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Ontogenetic and energetic consequences of an immunological challenge in Japanese Quail ( Coturnix coturnix japonica )

Ontogenetic and energetic consequences of an immunological challenge in Japanese Quail ( Coturnix coturnix japonica ). Imran Rafi Ahmed Punekar Dr. John Olson. Birds must devote energy to a wide variety of functions

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Ontogenetic and energetic consequences of an immunological challenge in Japanese Quail ( Coturnix coturnix japonica )

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  1. Ontogenetic and energetic consequences of an immunological challenge in Japanese Quail (Coturnix coturnix japonica) Imran Rafi Ahmed PunekarDr. John Olson

  2. Birds must devote energy to a wide variety of functions • Because the bird must strike a balance between devoting energy to growth, foraging, thermoregulation, digestion, and homeostasis So, Like any good budget-keeper…

  3. Derive an equation for total energetics: Etme = Em(maintenance) + Eb(biochemical) + Esda (specific dynamic action) + Etr(thermoregulation) + Eact(activity) + Ep(production) • Em + Eb = Ebmr • Precocial species have higher Em, higher Etr, and higher Eact

  4. Precocial Birds • Such as Japanese Quail (Coturnix coturnix japonica) hatch with a layer of down feathers, with open eyes, and are capable of locomotion • Incubation period is typically 17days • Naturally found across much of Europe and Asia, came to Japan as pets in the 11th century, became popular as poultry in the early 1900’s. • Introduced to the United States around 1870 • Clutch size: 5-12 eggs • Sexually mature after 40 days • Range from 100-180g at maturity, depending on sex • Optimally lay 200-300 eggs a year • Japanese Quail typically grow up in grassy fields, or near riverbanks • Are able to perform some thermoregulation • Are able to move at birth in order to forage for food, avoid predation, and find suitable habitats • Display significant growth and development during the post-hatch period

  5. In precocial species, less Energy is available for growth Simons & Pacifici, NC State Univ. http://www.cals.ncsu.edu:8050/course/zo501/Notes/Chapter%2018%202005.pdf#search='altricial%20species%20growth%20curve'

  6. Simons & Pacifici, NC State Univ. http://www.cals.ncsu.edu:8050/course/zo501/Notes/Chapter%2018%202005.pdf#search='altricial%20species%20growth%20curve'

  7. What happens if… • We add/increase a new energy cost? • Specifically, by eliciting an immune response which would possibly imbalance the equation and cause less energy to be devoted elsewhere Little is known about the ontogeny of the immune system in birds, much less about the energetic costs of mounting an immune response

  8. The Lab is performing an extensive experiment that is testing the hypotheses that • younger precocial Japanese Quail (Cotunix coturnix japonica) would be less effective in mounting an immune response than older birds That is, that the immune system develops with age and immunocompetence increases as the bird gets older (2) an immunological challenge will adversely affect maintenance, growth and development of birds by reallocating energy from that devoted to overall growth, maintenance of hematology, organ growth, and thermoregulation

  9. Dependent Variables • Hematology • Hematocrit Data • Blood Smears and Differential counts • Ig’s • Immunohistochemistry • Growth • Gross morphology • Organ growth • Metabolic: • Max VO2 during cold challenge • BMR • Biochemical • Catabolic enzyme activity

  10. This study concentrates on hematology and some growth factors • Predictions: Immunologically challenged birds will experience: - less growth and organ development, - altered concentrations of immune cells in the blood - exhibit different changes in hematology over time …when compared to control birds

  11. Experimental Design Ontogenetic changes in effect of immunological challenge 8-day chick Adult Controloil suspensionvehicle Controloil suspensionvehicle MS-bacin oil suspension MS-bacin oil suspension Pre-injection(1 h before ) Post-injection samples d 1 d 4 d 7 d 10 d 13 d 16 d 19 d 22 Baseline ontogenetic changes d 4 d 8 d 12 d 16 d 20 d 24 d 28 Ages d 32 d 36 d 40 d 44 d 48 d 52 d 56

  12. Hematology • Blood samples take from brachial vein before injection, 1d post injection, and every 3 days after that. • Organ Growth • Birds were raised, receiving blood sample battery above until 22d post injection after which they were sacrificed via cardiac puncture, dissected immediately, and organs harvested.

  13. RESULTS • Morphological DataBaseline Mass Growth Curve

  14. Morphological DataJuvenile Experimental Group Growth Curve

  15. Morphological DataAdult Experimental Group Growth Curve

  16. Hematocrit Data 60 50 40 30 20 0 5 10 15 20 25 60 50 40 30 20 0 5 10 15 20 25 Injection itself causes a profound decrease in Hemotcrit percentages in both experimental and control birds. Also, immunized birds also had a decrease in hematocrit values between 7-10 days, which is when the immune response to the vaccine is strongest, while control birds experienced smooth changes in hematocrit percentages 8d MS-bac Hematocrit, % 8d Control Time post-injection, d

  17. Hematocrit Data 60 50 40 30 20 0 5 10 15 20 25 60 50 40 30 20 0 5 10 15 20 25 Hematocrit values were much more variable in adult birds, but similar fluctuations were seen between both groups. In adults, we see a huge discrepancy between what happens to hematocrit levels after injection. In adult control specimens, we see the expected drop of similar magnitude to the drop seen in juvenile birds, but in the experimental group, hematocrit levels actually increase in all birds Adult MS-bac Hematocrit, % Adult Control Time post-injection, d

  18. Blood Smears • Few differences noted in ratio of white blood cells to red blood cells in smears stained with Giemsa in juvenile birds

  19. 10 8 6 4 2 0 MS C MS C MS C MS C 8 d(7d post) 8 d(22d post) Adult(7d post) Adult(22d post) Pectoralis index did not change with an immunological challenge. However, the size of the pectoralis does increase with age. Pectoralis Index (mean ± 1sd) Mass index, % body mass ANOVA with post hoc comparisons using student t-test : F= 3.7554 p=0.0217 d.f.=7

  20. 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 MS C MS C MS C MS C 8 d(7d post) 8 d(22d post) Adult(7d post) Adult(22d post) Gastrocnemius index did not change either with an immunological challenge nor with age. Gastrocnemius Index (mean ± 1sd) Mass index, % body mass ANOVA: F= 1.6507 p=0.2126 d.f.=7

  21. 4 3 2 1 0 MS C MS C MS C MS C 8 d(7d post) 8 d(22d post) Adult(7d post) Adult(22d post) Liver index did not change with an immunological challenge. However, the size of the liver does increase in adults. Liver Index (mean ± 1sd) Mass index, % body mass ANOVA with post hoc comparisons using student t-tests: F= 3.7558 p=0.0217 d.f.=7

  22. 1.2 1.0 0.8 0.6 0.4 0.2 0.0 MS C MS C MS C MS C 8 d(7d post) 8 d(22d post) Adult(7d post) Adult(22d post) Heart index did not change either in response to an immunological challenge nor with age. Heart Index (mean ± 1sd) Mass index, % body mass ANOVA: F= 0.5195 p=0.8037 d.f.=7

  23. 0.6 0.5 0.4 0.3 0.2 0.1 0.0 MS C MS C MS C MS C 8 d(7d post) 8 d(22d post) Adult(7d post) Adult(22d post) Pancreas index did not change with an immunological challenge. However, the size of the pancreas does decrease with age. Pancreas Index (mean ± 1sd) Mass index, % body mass ANOVA with post hoc comparisons using student t-tests: F= 4.5449 p=0.0108 d.f.=7

  24. 0.10 0.08 0.06 0.04 0.02 0.00 MS C MS C MS C MS C 8 d(7d post) 8 d(22d post) Adult(7d post) Adult(22d post) The size of the spleen, an important lymphoid organ, did not change with an immunological challenge. However, the size does increase with age between day 15 and day 30. Spleen (mean ± 1sd) Mass index, % body mass ANOVA with post hoc comparisons using student t-tests: F= 4.5449 p=0.0108 d.f.=7

  25. 0.20 0.15 0.10 0.05 0.00 MS C MS C MS C MS C 8 d(7d post) 8 d(22d post) Adult(7d post) Adult(22d post) The size of the Bursa Fabricius, another important lymphoid organ in birds, did not change with an immunological challenge. Like the thymus, however, the size does decrease significantly with age. Bursa fabricius (mean ± 1sd) Mass index, % body mass ANOVA with post hoc comparisons using student t-tests: F= 4.3942 p=0.0147 d.f.=7

  26. CONCLUSIONS • From this we conclude that: • Our hypothesis that young birds would exhibit less of an immune response than older birds was…. • Our hypothesis that the cost of immunological challenges would result in trade-offs in growth, organ development and maintenance of hematology was…

  27. Ongoing and Further Research • Comparison between the ontogeny of the immune system in a Precocial vs. Altricial species. • A comparison between the ontogeny of a T-cell mediated response vs. the ontogeny of a T-cell independent immune response. • An analysis of Substrate metabolism; determining what effect an immune stress has on substrate metabolism as well as on levels of endogenous stress hormones • In this experiment, we look at the effects of immunization on cold-hardiness and thermoregulatory capability, however a look at the opposite relationship, ie. that of cold stress on immuncompetence would be particularly interesting and helpful in the understanding of the effects of environmental stresses that exist in the wild.

  28. ACKNOWLEDGMENTS Special thanks to: The Olson Lab Crew: • Dr. John Olson • Doug Corwin • Lauren DiMenna • James “Rory” Tucker

  29. ACKNOWLEDGMENTS • Dr. Anil Bamezai • Dr. Norman Dollahon • Dr. Robert Curry And thanks to all the Faculty, staff, and students at the Biology Department at Villanova who helped in any way make this possible.

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