The effects of vaccination with keyhole limpet hemocyanin or oral administration of Salmonella enterica serovar Enteritidis on the growth performance of immunoglobulin knockout chickens.

The nutritional cost of activating B cell-mediated immunity is thought to be low in chickens. However, this assumption is incompletely characterized. Immunoglobulin knockout (Ig-KO) chickens lacking B cells and immunoglobulin may potentially be a robust model to investigate the nutritional cost of immunity. Specifically, comparing the growth of Ig-KO chickens to immunocompetent birds (WT) following activation of the immune system should indicate costs associated with B cell-mediated immunity. A total of 3 experiments were conducted to determine if (1) an antibody response to keyhole limpet hemocyanin (KLH), (2) an intra-abdominal injection with Salmonella enterica serovar Enteritidis (SE), or (3) an oral challenge of SE would affect body weight gain (BWG) in homozygous Ig-KO (KO) chickens and their immunocompetent counterparts. In Experiment 1, a significant genotype*vaccination status interaction was observed (P = 0.03) during the period from 0 to 6 D after initial vaccination in which KLH injection reduced BWG in WT birds, but not in KO birds. A genotype*vaccination status interaction was present (P = 0.002) from 0 to 7 D after the first booster in which KLH injection due to reduced BWG in KO birds, but not WT birds. In Experiment 2, both KO and immunocompetent birds lost body weight during the period from 0 to 2 D after the SE injection, with no significant differences due to genotype (P = 0.92). Experiment 3 demonstrated that KO birds gained less weight than immunocompetent birds, with a tendency for less weight gain after an initial challenge (P = 0.07) and significantly so after a secondary challenge (P = 0.03). The results from these experiments collectively demonstrate that B cell-mediated immunity can affect growth performance in chickens. Furthermore, these effects can either preserve or impair growth performance, likely via mechanisms related to the immune response rather than the discrete nutritional cost of B cell-mediated immunity.

Space allowance influences individually housed Holstein male calves' age at feed consumption, standing behaviors, and measures of immune resilience before and after step-down weaning.

Many dairy calves in the southwestern regions of the United States are raised in wooden hutches with 1.23 m2 of free space that house 3 calves individually. Producers claim that calves in hutch systems are not ready to wean and be placed in groups at the country-average age of 6 wk. Calves may remain in this individual housing system until as late as 10 wk of age. The objective of this study was to modify space allowance of hutches and evaluate weaning readiness using age at solid feed consumption, standing behaviors, and measures of immunity. Calves were randomly assigned at 4 d of age to conventional (CONV; 1.23 m2 of space; n = 18), moderate (MOD; 1.85 m2 of space; n = 17), or maximized (MAX; 3.71 m2 of space; n = 19) space allowance in hutches. These modifications also changed the number of calves housed per hutch from 3 (CONV) to 2 (MOD) and 1 (MAX). Calves were fed milk replacer via bottle twice daily until weaning and offered ad libitum feed throughout the experiment. Step-down weaning was initiated (Wi) at age 53 or 54 d by withdrawal of the p.m. bottle and was completed (Wc) 11 d later by removal of the a.m. milk replacer. Accelerometer data for standing behaviors were collected relative to Wi (3 consecutive days to represent -4 wk, -3 d, 3 d, and 3 and 5 wk). Blood samples were collected in the a.m. just before Wi (d 0) and at d 3, 11, 14, and 18 after Wi. Calves provided with more space (MOD and MAX) compared with CONV calves consumed feed at an earlier age and had slightly healthier erythrocytes, greater circulating glucose, and fewer circulating eosinophils. The CONV calves had haptoglobin (Hp) responses to the stressors of both Wi and Wc and had more IFN-γ from whole blood stimulated with phytohemagglutinin-P. The MAX calves had the least active neutrophils (phagocytosis and oxidative burst), but MOD calves' leukocytes secreted the most TNF-α from whole blood stimulated with lipopolysaccharide. Just before and after weaning, MAX calves spent more time per day in the standing position than CONV and MOD calves and had an Hp response to Wc, but MOD calves did not have an Hp response to Wi or Wc. Based on these results, MOD calves were the most ready for weaning; therefore, they potentially can be moved to group housing at an earlier age than CONV calves, thus improving animal welfare concerns over space allowance and individual housing.

Effects of immunomodulatory nutrients on growth performance and immune-related gene expression in layer chicks challenged with lipopolysaccharide.

Immunomodulatory nutrients alter the immune response to pathogens. This study was conducted to determine the effects of immunomodulatory nutrients on the immune response to a lipopolysaccharide (LPS) challenge of layer chicks fed supplemental corn oil (control; 3%), fish oil (3%), conjugated linoleic acid (CLA; 1%), lutein (0.05 g/kg), or vitamin E (90 I.U./ kg). Four-week-old layer chicks were allotted to 10 treatment groups arranged as a 2 × 5 factorial with 2 sexes and 5 dietary treatments (TRT; 5 replicate pens of 2 male and 2 female chicks per TRT). After a 2-week diet adaptation, all birds were injected intraperitoneal with 1.5 mg/kg BW LPS. Twelve hours post challenge, samples were collected. Fish oil fed birds had greater (P = 0.03) spleen weight (% final BW) than the CLA fed birds. In the liver, the fish oil TRT had higher (P = 0.040) IL-12 expression than the corn oil TRT, but the corn oil TRT had greater (P = 0.001) IL-4 expression than the CLA, lutein, and vitamin E TRT. There was a main effect of sex of the birds on growth parameters at 12 h post LPS challenge in which male birds had greater beginning BW (P < 0.001), final BW (P < 0.001), and greater 12-hour BW loss (P = 0.020) than the female birds, but not relative weight loss. There were also main effects of sex on immune-related gene expression with the females having greater gene expression than the males in the duodenal mucosal scrapings [IL-1ß, IL-12, and TLR-4 gene expression (P = 0.026, 0.011, and 0.002, respectively)]; liver [IL-10, IL-4, and iNOS gene expression (P = 0.017, 0.032, and 0.006, respectively)]; and spleen [IL-1ß, IL-10, IL-4, and iNOS gene expression (P = 0.001, 0.001, 0.001, and 0.005, respectively)]. Therefore, each immunomodulatory nutrient added to the diets of layer chickens resulted in different immune responses to an LPS challenge.

Space allowance influences individually housed Holstein bull calf innate immune measures and standing behaviors after castration at 3 weeks of age.

Dairy calves in the Southwest regions of the United States are typically raised individually in wooden hutches with 1.23 m2 of space. The objective of the study was to determine if increased space allowance in wooden hutches influences measures of innate immunity and behaviors of Holstein bull calves pre- and postcastration. Calves were randomly assigned at 4 d of age to conventional (CONV; 1.23 m2 of space; n = 18), moderate (MOD; 1.85 m2 space; n = 17), or maximized space allowance (MAX; 3.71 m2 space; n = 19) in hutches. Calves were surgically castrated at 24 d of age. Peripheral whole blood samples were collected at -1, +1, +5, and +12 d of castration. Accelerometer loggers (n = 16 calves per treatment) were used from -3 to +5 d of castration to assess standing behaviors. All calves decreased total standing duration the day of castration versus precastration. Overall, MAX spent the most time in the stand position postcastration versus CONV and MOD. Within treatments, MOD and MAX had increased plasma cortisol 1 d postcastration versus precastration. A treatment × time tendency was observed for cortisol at 12 d postcastration; MAX had the least circulating cortisol. A treatment × time tendency for circulating haptoglobin (Hp) was observed and Hp was greatest among CONV 1 d pre- and 12 d postcastration. Compared with precastration, CONV had increased Hp at 1, 5, and 12 d, whereas MOD had increased Hp at 5 d, and Hp remained similar within MAX. A treatment × time tendency for tumor necrosis factor-α (TNF-α) from lipopolysaccharide-stimulated whole blood was observed; at 1 d postcastration, MOD had the most TNF-α, whereas MAX had the least. Within MAX, calves had increased TNF-α from precastration to 5 d postcastration. A treatment × time interaction was observed for whole blood bactericidal activity against Escherichia coli (WB anti-E). The CONV tended to have the greatest WB anti-E at d -1, but at d 1 and 5 postcastration, CONV had the least WB anti-E. Overall, MAX had less intensity of neutrophil oxidative burst versus CONV and MOD. The lower response of neutrophil oxidative burst and slower Hp secretion after castration is indicative that the wound site likely had less microbial exposure. The findings of this study suggest that calves housed with more space are potentially at less risk of too much inflammation after castration, which may likely be due to the effects of increased space on hide cleanliness and increased standing time.

Liver mitochondrial oxygen consumption and proton leak kinetics in broilers supplemented with dietary copper or zinc following coccidiosis challenge.

Mitochondrial respiration was assessed in sixteen 7-day-old broilers as a subset of a larger study assessing the effects of Cu and Zn supplementation above requirements with a coccidiosis challenge on gain/feed ratio. The birds were selected from four treatments (four birds/treatment): a control diet (Cu 15 mg/kg and Zn 60 mg/kg) + coccidiosis challenge (CC), a Cu diet with 245 mg/kg Cu from tribasic copper chloride (TBCC) + CC, a negative control diet (Cu 15 mg/kg and Zn 60 mg/kg) - CC and a Zn diet with 2000 mg/kg Zn from ZnO. The diets were composed of 49% corn, 40% soybean meal, 6.2% vegetable oil (diet dry matter = 90.62%, crude protein = 21.37%, fat = 7.7%, metabolizable energy = 12.1 MJ/day) and were fed for 14 days. Birds were dissected, and approximately one gram of liver tissue was used for mitochondrial oxygen consumption and proton leak kinetics assays. Respiratory control ratio and mitochondrial proton leak assessed by calculating rates of oxygen consumption at 175mV membrane potential were greater for the negative control group, but there were no differences in average gain/feed among treatments. In summary, broilers that did not undergo coccidiosis challenge had lower proton leak and higher respiratory control ratio. However, the impact of supplementation of Cu and Zn above requirements did not appear to prevent changes in respiratory control ratio and proton leak kinetics with coccidiosis challenge.

Wooden hutch space allowance influences male Holstein calf health, performance, daily lying time, and respiratory immunity.

Dairy calves in the western United States are commonly raised individually in wooden hutches with a space allowance of 1.23m(2)/calf. Recent legislative initiatives in California and across the United States were passed regarding concern over space allowance for farm animals. The objective of this study was to determine if rearing male Holstein calves in wooden hutches modified to increase space allowance would influence measures of performance, lying time per day, health, and respiratory immunocompetence. At 4d of age, 60 calves were randomly assigned to 1 of 3housing treatments: (1) conventional housing (CONV; 1.23m(2)/calf), (2) 1.5 × CONV (MOD; 1.85m(2)/calf), or (3) 3 × CONV (MAX; 3.71m(2)/calf). Intakes of milk and solid feed were recorded daily and body weight was measured at 0, 3, 6, 10, and 12 wk of age. For the first 3 wk of the trial, calves were scored daily for fecal consistency, hydration, and hide cleanliness. In addition, calves were scored for respiratory health (i.e., nasal and eye discharge, ear position) until 7 wk of age. The total lying duration per day was recorded using data loggers at 3, 6, and 10 wk of age. Eight clinically healthy calves from each treatment were sensitized with subcutaneous ovalbumin (OVA) and then challenged with aerosolized OVA to assess calf respiratory immunity at 11 wk of age. Bronchoalveolar lavage fluid (BALF) was collected 4d after the OVA challenge and analyzed for leukocyte differentials and OVA-specific IgG, IgG1, IgA, and IgE. Calf average daily gain and body weight were positively associated with space allowance at approximately 3 wk before weaning and throughout postweaning, respectively. A greater space allowance decreased lying time after 46d. Space allowance did not influence fecal consistency, but there was a tendency for MAX calves to take 1d longer to recover from loose feces than MOD calves. The MAX calves had the fewest (%) observations with feces on their body compared with CONV or MOD. At 3 wk of age, peripheral eosinophil concentrations decreased with increased space allowance. However, observations (%) of eye discharge increased with greater space allowance. Among calves challenged with OVA, MOD calves had the least BALF OVA-IgE, and the percent of BALF eosinophils decreased with increased space allowance. Increased space allowance for calves raised in wooden hutches may improve some measures of calf performance, health, and respiratory immunocompetence.

Effects of conjugated linoleic acid and lutein on the growth performance and immune response of broiler chickens.

The effects of lutein and conjugated linoleic acid (CLA) on growth performance and immune response of broiler chickens were evaluated in the presence and absence of Salmonella lipopolysaccharide (LPS) immune challenge. Cobb chicks (360; 1 to 22 d of age) were used in a 3 × 2 factorial arrangement of CLA (0, 1, and 2%) and lutein (0 and 50 mg/kg) dietary levels. At d 8 and 15, birds were injected with BSA to assess IgY production. At d 20, birds were injected with LPS. Samples of liver, spleen, and duodenum were collected at 3 and 16 h post-LPS challenge for RT-qPCR analysis of RXRα, RXRγ, PPARα, PPARγ, TLR-4, IL-1ß, IL-2, IL-10, and IL-12 gene expression. CLA decreased BW, BW gain (BWG), and G:F from d 1 to 20, but these effects were reversed when lutein was included in the 1% CLA diet (P < 0.001). The production of IgY anti-BSA increased following a 2% CLA supplementation (P < 0.01). LPS increased the liver:BW ratio at 3 h post-injection (P < 0.001) and decreased BWG at 3, 16, and 40 h (P < 0.001). Lutein decreased plasmatic nitric oxide levels (P < 0.01). LPS downregulated PPARα mRNA in the duodenum (P = 0.02) and liver (P = 0.04), and PPARγ (P = 0.01) and RXRα (P = 0.08) in the spleen; these effects were not reversed by CLA or lutein as initially hypothesized. Although LPS upregulated IL-1ß (P = 0.02) and IL-12 (P = 0.07) expression, lutein downregulated these pro-inflammatory cytokines in the liver (P = 0.03 and P = 0.07, respectively). Lutein decreased splenic (P = 0.09) but increased hepatic (P = 0.06) TLR-4 mRNA. A dietary CLA supplementation of 2% increased hepatic RXRα (P = 0.10). In conclusion, CLA decreased broiler chicken growth performance, but lutein could prevent this negative effect (depending on CLA dose). Lutein had an anti-inflammatory effect, and a 2% CLA supplementation improved the humoral immune response.

Changes in the amount of lysine in protective proteins and immune cells after a systemic response to dead Escherichia coli: implications for the nutritional costs of immunity.

The nutritional demands of the immune system may result in tradeoffs with competing processes such as growth and reproduction. The magnitude of the nutritional costs of immunity is largely unknown. Thus, we examine the lysine content of the systemic components of the immune system in adult male chickens (Gallus gallus domesticus) in a healthy condition (maintenance) and following a robust Escherichia coli-specific immune response. Lysine was used as a metric, because it is found both in leukocytes and in protective proteins. The dynamics of subsets of leukocytes were monitored in primary and secondary immune tissues (thymus, bone marrow, and spleen) that would be expected to be involved in the response following iv injection of E. coli. The systemic immune system at maintenance has the same lysine content as 332 average-sized feathers, 16% of an egg, or 5.4% of a pectoralis muscle from an adult chicken. During the acute-phase response to E. coli, the additional lysine needed would equal 355 feathers, 17% of an egg, or 5.5% of a pectoralis muscle. The acute-phase proteins accounted for the greatest proportion of lysine in the immune system at maintenance and the proportion increased substantially during an acute-phase response. Hypertrophy of the liver required more lysine than all of the leukocytes and protective proteins that were produced during the acute-phase response. Size of the liver and levels of protein during the acute phase returned to normal during the time when the adaptive response began to utilize significant quantities of lysine. The catabolism would release a surfeit of lysine to provision the anabolic processes of the adaptive response, thus making proliferation of lymphocytes and production of immunoglobulins very cheap.

Effect of weaning and in-feed high doses of zinc oxide on zinc levels in different body compartments of piglets.

High doses of Zn are widely used for prevention and treatment of diarrhoea in weaning piglets; however, the mechanism of action of Zn against diarrhoea is still not well understood. The objective of this study was to evaluate whether weaning induces Zn deficiency in piglets. Eight litters of primiparous sows were selected for the experiment, and 3 piglets presenting similar weights were selected within each litter. Two of the three selected piglets from each litter were weaned at 21d of age and fed two different diets: a commercial control diet (WCt) and the same diet plus 2000 ppm of Zn as ZnO (WZn). The third selected pig from each litter was kept unweaned (Uw) with the sow and the rest of the litter. All 24 selected animals were killed at 28 d of age, and blood, gastrointestinal content, liver, pancreas and spleen were sampled for Zn, Fe and Cu analysis (mg/kg or L of sample). Data were analysed using anova including treatment as a fixed factor. Weaned pigs fed WCt diet presented a lower Zn concentration in plasma than Uw animals (0.76 ± 0.091 vs. 1.10 ± 0.099 mg/L, p = 0.05). Zinc levels in liver, pancreas and spleen were not affected by weaning. Total concentration of Zn was higher in gastrointestinal contents of weaned animals fed WCt diet than in Uw pigs (p ≤ 0.001 for stomach, jejunum, ileum, caecum and colon). Supplementation with high doses of ZnO increased levels of Zn in gastrointestinal content (p < 0.001), liver (p < 0.001) and pancreas (p < 0.001) compared to WCt diet. It also increased plasma Zn to non-deficient levels (1.32 ± 0.086), but the increase was not as marked as in other locations and final concentration was not different than that in Uw animals (p = 0.231). Weaning creates a Zn deficiency situation in weaned pigs as observed by plasma Zn concentrations. ZnO supplementation counteracts Zn deficiency.

Dynamics of the systemic components of the chicken (Gallus gallus domesticus) immune system following activation by Escherichia coli; implications for the costs of immunity.

The immune response is thought to be costly and deters from growth and reproduction, but the magnitude and sources of these costs are unknown. Thus, we quantified the changes in mass of leukocytes (CD4(+) and CD8(+) T cells, Bu1(+) IgM(+) and Bu1(+) IgG(+) B cells, monocytes/macrophages, heterophils and thrombocytes) and protective plasma proteins in systemic (non-mucosal) components of adult chickens injected intravenously with dead Escherichia coli. During the first day after E. coli injection most types of blood leukocytes decreased and α-1-acid glycoprotein increased. Specific IgM, specific IgY, total IgM, Bu1(+) lymphocytes in the spleen and bone marrow and thymic CD8(+) lymphocytes increased at 5d post-injection. Quantitatively, the increases in the weight of cells and antibodies due to E. coli were dwarfed by the increase in the weight of the liver and acute phase proteins. Thus the acute phase response was markedly more costly than the subsequent adaptive response. The weight of the cells and proteins of the systemic immune system prior to challenge was 0.14% of body weight. Following E. coli injection, the additional weight of the immune components and the hypertrophy of the liver resulted in a 3.6-fold increase in weight which is equivalent to 18.5% of a large egg.

Effect of dietary omega-3 fatty acids on red blood cell lipid composition and plasma metabolites in the cockatiel, Nymphicus hollandicus.

Although dietary n-3 fatty acids have been extensively studied in poultry, they have not yet been prospectively investigated in psittacines, despite potential benefits for preventing and treating atherosclerosis, osteoarthritis, and other chronic disease processes. The objectives of this study were to investigate the incorporation of dietary n-3 fatty acids into red blood cells (RBC) and to determine the effects of supplementation of psittacine diets with fish or flax oil on plasma lipids and lipoproteins in the cockatiel. Adult cockatiels were fed a custom-formulated diet containing either 4% (wt/wt, as-fed) beef tallow (CON), 3% fish oil + 1% tallow (FSH), or 3.5% flax oil + 0.5% tallow (FLX; n = 20 per diet group). Baseline measurements were obtained for RBC fatty acid composition, triacylglycerides (TAG), and cholesterol. After 8 to 13 wk on the study diets, plasma chemistry profiles, lipoprotein density profiles, and RBC fatty acid composition were determined. At 8 wk, total plasma cholesterol was least in FSH birds (P < 0.05) and TAG concentrations were less in FSH birds than FLX birds (P < 0.05). Total n-3 fatty acids, docosahexaenoic acid, docosapentaenoic acid, and eicosapentaenoic acid were markedly greater in the RBC of FSH birds than FLX or CON birds (P < 0.05). Alpha linolenic acid was greatest in FLX (P < 0.05). Initial and final BW, and nonlipid plasma chemistry values did not differ among diet groups. No adverse effects of dietary supplementation of cockatiels with 3.5% flax oil or 3% fish oil were observed during the 13-wk feeding period. Although fish and flax oils provided similar total n-3 PUFA to the diets, fish oil caused greater reductions in cholesterol and TAG, and greater total RBC n-3 incorporation. Thus, dietary modification of psittacine diets with long chain n-3 PUFA from fish oil appears safe and may be beneficial to these long-lived companion birds.

Evolution of zinc, iron, and copper concentrations along the gastrointestinal tract of piglets weaned with or without in-feed high doses of zinc oxide compared to unweaned littermates.

High doses of ZnO are used to treat diarrhea in weaning pigs but are also an environmental concern. The mechanism of action of Zn against diarrhea is still not well understood. The amount of solubilized Zn, the relation of Zn with Fe and Cu, and the concentration of these elements in the gastrointestinal tract (GIT) are key data to understanding its mechanism of action and optimize its use. Therefore, we studied the Zn, Fe, and Cu concentrations in the GIT content of unweaned, weaned, and ZnO-treated pigs. Eight litters were used and 3 piglets were selected within each litter. Two piglets from each litter were weaned at 21 d of age and fed a commercial control diet (WCt) or WCt + 2000 mg/kg of Zn as ZnO (WZn). The third pig was kept unweaned (Uw) with the sow. All 24 pigs were killed at 28 d of age, GIT contents were sampled, soluble and insoluble fractions of the content were separated, and Zn, Fe, and Cu concentrations were analyzed. Concentration of Zn increased 3 to 5 fold along the GIT (2 to 10 fold in the soluble fraction) for weaned pigs on WZn compared to WCt and Uw pigs (P < 0.01 in all cases). The proportion of total Zn that was present in the soluble fraction was 4 to 10 folds higher in jejunum, ileum, and cecum of Uw pigs than in those weaned (P < 0.01 in all cases) but was not affected by ZnO treatment. Concentration of Fe in the soluble fraction was higher for Uw pigs compared to weaned pigs along the GIT (P < 0.05 in all cases) even when concentration in total content was lower for Uw pigs in stomach (P = 0.001) and jejunum (P = 0.029). Concentrations of Cu were lower in Uw pigs than in weaned pigs along the GIT (P < 0.05 in all cases). Surprisingly, animals on WZn showed a 5 to 10 fold increase of Cu solubilized in distal parts of the GIT (cecum and colon; P < 0.001) compared to other groups. Differences in Zn, Fe, and Cu concentrations found among treatments will be useful in future studies for understanding mechanism of action of ZnO and optimizing its use in order to avoid environmental concerns.

Domestic cats convert [2H8]-ß-carotene to [2H4]-retinol following a single oral dose.

Many animals convert ß-carotene to retinol to meet their vitamin A (VA) requirement. However, this pathway is inefficient in many carnivores. This study quantified the plasma response to a single oral dose of [(2) H(8)]-ß-carotene in adult domestic cats, including measurement of [(2) H(4)]-retinol derived from the dose. Cats were fed with either a control diet containing adequate VA (n = 5) or a VA-devoid diet (n = 5) for 28 days. An oral dose of either 5 mg/kg body weight (BW) (n = 4) or 10 mg/kg BW (n = 6) of [(2) H(8) ]-ß-carotene was administered on day 28. Plasma samples were collected prior to dosing and at 6, 12, 24, 32, 48, 72, 120, 168 and 216 h post-dose. Plasma retinoids and ß-carotene were measured using HPLC and [(2) H(4)]-retinol by GC-ECNCI-MS (gas chromatography/electron capture negative chemical ionization/mass spectrometry). ß-carotene was undetectable in plasma prior to dosing. Post-dose, mean peak plasma ß-carotene was 0.37 ± 0.06 nmol/ml at 9.0 ± 1.8 h following the dose, while [(2) H(4) ]-retinol peaked at 3.71 ± 0.69 pmol/ml at 55.2 ± 16.3 h. The ratio per cent of total area under the curve for [(2) H(4)]-retinol compared with the ß-carotene response was 4.6 ± 2.6%. There was little effect of diet or dose on the ß-carotene or [(2) H(4)]-retinol responses. The appearance of [(2) H(4)]-retinol in plasma indicates that cats are capable of converting ß-carotene to active VA. Conversion efficiency was not calculated in this study, but it is likely inadequate to meet cats' VA requirement without the inclusion of preformed VA in the diet.

Retinyl palmitate does not have an adjuvant effect on the antibody response of chicks to keyhole limpet hemocyanin regardless of vitamin A status.

To determine whether supplementing retinyl palmitate (RP) to vitamin A-deficient or -adequate chicks during vaccination increased the antibody response, an experiment was conducted that used a 2-generational model. White Leghorn chicks were hatched from eggs that contained 1.06 ± 0.13, 1.41 ± 0.14, or 1.90 ± 0.13 µg of retinol/g of egg yolk. Upon hatch, chicks were allotted to a diet that contained 70, 100, or 130% of the vitamin A requirement (1,500 IU/kg). At 14 d of age, chicks were challenged with keyhole limpet hemocyanin (KLH), Freund's incomplete adjuvant (FIA), KLH and FIA, KLH and RP administered in the same injection, or KLH and RP administered at different injection sites. The RP dosage was 7,000 IU/kg of BW, and dosage of the KLH was 1 mg/kg of BW. Chicks were bled and rechallenged at 19 d of age, and a final blood draw was taken at 29 d of age. The IgM and IgG antibody responses to KLH were determined by ELISA, and agglutination was used to determine natural antibody titers. When compared with chicks fed diets that contained 70 and 100% of the vitamin A requirement, the chicks fed 130% of the vitamin A requirement had a lower primary IgM and IgG antibody response (P < 0.02) and the secondary IgG response was significantly lower (P = 0.004). However, administering RP together or separate from KLH did not significantly change the secondary IgM or IgG responses compared with chicks challenged with KLH and FIA (P < 0.01). Moreover, natural antibody titers were not different among dietary treatments (P = 0.91). In conclusion, administration of RP with KLH did not improve antibody response in chicks.

Lutein exposure, in ovo or in the diet, reduces parameters of inflammation in the liver and spleen laying-type chicks (Gallus gallus domesticus).

These trials examined whether the demonstrated effects of embryonic and dietary carotenoid exposure on the inflammatory immune response in fast growing chickens also occur in slow growing chickens. The systemic and local inflammatory responses of chicks were examined in two experiments with two in ovo lutein levels (C+, carotenoid replete; or C-, carotenoid-deplete), two dietary lutein levels (0 or 40 mg lutein/kg diet), and two inflammatory challenges [no exposure or lipopolysaccharide (LPS)-vaccinated]. At 24 h after LPS vaccination, spleen weight was not affected by diet or in ovo lutein, but liver weight increased from C+ eggs (p < 0.01), and in LPS-vaccinated chicks fed 0 mg lutein (p < 0.05), but not in chicks fed 40 mg lutein. Plasma carotenoids and liver carotenoids were reduced post-LPS (p < 0.05). Splenic IL-6 mRNA abundance was the greatest post-LPS in C- chicks fed 40 mg lutein vs. C+ chicks fed 40 mg lutein (p < 0.05). Hepatic IL-6, iNOS and TGFß and splenic iNOS and TGFß were not affected by in ovo or dietary lutein. The systemic and local inflammatory results are similar to those observed in fast growing chickens, and support that lutein-depleted birds have greater inflammatory responses.

The effect of light intensity on the behavior, eye and leg health, and immune function of broiler chickens.

Broilers are typically raised commercially in dim lighting. It has been suggested that providing brighter light intensity could improve health and provide opportunities for more normal behavioral rhythms. We examined the effects of 3 photophase light intensities (5, 50, and 200 lx) on activity patterns, immune function, and eye and leg condition of broilers (n = 753; 6 replicate pens/treatment). Broilers were reared with one of these intensities from 1 to 6 wk of age; photoperiod consisted of 16L:8D with 1 lx intensity during the scotophase. Broilers reared with 5 lx were less active (P = 0.023) during the day than 50 or 200 lx and showed less (P < 0.0001) change in activity between day and night than 50 or 200 lx. There was no difference between treatments for final BW (2.30 +/- 0.02 kg) or for most immune parameters (IgG primary and secondary responses to keyhole limpet hemocyanin, B and T lymphocyte proliferation, plasma lysozyme, haptoglobin, NO, whole blood killing of Escherichia coli and Staphylococcus aureus), but there was a trend (P = 0.072) for a greater IgM response in 50 lx (6.21 titer) than 5 lx (5.78 titer), with 200 lx (5.92 titer) intermediate. There was no effect of light intensity on back-to-front (1.13 +/- 0.01 cm) or side-to-side (1.48 +/- 0.01 cm) diameter of the eyes or on corneal radii (0.82 +/- 0.01 cm), but 5 lx (2.33 +/- 0.07 g) had heavier eyes (P = 0.002) than 50 lx (2.09 +/- 0.04 g) or 200 lx (2.11 +/- 0.04 g). There were no differences in gait score, although 200 lx broilers had more hock and footpad bruising (P = 0.038) but fewer erosions (P = 0.006) than 5 or 50 lx. Increased daylight intensity had little effect on broiler health but resulted in more pronounced behavioral rhythms.

Dietary ß-hydroxy-ß-methylbutyrate supplementation influences performance differently after immunization in broiler chickens.

Dietary addition of the leucine metabolite ß-hydroxy-ß-methylbutyrate (HMB) promotes growth in various species. In addition, HMB is described to enhance immune responses which might be associated with metabolic costs. We elaborated further on the role of HMB in growth, metabolism and immunity of meat-type chickens using the following parameters: zootechnical performance, blood chemistry and a specific immune responses after immunization with a human serum albumin (HSA)/Freund's (in) complete adjuvant combination. The chickens received commercial feeds either unsupplemented or supplemented with 300 mg HMB/kg feed. ß-hydroxy-ß-methylbutyrate-supplemented chickens were significantly heavier at 2 weeks of age but this difference was attenuated at later ages. Compared with their unsupplemented controls, cumulative feed conversion was significantly lower in HMB-supplemented chickens. There were no differences in blood chemistry between both dietary treatments. After immunization, HMB significantly attenuated the acute phase protein response at day 1 of post-immunization compared with that of their unsupplemented counterparts. After day 7 post-immunization, body weight gain of the immuno-challenged HMB-supplemented chickens was significantly depressed, but their specific anti-HSA IgG response was significantly enhanced compared with that of their immuno-challenged unsupplemented counterparts. The underlying mechanisms and signalling pathways for these phenomena need to be elucidated. Nevertheless, we are able to conclude that HMB is beneficial for performance under normal circumstances. On the other hand, HMB stimulates the immune response after an immunological challenge, though at the cost of reduced growth.

Constitutive immune defences correlate with life-history variables in tropical birds.

1. It has been suggested that immune defences are shaped by life history and ecology, but few general patterns have been described across species. We hypothesized that 'fast' life-history traits (e.g. short development times, large clutch sizes) would be associated with developmentally inexpensive immune defences, minimizing the resource demands of young animals' immune systems during periods of rapid growth. Conversely, 'slow' life histories should be associated with well developed antibody-mediated defences, which are developmentally costly. 2. We therefore predicted that 'fast-living' species would exhibit higher levels of complement proteins, a component of non-specific innate defence, but lower levels of constitutive ('natural') antibodies. Additionally, we predicted that constitutive immune defences in general would be higher in species with ecological characteristics that might increase exposure to pathogens, such as open nests, omnivorous diets, gregariousness, and closed forested habitat. 3. Across 70 Neotropical bird species, we found a strongly positive relationship between incubation period and natural antibody levels in adult birds, suggesting that longer developmental times might allow the production of a more diverse and/or more reactive adaptive immune system. Complement activity was positively, although weakly, correlated with clutch size, providing some support for the hypothesis that faster-living species rely more on innate defences, such as complement. Unexpectedly, solitary species had higher natural antibody titres than species that frequently join flocks. 4. Our results suggest that, despite probably widespread differences in the intensity and diversity of pathogen exposure, species-level variation in constitutive immune defences is understandable within the context of life-history theory.

Nutrition and the immune system.

1. Infectious diseases reduce productivity and diminish animal welfare. 2. Appropriate nutrition may aid in minimising the incidence of diseases by enhancing immunity. 3. An understanding of the pressures imposed by evolution that underlie poultry nutrition as well as those which underlie immunity provides focus to the field of nutritional immunology. Additional understanding is provided by knowing the specific cellular mechanisms by which diet affects immunity, and how these mechanisms pertain to specific nutrients and pathogens. 4. These approaches indicate that higher inclusion rates of all nutrients are not always better and that the traditional ideas of adding a surfeit of specific nutrients to improve immunity does not usually serve us well. 5. Understanding the nuances of nutrition and immunity is important for optimising bird health and productivity, and will be an important contributor towards fulfilling the consumer's conflicting demands for more natural production and better animal welfare.

Oestradiol, but not genistein, inhibits the rise in food intake following gonadectomy in cats, but genistein is associated with an increase in lean body mass.

The prevalence of obesity in domestic cats is increasing worldwide, and is strongly associated with gonadectomy. We have previously demonstrated the effectiveness of oestradiol in reducing food intake in both male and female neutered cats. This experiment was designed to test the hypothesis that oestradiol or genistein would prevent the increase in food intake following gonadectomy of male and female cats, and would prevent an increase in body fat mass. Three groups of eight cats each were surgically neutered then treated daily with either 0.5 mug oestradiol subcutaneously, 100 mg/kg genistein orally, or vehicle only. Effect of treatment on food intake, vaginal cytology and body weight were recorded, and body composition was assayed using the D(2)O isotopic dilution method. Neutering was followed by an increase in food intake, bodyweight and body fat mass in the control group, which were almost completely prevented by treatment with oestradiol (p < 0.001). Treatment with genistein had no effect on food intake or bodyweight increase, but was associated with a significant increase in lean body mass (p = 0.018), and significantly less body fat accumulation than the control group (p = 0.01). There were no significant differences in responses to treatment between sexes. These findings demonstrate the importance of gonadal oestrogen for the control of food intake in male and female cats, and suggest the provision of an oestrogenic compound could help prevent obesity following neutering. In addition, the findings of this study are consistent with observations in rodents of the efficacy of genistein in inhibiting adipogenesis and promoting lean body tissue development.