Immune adaptation in birds

Ewan Hilbrands Anna Lauxen Bart-Jan Mazenier Douwe Mul Patrick Werndlij

Immune adaptation in birds By Ewan Hilbrands, Anna Lauxen, Bart-Jan Mazenier, Douwe Mul and Patrick Werndlij. Professional assistance provided by dr. K.D. Matson

Table of contents
The standard avian immune system ........................................ 5 Measuring the immune system................................................ 7 Size of immune system organs ............................................. 7 White blood cells .................................................................. 8 Presence of antibodies & cytokines ..................................... 9 Energy cost of the immune system ........................................ 10 Difference in immune system between migrating and nonmigrating birds ....................................................................... 11 Why is there a difference in immune response? ................... 13 Conclusion .............................................................................. 18 References .............................................................................. 19

Introduction

Avian flu is set to become one of the most dangerous diseases in the world. With the Spanish flu (H1N1) of 1918 causing approximately 20 million deaths worldwide, and more recent reoccurrences of H1N1 strain viruses the topic seems more important than ever. Yet, there is still much unknown about the avian immune system. In this flyer, we will be discussing the avian immune system, and differences between migratory and non migratory birds; specifically the type of adaptation they have undergone through evolution.

The standard avian immune system

To understand differences in the immune system between migratory and non-migratory species we first have to get a basic grasp of the standard avian immune system. The recent years have shown remarkable progress in the field. Most of this research was performed in poultry. From this research we have learned that the avian immune system looks similar to that of most mammals, but there are some important differences. To explore these differences, we will split the immune system into two parts; humoral and cellular. The cellular immune system is regulated by the thymus, which is essential for the development of for example T-lymphocytes and macrophages. The humoral immune system, which is responsible for antibodies produced by B-lymphocytes is regulated by an organ known as the Bursa of Fabricius. This organ is strictly avian. Another important difference can be found in the development of T-cells in birds. While it is largely similar, researchers found a type of T-cell that had not been seen before in any other type of animal. The production and regulation of T-cells is done by the thymus, Bursa of Fabricius and spleen.

Over the past years, research has shown that the immune response in birds relies on lymphokines. These molecules, produced by lymphocytes, move to the site of infection and serve as a target for other parts of the immuneresponse. Not much research has been done on this subject, because scientists havent been able to clone the genetic code responsible for these lymphokines yet. Figure 1 presents an example of where in the body different organs that make up the immune system are located. It is notable that they are spread throughout the body rather than being concentrated in a specific region.

Figure 1. An overview of location of organs responsible for the immune response in birds.

Measuring the immune system

In order to objectively talk about differences in immune response we must establish methods to measure it. In this section, we will discuss three different methods that have been used in actual research.

Size of immune system organs

The first method we will discuss is looking at the size of the different organs mentioned earlier that are relevant to the immune system. Different species of birds will often have different sized organs proportional to body size, though there are some exceptions. Different situations call for differing amounts of evolution of the organs. The general expectation would be that larger organs indicate a strong capacity for immune response, which we will discuss later on.

Figure 2. Size comparison of Bursa of Fabricius. Left is normal sized.

White blood cells

The second method used by researchers is investigating the white blood cells; the concentration of white blood cells can be measured from a blood assay. If the concentration is lower than average, it would indicate that the immune system is weakened at the time of the assay. This can have multiple reasons, discussed later. Another more specific method of determining the immune systems fitness is by determining the ratio of lymphocytes and heterophils. If the heterophil count is high compared to the lymphocyte count, the system is considered suppressed. The advantage of using blood assays is that only a very small amount of blood is necessary for an assay (~150 L), which means specific animals can be studied multiple times over a longer duration.

Figure 3. Graphs showing relevance of heterophil to lymphocite ratio to presence of immunoglobulin. More immunoglobulin indicates a stronger immune system.

Presence of antibodies & cytokines

The third method to get an idea of the fitness of the immune system is by checking the concentration of antibodies and cytokines. The usual way to use this method is to first check a healthy birds concentrations, then inject it with LPS (Lipopolysaccharide), which elicits a strong immune response in animals. After the injection the concentrations can be measured again, giving a full picture of how well the immune system is functioning. The stronger the reaction, the healthier the immune system. In figure 3, the right hand graph gives a sample of measured immunoglobulin in animals. The higher the measured amount, the stronger the animal in question is with regards to immunity.

Energy cost of the immune system

It goes without saying that possessing an immune system costs energy. An organism that chooses to maintain it has to invest energy in a plethora of immune-cells: Lymphocytes, TCells, antibodies and more. Because energy is limited, it follows that not every process can run at maximum efficiency. Growth, reproduction and thermoregulation are all processes that need a lot of energy. Multiple experiments have shown that energy reallocation can have drastic effects on life expectancy: rats raised in germ-free conditions, eliminating the need for an immune system, lived longer than rats raised under normal conditions. In a different study, chicks raised in a germ-free environment weighed more, used more energy and proteins for growth, and the efficiency of those proteins went up. In another study, chickens, guinea fowls and swine were administered antibiotics, reducing the energy investment required in the immune system. This resulted in weight gain and increased nitrogen retention. It is clear that maintaining an immune system is costly, but vital. Research by T. Piersma in 1997 showed that most species try to stay in a habitat that is unfriendly to parasites, e.g. near the sea where the salt is unfavorable. This results in a larger budget of energy for other things, and generally increased fitness.

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Difference in immune system between migrating and non-migrating birds

In the previous section we discussed the energy costs associated with the immune system; in this section we'll be looking at some of the practical implications of these costs. As preparation for the migration flight, birds have been shown to convert approximately 50% of their lean body mass (non-fat tissue) into fat to serve as an energy store. Despite this preparation, it has been shown that this entire store is depleted over the course of the migratory flight (Barlein 1985; Moore and Kerlinger 1987). This drastic change in the bird's body gives us a good idea of the investment involved in migration. A logical conclusion to make from this is that other energy-consuming systems are reduced or shut down. While immunosuppresion may seem like a vital flaw in survival strategy, it has been shown that certain organs that play a role in the immune reponse are more developed than in non-migratory birds; specifically the spleen and bursa of Fabricius have been shown to be substantially bigger.

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While their organs have adapted to perform better during immunosuppression, a study in Swainson's thrushes (Catharus Ustulatus) by Owen and Moore (2008) has shown that the immune response is generally lower in birds that have prepared for migration. Conserving energy is one of the main benefits of immunosuppression, but it also serves to lower the impact of immunopathology during a period of stress. If an infection were to occur as a result of exercise, the resulting tissue damage would likely result in the bird's death. Overall, while at first glance it would seem wise to invest heavily in a strong immune system due to a larger range of risk factors, the constraints of energy limit the adaptability of birds.

Figure 4. Swainsons Thrush (Catharus Ustulatus)

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Why is there a difference in immune response?

As explained earlier, migratory birds have a more evolved set of immune organs. What has led to this evolution though? The difference can be explained through a multitude of factors, including climates, food availability and exposure to pathogens. Knowing that having and maintaining an immune system has energy costs associated with it, J.M Fair et al. have shown in their experiments that when birds are exposed to antigens their growth significantly decreases. See figure 5.

Figure 5. Growth of Japanese Quails in relation to exposure to antigens.

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When one looks at migration and the necessity of the immune system, a logical conclusion is that it would be less evolved due to the priority of everything else; as shown earlier, this is not correct. In fact, the relevant organs are more evolved. It seems likely that due to having more evolved organs the amount of energy required can be reduced. Another explanation could be that birds lose their Bursa of Fabricius when they become sexually mature. Therefore migratory birds have to adapt to a more diverse parasite fauna before the start of their first migration. As the biggest energy cost is already fulfilled before migration, the energy costs during migration might be uninfluenced. Another explanation can be found in the effect of temperature on immune response. Migratory birds are exposed to at least two different climates during their lifetime while residential birds are not. It could be that higher environmental temperatures make it easier for the immune system to function. This was researched by A.M. Henken et al. The results showed no effect on the immune system itself, but it did have an effect on food intake when exposed to antigens. It could be that because there is more food available to migratory birds over the course of a lifetime and because they dont encounter big temperature differences there is enough energy intake to maintain their immune system, while residential birds have less energy intake, leaving a smaller budget.

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Because migratory birds are more likely to encounter a larger variety of pathogens during their lifetime due to exposure to different regions, they need a more evolved immune system to keep themselves healthy. Overall, migration increases food availability and allows birds to stay closer to their preferred environmental temperature, but comes with the extra invest of energy to maintain the immune system.

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Figure 6. Table comparing the size of Bursa of Fabricius and spleen in similar migratory and non-migratory birds.

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Conclusion
Overall, research has shown that migratory birds have a more evolved immune system, though during migratory periods their immune response will still be lowered due to the immense energy requirement of migration. Much is still unclear about the exact workings of the avian immune system and the adaptations they make to prepare for migration. The threat of avian pathogens is a very real one, making it an interesting subject to study for likely years to come.

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References
J.M. Sharma; Overview of the avian immune system Overview of the avian immune system, 30 ( 1991 ) 1317 Bruce Glick; The Avian Immune System uit: Avian Diseases, 23 (1979) 282-289 Pter L. Pap , Csongor I. Vgsi, Jcint Tklyi, Gbor . Czirjk & Zoltn Barta, Variation in Haematological Indices and Immune Function During the Annual Cycle in the Great Tit Parus major, (2010), Ardea 98(1):105-112 Katrina G. Salvante, Techniques for Studying Integrated Immune Function in Birds (2013), The Auk, Vol. 123, No. 2 (Apr., 2006), pp. 575-586 A.P.Moller, J.Erritzoe, Host immune defenseand migration in birds,(1998), Paris, Evolutionary ecology, issue 12, p-945-953. Bairlein F (1985) Body weights and fat deposition of Palaearctic passerine migrants in the central Sahara. Oecologia 66:141146 Moore FR, Kerlinger P (1987) Stopover and fat deposition by North American wood-warblers (Parulinae) following spring migration over the Gulf of Mexico. Oecologia 74:4754

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A.P.Moller, J.Erritzoe, Host immune defense and migration in birds,(1998), Paris, Evolutionary ecology, issue 12, p-945-953. J.C. Owen, F. R. Moore, Swainson's thrushes in migratory disposition exhibit reduced immune function (2008) Trade-offs in evolutionary immunology: just what is the cost of immunity? Robert L. Lochmiller and Charlotte Deerenberg, 2000. J.M.Fair, E.S.Hansen,R.E.Ricklefs, Growth, Developmental stability and immune response in juvenile Japanese quails (Coturnix coturnix japonica),(1999) St. Louis, The royal society, issue 266, p-1735-1742. A.M.Henken, A.M.Groote Schaarsberg, M.G.Nieuwland, The effect of environmental temperature on immune response and metabolism of the young chicken. 3. Effect of environmental temperature on the humoral immune response following injection of sheep red blood cells, (1983), Wageningen, Poultry science, issue 62, p-59-67.

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