A biofilm is any group of microorganisms in which cells stick to each other and

often these cells adhere to a surface. These adherent cells are frequently embed
ded within a self-produced matrix of extracellular polymeric substance (EPS). Bi
ofilm extracellular polymeric substance, which is also referred to as slime (alt
hough not everything described as slimeis a biofilm), is a polymeric conglomerat
ion generally composed of extracellular DNA, proteins, and polysaccharides. Biof
ilms may form on living or non-living surfaces and can be prevalent in natural,
industrial and hospital settings.[2][3] The microbial cells growing in a biofilm
are physiologically distinct from planktonic cells of the same organism, which,
by contrast, are single-cells that may float or swim in a liquid medium.
Microbes form a biofilm in response to many factors, which may include cellular
recognition of specific or non-specific attachment sites on a surface, nutrition
al cues, or in some cases, by exposure of planktonic cells to sub-inhibitory con
centrations of antibiotics.[4][5] When a cell switches to the biofilm mode of gr
owth, it undergoes a phenotypic shift in behavior in which large suites of genes
are differentially regulated.[6]
Formation[edit]
An iridescent biofilm on the surface of a fish tank.
Formation of a biofilm begins with the attachment of free-floating microorganism
s to a surface. While still not fully understood, it is thought that the first c
olonists of a biofilm adhere to the surface initially through weak, reversible a
dhesion via van der Waals forces and hydrophobic effects.[7][8] If the colonists
are not immediately separated from the surface, they can anchor themselves more
permanently using cell adhesion structures such as pili. Hydrophobicity also pl
ays an important role in determining the ability of bacteria to form biofilms, a
s those with increased hydrophobicity have reduced repulsion between the extrace
llular matrix and the bacterium.[9]
Some species are not able to attach to a surface on their own but are instead ab
le to anchor themselves to the matrix or directly to earlier colonists. It is du
ring this colonization that the cells are able to communicate via quorum sensing
(QS) using products such as N-acyl homoserine lactone (AHL). Some bacteria are
unable to form biofilms as successfully due to their limited motility. Non-motil
e bacteria cannot recognize the surface or aggregate together as easily as motil
e bacteria.[9] Once colonization has begun, the biofilm grows through a combinat
ion of cell division and recruitment. Polysaccharide matrices typically enclose
bacterial biofilms. In addition to the polysaccharides, these matrices may also
contain material from the surrounding environment, including but not limited to
minerals, soil particles, and blood components, such as erythrocytes and fibrin.
[9] The final stage of biofilm formation is known as dispersion, and is the stag
e in which the biofilm is established and may only change in shape and size.
The development of a biofilm may allow for an aggregate cell colony (or colonies
) to be increasingly resistant to antibiotics. Cell-cell communication or quorum
sensing has been shown to be involved in the formation of biofilm in several ba
cterial species.[10]
Development[edit]
Five stages of biofilm development: (1) Initial attachment, (2) Irreversible att
achment, (3) Maturation I, (4) Maturation II, and (5) Dispersion. Each stage of
development in the diagram is paired with a photomicrograph of a developing P. a
eruginosa biofilm. All photomicrographs are shown to the same scale.
There are five stages of biofilm development (see illustration at right):
1.
Initial attachment.
2.
Irreversible attachment.
3.
Maturation I.
4.
Maturation II.
5.
Dispersion.

In conclusion, it appears that targeting QS to mitigate membrane biofouling is a

promising technology for the enhancement of efficiency and performance in membr
ane systems. Results from recent investigations verified the existence of a corr
elation between QS activity and membrane biofouling. Different strategies have b
een demonstrated to target QS activity, in order to mitigate membrane biofouling
. However, current validation methods for QS must be improved and optimized, wit
h its particular toxicity effects on bacterial viability. Furthermore, emergence
of bacterial resistance to QS inhibitors and its adverse effects must be evalua
ted in-depth. Future advances in membrane biofouling mitigation based on QS can
be expected from further fundamental research.[citation needed]
Dispersal[edit]
Dispersal of cells from the biofilm colony is an essential stage of the biofilm
life cycle. Dispersal enables biofilms to spread and colonize new surfaces. Enzy
mes that degrade the biofilm extracellular matrix, such as dispersin Band deoxyr
ibonuclease, may play a role in biofilm dispersal.[11][12] Biofilm matrix degrad
ing enzymes may be useful as anti-biofilm agents.[13][14] Recent evidence has sh
own that a fatty acid messenger, cis-2-decenoic acid, is capable of inducing dis
persion and inhibiting growth of biofilm colonies. Secreted by Pseudomonas aerug
inosa, this compound induces cyclo heteromorphic cells in several species of bac
teria and the yeast Candida albicans.[15] Nitric oxide has also been shown to tr
igger the dispersal of biofilms of several bacteria species[16][17] at sub-toxic
concentrations. Nitric oxide has the potential for the treatment of patients th
at suffer from chronic infections caused by biofilms.[18]
It is generally assumed that cells dispersed from biofilms immediately go into t
he planktonic growth phase. However, recent studies have shown that the physiolo
gy of dispersed cells from Pseudomonas aeruginosa biofilms is highly different f
rom those of planktonic and biofilm cells.[19][20] Hence, the dispersal process
is a unique stage during the transition from biofilm to planktonic lifestyle in
bacteria. Dispersed cells are found to be highly virulent against macrophages an
d Caenorhabditis elegans, but highly sensitive towards iron stress, as compared
with planktonic cells.[19]
Properties[edit]
Biofilms are usually found on solid substrates submerged in or exposed to an aqu
eous solution, although they can form as floating mats on liquid surfaces and al
so on the surface of leaves, particularly in high humidity climates. Given suffi
cient resources for growth, a biofilm will quickly grow to be macroscopic (visib
le to the naked eye). Biofilms can contain many different types of microorganism
, e.g. bacteria, archaea, protozoa, fungi andalgae; each group performs speciali
zed metabolic functions. However, some organisms will form single-species films
under certain conditions. The social structure (cooperation/competition) within
a biofilm depends highly on the different species present.[21]
Extracellular matrix[edit]
The biofilm is held together and protected by a matrix of secreted polymeric com
pounds called EPS. EPS is an abbreviation for either extracellular polymeric sub
stance or exopolysaccharide, although the latter one only refers to the polysacc
haride moiety of EPS. In fact, the EPS matrix consists not only of polysaccharid
es but also of proteins (which may be the major component in environmental and w
aste water biofilms) and nucleic acids. A large proportion of the EPS is more or
less strongly hydrated, however, hydrophobic EPS also occur; one example is cel
lulose which is produced by a range of microorganisms. This matrix encases the c
ells within it and facilitates communication among them through biochemical sign
als as well as gene exchange. The EPS matrix is an important key to the evolutio
nary success of biofilms. One reason is that it traps extracellular enzymes and
keeps them in close proximity to the cells. Thus, the matrix represents an exter
nal digestion system and allows for stable synergistic microconsortia of differe
nt species (Wingender and Flemming, Nat. Rev. Microbiol. 8, 623-633). Some biofi
lms have been found to contain water channels that help distribute nutrients and
signalling molecules.[22] This matrix is strong enough that under certain condi
tions, biofilms can become fossilized (Stromatolites).
Bacteria living in a biofilm usually have significantly different properties fro

m free-floating bacteria of the same species, as the dense and protected environ
ment of the film allows them to cooperate and interact in various ways. One bene
fit of this environment is increased resistance to detergents and antibiotics, a
s the dense extracellular matrix and the outer layer of cells protect the interi
or of the community. In some cases antibiotic resistance can be increased a thou
sandfold.[23] Lateral gene transfer is greatly facilitated in biofilms and leads
to a more stable biofilm structure.[24] Extracellular DNA is a major structural
component of many different microbial biofilms.[25]Enzymatic degradation of ext
racellular DNA can weaken the biofilm structure and release microbial cells from
the surface.
However, biofilms are not always less susceptible to antibiotics. For instance,
the biofilm form of Pseudomonas aeruginosa has no greater resistance to antimicr
obials than do stationary-phase planktonic cells, although when the biofilm is c
ompared to logarithmic-phase planktonic cells, the biofilm does have greater res
istance to antimicrobials. This resistance to antibiotics in both stationary-pha
se cells and biofilms may be due to the presence ofpersister cells.[26]
Habitat[edit]
Mats of bacterial biofilm color the hot springs inYellowstone National Park. The
longest raised mat area is about half a meter long.
Thermophilic bacteria in the outflow of Mickey Hot Springs,Oregon, approximately
20mm thick.
Biofilms are ubiquitous. Nearly every species of microorganism, not only bacteri
a and archaea, have mechanisms by which they can adhere to surfaces and to each
other. Biofilms will form on virtually every non-shedding surface in a non-steri
le aqueous (or very humid) environment.

Biofilms can be found on rocks and pebbles at the bottom of most streams
or rivers and often form on the surface of stagnant pools of water. In fact, bi
ofilms are important components offood chains in rivers and streams and are graz
ed by the aquatic invertebrates upon which many fish feed.

Biofilms can grow in the most extreme environments: from, for example, t
he extremely hot, briny waters of hot springs ranging from very acidic to very a
lkaline, to frozen glaciers.

In the human environment, biofilms can grow in showers very easily since
they provide a moist and warm environment for the biofilm to thrive. Biofilms c
an form inside water and sewagepipes and cause clogging and corrosion. Biofilms
on floors and counters can make sanitation difficult in food preparation areas.

Biofilms in cooling- or heating-water systems are known to reduce heat t

ransfer.[27]

Biofilms in marine engineering systems, such as pipelines of the offshor

e oil and gas industry,[28] can lead to substantial corrosion problems. Corrosio
n is mainly due to abiotic factors; however, at least 20% of corrosion is caused
by microorganisms that are attached to the metal subsurface (i.e., microbially
influenced corrosion).

Bacterial adhesion to boat hulls serves as the foundation for biofouling

of seagoing vessels. Once a film of bacteria forms, it is easier for other mari
ne organisms such as barnacles to attach. Such fouling can reduce maximum vessel
speed by up to 20%, prolonging voyages and consuming fuel. Time in dry dock for
refitting and repainting reduces the productivity of shipping assets, and the u
seful life of ships is also reduced due to corrosion and mechanical removal (scr
aping) of marine organisms from ships' hulls.

Biofilms can also be harnessed for constructive purposes. For example, m

any sewage treatment plants include a secondary treatment stage in which waste w
ater passes over biofilms grown on filters, which extract and digest organic com
pounds. In such biofilms, bacteria are mainly responsible for removal of organic
matter (BOD), while protozoa and rotifers are mainly responsible for removal of
suspended solids (SS), including pathogens and other microorganisms. Slow sand

filters rely on biofilm development in the same way to filter surface water from
lake, spring or river sources for drinking purposes. What we regard as clean wa
ter is effectively a waste material to these microcellular organisms.

Biofilms can help eliminate petroleum oil from contaminated oceans or ma

rine systems. The oil is eliminated by the hydrocarbon-degrading activities of m
icrobial communities, in particular by a remarkable recently discovered group of
specialists, the so-called hydrocarbonoclastic bacteria (HCB).[29]

Stromatolites are layered accretionary structures formed in shallow wate

r by the trapping, binding and cementation of sedimentary grains by microbial bi
ofilms, especially of cyanobacteria. Stromatolites include some of the most anci
ent records of life on Earth, and are still forming today.

Biofilms are present on the teeth of most animals as dental plaque, wher
e they may cause tooth decay and gum disease.

Biofilms are found on the surface of and inside plants. They can either
contribute to crop disease or, as in the case of nitrogen-fixing Rhizobium on ro
ots, exist symbiotically with the plant.[30] Examples of crop diseases related t
o biofilms include Citrus Canker, Pierce's Disease of grapes, and Bacterial Spot
of plants such as peppers and tomatoes.[31]

Biofilms are used in microbial fuel cells (MFCs) to generate electricity

from a variety of starting materials, including complex organic waste and renew
able biomass.[3][32][33]

Recent studies in 2003 discovered that the immune system supports bio-fi
lm development in the large intestine. This was supported mainly with the fact t
hat the two most abundantly produced molecules by the immune system also support
bio-film production and are associated with the bio-films developed in the gut.
This is especially important because the appendix holds a mass amount of these
bacterial bio-films.[34] This discovery helps to distinguish the possible functi
on of the appendix and the idea that the appendix can help reinoculate the gut w
ith good gut flora.
Taxonomic diversity[edit]
Many different bacteria form biofilms, including gram-positive (e.g. Bacillus sp
p, Listeria monocytogenes, Staphylococcus spp, and lactic acid bacteria, includi
ng Lactobacillus plantarum and Lactococcus lactis) and gram-negative species (e.
g. Escherichia coli, or Pseudomonas aeruginosa).[35]
Biofilms are formed by bacteria that colonize plants, e.g. Pseudomonas putida, P
seudomonas fluorescens, and related pseudomonads which are common plant-associat
ed bacteria found on leaves, roots, and in the soil, and the majority of their n
atural isolates form biofilms.[36] Several nitrogen-fixing symbionts of legumes
such as Rhizobium leguminosarum and Sinorhizobium meliloti form biofilms on legu
me roots and other inert surfaces.[36]
For other species in disease-associated biofilms see below.
Biofilms and infectious diseases[edit]
Biofilms have been found to be involved in a wide variety of microbial infection
s in the body, by one estimate 80% of all infections.[37] Infectious processes i
n which biofilms have been implicated include common problems such as bacterial
vaginosis, urinary tract infections, catheter infections, middle-ear infections,
formation of dental plaque,[38] gingivitis, coating contact lenses,[39] and les
s common but more lethal processes such as endocarditis, infections in cystic fi
brosis, and infections of permanent indwelling devices such as joint prostheses
and heart valves.[40][41] More recently it has been noted that bacterial biofilm
s may impair cutaneous wound healing and reduce topical antibacterial efficiency
in healing or treating infected skin wounds.[42] Early detection of biofilms in
wounds is crucial to successful chronic wound management. Although many techniq
ues have developed to identify planktonic bacteria in viable wounds, few have be
en able to quickly and accurately identify bacterial biofilms. Future studies ar
e needed to find means of identifying and monitoring biofilm colonization at the
bedside to permit timely initiation of treatment.[43]
It has recently been shown that biofilms are present on the removed tissue of 80
% of patients undergoing surgery for chronic sinusitis. The patients with biofil
ms were shown to have been denuded of cilia and goblet cells, unlike the control

s without biofilms who had normal cilia and goblet cell morphology.[44] Biofilms
were also found on samples from two of 10 healthy controls mentioned. The speci
es of bacteria from interoperative cultures did not correspond to the bacteria s
pecies in the biofilm on the respective patient's tissue. In other words, the cu
ltures were negative though the bacteria were present.[45]
Biofilms can also be formed on the inert surfaces of implanted devices such as c
atheters, prosthetic cardiac valves and intrauterine devices. [46]
New staining techniques are being developed to differentiate bacterial cells gro
wing in living animals, e.g. from tissues with allergy-inflammations.[47]
Research has shown that sub-therapeutic levels of -lactam anti iotics induce iof
ilm formation in Staphylococcus aureus. This su -therapeutic level of anti iotic
may result from the use of anti iotics as growth promoters in agriculture, or d
uring the normal course of anti iotic therapy. The iofilm formation induced y
low-level methicillin was inhi ited y DNase, suggesting that the su -therapeuti
c levels of anti iotic also induce extracellular DNA release.[48] Moreover, from
an evolutionary point of view, the creation of the tragedy of the commons in pa
thogenic micro es may provide advanced therapeutic ways for chronic infections c
aused y iofilms via genetically engineered invasive cheaters who can invade wi
ld-types cooperators of pathogenic acteria until cooperator populations go to ext
inction or overall population cooperators and cheaters go to extinction.[49]
Dental plaque[edit]
Dental plaque is an oral iofilm that adheres to the teeth and consists of many
species of oth acteria and fungi (such as Streptococcus mutans and Candida al
icans), em edded in salivary polymers and micro ial extracellular products. The
accumulation of microorganisms su jects the teeth and gingival tissues to high c
oncentrations of acterial meta olites which results in dental disease.[50]
The iofilm on the surface of teeth is frequently su ject to oxidative stress[51
] and acid stress.[52] Dietary car ohydrates can cause a dramatic decrease in pH
in oral iofilms to values of 4 and elow (acid stress).[52] A pH of 4 at ody
temperature of 37C causes depurination of DNA, leaving apurinic (AP) sites in DNA,
[53] especially loss of guanine.[54]
The Dental plaque iofilm can result in the disease dental caries if it is allow
ed to develop over time. An ecologic shift away from alanced populations within
the dental iofilm is driven y certain (cariogenic) micro iological population
s eginning to dominate when the environment favours them. The shift to an Acido
genic, aciduric, and cariogenic micro iological population develops and is maint
ained y frequent consumption of fermenta le dietary car ohydrate. The resulting
activity shift in the iofilm (and resulting acid production within the iofilm
, at the tooth surface) is associated with an im alance etween demineralization
and remineralisation leading to net mineral loss within dental hard tissues (en
amel and then dentin), the sign and symptom eing a carious lesion. By preventin
g the Dental plaque iofilm from maturing or y returning it ack to a non-cario
genic state, dental caries can e prevented and arrested.[55] This can e achiev
ed though the ehavioural step of reducing the supply of fermenta le car ohydrat
es (i.e. sugar intake) and frequent removal of the iofilm (i.e. tooth rushing).
A peptide pheromone quorum sensing signaling system in S. mutans includes the Co
mpetence Stimulating Peptide (CSP) that controls genetic competence.[56][57] Gen
etic competence is the a ility of a cell to take up DNA released y another cell
. Competence can lead to genetic transformation, a form of sexual interaction, f
avored under conditions of high cell density and/or stress where there is maxima
l opportunity for interaction etween the competent cell and the DNA released fr
om near y donor cells. This system is optimally expressed when S. mutans cells r
eside in an actively growing iofilm. Biofilm grown S. mutans cells are genetica
lly transformed at a rate 10- to 600-fold higher than S. mutans growing as freefloating planktonic cells suspended in liquid.[56]
When the iofilm, containing S. mutans and related oral streptococci, is su ject
ed to acid stress, the competence regulon is induced, leading to resistance to
eing killed y acid.[52] As pointed out y Michod et al., transformation in act
erial pathogens likely provides for effective and efficient recom inational repa
ir of DNA damages.[58] It appears that S. mutans can survive the frequent acid s

tress in oral iofilms, in part, through the recom inational repair provided y
competence and transformation.
Streptococcus pneumoniae[edit]
S. pneumoniae is the main cause of community-acquired pneumonia and meningitis i
n children and the elderly, and of septicemia in HIV-infected persons. When S. p
neumonia grows in iofilms, genes are specifically expressed that respond to oxi
dative stress and induce competence.[59] Formation of a iofilm depends on compe
tence stimulating peptide (CSP). CSP also functions as a quorum-sensing peptide.
It not only induces iofilm formation, ut also increases virulence in pneumoni
a and meningitis.
It has een proposed that competence development and iofilm formation is an ada
ptation of S. pneumoniae to survive the defenses of the host.[58] In particular,
the hosts polymorphonuclear leukocytes produce an oxidative urst to defend agai
nst the invading acteria, and this response can kill acteria y damaging their
DNA. Competent S. pneumoniae in a iofilm have the survival advantage that they
can more easily take up transforming DNA from near y cells in the iofilm to us
e for recom inational repair of oxidative damages in their DNA. Competent S. pne
umoniae can also secrete an enzyme (murein hydrolase) that destroys non-competen
t cells (fratricide) causing DNA to e released into the surrounding medium for
potential use y the competent cells.[60]
Biofilms in medicine[edit]
The rapidly expanding worldwide industry for iomedical devices and tissue engin
eering related products is already at $180 illion per year, yet this industry c
ontinues to suffer from micro ial colonization. No matter the sophistication, mi
cro ial infections can develop on all medical devices and tissue engineering con
structs.[61]
60-70% of nosocomial or hospital acquired infections are associated with the imp
lantation of a iomedical device.[61] This leads to 2 million cases annually in
the U.S., costing the healthcare system over $5 illion in additional healthcare
expenses.[61]
If an infection develops a iofilm, it ecomes even harder to treat. As the act
eria changes, it ecomes more resistant to anti iotics and the ody
s own host d
efenses.[61]
Biofilms in the food industry[edit]
Biofilms have ecome pro lematic in several food industries due to the a ility t
o form on plants and during industrial processes.[62] Bacteria can survive long
periods of time in water, animal manure, and soil, causing iofilm formation on
plants or in the processing equipment.[63] The uildup of iofilms can affect th
e heat flow across a surface and increase surface corrosion and frictional resis
tance of fluids.[64] These can lead to a loss of energy in a system and overall
loss of products.[64] Along with economic pro lems iofilm formation on food pos
es a health risk to consumers due to the a ility to make the food more resistant
to disinfectants [62] As a result, from 1996 to 2010 the Center for Disease Con
trol and Prevention estimated 48 million food orne illnesses per year.[62] Biofi
lms have een connected to a out 80% of acterial infections in the United State
s.[62]
In produce, microorganisms attach to the surfaces and iofilms develop internall
y.[62] During the washing process, iofilms resist sanitization and allow acter
ia to spread across the produce.[62] This pro lem is also found in ready to eat
foods ecause the foods go through limited cleaning procedures efore consumptio
n [62] Due to the perisha ility of dairy products and limitations in cleaning pr
ocedures, resulting in the uildup of acteria, dairy is suscepti le to iofilm
formation and contamination.[62][64] The acteria can spoil the products more re
adily and contaminated products pose a health risk to consumers. One acteria th
at can e found in various industries and is a major cause of food orne disease
is Salmonella.[65] Large amounts of salmonella contamination can e found in the
poultry processing industry as a out 50% of salmonella strains can produce iof
ilms on poultry farms.[62]Salmonella increases the risk of food orne illnesses w
hen the poultry products are not cleaned and cooked correctly. Salmonella is als
o found in the seafood industry where iofilms form from seafood orne pathogens

on the seafood itself as well as in water.[65] Shrimp products are commonly aff
ected y salmonella ecause of unhygienic processing and handling techniques[65]
The preparation practices of shrimp and other seafood products can allow for a
cteria uildup on the products.[65]
New forms of cleaning procedures are eing tested in order to reduce iofilm for
mation in these processes which will lead to safer and more productive food proc
essing industries. These new forms of cleaning procedures also have a profound e
ffect on the environment, often releasing toxic gases into the groundwater reser
voirs.[64]