Changes in expression of an antimicrobial peptide, digestive enzymes, and nutrient transporters in the intestine of E. praecox-infected chickens.

Coccidiosis is a major intestinal disease of poultry, caused by several species of the protozoan Eimeria. The objective of this study was to examine changes in expression of digestive enzymes, nutrient transporters, and an antimicrobial peptide following an Eimeria praecox challenge of chickens at days 3 and 6 post-infection. Gene expression was determined by real-time PCR and analyzed by one-way ANOVA. In the duodenum, the primary site of E. praecox infection, a number of genes were downregulated at both d3 and d6 post-infection. These genes included liver expressed antimicrobial peptide 2 (LEAP2), the cationic (CAT1), anionic (EAAT3), and L-type (LAT1) amino acid transporters, the peptide transporter PepT1 and the zinc transporter ZnT1. Other transporters were downregulated either at d3 or d6. At both d3 and d6, there was downregulation of B(o)AT and CAT1 in the jejunum and downregulation of LEAP2 and LAT1 in the ileum. LEAP2, EAAT3, and ZnT1 have been found to be downregulated following challenge with other Eimeria species, suggesting a common cellular response to Eimeria.

Chickens from lines artificially selected for juvenile low and high body weight differ in glucose homeostasis and pancreas physiology.

Artificial selection of White Plymouth Rock chickens for juvenile (day 56) body weight resulted in two divergent genetic lines: hypophagic low weight (LWS) chickens and hyperphagic obese high weight (HWS) chickens, with the latter more than 10-fold heavier than the former at selection age. A study was designed to investigate glucose regulation and pancreas physiology at selection age in LWS chickens and HWS chickens. Oral glucose tolerance and insulin sensitivity tests revealed differences in threshold sensitivity to insulin and glucose clearance rate between the lines. Results from real-time PCR showed greater pancreatic mRNA expression of four glucose regulatory genes (preproinsulin, PPI; preproglucagon, PPG; glucose transporter 2, GLUT2; and pancreatic duodenal homeobox 1, Pdx1) in LWS chickens, than HWS chickens. Histological analysis of the pancreas revealed that HWS chickens have larger pancreatic islets, less pancreatic islet mass, and more pancreatic inflammation than LWS chickens, all of which presumably contribute to impaired glucose metabolism.

Chickens from lines selected for high and low body weight show differences in fatty acid oxidation efficiency and metabolic flexibility in skeletal muscle and white adipose tissue.

OBJECTIVE: The Virginia lines of chickens have resulted from more than 55 generations of artificial selection for low (LWS) or high (HWS) juvenile body weight. We hypothesized that the relative hyperphagia and greater body weight in juvenile HWS chickens are associated with altered fatty acid oxidation efficiency and metabolic flexibility in tissues associated with energy sensing and storage, and relative cellular hypertrophy in white adipose tissue. METHODS: Hypothalamus, liver, pectoralis major, gastrocnemius, abdominal fat, clavicular fat and subcutaneous fat were collected from the juvenile (56-65 days old) LWS and HWS chickens for metabolic, gene expression and histological assays. RESULTS: The HWS chickens had reduced fatty acid oxidation efficiency in abdominal fat (P<0.0001) and reduced rates of oxidation in abdominal fat and gastrocnemius (P<0.0001) as compared with the LWS. There was reduced citrate synthase activity in white adipose tissue (P<0.0001) and greater metabolic inflexibility in skeletal muscle (P=0.006) of the HWS compared with the LWS. Greater pyruvate dehydrogenase kinase 4 (PDK4) and forkhead box O1A (FoxO1) mRNA were found in skeletal muscle and white adipose tissue of 56-day-old HWS than LWS. Expression of peroxisome proliferator-activated receptor γ (PPARγ) in all adipose tissue depots was greater (P<0.05) in LWS than in HWS chickens. The HWS chickens had larger (P<0.0001) and fewer (P<0.0001) adipocytes per unit area than the LWS. CONCLUSION: Compared with the LWS, the HWS chickens have impaired metabolic flexibility and fatty acid oxidation efficiency due to greater pyruvate dehydrogenase activity to accommodate the influx of acetyl-CoA from fatty acid oxidation in skeletal muscle and adipose tissue. These metabolic adaptations can be linked to differences in gene expression regulation, adipocyte cellularity and body composition between the lines, which may provide valuable insight into metabolic disorders in other species.

Immunological responses to Clostridium perfringens alpha-toxin in two genetically divergent lines of chickens as influenced by major histocompatibility complex genotype.

Chickens genetically selected for low (LA) or high (HA) antibody response to SRBC displayed a correlated change in MHC, so that LA chickens were 96% B13 and HA chickens were 96% B21. The LA line appears to be less susceptible to invasion by extracellular pathogens, whereas HA chickens are more resistant to infection by intracellular organisms. Resistance to Clostridium perfringens is one instance in which the lines do not follow their established trend of pathogen susceptibility, where during a clinical outbreak of necrotic enteritis, B21B21 genotypes experienced significantly less mortality than B13B13 genotypes. A study was carried out to assess immunological differences between LA and HA lines during exposure to C. perfringens α-toxin. Peripheral blood mononuclear cells were isolated from each genetic line, cultured with or without lipopolysaccharide (4 h), and exposed to varying concentrations of α-toxin (1; 10; 100; and 1,000 U/L) for 2 and 4 h. Evaluation of cellular proliferation, percentage of cytotoxicity, and immunological gene expression was carried out in a series of experiments. Cells isolated from HA chickens had significantly increased proliferation than those from LA chickens at low toxin levels (1 and 10 U/L) and significantly decreased proliferation at high toxin levels (100 and 1,000 U/L). Following exposure to lipopolysaccharide, the percentage of cytotoxicity was higher for LA than HA cells. In both assays, HA cells displayed superior performance following lipopolysaccharide-stimulation. Gene expression analysis of immune transcripts by quantitative real-time PCR revealed significantly upregulated expression of interferon (IFN)-γ, interleukin (IL)-8, IL-13 (2 h), IL-15, and CXCLi1 (4 h) in HA than LA chickens. Cells isolated from the LA line displayed significantly elevated expression of IL-2, IL-10, IL-13 (4 h), IL-16, IL-18, inducible nitric oxide synthase (iNOS), CXCLi1 (2 h), and lipopolysaccharide-induced tumor necrosis factor-α factor (LITAF) compared with the HA line. Clearly, these 2 genetic lines display highly divergent immune responses in regards to C. perfringens toxin exposure.

Immune responses to dietary beta-glucan in broiler chicks during an Eimeria challenge.

Escalating consumer concerns regarding pathogen resistance have placed the poultry industry under mounting pressure to eliminate the use of chemotherapeutic agents as feed additives. One possible alternative receiving increased attention is the use of immunomodulators such as ß-glucan. A study was conducted to investigate the effects of a yeast-derived ß-glucan (Auxoferm YGT) on broiler chick performance, lesion scores, and immune-related gene expression during a mixed Eimeria infection. Day-old chicks were fed diets containing 0, 0.02, or 0.1% YGT. On d 8 posthatch, one-half of the replicate pens were challenged with a mixed inoculum of Eimeria acervulina, Eimeria maxima, and Eimeria tenella. Measurements were taken and samples collected on d 4, 10, 14, and 21 posthatch. Dietary supplementation had no effect on performance or mortality. On d 14, 3 birds per pen (n = 24/treatment) were scored for intestinal coccidia lesions. Gross lesion severity was significantly reduced in birds supplemented with 0.1% YGT. On d 10, inducible nitric oxide synthase (iNOS) expression was downregulated in the jejunum of challenged birds fed 0.1% YGT. Expression of iNOS in the ileum was downregulated in the nonchallenged birds, but upregulated in the challenged birds fed 0.1% YGT on d 14. Interleukin (IL)-18 was upregulated in the jejunum of 0.1% YGT-treated birds. Interferon (IFN)-γ expression was decreased in challenged and nonchallenged birds fed 0.1% YGT. The IL-4 expression was downregulated in the nonchallenged birds with 0.1% YGT diet supplementation. The IL-13 and mucin-1 levels were also reduced due to ß-glucan supplementation. Mucin-2 expression was increased in the nonchallenged birds, but decreased in the infected birds fed 0.1% YGT. These results suggest that although Auxoferm YGT at doses of 0.02 and 0.1% does not influence performance, it significantly reduces lesion severity and is capable of altering immune-related gene expression profiles, favoring an enhanced T helper type-1 cell response during coccidiosis.