BUN, this is
unlikely in the current study as clean drinking water
was made available all times. Therefore it may be
reasonably suggested that the probable cause for
elevated BUN was the high CP content in TMR diet,
importantly animals showed no signs of toxicity during
the entire period of study in spite of high BUN levels.
This indicated that dairy cows can tolerate high BUN
levels without any ill effects.
4. Conclusions
In the current study, feeding Siamese neem leaves
and Zanthoxylum pods as feed additives in dairy cows

had no effect on DMI when rice straw was the main
source of fiber, however, there was significant (P <
0.05) effect on DM and EE digestibility. The DMI
was lower in higher dose levels of Siamese neem and
Zanthoxylum compared to lower dose levels, but the
DMD was higher in higher dose levels of Siamese
neem and Zanthoxylum. Highest DMD (84.58%) was
noticed in Zanthoxylum at 0.5 g kg-1 DM. This
probably was due to the increased salivation caused
through tingling and numbing sensation in oral
cavity by sanshool compounds of Zanthoxylum, which
aided in buffering ruminal pH thereby favoring
digestibility in the rumen. The milk yields were not
significant; however, when compared within plant at
two dose levels higher doses tended to promote higher
milk yields, which were due to increased DMD, in
spite of low DMI at the same dose levels. Therefore,
Siamese neem and Zanthoxylum are potential plants
for use as additives in dairy production system.
Acknowledgments
The study was funded by Thailand International
Development
Cooperation
Agency
(TICA)
and
supported in part by Thermo-tolerant Dairy Cattle

Research Group, and Food and Functional Food
Research Cluster, Faculty of Agriculture, Khon Kaen
University.
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management [Online], 2002, Retrieved Apr. 3, 2011,
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Bov. Practitioner 18 (1983) 114-118.

DAVID
PUBLISHING
Feeding Effect of Triticale Fodder as Replacement of

Straw on Production Performance of Dairy Cows
Nathu Ram Sarker1, Mohammad Asaduzzaman1, Khan Shahidul Huque1, Mohammad Toyebur Rahman2, Nazrul
Islam2, Mohammad Enamul Haque3 and Stephen R. Waadington3
1. Animal Production Research Division, Bangladesh Livestock Research Institute, Savar, Dhaka 1341, Bangladesh
2. Livestock Officer, Department of Livestock Services, Krishi Kamar Sarak, Dhaka 1207, Bangladesh
3. Programm Officer and CIMMYT Regional Agronomist, CIMMYT Bangladesh Office, Uttara, Dhaka, Bangladesh
Received: July 27, 2012 / Published: January 20, 2013.

Abstract: Twenty lactating cows of two to five parity having an average live weight of 290.00 to 330.00 kg and an average milk
production of 4.54 kg/head/day to 4.66 kg/head/day were selected from the Central Cattle breeding Station and Dairy Farm. The four
dietary treatment were: S100T0 (Straw 100: Triticale 0 as control); S50T50 (Straw 50: Triticale 50); S25T75 (Straw 25: Triticale 75) and
S0T100 (Straw 0: Triticale 100). It was observed that the roughage dry matter intake (DMI) (2.61 0.07 kg) in percent live weight was
significantly (P < 0.05) higher in S25T75 and the lowest (1.76 0.018 kg) in the control group (S100T0) and the differences were
significant among the dietary treatments except S50T50 and S0T100. The total DMI was significantly (P < 0.05) higher in S0T100 (13.36
0.13 kg) followed by S25T75, S50T50 and S100T0, respectively. The digestibility of crude protein (CP) was slightly higher in S25T75
(75.48 0.96) compared to S0T100 (75.31 1.45) and the difference was non-significant (P > 0.05). Milk production was
significantly (P < 0.05) the highest in S50T50 and the lowest in S100T0 followed by S0T100, S25T75, respectively. The percent increased
in milk yield was also the highest in S50T50 (67.68%) and the lowest in S100T0 (28.85%). The 4% fat corrected milk was also
significantly (P < 0.05) higher in S50T50 followed by S0T100, S25T75 and S100T0, respectively. Therefore, it can be concluded that
triticale and straw at a ratio of 50:50 may be fed for better production performance of dairy cows.
Key words: Triticale green fodder, production performance, replacement, milk yield.
1. Introduction
Feed scarcity is one of the major problem and most
limiting factor in livestock and poultry production in
Bangladesh, especially, during lean season from
March to May for ruminants livestock and throughout
the year for poultry. It has been estimated that in each
year, Bangladesh requires about 27 million tons of
roughage to achieve reasonable production levels from
its current herds. But, the existing supply meets only
50% of the total demand [1]. Imports to meet the gaps
are neither always feasible nor economic and seasonal
feed shortages mean high fodder prices and as a
Corresponding author: Nathu Ram Sarker, Senior Scientist,

research fields: animal nutrition, fodder production,
preservation & utilization. E-mail: sarkernr62@yahoo.com.
results poor in livestock production performance. The

situation is mostly aggravated during lean season and
in areas recurrently affected by drought and salinity.
During these periods, the quantity of fodder is
insufficient and its quality in terms of protein and
energy is very low. The challenge for researchers
working with the small-scale farmers is to identify
fodder technologies that matching the existing
cropping pattern of small holder farmers without
affecting major changes in inputs and risks.
Overcoming this challenge is the key to satisfy the
needs and aspirations of many farmers who want to
rear dairy animals and poultry and its productivity
through improvement of cropping systems. It was
observed that small-scale dairy and poultry production
Feeding Effect of Triticale Fodder as Replacement of Straw on Production Performance of Dairy Cows
have preferred option for small-scale farm households

as an income generating activity and employment
opportunities. Based on the research and field
experiments, it was revealed that triticale, is a crop
with good potential to meet the demands of quality
fodder as well as grain for a significant number of
farmers. Most triticale production is utilized as a feed
grain forage, or both in animal feeding, including
poultry, monogastrics, and ruminants.
The transformation of triticale from a scientific
curiosity to a viable crop in the course of a few
decades has been a remarkable achievement in plant
breeding. However, several grain and non-grain
factors have caused triticale to fail as a commercial
food grain. Overenthusiastic promotion of triticale as a
great nutritious new grain in the early 1970s
disappointed those who attempted to exploit it,
commercially, greatly damaging the image of a
cereal that was still far from having more stable and
acceptable attributes. Global wheat surpluses, lack of
year to year consistency in the composition of triticale
grain, absence of official triticale grading systems, and
lack of proper promotion are additional factors that
have not permitted the formation of the farmer
industry-consumer chain necessary for triticale to
become established as a commercial food grain. This
resulted in disappointment for both farmers and
researchers in developed and developing countries.
Despite this, efforts to resolve the basic problems of
triticale continued. As a consequence, the areas under
triticale production worldwide during the 1986-1992
periods increased from 1 million to nearly 2.5 million
hectares. At present, most triticale cultivation is in
Europe (78%), followed by North America (7%),
Africa (6%), Latin America (5%), and Australia and
New Zealand (4%). Except for a few planted areas in
China, the crop is not commercially grown in Asia.
Active research in enhancing the productivity and end
product quality and promotion of triticale is underway
in more than 30 countries [2].
Farmers in every part of the world have adopted
73
new techniques and accepted new crops that are

considered profitable and consistent with their
circumstances. The first factors, which favored
farmers adoption of triticale, were its superior
performance under unfavorable production conditions
including acidic soils, severe disease or insect
pressures, or drought. Second, it had the ability to
produce higher biomass and high regrowth capacity
after grazing and ability to grow better under
relatively cool temperatures, making it an excellent
forage crop. Third, and equally important, was the
usefulness of triticale as a feed grain mainly for
monogastric animals.
Interest in triticale has developed around two areas
of potential use for the grain. The first area of interest
is for use as a feed grain because it has proven to be a
good source of protein, amino acids and B vitamins. It
has shown promise as both a forage crop and as an
alternative protein source in formulated rations for
monogastrics, ruminants and poultry. The second area
of interest for triticale is in developing the grain as a
food grain cereal that would exhibit unique baking
traits. As a food grain, Triticale has also been
recognized as a hardy crop capable of helping combat
world hunger.
The versatility that triticale offers as a grain, a
forage, for straw, and as a cover crop adds to the
economic viability that sustains the interest in the crop.
Triticale will likely continue to experience increased
levels of production if it is supported with solid
research in genetics, production and utilization.
Triticale a cross cereal crop of wheat and rye
appears to be a particularly promising new crop for
farmers, because of its high yield potential, stress
tolerance (especially drought tolerance), and disease
resistance. Triticale is likely to become an
increasingly important cereal that may, in time, even
supplant wheat or maize in some areas [3].
Triticale possesses nutritional qualities very similar
to those of wheat, though it has a higher lysine content
and a better mineral balance. Triticales good protein
74
digestibility and its high content of essential amino

acids make it a suitable substitute for the most cereal
grains used in ruminant and non-ruminant diets.
Triticale contained some anti-nutritional factors such
as trypsin and chymotrypsin inhibitors that may
depress feed intake [4]. Triticale also exhibits special
qualities as a forage crop. It has higher protein content
than oats and produces higher forage and silage yields
than oats, barley, wheat, or rye [5]. Supplementation
is necessary to meet the production needs of
ruminants fed with crop residues. Supplementation
may affect the metabolism of both body and rumen
microbes of ruminants. The principal objectives of
supplementation of fibrous crop residues are to
optimize animal productivity through improved
utilization of the residues by the animal and to meet
the requirements of animal for production. Research
works on supplementation of rice straw by legume
forage have been conducted by the several workers
but very little or no work on utilization of triticale
green fodder supplementation with straw based diet.
Based on the above principles, the effect of triticale
green fodder supplementation to straw based diet of
lactating cows was investigated with the following
objectives: 1) to determine the optimum level of
supplementation of triticale green fodder to a rice straw
diet of dairy cows; 2) to determine effect of feeding
triticale green fodder on milk production and its quality.

2.1 Study Area
The study was conducted at Central Cattle Breeding
Centre and Dairy Farm under the Department of
Livestock Services (DLS) in collaboration with
Bangladesh Livestock Research Institute, Savar, Dhaka.
2.2 Experimental Design
A total of 20 lactating crossbred (F Deshi) cows
of similar lactation period and with average body
weight ranges from 290.00 to 330.00 kg were selected
for the trial. All the experimental cows were
distributed in the respective dietary treatments on the

basis of individual milk production not on the basis of
body weight. All the experimental animals were
selected from Savar Dairy Farm. Homogeneous
productive (group average milk production of animal
varies from 4.54 kg/head/day to 4.66 kg/head/day)
lactating cows of early lactation were randomly
allotted a total of four dietary treatments having five
replications in each dietary treatment. The design of
the experiment was organized in such way that in one
tail, the design of experiment had four straw based
diets which were supplemented with 0%, 50%, 75%
and 100% of triticale green fodder on DM basis and
all the four diets were assigned to four treatment
groups of lactating cows having five cows in each
group. On the other tail of the experiment, the
concentrate portion of other three straw based diets
were replaced by 25%, 50%, 100% triticale green
fodder. Therefore, a total of four treatment groups
including control in one tails were organized a
Complete Randomized Design (CRD) and other three
treatment groups excluding control were arranged in
Complete Randomized Design separately to determine
the treatment effect of supplementation and
replacement of concentrate by triticale and also data
were analyzed for determining the differences among
the treatment means. All cows were allowed to
concentrate mixture as per actual requirement of milk
production in each treatment group. The experimental
layout and composition of concentrate mixture are
presented in Tables 1 and 2.
2.3 Cultivation of Triticale Fodder
A total of 1.75 hectare of land under Bangladesh
Livestock Research Institute research farm was taken
for triticale cultivation as green fodder. The standard
agronomical practices were followed for cultivation of
triticale fodder. The triticale fodder was cut from the
field at the age of 50 days and feeding to the
experimental cows. Cut and carry method was
followed for feeding the animals. Daily requirements
of triticale were collected every day.
Table 1
Experimental layout.
Treatment groups
S50T50 S25T75 S0T100
S100 (T0)
(T1)
(T2)
(T3)
Rice Straw
100
50
25
0
Triticale green fodder
0
50
75
100
Concentrate*
100
100
100
100
No. of animals
5
5
5
5
*Supplied from the savar dairy farm, DLS. S = Straw, C =
100% means concentrate mixture was supplied to cows as per
milk production.
Feedstuff
Table 2 Composition of concentrate mixture used in the

feeding trial at Savar dairy farm
Ingredients
Maize grain
Wheat bran
Khesari bran
Soybean meal
Dicalcium phosphate powder (DCP)
Common salt
Total
Percent of concentrate
mixture
13.00
54.00
15.00
15.00
1.5
1.5
100
2.4 Feeding and Management

To accustom the test diets with experimental
animals, an adjustment period for 15 days was
followed before starting the actual feeding experiment.
De-worming and spraying against mites and others
external parasites were done before starting the
experiment. During the feeding trial, the animals were
confined in their respective feeding stalls for all the
times. During the adjustment period, the amounts of
feed offered to the dairy cows were gradually
increased until they reached to a constant level of
forage intake required for each treatment. The animals
were fed roughage diet thrice in a day and concentrate
was supplied in the morning and evening before
milking as per dietary treatment groups. Fresh clean
drinking water was supplied ad lib. at all times. The
feeding experiment was conducted for period of 45
days. All the animals were weighed at the beginning
and at the end of each of 15 days for calculation of
live weight changes.
2.5 Digestibility Trial
A digestibility trial was conducted at the mid of the
75
experimental period to determine the digestibility of

nutrients. On the 28th day of the feeding trial was
conducted and collection of fecal output was started.
During the digestibility trial, data were recorded on
the daily amount of feed offered, residues left and
faces voided. The total amount of faces voided for 24
h by the individual animal was collected quantitatively
at 8 am daily. It was weighed and representative
samples of each animal were drown in a polythene
bag and 1/20th the fresh daily sample was taken for
dry matter and crude protein analysis. The faecal
samples were collected daily and stored in freezer
(-20 C) for their further chemical analysis. Faces
samples, five days total collection were aggregated,
mixed and 10% sample was used for the chemical
analysis.
2.6 Data Collection
Feed offered and feed refusal were recorded daily to
determine voluntary DM intake. Daily milk yield of
cow was weighed individually and recorded. Animals
were also weighed and recorded individually at
fortnightly basis by using floor analog balance. Milk
samples were collected weekly basis immediately
after milking of cows and stored in a deep freeze for
future chemical analysis. The fat percentage of milk
sample was determined at same day of collection. The
representative samples of triticale green, straw,
concentrate mixture and refusals were also collected
for chemicals analysis. Milk samples were collected in
weekly basis in the morning was analyzed for butter
fat, crude protein, acidity and solids-not-fat (SNF)
contents.
2.7 Chemical Analysis
The proximate and other components of feed and
fecal materials were done by following the methods
described [6]. Feed DM was determined by oven
drying at 105 C for 48 h. Ash was determined by
incinerating 5 g of air-dried sample at 550 C for 16 h.
Acid detergent fibre was determined Van Soest
method using Labcono [7] (Model 30002 Hot
76
Extractor). Kjeldhal method was used for determining

the Nitrogen (N) content of feed, faeces and milk and
the crude protein content were estimated as N 6.25.
Gerber method was used for determining the fat
percentage of milk samples.
The Kjeldhal method was used for determining the

Nitrogen (N) content of milk and the milk protein
content were estimated as N 6.25 and milk fat
percentage was determined by the Gerber method.
The collected data will be analyzed through
Completely Randomized Design (CRD) and analysis
of variance was done to determine the treatment
effects. Collected data were analyzed statistically by
using Compare Means (CM) procedure of One-Way
Analysis of variance (ANOVA) [8]. Post Hoc
Multiple Comparisons of SPSS 11.5 for Windows
(SPSS Inc. 2002) [9].

3.1 Chemical Composition of the Feeds
The chemical composition of triticale green fodder,
paddy straw and concentrate mixture are presented in
Table 3. The average proximate component of triticale
fodder such as DM, CP, organic matter (OM) and ash
contents were 23.45%, 11.70%, 93.44% and 6.02%,
respectively. Similarly, the chemical composition of
paddy straw was used in the experiment containing
90.92%, 3.35%, 93.44% and 6.06% DM, CP, OM and
respectively. The concentrate
mixture was
Table 3 Chemical compositions (% DM basis) of triticale

green fodder, paddy straw and concentrate mixture.
Triticale
fodder
Dry matter (DM %)
23.45
Crude protein (CP)
11.70
Acid detergent fibre (ADF) 44.90
Ash
6.02
Organic matter (OM)
93.98
Parameters
Paddy
straw
90.92
3.35
44.77
6.06
93.44
and Dairy Farm, contained DM, CP, OM and ash

89.87%, 20.83%, 90.23% and 9.77%, respectively.
The ADF contents of triticale fodder, paddy straw and
concentrate mixture were 44.90%, 44.77% and
15.22%, respectively.
2.8 Analysis of Milk Fat and Protein
ash,
prepared and supplied by the Central Cattle Breeding
Concentrate
mixture
89.87
20.83
15.22
9.77
90.23
3.2 Dry Matter (DM) Intake

Average intake of dry matter (DM), estimated
metabolizable
energy
(ME)
and
estimated
metabolizable protein (MP) by the lactating cows is
shown in Table 3. Overall average daily total
roughage DM intake was increased with the increase
in supplementation of triticale green fodder. It reveals
that DM intake was significantly (P < 0.05) differ
among the treatment groups except 75% and 100%
triticale supplementation but the difference was
significant (P < 0.05) between S100T0 and S50T50
dietary treatments and between S100T0 and S25T75;
S100T0 and S0T100, respectively. Similarly, DM intake
was significantly (P < 0.05) differ between S50T50 and
S25T75; S50T50 and S0T100, respectively. The roughage
DM intake (2.61 0.07 kg) in percent live weight was
significantly (P < 0.05) higher in S25T75 treatment
group and lowest (1.76 0.018 kg) in control group
(S100T0) and differences were significant among the
dietary treatments groups except S50T50 and S0T100
treatments. The concentrate was supplied as per
requirement of the milk production of the animals.
Initially, concentrate requirement as per group average
of milk production was statistically differed among
the treatment groups but later on due to the increased
milk production, the requirement of concentrate
mixture among groups were differ significantly. The
study reveals that the average concentrate DM intake
was significantly higher (5.09 0.08 kg d-1) in S0T100
treatment group followed by S50T50, S25T75 and S100T0
treatment groups, respectively. It also indicates that
average concentrate DM intake between S100T0 and
S25T75 treatment groups (4.47 0.09 kg) vs 4.48
0.07 kg) was not differed significantly (P > 0.05). On
the other hand, the average concentrate DM intake

among S100T0, S50T50 and S0T100 treatment groups was
differ significantly (P < 0.05). The values of average
concentrate DM intake for the treatment groups S100T0,
S50T50 and S0T100 were 4.47 0.09, 4.69 0.09 and
5.09 0.08 kg, respectively.
The average total intake of dry matter was
significantly (P < 0.05) higher in S0T100 dietary group
(13.36 0.13 kg) followed by S25T75, S50T50 and
S100T0 dietary treatment groups, respectively. The total
DM intake in percent live weight was significantly (P
< 0.05) higher (4.02% 0.04%) in S25T75 treatment
group followed by S0T100, S50T50 and S100T0 dietary
treatment groups, respectively. The average values of
DM intake in percent live weight for S100T0, S50T50,
S25T75 and S0T100 treatments groups were 3.21%
0.03%, 3.55% 0.03%, 4.02% 0.04% and 3.82%
0.04%, respectively (Table 4). The total DM intake in
percent live weight was significantly (P < 0.05)
among S100T0, S50T50 and S25T75, but the difference
was statistically non-significant (P > 0.05) between
S50T50 and S0T100 treatment groups. It was reported
[10] that dry matter intake by the lactating cows of
developing countries between 250 and 300 kg body
weight was 6.4-7.3 kg d-1.
The DM intake on the basis of per kg metabolic
body size was significantly (P < 0.05) higher in S25T75
dietary treatment group (170 0.002 g-1 W0.75 kg)
followed by S0T100, S50T50 and S100T0, respectively.
The DM intake g-1 W0.75 was significantly differed
between S25T75 and S0T100 treatment groups and
among S50T50, S25T75 and S0T100 treatment, but there
was no significant difference between S100T0 and
S50T50 treatment groups.
Total estimated metabolizable energy (ME MJ d-1)
intake by the lactating cows of different dietary
treatment groups were 89.15 1.39, 108.27 0.99,
109.26 0.88 and 93.54 0.92 for S100T0, S50T50,
S25T75 and S0T100, respectively. The metabolizable
energy intake was significantly higher in S50T50 and
S25T75 treatment groups, but the difference between
77
two treatments was non-significant (P > 0.05). While

on the other hand, ME intake was significantly (P <
0.05) differ between S100T0 and S50T50; S100T0 and
S0T100 treatment groups. Similarly, the difference was
significant between S50T50 and S0T100; S25T75 and
S0T100 treatment groups, respectively. The ME intake
kg-1 W0.75 by lactating cows was highest in S25T75
dietary treatment group followed by S50T50, S100T0 and
S0T100 treatment groups, respectively. The values of
ME intake kg-1 W0.75 among different treatment groups
were 1.21 0.001, 1.32 0.008, 1.45 0.013 and
1.16 0.001 for S100T0, S50T50, S25T75 and S0T100 for
treatment groups, respectively [11]. Suggested that the
maintenance requirement of lactating cows is slightly
higher than non-lactating dairy animals. This may be
100 kcal ME kg-1 W0.75 for the non-lactating animal
and 117 kcal ME kg-1 W0.75 for the lactating cows.
The total intake of estimated metabolizable protein
(MP g d-1) by the lactating cows feeding triticale
fodder supplementation with straw based diets reveals
that the highest MP intake was observed in S25T75
diets followed by S50T50, S0T100 and S100T0,
respectively. It also found that MP g day-1 was
non-significant between S50T50 and S25T75 treatment
groups but the difference was significant (P < 0.05)
between S100T0 and S50T50; S100T0 and S0T100; S25T75
and S0T100, respectively. The values of MP intake g
kg-1 W0.75 were 11.62 0.12, 12.76 0.09, 13.96
0.13 and 11.09 0.11 for S100T0, S50T50, S25T75 and
S0T100 treatment groups, respectively. The difference
in MP intake g kg-1 W0.75 was significant among the
treatment groups except S100T0 and S0T100 treatments.
3.3 Production Performance
The production performance of cows fed straw
based diet supplemented with triticale fodder is
presented in Table 5. It indicates that initial milk
production was almost similar in all dietary treatment
groups, but the final milk production was significantly
(P < 0.05) the highest in S50T50 treatment group and
lowest in S100T0 followed by S0T100, S25T75 treatment
78
Table 4
Nutrient intake by lactating cows
Parameters
S100T0 (Mean SE) S50T50 (Mean SE) S25T75 (Mean SE) S0T100 (Mean SE) Sig.
Roughage dry matter intake (kg d-1)
*
5.38a 0.07
7.77b 0.04
8.25c 0.07
8.27c 0.13
Roughage dry matter intake as % of live 1.76a 0.018
*
2.24b 0.03
2.61c 0.07
2.37b 0.04
wt.
Concentrate dry matter intake (kg d-1) 4.47a 0.09
*
4.69b 0.09
4.48a 0.07
5.09c 0.08
-1
Total dry matter intake (kg d )
*
9.86a 0.15
12.45b 0.08
12.73b 0.09
13.36c 0.13
Total dry matter intake as % of live wt. 3.21a 0.03
*
3.55b 0.03
4.02c 0.04
3.82b 0.04
Dry matter intake (g/kg W0.75 d-1)
*
130a 0.001
150a 0.009
170c 0.002
160b 0.002
ME intake (MJ d-1)
*
89.15a 1.39
108.27c 0.99
109.26c 0.88
93.54b 0.92
Total MP intake (g d-1)
*
855.79a 13.44
1039.39c 9.46
1048.89c 8.52
898.05b 8.82
0.75 -1
ME intake (MJ/kg W d )
*
1.21a 0.001
1.32b 0.008
1.45c 0.013
1.16a 0.001
MP intake (g/kg W0.75 d-1)
*
11.62a 0.12
12.76 b 0.09
13.96c 0.13
11.09a 0.11
SE: Standard error of mean; NS: Non-significant; *Significant at 5% level; Means with different letters in the same row differ
significantly. (S100T0: 100% Rice Straw + Zero triticale fodder; S50T50: 50% Rice Straw + 50% Triticale Fodder; S25T75: 25% Rice
Straw + 75% Triticale Fodder and S0T100: 100% Triticale Fodder; In addition to the roughages, concentrate mixture was supplied as
per milk production of the cows in all treatment groups ).
Table 5
Production performances of lactating cows fed straw based diet supplemented with triticale green fodder.
Parameters
S100T0 (Mean SE) S50T50 (Mean SE)
Average initial milk yield (L d-1)
4.54 0.74
4.58 0.95
Average final milk yield (L d-1)
5.85d 1.28
7.68a 1.09
Milk yield increase (Ld -1)
1.31d
3.10a
Per cent of milk yield increase (%) 28.85d
67.68a
Initial body weight (kg)
290.00 23.32
330.00 27.82
Final body weight (kg)
318.00 22.00
377.00 27.29
Daily live weight gain(kg d-1)
0.65d 0.05
1.12a 0.09
SE: standard error of mean; NS: Non-significant; *Significant at 5% level;
significantly.
groups, respectively. The difference in milk

production between S25T75 and S0T100 was differed
significantly (P < 0.05). It also reveals that average
milk production increased L d-1 was the highest in
S50T50 treatment group (3.10 L d-1) and the lowest in
S100T0 treatment group (3.10 L d-1) and difference
between the treatment was significant (P < 0.05). In
other two treatment groups, the milk production was
increased by 2.05 and 2.34 L d-1 for S25T75 and S0T100
treatment groups, respectively. The study revealed that
cows received 50% straw and 50% (S50T50) triticale
fodder produced the highest amount of milk compared
to other treatment groups. This highest milk
production may due to the synergistic effect of straw
and triticale green fodder and better nutrients flow for
milk production. In terms of per cent increased in milk
yield was also the highest in S50T50 treatment group
S25T75 (Mean SE)

4.60 0.84
6.65c 0.50
2.05c
44.56c
303.00 15.62
333.00 16.04
0.75c 0.02
Means with different
S0T100 ( Mean SE) Sig.

NS
4.66 0.89
*
7.00b 0.44
2.34b
*
50.21b
*
NS
333.00 20.28
*
370.00 18.00
*
0.89b 0.13
letters in the same row differ
(67.68%) and the lowest in S100T0 treatment group

(28.85%). The differences in percent in milk
production among the treatment groups were differed
significantly (P < 0.05).
The initial average body weight of lactating cows
used in the experiment ranges from 290.00 23.32 to
333.00 20.28 kg. It mentioned here that selection of
cows was done based on milk production of the
animals. During selection, the average Body
Condition Score (BCS) of the experimental was 2.0.
Therefore, all the animals were weight gained in
addition to milk production. The nutrients supplied to
the cows through diet utilized for milk production and
additional nutrients were distributed in the tissue
levels and utilized for the muscle growth. The results
indicate that the highest body weight was observed in
S50T50 treatment group (1.12 0.09 kg d-1) and the
3.4 Milk Quality
10
Milk Yield (Litre)
lowest in S100T0 treatment group (0.65 0.05 kg d-1).

Further, it was also found that the body weight gain
was significantly (P < 0.05) differed among the
treatment groups (Table 5). The milk production trend
of different dietary treatment groups are shown in Figs.
1-4 and percent increased are shown in Figs. 5-7.
Milk production trend of different treatment groups
of lactating cows fed triticale fodder supplementation
with straw based diets.
6
4
y = -0.0008x + 7.0266
R2 = 0.0003
9 13 17 21 25 29 33 37 41
Lactation Period (d)
Fig. 4 Group average milk yield of 100% triticale fodder

+ 100% concentrate fed cows.
1.12
1.2
Live wt. gain (kg/d)
Milk Yield (Litre)
The effect of triticale feeding with straw based diet

on milk quality is shown in Table 6. It reveals that the
7
6
5
4
3
2
1
0
y = -0.0082x + 6.0316
R2 = 0.1251
0.89
1
0.8
0.75
0.67
0.6
0.4
0.2
0
Group A Group B Group C Group D
Group categories
9 13 17 21 25 29 33 37 41
Fig. 5
Live wt. gain performance.
Lactation Period (D)
Milk yield increased (l/d)
Average milk yield of cows fed 100% straw + 100%
Milk Yield ( Litre)
Fig. 1
conc.
10
8
6
y = 0.0245x + 7.0322
R2 = 0.2089
4
2
0
1
3.5
3.09
3
2.5
2.09
1.5
1.31
1
0.5
0
Group A Group B Group C Group D
Group Categories
Fig. 6 Milk production performance.
% increasd of milk yield
Milk Yield (Litre)
10
8
6
y = 0.021x + 6.1328
R2 = 0.1661
4
2
0
1
9 13 17 21 25 29 33 37 41
80
67.47
60
40
45.44
20
0
Group B
Group C
Categories
Fig. 7
50.21
28.85
Group A
Fig. 3 Group average milk yield of 25% strew + 75%

triticale fodder + 100% concentrate fed cows.
2.34
9 13 17 21 25 29 33 37 41
Fig. 2 Average milk yield of cows fed 50% straw + 50 %

Concentrate + 100% conc.
79
Percent milk yield increased.
Group D
80
Table 6
Feeding triticale fodder with straw based diet on milk quality of lactating cows.
Parameters
S100T0 (Mean SE) S50T50 (Mean SE) S25T75 (Mean SE) S0T100 ( Mean SE) Sig.
Fat (%)
*
3.69d 0.08
4.60a 0.09
4.12b 0.12
4.05c 0.19
Milk protein (%)
*
3.27d 0.07
3.60a 0.06
3.50b 0.07
3.39c 0.09
Solids-not-fat (%)
NS
8.14 0.12
8.33 0.06
8.26 0.09
8.05 0.09
Total solids (%)
*
11.91c 0.10
12.53a 0.09
12.36b 0.16
12.19b 0.24
Water (%)
NS
88.03 0.09
87.38 0.11
87.64 0.20
87.87 0.26
Specific gravity
NS
1.029 0.0003
1.029 0.0003
1.029 0.0003
1.029 0.0004
Milk yield (L d-1)
*
5.85d 0.44
7.51a 0.09
6.59c 0.09
7.00b 0.09
4% FCM
*
5.58d 0.04
8.18a 0.10
6.71c 0.10
7.06b 0.09
SE: standard error of mean, *significant at 5% level; Means with different letters in the same row differ significantly; NS: Non-significant.
(S100T0: 100% Paddy Straw + zero triticale + Concentrate, S50T50: 50% Paddy Straw + 50% Triticale Fodder + Concentrate; S25T75: 25%
Paddy Straw + 75 % Triticale Fodder + concentrate and S0T100: Zero straw + 100% Triticale Fodder + Concentrate).
significantly (P < 0.05) of the highest (4.60 0.09) fat

percent was observed in S50T50 treatment and the
lowest (3.69 0.08) in S100T0 treatment group and the
difference was significantly (P < 0.05) between the
treatment groups. The values for fat percent in S25T75
and S0T100 treatment groups were 4.12 0.12 and 4.05
0.19, respectively and the difference was also
significant (P < 0.05) between them. The SNF, SP and
percent water did not differ significantly (P > 0.05)
among the treatment groups. Further, it observed that
percent of total solids (TS) in milk was differed
significantly (P < 0.05) among the treatment groups.
The significantly (P < 0.05) highest (12.53 0.09) TS
was found in S50T50 treatment group and the lowest
(11.91 0.10) in S100T0 treatment group. The values
for TS in S25T75 and S0T100 treatment groups were
12.36 0.16 and 12.19 0.24, respectively. The milk
protein was significantly (P < 0.05) higher in S50T50
followed by S25T75, S0T100 and S100T0, respectively
and the differences were significant among the
treatment groups. The 4% fat corrected milk was also
significantly (P < 0.05) higher in S50T50 treatment
group followed by S0T100, S25T75 and S100T0 treatment
groups, respectively (Table 6).
3.5 Digestibility of Nutrients
The co-efficient digestibility of nutrients of
different diets fed lactating cows is presented in Table
7. It is indicated that the highest co-efficient of DM
digestibility (73.29 0.80) was found in S25T75
treatment group and the lowest in S100T0 treatment

group (64.98 2.25) and the difference was
significant (P < 0.05) between the treatment groups.
Further, the co-efficient of DM digestibility of S50T50
and S25T75 treatment groups were 66.96 0.69 and
73.29 0.80, respectively and there was a significant
(P < 0.05) difference between them. Though, there
was a non-significant (P > 0.05) difference of
co-efficient DM digestibility between S25T75 and
S0T100 treatment groups but little higher DM
digestibility was observed in 100% triticale
supplemented diet compared to 75% triticale group.
Similarly, the values of OM digestibility were 70.04
1.92, 70.23 0.64, 76.31 0.72 and 76.79 0.88 for
S100T0, S50T50, S25T75 and S0T100 treatment groups,
respectively. The co-efficient of digestibility of OM
did not differ significantly (P > 0.05) between S100T0
and S50T50; S25T75 and S0T100 treatment groups,
respectively. The values for co-efficient of
digestibility of ADF were 62.32 2.31, 64.02 0.71,
72.66 0.79 and 70.63 1.64 in S100T0, S50T50, S25T75
and S0T100 dietary treatment groups, respectively. It
also reveals that the ADF digestibility co-efficient was
significantly (P < 0.05) differ among the treatment
groups. The co-efficient of digestibility of CP was
slightly higher in S25T75 (75.48 0.96) treatment
group compared to S0T100 treatment group (75.31
1.45) and difference between the treatments was
non-significant (P > 0.05). On the other hand,
co-efficient of digestibility of CP in S100T0 and S50T50
Table 7
81
Co-efficient digestibility of triticale fodder supplemented with straw based ration fed lactating cows.
Parameters
DM (%)
OM (%)
ADF (%)
CP (%)
Total ash (%)
S100T0 (Mean SE)

64.98c 2.25
70.04b 1.92
62.32d 2.31
69.34b 1.94
22.80c 4.20
S50T50 (Mean SE)

66.96b 0.69
70.23b 0.64
64.02c 0.71
65.08c 1.68
26.66b 1.76
S25T75 (Mean SE)

73..29a 0.80
76.31a 0.72
72.66a 0.79
75.48a 0.96
35.58a 1.93
S0T100 (Mean SE)

73.64a 1.00
76.79a 0.88
70.63b 1.64
75.31a 1.45
35.07a 2.33
Sig.
*
*
*
*
*
SE: standard error of mean; *Significant at 5% level.
treatment groups were 69.34 1.94 and 65.08 1.68,

respectively and the difference was significant (P <
0.05) between the treatment groups. This similar
digestibility of DM, OM and CP was observed by
Sarker et al. [12] who compared the DM, OM and CP
digestibility of triticale fodder with triticale hay in
dairy cows. The values for digestibility co-efficient of
ash were 22.80 4.20, 26.66 1.76, 35.58 1.93 and
35.07 2.33 for S100T0, S50T50, S25T75 and S0T100
treatment groups, respectively. The data indicates that
with the increased level of triticale fodder
supplementation, digestibility co-efficient was also
increased. This due to fact that the total minerals
content of rice straw may be more liginified compared
to triticale fodder. Therefore, there was a increasing
trend of digestibility co-efficient of ash among the
different treatment groups. Sutton et al. [13] fed grass
silage and urea treated whole-crop wheat silage was
mixed with the grass silage to replace the 0.0, 0.33,
and 0.67 dry matter of the grass silage and found the
effect of the inclusion of urea treated whole-crop
wheat silage on the overall digestibility coefficients
was significant (P 0.05). The addition of the
UWCWS in the diet decreased the digestibility of the
DM, OM, ADF, and NFE but effect on the protein
digestibility was non significant.
50:50 for harvesting better yield from dairy cows and

as well as proper utilization of straw and reduce the
cost of production by saving extra fodder for future
use.
Acknowledgments
The authors are gratefully to Mr. Nur Islam,
Director (Production), Dr. Santi Ranjan Das, Deputy
Director, Mr. Kazi Asraful Alam, Buyer Officer and
Mr. Md. Shafiqur Rahman, Scientific Officer, Dairy
Section of Central Cattle Breeding and Dairy Farm
under the Department of Livestock Services, for their
heartfelt cooperation in facilitating this experiment by
providing dairy cows. The authors also acknowledge
the contribution of CIMMYT for providing financial
help through DANIDA to carryout this work smoothly
and timely. Authors are also thankful to all milkman
and Goalas for their co-operation, contribution and
help by timely milking the cows and helping data
recording during the whole period of the study.
References
[1]
[2]
4. Conclusions
Triticale fodder is good roughage that may be used
for dairy cows without any adverse effect on intake
and digestibility. The high digestibility of triticale
fodder suggesting that this is promising forage may be
used as supplementation with straw based at a ratio of
[3]
[4]
[5]
M. Saadullah, Smallholder dairy production and

marketing-opportunities and constraints, in: D.
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[8] G.W. Snedecor, W.G. Cochran, Statistical Methods, 7th
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[10] L. Kearl, Nutrient requirements of ruminants in
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[11] A.J. van Es, Nutrient requirements of dairy cattle, in: P.V.
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the 1st Int. Symp. on Feed Composition, Animal Nutrient
Requirements and Computerization of Diets, Utah Agr.
Exp. Sta., Utah State Univ. Logan, 1976, pp. 512-518.
[12] N.R. Sarker, M. Asaduzzaman, K.S. Huque, M.T.
Rahman, M.E. Haque, S.R. Waadington, Production
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Research 18 (1-2) (2011) 20-25.
[13] J.D. Sutton, K. Aston, D.E. Beever, M.S. Dhanoas, Milk
production from silage based diets: Effects of
high-protein concentrates for lactating heifers and cows
on intake, milk production and milk nitrogen fractions,
Animal Sci. 62 (1996) 207-215.

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engineering learning outcomes are limited and,

define common accreditation procedures, also in

Biosystems Engineering Curricula in Europe

criteria to be used for professional and academic

Agricultural and Bioprocess Engineering (Tables 1

Courses not included

Agricultural/Biological Sciences 9-11 %

General courses (adopted from E4-TN)

Biosystems Engineering Curricula in Europe

Agricultural/Biological Sciences part of the core curricula :

promoted, in August 2005, the dissemination proposal

develop a web-based database, containing the

courses or modules (set of courses) of the study

to specify the percentage of non-technical subjects

database will be continuously updated and made

available using a specific authorisation;

enhance the USAEE web-site, by creating links

with organisations, other related thematic networks

Agriculture and Related Sciences-ICA, Training &

Biosystems Engineering Curricula in Europe

quality assessment and employability;

2. From Agricultural Engineering to

Biosystems Engineering Curricula in Europe

study programs and the European doctorate in

define the emerging biosystems engineering

3. A Case of EU-US Cooperation in

Biosystems Engineering Curricula in Europe

The objectives of the POMSEBES project, which

courses/topics and disseminating these courses into

The highest achievements of ERABEE-TN, which

areas of application and a common definition of

student course load.

programs in Europe benefits from the results of the

G. Pellizzi, P. Febo, Degree study programs in

Biosystems Engineering Curricula in Europe

and Extension 1 (3) (1994) 59-68.

Studies in Engineering for Agriculture, Food and Rural

Journal of Agricultural Science and Technology A 3 (2013) 10-19