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At least 19 records

Effects of Wind Energy Development on Nesting Ecology of Greater Prairie-Chickens in Fragmented Grasslands: Wind Power and Prairie-Chickens

Wind energy is targeted to meet 20% of U.S. energy needs by 2030, but new sites for development of renewable energy may overlap with important habitats of declining populations of grassland birds. Greater Prairie-Chickens (Tympanuchus cupido) are an obligate grassland bird species predicted to respond negatively to energy development. We used a modified before–after control–impact design to test for impacts of a wind energy development on the reproductive ecology of prairie-chickens in a 5-year study. We located 59 and 185 nests before and after development, respectively, of a 201 MW wind energy facility in Greater Prairie-Chicken nesting habitat and assessed nest site selection and nest survival relative to proximity to wind energy infrastructure and habitat conditions. Proximity to turbines did not negatively affect nest site selection (β = 0.03, 95% CI = -1.2–1.3) or nest survival (β = -0.3, 95% CI = -0.1). Instead, nest site selection and survival were strongly related to vegetative cover and other local conditions determined by management for cattle production. Integration of our project results with previous reports of behavioral avoidance of oil and gas facilities by other species of prairie grouse suggests new avenues for research to mitigate impacts of energy development.

54 ENVIRONMENTAL SCIENCES↗

Defatted Microalgae-Mediated Enrichment of n–3 Polyunsaturated Fatty Acids in Chicken Muscle Is Not Affected by Dietary Selenium, Vitamin E, or Corn Oil

We previously showed enrichments of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) in broiler chicks fed defatted microalgae. The aims of this study were to determine 1) if the enrichments affected meat texture and were enhanced by manipulating dietary corn oil, selenium, and vitamin E concentrations and 2) how the enrichments corroborated with hepatic gene expression involved in biosynthesis and oxidation of EPA and DHA. Day-old hatching Cornish Giant cockerels (n= 216) were divided into 6 groups (6 cages/group and 6 chicks/cage). Chicks were fed 1 of the 6 diets: a control diet containing 4% corn oil, 25 IU vitamin E/kg, and 0.2 mg Se/kg (4CO), 4CO + 10% microalgae (defatted Nannochloropsis oceanica; 4CO+ MA), 4CO+ MA – 2% corn oil (2CO+MA), 2CO+MA + 75 IU vitamin E/kg (2CO+MA+E), 2CO+MA + 0.3 mg Se/kg (2CO+MA+Se), and 2CO+MA+E + 0.3 mg Se/kg (2CO+MA+E+Se). After 6 wk, fatty acid profiles, DHA and EPA biosynthesis and oxidation, gene expression, lipid peroxidation, antioxidant status, and meat texture were measured in liver, muscles, or both. Compared with the control diet, defatted microalgae (4CO+MA) enriched (P< 0.05) DHA and EPA by ≤116 and 24 mg/100 g tissue in the liver and muscles, respectively, and downregulated (41–76%, P< 0.01) hepatic mRNA abundance of 4 cytochrome P450 (CYP) enzymes (CYP2C23b, CYP2D6, CYP3A5, CYP4V2). Supplemental microalgae decreased (50–82%, P< 0.05) lipid peroxidation and improved (16–28%, P< 0.05) antioxidant status in the liver, muscles, or both. Furthermore, the microalgae-mediated enrichments in the muscles were not elevated by altering dietary corn oil, vitamin E, or selenium and did not affect meat texture. The microalgae-mediated enrichments of DHA and EPA in the chicken muscles were associated with decreased hepatic gene expression of their oxidation, but were not further enhanced by altering dietary corn oil, vitamin E, or selenium.

59 BASIC BIOLOGICAL SCIENCES↗

Chicken fat for catalysis: a scaffold is as important for molecular complexes for energy transformations as it is for enzymes in catalytic function

Metalloenzymes are remarkably active, specific, and efficient catalysts.Mimicking the active sites with organometallic complexes based on crystal structures has been a focus for many synthetic chemists. These surrogates have provided significant mechanistic insight into both the molecular complexes as well as the enzymes, however, one of the principle findings has been that just reproducing the active site is not adequate in reproducing the catalytic function. Specifically, the scaffold surrounding the active site plays a critical role in many aspects of catalysis, from positioning reactants optimally to positioning functional groups for reaction to creating the ideal environment for catalysis. Each of these can be controlled with both static positioning as well as small and large dynamics that can activate or control a specific part of the reaction. In this review, we will discuss the effect of organic/peptidic scaffolds, along with medium as an extension of the scaffold, and their varying impacts on catalytic reactivity.

Laureanti, Joseph A.↗

Environmental Impacts of Wind Power Development on the Population Biology of Greater Prairie-Chickens

1. We investigated the impacts of wind power development on the demography, movements, and population genetics of Greater Prairie-Chickens (Tympanuchus cupido) at three sites in northcentral and eastern Kansas for a 7-year period. Only 1 of 3 sites was developed for wind power, the 201MW Meridan Way Wind Power Facility at the Smoky Hills site in northcentral Kansas. Our project report is based on population data for prairie chickens collected during a 2-year preconstruction period (2007-2008), a 3-year postconstruction period (2009-2011) and one final year of lek surveys (2012). Where relevant, we present preconstruction data from our field studies at reference sites in the northern Flint Hills (2007-2009) and southern Flint Hills (2006-2008). 2. We addressed seven potential impacts of wind power development on prairie chickens: lek attendance, mating behavior, use of breeding habitat, fecundity rates, natal dispersal, survival rates, and population numbers. Our analyses of pre- and postconstruction impacts are based on an analysis of covariance design where we modeled population performance as a function of treatment period, distance to eventual or actual site of the nearest wind turbine, and the interaction of these factors. Our demographic and movement data from the 6-year study period at the Smoky Hills site included 23 lek sites, 251 radio-marked females monitored for 287 bird-years, and 264 nesting attempts. Our genetic data were based on genotypes of 1,760 females, males and chicks that were screened with a set of 27 microsatellite markers that were optimized in the lab. 3. In our analyses of lek attendance, the annual probability of lek persistence during the preconstruction period was ~0.9. During the postconstruction period, distance to nearest turbine did not have a significant effect on the probability of lek persistence. However, the probability of lek persistence increased from 0.69 at 0 m to 0.89 at 30 km from turbines, and most abandoned lek sites were located <5 km from turbines. Probability of lek persistence was significantly related to habitat and number of males. Leks had a higher probability of persistence in grasslands than agricultural fields, and increased from ~0.2 for leks of 5 males, to >0.9 for leks of 10 or more males. Large leks in grasslands should be a higher priority for conservation. Overall, wind power development had a weak effect on the annual probability of lek persistence. 3. We used molecular methods to investigate the mating behavior of prairie chickens. The prevailing view for lek-mating grouse is that females mate once to fertilize the clutch and that conspecific nest parasitism is rare. We found evidence that females mate multiple times to fertilize the clutch (8-18% of broods, 4-38% of chicks) and will parasitize nests of other females during egg-laying (~17% of nests). Variable rates of parentage were highest in the fragmented landscapes at the Smoky Hills field site, and were lower at the Flint Hills field site. Comparisons of the pre- and postconstruction periods showed that wind energy development did not affect the mating behaviors of prairie chickens. 4. We examined use of breeding habitats by radio-marked females and conducted separate analyses for nest site selection, and movements of females not attending nests or broods. The landscape was a mix of native prairie and agricultural habitats, and nest site selection was not random because females preferred to nest in grasslands. Nests tended to be closer to turbines during the postconstruction period and there was no evidence of behavioral avoidance of turbines by females during nest site selection. Movements of females not attending nests or broods showed that females crossed the site of the wind power development at higher rates during the preconstruction period (20%) than the postconstruction period (11%), and that movements away from turbines were more frequent during the postconstruction period. Thus, wind power development appears to affect movements in breeding habitats but not nest site selection of female prairie chickens during the breeding season. 5. We tested the effects of wind power development on five components of female fecundity: timing of clutch initiation, clutch size of first nests and renests, nest survival, and hatchability of eggs. Average date of clutch initiation was 26 April, clutch size was 12.7 and 10.6 eggs for first nests and renests, probability of nest survival was low at 0.18, but egg hatchability was high at 0.79. Wind power development had no impact on reproductive effort or nesting success, and all five components of fecundity were not related to treatment period or distance to turbine. Nest survival was the main factor limiting reproductive output of female prairie chickens and most losses were due to predation. Daily nest survival was strongly related to vegetative cover at the nest. Changes to rangeland management practices that would double nesting cover from 2.5 to 5 dm would triple the probability of nest survival from 0.17 to 0.52. Grass and forb cover had weak positive effects on daily nest survival whereas shrub cover, proximity to woodlands, and recent rainfall had negative effects. Reproductive performance of prairie chickens is low in managed rangelands in northcentral Kansas and efforts to improve range conditions and reduce predator activity would aid recovery of prairie chicken populations. 6. We used molecular methods to investigate patterns of natal dispersal in prairie chickens. High rates of nest failure limited the number of young that we could sample. Direct detections of natal dispersal were limited because survival of newly hatched chicks to become adults were low and because we were unable to detect dispersal distances outside of our study area. Direct observations of natal movements were limited and were inadequate to make conclusions about the potential impacts of wind energy development on natal dispersal. Spatial correlograms of genetic distance among males at leks were a more sensitive measure of population structure, and indicated a weak effect of wind energy development on the spatial genetic structure of prairie chickens. 7. We tested the effects of wind power development on female survival with time-to-event models, and on residual body mass of males with analyses of covariance. Distance to turbine and the interaction of distance and treatment period had no effect on female survival. Contrary to predictions of negative impacts of wind power development, the probability of female was lowest during the preconstruction period (0.274) and increased significantly during the postconstruction period (0.543). Inspection of hazard functions indicated that the difference in annual survival could be attributed to a higher risk of mortality during the lekking season in the preconstruction period. We suggest that wind power development may have improved ecological conditions for prairie chickens by disrupting the foraging behavior of diurnal raptors that kill prairie chickens at lek sites. In support of this idea, raptor kills tended to be farther from turbines during the postconstruction period whereas mammalian kills were closer. Analyses of the major causes of mortality did not support our hypothesis because the odds of raptor predation were greater after development was completed. Most mortality losses of radio-marked females were due to predation, and losses to collision mortality or harvest were rare events. Low rates of natural mortality during fall and winter imply that harvest is likely to be additive mortality in prairie chickens. Wind energy development reduced the residual body mass of male Greater Prairie-Chickens at lek sites near turbines. Low values of residual body mass could have a negative impact on individual survival or fecundity rates, or may be related to predation risk and flight performance of males displaying at open lek sites. 8. We tested for impacts of wind power development on population numbers of prairie chickens with monitoring of male numbers at leks, and with genetic measures of population structure. Lek counts indicated that wind power development did not affect the population size of prairie chickens. Peak counts of males at leks were recorded the first year after construction was completed and the highest rates of population change were observed during the interval when the wind power facility was constructed. Population numbers of prairie chickens near and distant from turbines appeared to covary in parallel, probably because bird numbers were linked by dispersal movements. Estimates of population viability based on genetic diversity, effective population size and rates of population exchange did not show annual changes and were unaffected by wind development during our study. Estimates of relatedness among males at the same and different leks suggested that wind power development has either reduced dispersal rates or changed settlement patterns, leading to higher rates of relatedness among males displaying at the same lek site. 9. Greater Prairie-Chickens were not strongly affected by wind power development in Kansas. Negative impacts of wind power development included a trend for reductions in lek persistence near turbines, behavioral avoidance of turbines by females during their breeding season movements, and changes in the genetic structure of males at leks that were consistent with reduced dispersal or recruitment rates. We found no impacts of wind power development on nest site selection, female reproductive effort or nesting success, or population numbers. Positive impacts of wind power development included an increase in female survival rates. We hypothesized that the unexpected increase in female survival was related to changes in trophic interactions and disruption of the foraging behavior of raptors that kill prairie chickens at lek site. 10. Research funding for this project included a grant from the 20% Wind by 2030 Program of the Department of Energy (this final report), and grants from the Kansas Department of Wildlife, Parks and Tourism, the National Fish and Wildlife Federation, and initial funding from the National Wind Coordinating Collaborative. Research products from data collected during the preconstruction period have included five peer-reviewed research articles and two PhD dissertations at Kansas State University. Additional manuscripts are in review for possible publication in 2013-2014.

17 WIND ENERGY↗

Oral lipase activities and fat-taste receptors for fat-taste sensing in chickens

Highlights: • We cloned a fat-taste receptor candidate gene, CD36, from chickens. • Fat-taste receptor genes, GPR120 and CD36, were expressed in chicken oral tissues. • Several lipase genes were expressed in chicken oral tissues. • Chicken oral tissues contained functional lipases. It has been reported that a functional fat-taste receptor, GPR120, is present in chicken oral tissues, and that chickens can detect fat taste in a behavioral test. However, although triglycerides need to be digested to free fatty acids to be recognized by fat-taste receptors such as GPR120, it remains unknown whether lipase activities exist in chicken oral tissues. To examine this question, we first cloned another fat-taste receptor candidate gene, CD36, from the chicken palate. Then, using RT-PCR, we determined that GPR120 and CD36 were broadly expressed in chicken oral and gastrointestinal tissues. Also by RT-PCR, we confirmed that several lipase genes were expressed in both oral and gastrointestinal tissues. Finally, we analyzed the lipase activities of oral tissues by using a fluorogenic triglyceride analog as a lipase substrate. We found there are functional lipases in oral tissues as well as in the stomach and pancreas. These results suggested that chickens have a basic fat-taste reception system that incorporates a triglycerides/oral-lipases/free fatty acids/GPR120 axis and CD36 axis.

60 APPLIED LIFE SCIENCES↗

Role of cytochrome P450 hydroxylase in the decreased accumulation of vitamin E in muscle from turkeys compared to that from chickens

Turkeys and chickens reared to 5-weeks of age and fed diets with feedstuffs low in endogenous tocopherols were examined. Treatments included feed supplemented with RRR (natural source vitamin E) alpha tocopheryl acetate (AcT, 35 mg/kg feed) and all-racemic (synthetic vitamin E) AcT (10 and 58 mg/kg feed). AcT ingestion per kg body weight was statistically similar between chicken and turkey only when the RRR-AcT containing diet was fed. Metabolites of alpha tocopherol (AT) and gamma tocopherol (GT) were measured in bile. When fed the RRR AcT diet, AT- and GT-metabolites were 8.3 and 5.3-fold elevated in turkey compared to chicken, respectively (p<0.001). When fed all-racemic AcT at 58 mg/kg feed, AT- and GT-metabolites were 2.2 and 2.4-fold elevated in turkey compared to chicken, respectively (p<0.01) with AcT ingestion per kg body weight 1.29-fold higher in the chickens (p<0.001). Turkey cytochrome P450 2C29 (CYP2C29) was increased relative to its chicken ortholog based on both RNA-Seq (p<0.001) and activity-based protein profiling (p<0.01) of liver tissue. Turkey CYP3A9 and CYP4F22 were increased compared to their chicken orthologs using as determined by Activity- Based Protein Profiling (p<0.05). The 3A and 4F family are noted as tocopherol hydroxylases in mammals. Alpha tocopherol concentrations in plasma, liver and muscle from turkey were 1.7 to 4.7-fold lower than the respective tissues from chicken (p<0.05). Lipid oxidation occurred more rapidly in turkey thigh compared to chicken thigh (p<0.05). These studies suggest that elevated tocopherol metabolism by cytochrome P450 hydroxylases in turkey liver contribute to the decreased accumulation of tocopherols in turkey tissues compared to that of chicken.

60 APPLIED LIFE SCIENCES↗

Lead exposure to children from consumption of backyard chicken eggs

Highlights: • Backyard chicken eggs from most households providing egg samples (98%) contained detectable concentrations of lead. • IIn prediction models, backyard egg consumption increased overall blood lead levels in children • Infants < 1 year of age experience the greatest increase in predicted blood lead levels from backyard egg consumption. • Mean household egg lead concentration was strongly positively correlated with coop soil lead concentrations. Backyard chicken ownership is rapidly increasing in urban areas in the United States, largely as a way to provide eggs for household consumption. Despite elevated levels of environmental lead contamination in many US cities, the role of backyard chicken eggs as a pathway for lead exposure, particularly for children, has received limited scrutiny. To characterize lead exposure from consumption of backyard chicken eggs for children and predict related effects on blood lead level (BLL), we conducted a cross-sectional study of backyard chicken owners in the Greater Boston area (n = 51). We interviewed participants regarding egg consumption by household members and collected backyard eggs (n = 201) and coop soil samples (n = 48) for analysis. Inductively coupled plasma mass spectrometry (ICP-MS) was used to evaluate lead concentration in homogenized eggs and an X-ray fluorescence (XRF) portable device was used to assess soil lead levels in the laboratory. We used the USEPA's Integrated Exposure Uptake Biokinetic Model for Lead in Children (IEUBK) to assess the relative contribution of backyard egg consumption to aggregate BLL in children. Four scenarios were developed in the IEUBK model to address variability in egg consumption rates and egg lead contamination. Lead was detected in egg samples from 98% of the households that provided egg samples. Mean household lead concentration was 0.10 μg/g (SD: 0.18). Egg lead concentrations ranged from below the limit of detection (0.0014 μg/g) to 1.798 μg/g (<1.4–1198 ppb). Egg lead levels were strongly positively correlated with lead concentration in coop soil (r = 0.64; p < 0.001). In modeled scenarios where a child < 7 years frequently ate eggs highly contaminated with lead, BLLs are predicted to increase by 0.9–1.5 μg/dL. In three other scenarios reflecting more moderate egg lead contamination and consumption rates, BLLs were predicted to increase from 0.1 to 0.8 μg/dL. Consumption of backyard chicken eggs can contribute to lead exposure in children. Soil lead remediation prior to chicken ownership may reduce lead exposure from backyard eggs.

54 ENVIRONMENTAL SCIENCES↗

Pathogenicity in Chickens and Turkeys of a 2021 United States H5N1 Highly Pathogenic Avian Influenza Clade 2.3.4.4b Wild Bird Virus Compared to Two Previous H5N8 Clade 2.3.4.4 Viruses

Highly pathogenic avian influenza viruses (HPAIVs) of subtype H5 of the Gs/GD/96 lineage remain a major threat to poultry due to endemicity in wild birds. H5N1 HPAIVs from this lineage were detected in 2021 in the United States (U.S.) and since then have infected many wild and domestic birds. We evaluated the pathobiology of an early U.S. H5N1 HPAIV (clade 2.3.4.4b, 2021) and two H5N8 HPAIVs from previous outbreaks in the U.S. (clade 2.3.4.4c, 2014) and Europe (clade 2.3.4.4b, 2016) in chickens and turkeys. Differences in clinical signs, mean death times (MDTs), and virus transmissibility were found between chickens and turkeys. The mean bird infective dose (BID 50 ) of the 2021 H5N1 virus was approximately 2.6 log 10 50% embryo infective dose (EID 50 ) in chickens and 2.2 log 10 EID 50 in turkeys, and the virus transmitted to contact-exposed turkeys but not chickens. The BID 50 for the 2016 H5N8 virus was also slightly different in chickens and turkeys (4.2 and 4.7 log 10 EID 50 , respectively); however, the BID 50 for the 2014 H5N8 virus was higher for chickens than turkeys (3.9 and ~0.9 log 10 EID 50 , respectively). With all viruses, turkeys took longer to die (MDTs of 2.6–8.2 days for turkeys and 1–4 days for chickens), which increased the virus shedding period and facilitated transmission to contacts.

60 APPLIED LIFE SCIENCES↗

Epidemiological investigation of coccidiosis and associated risk factors in broiler chickens immunized with live anticoccidial vaccines in China

Coccidiosis is a costly intestinal disease of chickens caused by Eimeria species. This infection is associated with high mortality, reduced feed efficiency, and slowed body weight gain. The diagnosis and control of coccidiosis becomes challenging due to the fact that chickens can be infected by seven different Eimeria species and often occur mixed-species co-infections. Grasping the epidemiology of Eimeria species is crucial to estimate the efficiency of poultry management. This study aimed to explore the distribution of Eimeria species in broiler chickens in China after administering live anticoccidial vaccines. A total of 634 samples were obtained, and the survey results showed that the prevalence of Eimeria was 86.12% (546/634), and the most common species were E. acervulina (65.62%), E. necatrix (50.95%), E. mitis (50.79%), E. tenella (48.42%), and E. praecox (41.80%). Most samples indicated mixed-species infections (an average of 3.29 species per positive sample). Notably, 63.98% of samples contain 3 to 5 Eimeria species within a single fecal sample. The most prevalent combinations were E. acervulina – E. tenella (38.96%) and E. acervulina – E. necatrix (37.22%). Statistical analysis showed that flocks vaccinated with trivalent vaccines were significantly positive for E. necatrix in grower chickens (OR = 3.30, p < 0.05) compared with starter chickens, and tetravalent vaccinated flocks showed that starter chickens demonstrated a higher susceptibility to E. tenella – E. brunetti (OR = 2.03, p < 0.05) and E. acervulina – E. maxima (OR = 2.05, p < 0.05) compared with adult chickens. Geographically, in the case of tetravalent vaccine-immunized flocks, a substantial positive association was observed between E. necatrix infection rates and flocks from eastern (OR = 3.88, p < 0.001), central (OR = 2.65, p = 0.001), and southern China (OR = 3.17, p < 0.001) compared with southwestern China. This study also found a positive association between E. necatrix (OR = 1.64, p < 0.05), E. acervulina (OR = 1.59, p < 0.05), and E. praecox (OR = 1.81, p < 0.05) infection and coccidiosis occurrence compared with non-infected flocks in tetravalent vaccinated flocks. This molecular epidemiological investigation showed a high prevalence of Eimeria species in the field. The emergent species, E. brunetti and E. praecox , might be incorporated into the widely-used live vaccines in the future. These insights could be useful in refining coccidiosis control strategies in the poultry industry.

Liao, Shenquan↗

Co-Pyrolysis of Chicken and Cow Manure

Conventional pyrolysis is a relatively simple way for biochar production; however, a single biomass feedstock approach is not promising for the development of multiple properties of biochar used in various applications. Here, this study provides a detailed comparison in terms of pyrolysis kinetics between the cow manure and chicken manure, in addition to the co-pyrolysis by mixing the two livestock at different ratios. Eight different heating rates were tested, 5–40 °C/min with 5 °C/min step, on both livestock using the nitrogen as a gas agent with a flowrate of 50 ml/min. The initial results show that for the slow heating rates, 5 °C/min, the thermal degradation of the cow manure is different compared to that obtained from chicken manure. For the cow manure, the first peak, associated with hemicellulose decomposition, is higher than the second peak, associated with cellulose decomposition, which is the other way around for chicken manure. At 5 °C /min, the hemicellulose decomposition took place at 250 °C and 300 °C for the chicken manure and cow manure, respectively. The cellulose decomposition was started at 300 °C for chicken manure and 470 °C for cow manure. The lowest heat rate was selected for the co-pyrolysis case study to allow a quasi-equilibrium state to avoid measurement errors and heat transfer limitation in terms of thermal lag. Co-pyrolysis is studied with different blend ratios between the chicken and cow manure, where the 40% cow manure shows a positive result in terms of keeping an exothermic reaction over the co-pyrolysis process.

09 BIOMASS FUELS↗

Efficacy of commercial recombinant HVT vaccines against a North American clade 2.3.4.4b H5N1 highly pathogenic avian influenza virus in chickens

The outbreak of clade 2.3.4.4b H5 highly pathogenic avian influenza (HPAI) in North America that started in 2021 has increased interest in applying vaccination as a strategy to help control and prevent the disease in poultry. Two commercially available vaccines based on the recombinant herpes virus of turkeys (rHVT) vector were tested against a recent North American clade 2.3.4.4b H5 HPAI virus isolate: A/turkey/Indiana/22-003707-003/2022 H5N1 in specific pathogen free white leghorn (WL) chickens and commercial broiler chickens. One rHVT-H5 vaccine encodes a hemagglutinin (HA) gene designed by the computationally optimized broadly reactive antigen method (COBRA-HVT vaccine). The other encodes an HA gene of a clade 2.2 virus (2.2-HVT vaccine). There was 100% survival of both chicken types COBRA-HVT vaccinated groups and in the 2.2-HVT vaccinated groups there was 94.8% and 90% survival of the WL and broilers respectively. Compared to the 2.2-HVT vaccinated groups, WL in the COBRA-HVT vaccinated group shed significantly lower mean viral titers by the cloacal route and broilers shed significantly lower titers by the oropharyngeal route than broilers. Virus titers detected in oral and cloacal swabs were otherwise similar among both vaccine groups and chicken types. To assess antibody-based tests to identify birds that have been infected after vaccination (DIVA-VI), sera collected after the challenge were tested with enzyme-linked lectin assay-neuraminidase inhibition (ELLA-NI) for N1 neuraminidase antibody detection and by commercial ELISA for detection of antibodies to the NP protein. As early as 7 days post challenge (DPC) 100% of the chickens were positive by ELLA-NI. ELISA was less sensitive with a maximum of 75% positive at 10DPC in broilers vaccinated with 2.2-HVT. Both vaccines provided protection from challenge to both types of chickens and ELLA-NI was sensitive at identifying antibodies to the challenge virus therefore should be evaluated further for DIVA-VI.

60 APPLIED LIFE SCIENCES↗

Chicken mannose binding lectin has antiviral activity towards infectious bronchitis virus

Mannose binding lectin (MBL) is a collagenous C-type lectin, which plays an important role in innate immunity. It can bind to carbohydrates on the surface of a wide range of pathogens, including viruses. Here we studied the antiviral effect of recombinant chicken (rc)MBL against Infectious Bronchitis Virus (IBV), a highly contagious coronavirus of chicken. rcMBL inhibited in a dose-dependent manner the infection of BHK-21 cells by IBV-Beaudette, as detected by immunofluorescence staining of viral proteins and qPCR. ELISA and negative staining electron microscopy showed that rcMBL bound directly to IBV, resulting in the aggregation of viral particles. Furthermore, we demonstrated that MBL bound specifically to the spike S1 protein of IBV which mediates viral attachment. This subsequently blocked the attachment of S1 to IBV-susceptible cells in chicken tracheal tissues as shown in protein histochemistry. Taken together, rcMBL exhibits antiviral activity against IBV, based on a direct interaction with IBV virions. - Highlights: • Recombinant chicken MBL (rcMBL) has antiviral activity against IBV-Beaudette in cell culture. • rcMBL aggregates IBV virus-particles. • rcMBL binds through its carbohydrate recognition domain to the S1 spike protein of IBV. • rcMBL can prevent binding of IBV-S1 to chicken tracheal tissues.

60 APPLIED LIFE SCIENCES↗

Genetics of adaptation in modern chicken

We carried out whole genome resequencing of 127 chicken including red jungle fowl and multiple populations of commercial broilers and layers to perform a systematic screening of adaptive changes in modern chicken ( Gallus gallus domesticus ). We uncovered >21 million high quality SNPs of which 34% are newly detected variants. This panel comprises >115,000 predicted amino-acid altering substitutions as well as 1,100 SNPs predicted to be stop-gain or -loss, several of which reach high frequencies. Signatures of selection were investigated both through analyses of fixation and differentiation to reveal selective sweeps that may have had prominent roles during domestication and breed development. Contrasting wild and domestic chicken we confirmed selection at the BCO2 and TSHR loci and identified 34 putative sweeps co-localized with ALX1 , KITLG , EPGR , IGF1 , DLK1 , JPT2 , CRAMP1 , and GLI3 , among others. Analysis of enrichment between groups of wild vs. commercials and broilers vs. layers revealed a further panel of candidate genes including CORIN , SKIV2L2 implicated in pigmentation and LEPR , MEGF10 and SPEF2 , suggestive of production-oriented selection. SNPs with marked allele frequency differences between wild and domestic chicken showed a highly significant deficiency in the proportion of amino-acid altering mutations (P<2.5×10 -6 ). The results contribute to the understanding of major genetic changes that took place during the evolution of modern chickens and in poultry breeding.

60 APPLIED LIFE SCIENCES↗

Natural Selection Footprints Among African Chicken Breeds and Village Ecotypes

Natural selection is likely a major factor in shaping genomic variation of the African indigenous rural chicken, driving the development of genetic footprints. Selection footprints are expected to be associated with adaptation to locally prevailing environmental stressors, which may include diverse factors as high altitude, disease resistance, poor nutrition, oxidative and heat stresses. To determine the existence of a selection footprint, 268 birds were randomly sampled from three indigenous ecotypes from East Africa (Rwanda and Uganda) and North Africa (Baladi), and two registered Egyptian breeds (Dandarawi and Fayoumi). Samples were genotyped using the chicken Affymetrix 600K Axiom R Array. A total of 494,332 SNPs were utilized in the downstream analysis after implementing quality control measures. The intra-population runs of homozygosity (ROH) that occurred in >50% of individuals of an ecotype or in >75% of a breed were studied. To identify inter-population differentiation due to genetic structure, FST was calculated for North- vs. East-African populations and Baladi and Fayoumi vs. Dandarawi for overlapping windows (500 kb with a step-size of 250 kb). The ROH and FST mapping detected several selective sweeps on different autosomes. Results reflected selection footprints of the environmental stresses, breed behavior, and management. Intra-population ROH of the Egyptian chickens showed selection footprints bearing genes for adaptation to heat, solar radiation, ion transport and immunity. The high-altitude-adapted East-African populations’ ROH showed a selection signature with genes for angiogenesis, oxygen-heme binding and transport. The neuroglobin gene (GO:0019825 and GO:0015671) was detected on a Chromosome 5 ROH of Rwanda–Uganda ecotypes. The sodium-dependent noradrenaline transporter, SLC6A2 on a Chromosome 11 ROH in Fayoumi breed may reflect its active behavior. Inter-population FST among Egyptian populations reflected genetic mechanisms for the Fayoumi resistance to Newcastle Disease Virus (NDV), while FST between Egyptian and Rwanda–Uganda populations indicated the Secreted frizzled related protein 2, SFRP2, on Chromosome 4, that contributes to melanogenic activity and most likely enhances the Dandarawi chicken adaptation to high-intensity of solar radiation in Southern Egypt. These results enhance our understanding of the natural selection forces role in shaping genomic structure for adaptation to the stressful African conditions.

59 BASIC BIOLOGICAL SCIENCES↗

Supranutrition of microalgal docosahexaenoic acid and calcidiol improved growth performance, tissue lipid profiles, and tibia characteristics of broiler chickens

Abstract Background Docosahexaenoic acid (DHA) and calcidiol could be enriched in chicken for improving public nutrition and health. It remains unclear if supranutritional levels of DHA and calcidiol impair growth performance or metabolism of broiler chickens. This study was to determine singular and combined effects of high levels of supplemental DHA-rich microalgal biomass or oil and calcidiol on growth performance, concentrations of triglycerides, cholesterol, and nonesterfied fatty acids in plasma, liver, breast, and thigh, and biophysical properties of tibia. Methods In Exp. 1, 144 day-old Cornish chicks were divided into 4 groups (6 cages/treatment, 6 birds/cage), and were fed a corn-soybean meal basal diet (BD), BD + 10,000 IU calcidiol/kg (BD + Cal), BD + 1% DHA-rich Aurantiochytrium (1.2 g DHA/kg; BD + DHA), and BD + Cal + DHA for 6 weeks. In Exp. 2, 180 day-old chicks were divided into 5 groups, and were fed: BD, BD + DHA (0.33% to 0.66% oil, 1.5 to 3.0 g DHA/kg), BD + DHA + EPA (1.9% to 3.8% eicosapentaenoic acid-rich Nannochloropsis sp. CO18, 0.3 to 0.6 g EPA/kg), BD + DHA + calcidiol (6000 to 12,000 IU/kg diet), and BD + DHA + EPA + Cal for 6 weeks. Results Birds fed BD + Cal diet in Exp. 1 and BD + DHA + EPA diet in Exp. 2 had higher ( P < 0.05) body weight gain (10%–11%) and gain:feed ratio (7%), and lower ( P < 0.05) total cholesterol and triglyceride concentrations in plasma (18%–54%), liver (8%–26%), breast (19%–26%), and thigh (10%–19%), respectively, over the controls. The two diets also improved ( P < 0.05) tibial breaking strength (8%–24%), total bone volume (2%–13%), and (or) bone mineral density (3%–19%) of chickens. Conclusion Supranutrition of dietary calcidiol and DHA alone or together did not produce adverse effects, but led to moderate improvements of growth performance, lipid profiles of plasma and muscle, and bone properties of broiler chickens.

60 APPLIED LIFE SCIENCES↗

Synthesis of carbon nanofibres from waste chicken fat for field electron emission applications

Highlights: • Waste chicken fat is used as a starting material to produce CNFs via TCVD method. • High heating rate applied resulted in aggregation of catalyst particles. • Aggregated catalyst produced sea urchin-like CNFs with amorphous nature. • The as-grown CNFs presented a potential for field electron emission applications. - Abstract: Carbon nanofibres (CNFs) with sea urchin-like morphology were synthesised from waste chicken fat precursor via catalytic thermal chemical vapour deposition method at 750 °C. The CNFs showed amorphous structures under high-resolution transmission electron microscopy, micro-Raman spectroscopy and X-ray diffraction examination. X-ray photoelectron spectroscopy analysis confirmed that the core of the sea urchin-like CNFs was composed of Fe{sub 3}C formed within the first 20 min of synthesis time. The growth of amorphous CNFs from agglomerated Fe{sub 3}C particles was favourable due to the high heating rate applied during the synthesis. Field electron emission examination of the CNFs indicated turn-on and threshold field values of 5.4 and 6.6 V μm{sup −1} at current density of 1 and 10 μA cm{sup −2}, respectively. This study demonstrates that waste chicken fat, a low-cost and readily available resource, can be used as an inexpensive carbon source for the production of CNFs with a potential application in field electron emitters.

77 NANOSCIENCE AND NANOTECHNOLOGY↗

Tris(1,3-dichloro-2-propyl) phosphate perturbs the expression of genes involved in immune response and lipid and steroid metabolism in chicken embryos

We previously demonstrated that in ovo exposure to the flame retardant tris(1,3-dichloro-2-propyl) phosphate (TDCPP) decreased plasma thyroxine levels, reduced growth parameters, and decreased gallbladder size in chicken embryos. In the current study DNA microarrays were used to evaluate global mRNA expression in liver tissue of male chicken embryos that exhibited the above mentioned effects. Injected doses were dimethyl sulfoxide vehicle control, 7.6 or 45 μg TDCPP/g egg. TDCPP caused significant changes in the expression of five genes at the low dose and 47 genes at the high dose (False Discovery Rate p ≤ 0.1, fold change ≥ 1.5). The gene expression analysis suggested a compromised immune function, a state of cholestatic liver/biliary fibrosis, and disrupted lipid and steroid metabolism. Circulating bile acid levels were elevated, which is an indication of liver dysfunction, and plasma cholesterol levels were reduced; however, hepatic bile acid and cholesterol levels were unaltered. Interactome analyses identified apolipoprotein E, hepatocyte nuclear factor 4 alpha, and peroxisome proliferator-activated receptor alpha as key regulatory molecules involved in the effects of TDCPP. Our results demonstrate a targeted effect of TDCPP toxicity on lipid metabolism, including cholesterol, that helps explain the aforementioned phenotypic effects, as chicken embryos are highly dependent on yolk lipids for growth and maintenance throughout development. Finally, our results are in concordance with the literature that describes TDCPP as a cancer-causing agent, since the majority of dysregulated genes were involved in cancer pathways. - Highlights: • TDCPP dysregulates genes involved in immune function and lipid metabolism. • A targeted effect of TDCPP toxicity on cholesterol metabolism is apparent. • A state of cholestatic liver fibrosis is suggested by the expression profile. • Elevated plasma bile acids suggest that TDCPP causes liver dysfunction.

60 APPLIED LIFE SCIENCES↗