This study was to determine (1) if genetic selection for high (HWS) or low (LWS) body weight in chickens has altered the hypothalamic AMP-activated protein kinase (AMPK) system and (2) if this alteration contributes to the dissimilar feeding response to various appetite modulators between HWS and LWS lines. Compared to HWS, LWS chickens had higher levels of AMPK α and acetyl-CoA carboxylase (ACC) phosphorylation, which was caused by upregulation of the upstream factor calcium/calmodulin-dependent protein kinase kinase 2 (CAMKK β). There was greater mRNA expression of carnitine palmitoyltransferase I (CPT1), leptin receptor (LEPR) and neuropeptide Y (NPY) and less mRNA expression of ACC α, fatty acid synthase (FAS), fat mass and obesity associated gene (FTO), pro-opiomelanocortin (POMC) and orexin in LWS than HWS chickens. At 5 days of age, intracerebroventricular (ICV) injection of AICAR, 5-amino- 4-imidazolecarboxamide riboside, caused a quadratic dose-dependent decrease in food intake in LWS but not HWS chicks. Compound C, (6-(4-(2-piperidin-1-yl-ethoxy)-phenyl))-3-pyridin-4-yl-pyrazolo(1,5-a)-pyrimidine, caused a quadratic dose-dependent increase in food intake in HWS but not LWS chicks. The anorexigenic effect of AICAR in LWS chicks and orexigenic effect of Compound C in HWS chicks resulted from either activation or inhibition of other kinase pathways separate from AMPK. There is a lower threshold for the anorexigenic effect of ghrelin in LWS than HWS chicks, which was associated with differential hypothalamic AMPK signaling. ICV injection of ghrelin inhibited corticotrophin-releasing hormone (CRH), 20-hydroxysteroid dehydrogenase (20HSD), glucocorticoid receptor (GR), CPT1 and FTO expression in LWS but not HWS chicks. Additionally, the hypothalamic mRNA level of ghrelin was significantly higher in LWS than HWS chicks, which may also contribute to the differential threshold response to ghrelin in these two lines. Obestatin caused a linear dose-dependent increase in food intake in HWS but not LWS chicks. The orexigenic effect of obestatin in HWS chicks was not associated with altered AMPK. Obestatin inhibited LEPR and FTO expression in HWS but not LWS chicks. Thus, selection for body weight may alter the hypothalamic response to ghrelin by the AMPK pathway, CRH pathway, CPT1 and FTO, and to obestatin by LEPR and FTO. / Ph. D.

Amino acids are assimilated by membrane-associated transporters into and out of enterocytes either in their free form or in the form of peptides. The peptide transporter, PepT1, is thought to be the major facilitator of peptide transport in the enterocyte. It is unknown if the peptide transporters and free amino acid transporters operate in a compensatory fashion to regulate the amino acid balance within the enterocyte. Therefore, the objective was to examine the regulatory balance between PepT1 and other peptide and free amino acid transporters in enterocytes. The Mouse Small Intestinal Epithelial (MSIE) cells are conditionally immortalized. It was found that MSIE cells express BoAT1, CAT1, CAT2, LAT1, y+LAT1, and y+LAT2, but not PepT1, EAAT3, Bo,+AT, or LAT2, making this model similar to the basolateral membrane of enterocytes. Growing MSIE cells at high temperatures did not affect the nutrient transporter gene expression profile of these cells. Thus, the human colon carcinoma (Caco-2) cell line was used as a small intestinal in vitro model for this study. These cells express PepT1, HPT1, PTR3 EAAT1, EAAT3, rBAT, Bo,+AT CAT1, LAT1, y+LAT1, y+LAT2, ABCC3, ABCC4, which increased from D0 to D21 post confluency, indicating cell maturation. In Caco-2 cells, PepT1 gene silencing was induced in Caco-2 cells. Despite an reduction of PepT1 gene (82%, P < 0.05) protein (96%), no significant difference in any peptide (HPT1, PTR3, ABCC3, ABCC4) or free amino acid transporters (EAAT1, EAAT3, rBAT, Bo,+AT, BoAT1, CAT1, CAT2, LAT1, LAT2, y+LAT1, y+LAT2) between Caco-2 cells treated with PepT1 siRNA and Caco-2 cells treated with Control siRNA was observed. These results suggest no compensation at the gene expression level of these transporters in response to a reduction of PepT1. To account for the limitations of an in vitro and PepT1 kockout mouse model, transgenic chicken models were pursued. Potential cPepT1 overexpressing, cPepT1 shRNA or control shRNA expressing G0 chickens were generated by embryo injection of pseudolentiviral particles followed by ex ovo egg culture. Overall, 9 potential G0 cPepT1 overexpressing chickens, 15 potential G0 cPepT1 shRNA expressing chickens, and 4 potential G0 control shRNA expressing chickens were generated. / Ph. D.

Escalating consumer concerns have placed the poultry industry under mounting pressure to reduce the use of chemotherapeutic agents as feed additives. One possible alternative receiving increased attention is the use of immunomodulators such as β-glucan. A pilot study evaluated the effects of a yeast derived β-glucan (Auxoferm YGT) on growth performance and immune response of broiler chickens. Day-old chicks were fed a diet containing 0, 0.02, or 0.1% yeast β-glucan. On days 7 and 14 post-hatch, body weight and relative immune organ weights were measured, peripheral blood was collected to determine heterophil to lymphocyte (H:L) ratios, and small intestinal sections were sampled to evaluate relative gene expression. The addition of β-glucan had no influence on growth. Dietary β-glucan supplementation modulated the expression of interleukin (IL)-8, IL-18, interferon (IFN)-γ and inducible nitric oxide synthase (iNOS) in the small intestine. A subsequent study was conducted to investigate the effects of dietary β-glucan on broiler chick (1440 birds) performance and immune response during a mixed Eimeria infection (day 8 of age). Measurements were taken and samples collected on days 4, 7, 10, 14 and 21 post-hatch. The results from this study show that β-glucan supplementation did not negatively impact performance. The addition of β-glucan to the diet resulted in reduced gross lesion severity and increased H:L ratios. The gene expression results suggest that β-glucans are capable of skewing the host immune response toward aTh1 mediated response and consequently down-regulating the Th2 mediated response. / Master of Science

PepT1, PepT2 and PHT1 are all members of the proton-coupled oligopeptide transporter family, which are important in the transport of amino acids in peptide form. PepT1 acts as a low affinity/high capacity transporter and PepT2 as a high affinity/low capacity transporter for di- and tri-peptides. PHT1 transports di- and tri-peptides as well as histidine. The objective of this study was to profile PepT1, PepT2 and PHT1 mRNA expression in the proventriculus, duodenum, jejunum, ileum, ceca, large intestine, brain, heart, bursa of Fabricius, lung, kidney, and liver in layer chicks on embryonic days 18 and 20 and days 1, 3, 7, 10, and 14 post-hatch. Absolute quantification real-time PCR was used to measure gene expression. PepT1 expression was greatest in the duodenum, jejunum and ileum. Over time, PepT1 expression increased in the duodenum, jejunum, ileum and large intestine and decreased in the ceca. PepT2 expression was greatest in the brain, aiding in neuropeptide homeostasis, and the kidney, aiding in the reabsorption of substrates. Over time, PepT2 expression increased in the bursa of Fabricius and decreased in the proventriculus, duodenum, jejunum and liver. In the small intestine during embryogenesis, PepT2 may function to transport di- and tri-peptides prior to the induction of PepT1. PHT1 expression was expressed in all tissues analyzed. Over time, PHT1 expression increased in the jejunum, large intestine, brain and liver and decreased in the proventriculus. The uptake of peptides in the developing chick is regulated by peptide transporters that are expressed in a tissue- and development-specific manner. / Master of Science

The chicken yolk sac and small intestine are connected through the yolk stalk and share many biological similarities. During the embryonic stage, the extra-embryonic yolk sac helps the embryo to absorb nutrients primarily in the last two weeks of incubation. The chicken yolk sac physically moves yolk contents from the yolk sac to the small intestine at the end of embryogenesis. This is the time when the small intestine replaces the yolk sac in assimilating nutrients for the embryo and later for the posthatch chicken. Additionally, both chicken small intestinal epithelia and the yolk sac secrete beta defensins for promoting intestinal health. Since there are heterogeneous cell types along the mammalian intestinal villus, which are derived from the intestinal stem cells in the crypts, we investigated if cells of the chicken yolk sac and small intestine have the same ontogeny as mammalian intestinal epithelial cells. In this dissertation, we mainly focused on the spatial expression of nutrient transporters (PepT1 and SGLT1), intestinal stem cell markers (Lgr5 and Olfm4), and avian beta defensins in the chicken yolk sac and small intestine during the embryonic and early posthatch stages. RNAscope in situ hybridization was used to identify the distribution of cells expressing PepT1 mRNA in both the chicken yolk sac and small intestine. PepT1 mRNA was found to be expressed by epithelial cells in both the yolk sac and small intestine. In the yolk sac, PepT1 mRNA was uniformly distributed in each endodermal epithelial cell along the villus-like structure. The pattern of PepT1 mRNA expression observed in the chicken yolk sac during the last 10 days of incubation revealed that PepT1 mRNA was increased from e11 to e13, and decreased from e15 to day of hatch. The peak of PepT1 mRNA expression was between e13 and e15, when the yolk sac reaches maximum absorptive area and the growth of the chicken embryo is at its fastest rate. However, the expression of PepT1 mRNA in the intestine was only detected in columnar enterocytes along the villus and not in goblet cells or cells in the crypts. The immunofluorescence assay confirmed that PepT1 protein was located at the brush border membrane of the enterocytes and that protein expression of PepT1 was restricted to the intestinal epithelial cells from approximately the middle to the tip of the villus. In order to identify intestinal stem cells, we used the known mammalian stem cell markers, Lgr5 and Olfm4. Both Lgr5 and Olfm4 are specifically expressed by cells in the chicken intestinal crypts, suggesting that they can be used as biomarkers for chicken intestinal stem cells. Dual labelling of PepT1 and Olfm4 mRNA on the same chicken intestinal sample revealed that there was a gap between PepT1-expressing enterocytes and Olfm4-expressing intestinal stem cells. The cells in this gap were presumably transit amplifying (TA) cells. Additionally, we also found that the TA cell zone along the intestinal villus was reduced during chicken growth. This TA cell population could be clearly detected at day of hatch and d1 posthatch but not later. The expression of SGLT1 mRNA was localized to yolk sac endodermal epithelial cells and showed a sharp increase at the end of incubation. This increase of SGLT1 mRNA coincided with the increase in glucose in the yolk, indicating that the chicken embryo needs glucose as energy for hatching. The mRNA expression profiles of various avian beta defensins have been examined by qPCR and in situ hybridization to investigate the immune function of the yolk sac and small intestine. We found that AvBD10 mRNA showed the highest expression level in the yolk sac and was expressed predominantly in the yolk sac endodermal epithelial cells. Additionally, the expression of AvBD10 mRNA showed a development-specific pattern, which increased from e9 to e11, and decreased from e13 towards day of hatch. The expression patterns of AvBD1, 2, and 7 mRNA were similar to each other. These three genes were found to be expressed by chicken heterophils distributed in the yolk sac blood islands and small intestinal blood vessels. Only a subset of heterophils, which might be activated, were able to express AvBD1, 2, and 7 mRNA. In the intestine, the expression of AvBD10 mRNA was localized to cells along the villus at e19 and day of hatch, but later to only a few cells located above the intestinal crypts. In summary, the endodermal epithelial cells are responsible for the absorptive and immune functions of the chicken yolk sac. The yolk sac mesoderm is critical for embryonic hematopoiesis and innate immunity. The chicken small intestinal epithelial cells are derived from the intestinal stem cells in the crypts. These epithelial cells have different cell types, which are functioning to absorb nutrients and secrete antimicrobial peptides. / Ph. D. / The chicken yolk sac and small intestine are connected to each other and share many biological similarities. Both chicken small intestinal and yolk sac epithelia play critical roles for nutrient absorption and immune defense. In this dissertation, the mRNA for nutrient transporters such as the peptide transporter, PepT1 and the sodium-glucose co-transporter, SGLT1 were found to be expressed by absorptive epithelial cells in both the yolk sac and small intestine. Additionally, both intestinal and yolk sac epithelial cells expressed avian beta defensins (AvBDs), which are important chicken host defense peptides. In the small intestine, there are a number of differentiated cell types that originate from stem cells in the crypt that express the known mammalian stem cell markers, Olfm4 and Lgr5 mRNA. However, in the chicken yolk sac, only the stem cell marker Lgr5 mRNA was expressed by endothelial cells. In summary, the yolk sac epithelial cells are responsible for the absorptive and immune functions for the embryonic stage. The chicken small intestinal epithelial cells are derived from the intestinal stem cells in the crypts. These epithelial cells have different cell types, which function to absorb nutrients and secrete antimicrobial peptides.

Macrophage migration inhibitory factor (MIF) has been implicated in mediating both innate and adaptive immune responses in inflammatory and infectious diseases. The sequence and structure of MIF is highly conserved across the avian phylogeny, which underlies high sequence homology and functional similarities between turkey and chicken MIFs. Turkey MIF (TkMIF) inhibited cell migration and promoted cell proliferation with production of inflammatory mediators, comparable to the biological properties of chicken MIF (ChMIF), thus indicating the biological cross-reactivity between turkey and chicken MIFs. This study identified the cell surface receptor(s) that could bind ChMIF and the biological roles triggered by such interactions. In addition to CD74, a previously identified receptor, CXCR4 also interacts with ChMIF. Moreover, the formation of receptor complexes was shown between CXCR4 and CD74. MIF signaling through CXCR4 and CD74 led to cell chemotaxis and proliferation activity as well as intracellular calcium influx. Intriguingly, Eimeria MIF (EMIF), a homologue secreted following parasitic infection, also interacted with CD74 leading to comparable biological functions to those of ChMIF. Given such observations, we hypothesized that CXCR4 and CD74 are receptors for ChMIF leading to the functional consequences similarly manifested by EMIF interaction with the corresponding receptors. EMIF, predominantly secreted from the invasive merozoite stage, may help the parasite exploit the host immune response by interacting with common ChMIF receptors. This may lead to functional mimicry thus provoking the question of whether EMIF would modulate the biological functions of ChMIF to manipulate the host defense that allows more efficient invasion of the host. To evaluate this concept, a transgenic E. tenella lacking MIF was generated by in vivo passage of E. tenella transfected with a CRISPR plasmid targeting EMIF. Although not fully disrupted, reduction of EMIF expression was observed in the transgenic E. tenella itself as well as in inoculated cells, which resulted in enhanced survival of host cells. Herein, we achieved a better characterization of the functional roles of both avian and parasite MIFs underlying the interaction with common host receptors, along with the essential role of parasite MIF promoting host cell death during parasitic infection. / PHD / When animals get infected or injured, their immune system senses invading pathogens or damaged tissues as danger signals, which often elicits the production of inflammatory mediators. These are chemical messengers secreted mostly by immune cells that initiate cellular communication and infiltration of immune cells to the infection/damaged site leading to inflammatory responses to eliminate the infectious agents and repair damaged tissues. Among many inflammatory mediators, macrophage migration inhibitory factor (MIF) is involved in inflammatory and immune response by regulating cell migration. Interestingly, MIF is secreted by Eimeria parasites (that cause the costly coccidiosis disease in poultry) as well as by chickens (host animal) after infection with this pathogen. Toward a better understanding of the impacts of both avian and parasite MIFs on the host immune response, three specific studies were completed. First, MIF displayed high degree of gene sequence identity and functional similarity between chicken and turkey, supporting the evolutionarily conservation of MIF across birds. The second study identified the MIF receptors and their complexes, which engage in the biological functions of chicken MIF. Through binding to these cell surface receptors, chicken MIF can regulate cell migration and proliferation with calcium release. Intriguingly, Eimeria MIF secreted after parasitic infection is able to bind the same receptors leading to comparable biological functions to those of chicken MIF. Lastly, the role of Eimeria MIF was further evaluated by disrupting its gene in the parasite. Although not fully disrupted in the transgenic parasites, its expression was decreased resulting in enhanced survival of host cells, thus suggesting a deleterious effect of Eimeria MIF on the host, as well as its potential as a therapeutic target to control coccidiosis in poultry.

Feeding behavior is a complex trait that is regulated by various hypothalamic neuropeptides and neuronal populations (nuclei). Understanding the physiological regulation of food intake is important for improving nutrient utilization efficiency in agricultural species and for understanding and treating eating disorders. Knowledge about appetite in birds has agricultural and biomedical relevance and provides evolutionary perspective. I thus investigated hypothalamic molecular mechanisms associated with appetite in broilers, layers, chicken lines selected for low (LWS) or high (HWS) body weight, and Japanese quail, which provide a unique perspective to understanding appetite. Broiler-type chicks have been genetically selected for rapid growth and consume much more feed than do layer-type chicks which have been selected for egg production. Long-term selection has caused the LWS chicks to have different severities of anorexia while the HWS chicks become obese, thus making these lines a valuable model for metabolic disorders. Lastly, the Japanese quail have not undergone as extensive artificial selection as the chicken, thus this model may provide insights on how human intervention has changed the mechanisms that regulate feeding behavior in birds. This research involved applying a variety of different treatments including fasting and refeeding, diets differing in macronutrient composition, and/or central administration of neuropeptide Y, xenopsin, neuropeptide K, oxytocin, mesotocin, gonadotropin-inhibitory hormone, and prolactin-releasing peptide, after which I measured feeding behavior and various aspects of hypothalamic physiology. I measured nuclei activation in hypothalamic appetite-associated regions including the lateral hypothalamus, paraventricular nucleus, ventromedial hypothalamus, dorsomedial nucleus, and arcuate nucleus and I measured gene expression of various appetite-associated factors in the whole hypothalamus and individual nuclei. These data provided information about the regions of the brain involved in mediating effects on appetite and the molecular pathways involved in the effect on appetite. There were differences in dose threshold sensitivity to various injected factors in the different stocks, differential responses to fasting and refeeding, and differences in nuclei and genes that were activated in response to the various treatments. These data provide valuable insights on the molecular mechanisms that are associated with the short-term regulation of feeding behavior and pathways that may be genetically stock-dependent. / PHD / Poultry production and welfare may be enhanced by regulating food intake of chickens during specific stages of growth, resulting in improved nutrient utilization efficiency. For example, broiler breeders are feed restricted to achieve target weights in order to prevent obesity and other disorders and improve performance. To affect the appetite of chickens, an understanding of the neural mechanisms mediating food intake is needed but there is a lack of information in this area. Thus, the purpose of this dissertation was to elucidate some of these mechanisms that control appetite regulation in poultry using chickens and quail as models. Several neuropeptide-associated pathways were studied and appetite-related molecular mechanisms were elucidated for neuropeptides K and Y, oxytocin, mesotocin, gonadotropin-inhibitory hormone, prolactin-releasing peptide, xenopsin, different macronutrient composition diets, and fasting and refeeding.

Typescript (photocopy). / Digitized by Kansas Correctional Industries

La empresa Industrias Surco S.R.Ltda., tiene más de 20 años de experiencia fabricando equipos gastronómicos para restaurantes, cuentan con una planta de 1 200 m2 en el Parque Industrial de Villa El Salvador en Lima. Industrias Surco, ha patentado en Indecopi, el Horno Pollero Ecológico, además tiene registrada su marca en Estados Unidos, cumpliendo las normativas de fabricación americana. Las estrategias que utilizará la empresa para potenciar sus exportaciones serán: • Equipos para la industria gastronómica con alto valor agregado. • Incrementar la participación en ferias comerciales. • Aprovechar el potencial de demanda del mercado americano y la imagen ganada por la gastronomía peruana, además del consumo de pollo rostizado. • Implementar una campaña promocional para llegar a los clientes en el mercado americano, aprovechando el TLC. • Posicionar a la empresa como un proveedor de clase mundial. Para ello, la empresa participará como visitante en la Feria Hotel Motel and Restaurant Supply Show y luego como expositor en la Feria International Restaurant & Foodservice Show of New York, cumpliendo con los siguientes objetivos: • Incrementar las exportaciones en al menos US$ $28.000 en el 2019. • Obtener como mínimo dos (02) clientes en Estados Unidos en el 2019. • Participación en ferias internacionales en Estados Unidos. Para estos objetivos se obtendrá recursos no reembolsables del Programa de Apoyo a la Internacionalización - PAI. La participación de Industrias Surco en las Ferias en Estados Unidos será el inicio del crecimiento de sus exportaciones y de la consolidación como empresa peruana líder. / Industrias Surco S.R.Ltda., Has more than 20 years of experience manufacturing gastronomic equipment for restaurants, has a plant of 1,200 m2 in the Industrial Park of Villa El Salvador in Lima. Industrias Surco, has patented in Indecopi, the Ecological Pollero Oven, also has its trademark registered in the United States, complying with American manufacturing regulations. The strategies that the company will use to boost its exports will be: • Equipment for the gastronomic industry with high added value. • Increase participation in trade fairs. • Take advantage of the demand potential of the American market and the image gained by Peruvian cuisine, in addition to the consumption of roasted chicken. • Implement a promotional campaign to reach customers in the American market, taking advantage of the FTA. • Position the company as a world class supplier. For this purpose, the company will participate as a visitor at the Hotel Motel and Restaurant Supply Show and then as an exhibitor at the International Restaurant & Foodservice Show of New York, meeting the following objectives: • Increase exports by at least US $ 28,000 in 2019. • Obtain at least two (02) customers in the United States in 2019. • Participation in international fairs in the United States. For these objectives, will be obtained from the Programa de Apoyo a la Internacionalización - PAI, non-reimbursable resources. Industrias Surco's participation in the Fairs in the United States will be the beginning of its export growth and consolidation as a leading Peruvian company. / Tesis

Thesis (MSc. Agriculture (Animal Production)) -- University of Limpopo, 2014 / Two experiments were conducted to determine the effect of moringa seed meal supplementation on productivity and carcass characteristics of Ross 308 broiler chickens. The first experiment determined the effect of moringa seed meal supplementation on productivity of Ross 308 broiler chickens aged one to 21 days. Two hundred and fifty unsexed day-old Ross 308 broiler chicks were randomly allocated to five dietary treatments, replicated five times, and each replication having 10 chickens. A completely randomized design was used. The chickens were fed on a grower diet supplemented with 0 (M0), 5 (M5), 10 (M10), 15 (M15) and 20 (M20) g of moringa seed meal/bird/day. Moringa seed meal supplementation had no effect (P>0.05) on feed intake, metabolisable energy intake, nitrogen retention, feed conversion ratio and live weight of unsexed Ross 308 broiler chickens. Moringa seed meal supplementation improved (P<0.05) growth rates of unsexed Ross 308 broiler chickens aged one to 21 days. A moringa seed meal supplementation level of 13.3 g/kg DM feed optimized growth rate of Ross 308 broiler chickens aged one to 21 days. The second experiment determined the effect of moringa seed meal supplementation on productivity and carcass characteristics of female Ross 308 broiler chickens aged 22 to 42 days. The chickens weighing 558 ± 10 g/bird were randomly allocated to five treatments with five replications having 10 birds. The chickens, aged 21 days, were allocated to the treatments in a completely randomized design. The chickens were fed on a grower diet supplemented with 0 (FM0), 5 (FM5), 10 (FM10), 15 (FM15) and 20 (FM20) g of moringa seed meal per kg DM. Moringa seed meal supplementation had no effect (P>0.05) on feed intake, growth rate, feed conversion ratio, live weight, metabolisable energy intake, carcass weight, breast meat weight, abdominal fat pad weight, liver weight, heart weight, thigh weight, meat flavour, juiciness and tenderness of female Ross 308 broiler chickens. However, moringa seed meal supplementation improved (P<0.05) nitrogen retention and gizzard weights of female Ross 308 broiler chickens. vi It was concluded that moringa seed meal supplementation improved growth rate of unsexed Ross 308 broiler chickens aged one to 21 days. Similarly, moringa seed meal supplementation increased nitrogen retention and gizzard weights of female Ross 308 broiler chickens aged 22 to 42 days.