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Selective breeding of chickens for high growth rate and other production traits has led to the modern commercial broiler, a bird that has the genetic potential for reaching an average body weight of 2.7kg within 6 weeks of hatch. However, the breeding stock for modern broilers has to be feed controlled in order to lay large numbers of viable hatching eggs. Broiler breeders, when fed ad libitum, have a propensity to produce internal ovulations, double-yolked, misshapen or shell-less eggs. This is due to the release of multiple ova at ovulation, which results in a significant loss of production. Feed control has been shown to mitigate this effect but welfare concerns have been raised as to the side-effects for the birds. The main objective of this research was to determine the genetic basis for the regulation of ovarian follicle selection and its dysfunction in ad libitum-fed broiler breeders, and how this might be addressed by genetic selection to limit the impact on the management and welfare of future broiler breeders. A multi-layered statistical, expression profiling and cluster analysis of ovarian gene expression data from a microarray study was carried out to identify candidate genes for further study.Key stages of development were investigated for feed restricted and ad libitum-fed broiler breeders. Several gene candidate genes were validated by qPCR in a comparison of different ovarian tissues in layer type hens for subsequent analysis in broiler breeders. Sequencing of the founders of an Advanced Intercross Line (AIL) of commercial broiler breeders and White Leghorn layers was performed covering 3 regions of each of the primary candidate genes in order to identify genetic variation that could account for differences in follicle number between broilers and layers. Expression data from a microarray study highlighted a number of potential candidate genes for regulation of follicle development. One of these genes, Platelet Derived Growth Factor Receptor Like (PDGFRL), shares significant sequence homology with the active domains of Platelet Derived Growth Factor Receptor β. Expression profiling in layers showed peak PDGFRL expression in 5-6 mm follicles and the F2 follicle (P <0.001). PDGFRL was also up-regulated in response to ad libitum feeding in broiler breeders in 6-8 mm follicles (P<0.016), the point at which follicle selection and recruitment is considered to occur. In addition to this, while PDGFRL expression remains relatively constant between tissues under ad libitum conditions, it shows a clear reduction in expression (P <0.001) in prehierarchical follicles relative to the stroma and the F1 follicle under feed restriction. This observation is consistent with results from the original microarray study. Sequencing of the AIL Founders highlighted several SNPs in the broiler that have the potential to be used as markers for incorporation into commercial selection programs. EST alignment in preparation for targeted sequencing of PDGFRL also highlighted three potential forms of the protein, each with a different 5’ starting sequence. Initial investigation has shown all three to be expressed in ovarian follicles. QPCR in a panel of 13 tissues shows marked differences between the 3 variants, implying different and perhaps specialised roles for each. The PDGFR family has a potential role in steroidogenesis, and the expression profiling, combined with the clear effect on expression from ad libitum feeding in broiler breeders, suggest that PDGFRL is a strong candidate for involvement in the regulation of follicle development GDF9, shown to be associated with multiple ovulation in sheep, and FSH receptor, a mediator of neuroendocrine signalling to the ovary, were also investigated. They behaved as expected in layer type birds but both showed significant differential expression (P = 0.005 and 0.018 respectively) as a result of ad libitum feeding in broiler breeders. Though these two genes have been extensively investigated, these are previously unobserved effects. SNPs have also been identified in these genes which have the potential to be used as markers for incorporation into commercial selection programs. To fully exploit these results, additional investigation is recommended to confirm these results in commercial populations and to determine how they can be employed to best effect.

The objective of this study was to examine the effects of rearing temperature on the meat yield of broilers. Four experiments were conducted. Experiment 1 examined meat yield at different temperatures and on different diets when birds were grown to a fixed age. Experiments 2 and 3 were aimed at the effects of temperature and of the interaction between temperature and stocking density respectively on the meat yield of broilers when killed at a specified liveweight (i.e. allowing the rearing period to vary). Experiment 4 was designed to assess whether depression of growth rate at high temperature affected the meat yield in broilers.

The pathogenesis of <i>C. jejuni</i> in broiler chickens is still poorly understood despite the importance of poultry meat as a source of infection in humans. The overall objective of this project was to understand the role of flagella and Campylobacter invasion antigens in mucosal and systemic colonization, and to evaluate the vaccine potential of <i>C. jejuni</i> paralyzed flagella mutants. As a first step to track <i>C. jejuni in vivo</i>, a Green Fluorescent Protein (GFP) reporter system that is constitutively expressed was constructed. The system was transformed into different <i>C. jejuni</i> strains and isolates, and their mucosal and systemic spreading was studied over the period of 7 days. <i>C. jejuni</i> NCTC11168V1 and V26 share the same background but differ in their ability to colonize chickens. <i>C. jejuni</i> 81-176 and K2-55 share the same genetic background but K2-55 has an insertion mutation in <i>pflA</i> gene that produced paralyzed flagella. Although the K2-55 flagella remained intact structurally, it did not secret <i>Campylobacter</i> invasion antigens (Cia). The reporter system was stable in all of these strains both <i>in vitro</i> and <i>in vivo</i>. Fluorescent bacteria were visualized successfully using fluorescent and confocal microscopes. C. jejuni NCTC11168V1 and 81-176 were detected in the intestinal tract and in the liver and spleen of more than 30% of the challenged birds, while V26 and K2-55 were only detected in the intestinal tract. <i>C. jejuni</i> 81-176 and K2-55 did not spread systemically to the spleen and liver of BALB/c mice challenged using the same approach, although they colonized the ceca.<p> A live attenuated vaccine based on <i>C. jejuni</i> K2-55 protected broiler chickens from <i>C. jejuni</i> 81-176 challenge in chickens following streptomycin treatment of drinking water. The same vaccine had no significant protection against a heterolgous <i>C. jejuni</i> NCTC11168V1 strain challenge. The vaccine was a poor stimulator of secretory IgA.<p> Macrophage-like HD11 cells inflammatory response to the presence of <i>C. jejuni</i> K2-55 was not significantly different from their response to wild-type 81-176 when measured by qRT-PCR. The lack of Cia secretion and motility had no effect on expression of IL-1â, IL-2, IL-6, IL-8, IL, IL-10, IL-12â, or TLR5. A <i>flgK</i> mutant expressing the flagella up to the hook had a significantly lower expression of these genes.

The pathogenesis of <i>C. jejuni</i> in broiler chickens is still poorly understood despite the importance of poultry meat as a source of infection in humans. The overall objective of this project was to understand the role of flagella and Campylobacter invasion antigens in mucosal and systemic colonization, and to evaluate the vaccine potential of <i>C. jejuni</i> paralyzed flagella mutants. As a first step to track <i>C. jejuni in vivo</i>, a Green Fluorescent Protein (GFP) reporter system that is constitutively expressed was constructed. The system was transformed into different <i>C. jejuni</i> strains and isolates, and their mucosal and systemic spreading was studied over the period of 7 days. <i>C. jejuni</i> NCTC11168V1 and V26 share the same background but differ in their ability to colonize chickens. <i>C. jejuni</i> 81-176 and K2-55 share the same genetic background but K2-55 has an insertion mutation in <i>pflA</i> gene that produced paralyzed flagella. Although the K2-55 flagella remained intact structurally, it did not secret <i>Campylobacter</i> invasion antigens (Cia). The reporter system was stable in all of these strains both <i>in vitro</i> and <i>in vivo</i>. Fluorescent bacteria were visualized successfully using fluorescent and confocal microscopes. C. jejuni NCTC11168V1 and 81-176 were detected in the intestinal tract and in the liver and spleen of more than 30% of the challenged birds, while V26 and K2-55 were only detected in the intestinal tract. <i>C. jejuni</i> 81-176 and K2-55 did not spread systemically to the spleen and liver of BALB/c mice challenged using the same approach, although they colonized the ceca.<p> A live attenuated vaccine based on <i>C. jejuni</i> K2-55 protected broiler chickens from <i>C. jejuni</i> 81-176 challenge in chickens following streptomycin treatment of drinking water. The same vaccine had no significant protection against a heterolgous <i>C. jejuni</i> NCTC11168V1 strain challenge. The vaccine was a poor stimulator of secretory IgA.<p> Macrophage-like HD11 cells inflammatory response to the presence of <i>C. jejuni</i> K2-55 was not significantly different from their response to wild-type 81-176 when measured by qRT-PCR. The lack of Cia secretion and motility had no effect on expression of IL-1â, IL-2, IL-6, IL-8, IL, IL-10, IL-12â, or TLR5. A <i>flgK</i> mutant expressing the flagella up to the hook had a significantly lower expression of these genes.

Four research experiments were conducted to evaluate the influence of dietary enzyme inclusion (phytase and NSPase) on broiler performance parameters, processing yields, and bone mineralization. In Experiment 1, a 35-day grow out trial was conducted to investigate the effect of three commercially available phytase enzymes on growth performance and bone mineralization in phosphorus deficient corn/soy based diets. Increasing the level of available phosphorus (aP) in the control diets resulted in improved bird performance and bone ash data. The presence of dietary phytase in phosphorus deficient diets resulted in improvements in growth parameters and bone mineralization. Regression analysis confirmed that phytase supplementation can potentially increase the bioavailability of phosphorus in broiler diets up to 0.15 to 0.20 percent, however, the responses varied according to the enzyme used and inclusion level. In Experiment 2, a 42-day grow out trial was conducted to analyze the effects of NSPase inclusion on broiler performance and processing parameters when supplemented in diets with varying protein and energy concentrations. Reduced protein and energy levels reduced bird performance throughout the trial. The inclusion of both NSPase enzymes resulted in improvements in feed conversion throughout the starter and grower periods (day 26 of age). The results from this trial showed that NSPase inclusion can improve broiler performance and processing parameters. In Experiments 3 and 4, a battery trial and a floor trial were conducted to determine the effects of phytase and NSPase enzyme co-administration on growth and bone ash in low phosphorus diets. Increasing the level of available phosphorus resulted in increased bird performance and bone ash. The inclusion of phytase enhanced bird performance and bone mineralization. NSPase inclusion in diets containing low levels of phytase had improvements in bird performance during early stages of growth. The enhanced effects associated with dual administration of phytase and NSPase were not observed in a full grow out trial during later stages of growth. These four experiments indicate that phytase and NSPase enzyme inclusion in broiler diets have the ability to enhance bird performance, processing yield, and bone mineralization.

This research program included a series of experiments to investigate the effect of starter diet protein level on the performance of broilers vaccinated with Coccivac®-B and subsequently challenged with a mixed species Eimeria challenge compared to nonvaccinated broilers. Pre-challenge performance data indicates that vaccination may decrease body weights and increase feed conversion ratio (FCR) with vaccination. The time period associated with the observed effects is between 13 to 17 d of age. This reduction in performance of vaccinated broilers versus non-vaccinated broilers was eliminated by the conclusion of the experiments (27 d) in the higher protein diets. Vaccination was effective at generating protective immunity against the Eimeria challenge evidenced by significantly increased body weight gains, improved feed conversions, reduced post-challenge mortality, and reduced lesion development in vaccinated broilers compared to non-vaccinated. The final experiment included the comparison of Coccivac®-B to Bio-Cox® (salinomycin) for controlling field strain Eimeria in broilers reared on two different dietary rations varying in protein concentration. Diet A had a lower protein concentration than Diet B. On day 14, Eimeria collected from commercial broiler farms in Texas were spray applied to the litter in all pens. Broilers reared on Diet B were heavier at Day 40 while body weights at day 50 were similar for all groups. Broilers fed Diet B had lower FCR during the starter and finisher diets. Broilers fed salinomycin had lower FCR for the starter and grower diets while vaccinated broilers had lower FCR during the withdrawal period. Cumulative FCR for the entire grow out period were similar for all groups. These data indicate that vaccination can be utilized as an anticoccidial preventive and are suggestive that reduced protein concentration of starter diets can lead to significant losses in broiler performance when utilizing a vaccination program to prevent coccidiosis. Feeding an appropriately formulated diet while vaccinating broilers with Coccivac®-B as an alternative to the use of salinomycin yields at least equivalent if not elevated performance in the presence of field-strain Eimeria during grow-out with no effect on the cost of production.

Thesis (doctoral)--Universität, Hohenheim, 2000.

The aim of the four studies reported in this thesis was to evaluate the effects of the inclusion of lupins in the diets for broiler chicks, and to evaluate any effect of enzyme supplementation on performance of birds fed lupin-based diets.  Five types of lupin were used in the studies.  Two determinate cultivars of <i>Lupinus albus, </i>Lucyane and Ludet, and two dwarf cultivars of <i>Lupinus albus, </i>Luniverse and Lucille.  One cultivar of <i>L. luteus </i>cv. Wodjil was also used.  The nutritive value of the lupins was determined by two precision feeding studies.  The value of including the lupins in diets for broilers was evaluated using two growth trails.  One trial was conducted using birds grown from 7-28 days and the other using birds grown from 0-13 days.  The use of sialic acid as a measure of endogenous losses was also investigated.  Effects of the diets on microflora in the birds gastro-intestinal tract were determined using GC profiling of the caecal contents. The five lupin types used in the study had protein levels that were similar to soyabean meal. Results of all studies indicated that the different cultivars of lupin behaved very differently, and therefore need to be considered separately in terms of which enzyme to use and levels of supplementation.  It cannot be assumed that all lupins will respond in the same way to enzyme supplementation.  The results of both growth studies showed that the two types of lupin cultivar, determinate and dwarf, respond differently to enzyme supplementation.  This is possibly due to the different routes of plant breeding. It was clear from the study that the effects of lupin inclusion, enzyme supplementation and the interactions between the two are very complex.  Further investigations of the mechanisms behind the effects are recommended.