Progress 10/01/08 to 09/30/13
Outputs
Target Audience: (1) Scientific peers, through presentations at meetings and peer-reviewed publications (2) high school and undergraduate researchers that were involved in the project Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? PhD student Tess Killpack was trained. Numerous undergraduate researchers and high school researchers were trained. How have the results been disseminated to communities of interest? Yes, many of the papers were published. A few more are submitted but under reveiw. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
All goals were met. The first study examined the ontogeny of specific adaptive immune response in altricial zebra finches Taeniopygia guttata, using repeated vaccinations with non-infectious keyhole limpet hemocyanin (KLH) antigen. We found that capacity for adaptive antibody response developed after the first week post-hatch and that strength of adaptive antibody response significantly increased with age throughout the nestling period. However, we found that mature adult secondary antibody response level was not achieved in zebra finches prior to fledging (21d). Because innate immune functions purportedly develop rapidly post-hatch, prior to adaptive functions, we then examined developmental patterns of constitutive innate and adaptive immune indices in house sparrows Passer domesticus (fledging age: 14-15d). Lysozyme activity significantly decreased with age, likely representing catabolism of maternal lysozyme from the egg albumen. Levels of total IgY (indexing adaptive immune function), as well as agglutination and lysis (indexing constitutive innate immune function), increased throughout the nestling period, but were significantly below levels found in fully-grown birds near fledging (12d). There were no significant differences between hatch-year birds (2-3 months) and adults in these measures, indicating that full maturation occurs early post-fledging. After examining the ontogeny of immune function throughout the nestling period, we examined how immune function is altered by food restriction and whether trade-offs occur between growth and development and immune function. The third study examined the phenomenon of compensatory growth in zebra finches following food restriction during the nestling period. While compensatory growth allows birds to fledge at a suitable size and time, it may have persistent detrimental effects on the development of tissues and of complex systems such as the adaptive immune system. Compensatory body mass, but not tarsus and culmen, growth was observed in previously food restricted zebra finches. No impact on nestling adaptive antibody response to KLH was observed post-restriction. Reductions in tissue maturity, indexed by water content, were observed following compensatory growth, indicating a decoupling of chronological age and physiological age as a result of accelerated growth in food restricted birds. While we saw no impact of food restriction on immune function after re-feeding, we were also interested in examining whether trade-offs occur between growth and immune function during food restriction. In the final study, we imposed food restriction on early and late-stage house sparrow nestlings and measured multiple indices of immune function, growth, and development. During food restriction, levels of the acute phase protein haptoglobin (Hp), an inducible component of the innate immune system, were reduced compared with controls. Food restriction did not significantly impact complement-mediated lysis, a constitutive component of immune function, or circulating IgY antibody levels, an index of adaptive humoral function. Food restriction resulted in significant reductions in alimentary organ, heart, and flight muscle masses. Reductions in muscle maturity (indexed by tissue water content and mean total citrate synthase enzyme activity) were also observed with early food restriction and were not recovered with re-feeding. Findings from this study suggest that immune function, like organ growth, appears to be flexible to resource supply during the nestling period, and that growth may be prioritized or relatively more costly than immune function during this time.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Killpack, T.L., Y. Oguchi, and W.H. Karasov (2013) Ontogenetic patterns of constituitive innate immune parameters in altricial house sparrow nestlings. Journal of Avian Biology 44:513-520.
- Type:
Other
Status:
Published
Year Published:
2013
Citation:
Killpack, T. L., E. Carrel, W. H. Karasov. 2013. Impact of food restriction on immune function in altricial house sparrow nestlings. Integrative and Comparative Biology 53:E110-E110.
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2013
Citation:
Killpack, T.L. (2013) Immune Capacity Development and Factors that Influence the Strength of Immune Response in Growing Altricial Birds. PhD dissertation, University of Wisconsin-Madison
- Type:
Journal Articles
Status:
Submitted
Year Published:
2013
Citation:
Killpack, T.L., E. Carrel, and W.H. Karasov. Energetic costs of immune function in growing altricial house sparrow nestlings. Submitted to Physiological and Biochemical Zoology
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2014
Citation:
Killpack, T.L., D. Nan Tie, and W.H. Karasov (2014) Compensatory growth in zebra finches and its impact on body composition and adaptive immune function. The Auk (in press)
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Progress 01/01/12 to 12/31/12
Outputs
OUTPUTS: ACTIVITIES: (1) In food restricted nestling, house sparrows we tested for trade-offs in immune function and, particularly, that immune function would be most reduced in a defense considered costly, like the acute phase response to injected lipopolysaccharide (LPS), than one considered less costly, like complement-mediated lysis. We tested birds both early in the nestling period, when growth demands are high, and late in the nestling period, when growth has reached a plateau. We also examined the long-term effects of early food restriction (FR) on birds re-fed and examined late in the nestling period. (2) Graduate student Killpack taught upper-level undergraduate Physiological Animal Ecology course (Fall 2012), including material on growth and immune function, influenced by research efforts. (3) Killpack and Karasov mentored high school and undergraduate students in laboratory research associated with experiments listed above: One high school student, summer 2012 from Madison Metropolitan School District High School Science Research Program; One undergraduate, summer 2012 from UW-PEOPLE/CALS Summer Internship Program for minority and first-generation high school students (2010, 2012). EVENTS Graduate student Killpack presented research at 3 conferences: (1) Killpack, TL; Karasov, WH. 2012. Post-hatching ontogeny of adaptive antibody response to two distinct antigens in captive zebra finches. Oral presentation at the annual meeting of the Society for Integrative and Comparative Biology, Charleston, SC. (2) Nan Tie, D.E; Killpack, TL; Karasov, WH. 2012. Effect of food restriction and realimentation on growth, immune response, and body composition in Zebra finches. Oral presentation at the annual meeting of the Society for Integrative and Comparative Biology, Charleston, SC. (3) Killpack, TL; Oguchi, Y; Karasov, WH. 2012. Ontogeny of constitutive innate immune function in free-living altricial house sparrow nestlings. Poster presentation at the annual meeting of the Society for Integrative and Comparative Biology, Charleston, SC. PARTICIPANTS: Graduate students trained - Tess Killpack, Tawnya Cary; Collaborations: Dr. Erik K. Hofmeister, USGS National Wildlife Health Center, Madison, WI 53711. TARGET AUDIENCES: Our research will be of interest to the animal production industry, especially those interested in possible alternatives for regulating the gastrointestinal tract's immune functions without compromising food/growth efficiency. Also, our research will be of interest to those interested in health in wildlife, as well as in humans and their domesticated animals, especially as regards West Nile virus. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
CHANGE IN KNOWLEDGE: Altricial passerines hatch in a relatively undeveloped state and grow more rapidly than the precocial species on which estimates of immune function development have been based. We have examined age-related changes in constitutive, innate immune function in free-living house sparrows, as well as adaptive antibody response to a model antigen in captive zebra finches. These studies demonstrated that immune function is limited in nestlings and does not reach mature adult levels in altricial birds until after fledging, which will help define when these birds may be particularly vulnerable to infection during the nestling and post-fledging periods. The field of ecological immunology has proposed a trade-off between investment in important life history components (growth, development, and reproduction) and purportedly costly immune defense in hosts. It is unclear how flexible immune function is to changing food supply during the nestling period of altricial birds. We have examined whether trade-offs occur in resource allocation to growth, development and immune response when house sparrows are food restricted during the nestling period, and whether these trade-offs are reversible upon re-feeding. We have also studied the long-term impacts of compensatory growth on innate and adaptive immune function in captive zebra finch nestlings. These findings have broad ecological implications given that food restriction is a common occurrence for nestling birds, especially with changing global climate patterns. Experiments led to new methods for stimulating and measuring immune response in small altricial nestlings which were not previously used in our laboratory, or in some cases, in the published literature. These methods include vaccination protocols, protocols for measuring innate immune responses (natural antibody-mediated agglutination, complement-mediated lysis, lysozyme, and haptoglobin), and protocols for measuring adaptive antibody responses (antigen-specific ELISAs of nestling antibodies).
Publications
- Killpack, TL. and Karasov, WH. 2012. Ontogeny of adaptive antibody response to a model antigen in captive altricial zebra finches. PLoS ONE. 7(10): e47294. doi:10.1371/journal.pone.0047294.
- Killpack, TL. and Karasov, WH. 2012. Growth and development of house sparrows (Passer domesticus) in response to chronic food restriction throughout the nestling period. Journal of Experimental Biology. 215:1806-1815.
- Killpack, TL; Karasov, WH. 2012. Post-hatching ontogeny of adaptive antibody response to two distinct antigens in captive zebra finches. Integrative and Comparative Biology. 52: E93.Supplement 1. Oral presentation at the annual meeting of the Society for Integrative and Comparative Biology, Charleston, SC.
- Nan Tie, D.E; Killpack, TL; Karasov, WH. 2012. Effect of food restriction and realimentation on growth, immune response, and body composition in Zebra finches. Integrative and Comparative Biology. 52: E128 Supplement 1. Oral presentation at the annual meeting of the Society for Integrative and Comparative Biology, Charleston, SC.
- Killpack, TL; Oguchi, Y; Karasov, WH. 2012. Ontogeny of constitutive innate immune function in free-living altricial house sparrow nestlings. Integrative and Comparative Biology. E275. Poster presentation at the annual meeting of the Society for Integrative and Comparative Biology, Charleston, SC.
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Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: Activities (1) We collected house sparrow nestlings at 3, 6, 9, or 12 d. Body mass and skeletal lengths were measured and blood samples were taken for hematocrit and for analysis of 3 measures: natural antibody-mediated agglutination, complement-mediated lysis, and bacterial killing activity of lysozyme. Adults were sampled for comparison to nestlings. (2) We hypothesized that food restricted zebra finch nestlings would display compensatory growth, no difference in structural growth, increased tissue water content (an index of functional maturity), decreased lipid content, and reduced immune function compared to nestlings fed ad libitum for the duration of the study. Experimental broods were fed ad libitum till 6 days of age when a food restriction of 70% ad libitum was implemented for 3 days where upon 9 days of age ad libitum was restored. Body mass, structural measurements, and food consumption were taken daily until day 33 days of age when the nestlings were euthanized for analysis of body composition (water/lipid content). Control and experimental broods were tested for innate immune response to LPS and adaptive immune response to KLH. (3) We predicted that antibody responsiveness in altricial nestlings increases post-hatch and reaches adult levels by fledging age, and that nestlings generate more robust antibody responses to larger injected antigens. Nestling zebra finches were injected at 7 days (7d group) or 14 days (14d group) post-hatch with keyhole limpet hemocyanin (KLH; antigen size: ~400kDa) and West Nile Virus (WNV; antigen size ~50kDa) antigens, and boosted seven days later. Adults were vaccinated in the same manner. Induced KLH- and WNV-specific IgG response was measured using ELISA. Dissemination (1) Gillian-Daniel, D; Killpack, T; Malkepour, S; Shin, H. 2011. The achievement gap at UWMadison: Exploring solutions. UW Madison Center for the First-Year Experience, 2011 Conference, Madison, WI. (2) Killpack, TL, Stone, K, and Karasov, WH. 2011. Body composition and developmental maturity in chronically food restricted nestling House sparrows (Passer domesticus). Poster presentation at the annual meeting of the Society for Integrative and Comparative Biology, Salt Lake City, UT. (3) Killpack, TL and K Stone. 2010. Impact of chronic food restriction on the growth and development of nestling House sparrows. Oral presentation at the First annual meeting of the Northern Integrative Biology Society, Decorah, IA. (4) Killpack, TL and WH Karasov. 2010. Patterns of Gut Size and Digestive Enzyme Activity with Chronic Food Restriction in Nestling House Sparrows. Poster presentation at 2010 APS Intersociety Meeting: Global Change & Global Science: Comparative Physiology in a Changing World. Westminster, CO. Killpack, TL, Singh, N, and Karasov WH. 2010. Effect of chronic food restriction on gut morphology and digestive enzymes in nestling House sparrows. Oral presentation at the annual meeting of the Society for Integrative and Comparative Biology, Seattle, WA. PARTICIPANTS: PhD students Tess Killpack and Tawnya Cary were involved in developing immune assays, performing experiments on birds, analyzing data, and preparing material for dissemination. Undergraduate researchers Dan Nan Tie and Yushi Oguchi were involved in experiments with birds, analyzing data, and preparing material for dissemination. Dr. Bill Karasov supervised activities and was involved in analyzing data and preparing material for dissemination. TARGET AUDIENCES: Primary target audiences so far are scientific peers, reached through presentations at meetings and publications. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Main outputs so far are in advancing knowledge about ontogeny of immune function in altricial birds. Findings are significant enough to be published, and manuscripts are submitted or in preparation.
Publications
- Killpack, TL, Stone, K, and Karasov, WH. 2011 Body composition and developmental maturity in chronically food restricted nestling House sparrows (Passer domesticus). Integrative and Comparative Biology. Abstract, In press.
- Killpack, TL and WH Karasov. 2010. Patterns of Gut Size and Digestive Enzyme Activity with Chronic Food Restriction in Nestling House Sparrows. The Physiologist 53:32. Abstract.
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Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: We published one abstract on our pilot study, PARTICIPANTS: Graduate student trained - Tess Killpack, Tawnya Cary; Collaborations: Dr. Erik K. Hofmeister, USGS National Wildlife Health Center, Madison, WI 53711 TARGET AUDIENCES: Target audience: Our research will be of interest to the animal production industry, especially those interested in possible alternatives for regulating the gastrointestinal tract's immune functions without compromising food/growth efficiency. Also, our research will of interest to those intreseted in health in wildlife as well as in humans and their domesticated animals, especially as regards West Nile virus. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
House sparrow nestlings were raised in the lab on age-specific meal sizes (controls) or 75% of age-specific size (food restricted) and dissected and analyzed for composition at day 6, 9, and 12 post-hatch. Generally, skeletal lengths were not significantly different between treatment groups. Food restricted birds at days 9 and 12 had significantly smaller body and lean dry muscle masses, and significantly higher muscle water content, compared to same-aged controls. Lean dry liver mass and liver lipid content were significantly lower in restricted birds. Intestine mass was significantly lower in food-restricted birds though mass-specific digestive enzymatic capacity was not different from controls. Food restricted birds at day 12 had reduced lysozymeactivity and hematocritvalues. From these data, it appears that altricial birds chronically restricted throughout the nestling period maintain structural growth at the expense of muscle and organ development and body condition.
Publications
- Killpack, T.; Singh, N.; Karasov, W.H. (2010) Effect of chronic food restriction on gut morphology and digestive enzymes in nestling House sparrows. Integrative and Comparative Biology 50: E90-E90.
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: We published a paper showing that in young chickens (age 28 days) intramuscular (IM) vaccination with envelope protein from West Nile virus (WNV E protein) is protective against WNV infection, and results in a significantly different antibody production profile compared to both control birds and birds orally vaccinated with WNV E protein. We completed a pilot study for our project on ontogeny of immune function in nestling birds and published an abstract showing that response of the digestive organs of altricial nestlings facing chronic food restriction is similar to the response found in food-restricted adults. PARTICIPANTS: Graduate student trained - Tess Killpack, Carol Fassbinder-Orth; Collaborations: Dr. Erik K. Hofmeister, USGS National Wildlife Health Center, Madison, WI 53711 TARGET AUDIENCES: Target audience: Our research will be of interest to the animal production industry, especially those interested in possible alternatives for regulating the gastrointestinal tract's immune functions without compromising food/growth efficiency. Also, our research will of interest to those intreseted in health in wildlife as well as in humans and their domesticated animals, especially as regards West Nile virus. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
West Nile virus (WNV) causes morbidity and mortality in humans, horses and many bird species in North America. We investigated the use of a recombinant WNV envelope (E) protein as both an oral and intramuscular (IM)-delivered vaccine. White leghorn cockerels (age 28 days) were divided into four treatment groups: low oral (20 microgram E protein/dose), high oral (100 microgram E protein/dose), IM (intramuscular; 20 microgram E protein/dose), and Non-vaccinated WNV control (0 micro gram E protein/dose). Chicks were given a priming vaccination at age 28 d and booster vaccinations two and four weeks later. Two weeks following the final vaccination (70 day old), birds were infected subcutaneously with 105 plaque-forming units (PFUs) of the NY99 strain of WNV. Serum was collected every other day from each bird on 1-8 days post infection (dpi), and again on 10, 14 and 21 dpi to assess viremias and antibody production. WNV viremia levels were significantly lower in the IM group than both oral groups and the control group (P< 0.001). Total WNVEP IgG production was significantly greater in the IM group compared to all other treatment groups (P<0.001), and was first detected one week after the second WNV E protein vaccination. The IM group produced significantly lower levels of WNVNS1 IgG compared to all other groups post WNV infection (P<0.003). The results of this study indicate that IM vaccination of chickens with E protein is protective against WNV infection, and results in a significantly different antibody production profile compared to both control birds and birds orally vaccinated with WNV E protein. In adult birds, food restriction typically results in reductions in size of assimilation organs such as intestine and liver and in total enzymatic capacity, though mass-specific enzyme activities may not be altered. Food restriction due to periodic food shortage can occur during chick development, although their response has been little studied. We hypothesized that masses and biochemical capacities of assimilation organs are maintained constant in developing nestling birds, despite food restriction, to support demands of growth and development. House sparrow (Passer domesticus) nestlings were raised in the laboratory from day 3 to day 25 post-hatch and fed either age-specific meal sizes (control group) or meal sizes 75% of age-specific amounts (restricted group). On day 25, body and organ masses were measured and intestines were stored for digestive enzyme assays. Chronically restricted nestlings had significantly lower body mass (by 14%). Intestine mass and length were significantly lower in restricted birds (by 17% and 9%, respectively), even when corrected for differences in body mass. Liver mass was also significantly lower (by 18%) in restricted birds. There was no significant difference between groups in maltase or aminopeptidase-N activity per unit intestine mass. Results so far show that response of the digestive organs of altricial nestlings facing chronic food restriction is not different from findings in restricted adults.
Publications
- 1 Fassbinder-Orth, C.A., E.K. Hofmeister, C.Weeks-Levy and W.H. Karasov (2009) Oral and parenteral immunization of chickens (Gallus gallus) agains West Nile virus with recombinant envelope protein. Avian Diseases: Vol. 53, No. 4, pp. 502-509 (doi: 10.1637/8688-031009-Reg.1) 2 Killpack, T.; Singh, N.; Karasov, W.H. (2010) Effect of chronic food restriction on gut morphology and digestive enzymes in nestling House sparrows. Integrative and Comparative Biology (in press).
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