T. J. Applegate

Influence of Stressors on Normal Intestinal Microbiota, Intestinal Morphology, and Susceptibility to Salmonella Enteritidis Colonization in Broilers

In modern poultry production systems, environmental stressors may influence bird performance and susceptibility to pathogens such as Salmonella Enteritidis. Experiments were conducted to determine the influence of 24-h feed withdrawal and 24-h exposure to high temperature (30 degrees C) on intestinal characteristics of broilers. Attachment of Salmonella Enteritidis to ileal tissue was determined using an in vitro ileal loop assay. Changes in commensal intestinal microbial populations were determined using denaturing gradient gel electrophoresis, and alterations in ileal morphology were determined histologically. Ex vivo attachment of Salmonella Enteritidis to ileal tissues increased by 1.5 logs (9.05 log10 vs. 7.59 log (10) Salmonella Enteritidis/g of ileal tissue; P = 0.0006) in broilers fasted for 24 h. Similarly, ileal tissues from birds subjected to 30 degrees C for 24 h had increased ex vivo attachment of Salmonella Enteritidis (8.77 log(10) vs. 8.50 log(10) Salmonella Enteritidis/g of ileum; P = 0.01) compared with birds held at 23 degrees C. Exposure to 30 degrees C for 24 h also altered microbial community structure in the ileum and cecum. Subjecting birds to 30 degrees C for 24 h reduced crypt depth (6.0 vs. 7.8 microm, respectively; P = 0.002), but had no effect on villus height or villus:crypt ratio. This research shows that acute stressors in poultry production systems can cause changes in the normal intestinal microbiota and epithelial structure, which may lead to increased attachment of Salmonella Enteritidis.

Modulation of Intestinal Functions Following Mycotoxin Ingestion: Meta-Analysis of Published Experiments in Animals

Mycotoxins are secondary metabolites of fungi that can cause serious health problems in animals, and may result in severe economic losses. Deleterious effects of these feed contaminants in animals are well documented, ranging from growth impairment, decreased resistance to pathogens, hepato- and nephrotoxicity to death. By contrast, data with regard to their impact on intestinal functions are more limited. However, intestinal cells are the first cells to be exposed to mycotoxins, and often at higher concentrations than other tissues. In addition, mycotoxins specifically target high protein turnover- and activated-cells, which are predominant in gut epithelium. Therefore, intestinal investigations have gained significant interest over the last decade, and some publications have demonstrated that mycotoxins are able to compromise several key functions of the gastrointestinal tract, including decreased surface area available for nutrient absorption, modulation of nutrient transporters, or loss of barrier function. In addition some mycotoxins facilitate persistence of intestinal pathogens and potentiate intestinal inflammation. By contrast, the effect of these fungal metabolites on the intestinal microbiota is largely unknown. This review focuses on mycotoxins which are of concern in terms of occurrence and toxicity, namely: aflatoxins, ochratoxin A and Fusarium toxins. Results from nearly 100 published experiments (in vitro, ex vivo and in vivo) were analyzed with a special attention to the doses used.

Standardized Ileal Amino Acid Digestibility of Plant Feedstuffs in Broiler Chickens and Turkey Poults Using a Nitrogen-Free or Casein Diet

The standardized ileal amino acid digestibility (SIAAD) of 5 plant-based ingredients was determined in 5- and 21-d-old broiler chicks and turkey poults. Two diets (N-free diet, NFD, and a highly digestible protein diet 10% casein, HDP) were fed to determine ileal endogenous amino acid flow, and each was used for endogenous amino acid correction for the plant-based ingredients. The plant-based ingredients evaluated were 2 samples of corn distillers dried grains with solubles (light and dark DDGS), canola meal, corn, and soybean meal. Each diet was fed for 5 d to 6 replicate cages containing 30 or 8 birds, which were sampled on d 5 or 21, respectively. For chicks, apparent ileal amino acid digestibility increased with age for all the feed ingredients. After standardization, SIAAD increased with age when chicks were fed DDGS and corn, but not soybean meal or canola meal. For all the feed ingredients evaluated, SIAAD values for both methods of standardization were not different at d 5. However, the HDP method resulted in greater SIAAD values for most of the amino acids in light DDGS, canola meal, and corn (d 21). In poults, the apparent ileal amino acid digestibility values increased with age for all the feed ingredients except the dark DDGS and canola meal. After standardization, there was no effect of age on amino acid digestibility in all the feed ingredients evaluated, with the exception of corn. Within a particular age (d 5 or 21), both methods of standardization resulted in similar amino acid digestibility values except for corn, for which the SIAAD values were greater (NFD method) in 11 amino acids. Results from this study showed that age-appropriate endogenous amino acid standardization is necessary and, in some cases, results in no age-associated differences in amino acid digestibility. With the exception of corn, standardization of amino acid digestibility with ileal endogenous amino acid flow from birds fed an NFD or HDP did not differ for most plant feedstuffs.

Intestinal mucin dynamics: Response of broiler chicks and White Pekin ducklings to dietary threonine

Mucin dynamics may be particularly sensitive to a Thr deficiency due to the high concentration and structural importance of Thr in the mucin protein backbone. Intestinal mucin secretion, expression of mucin gene (MUC2), and histological characteristics were investigated in male broilers and White Pekin ducklings offered diets containing 3.3, 5.8, or 8.2 g of Thr/kg in 4 studies. Seventy-two birds of each species were fed a standard broiler starter diet from 1 to 14 d of age followed by assignment to 3 dietary treatments in a randomized complete block design for a 7-d feeding trial in experiment 1 (broilers) and experiment 2 (ducklings). The dietary treatments consisted of an isonitrogenous corn-soybean meal-based diet with the addition of crystalline amino acids and graded levels of Thr. Dietary treatments contained 3.3, 5.8, or 8.2 g of Thr/kg. Dietary formulation and experimental design for experiments 3 (broilers) and 4 (ducklings) were similar to experiments 1 and 2 except that birds were fed 3.3 or 8.2 g of Thr/kg for durations of 7 or 14 d. For chicks, increased dietary Thr resulted in higher levels of intestinal crude mucin excretion in experiment 1 (P=0.04) but not in experiment 3, whereas intestinal sialic acid excretion increased in experiment 3 (P=0.02) but not in experiment 1. Furthermore, there was no effect of Thr on intestinal goblet cell density or MUC2 mRNA abundance for broilers. For ducklings, there was an increase in intestinal crude mucin excretion in both experiments (P<0.05) as dietary Thr increased, although there was no effect of Thr on intestinal sialic acid excretion. There was a tendency for an increase in intestinal goblet cell density (cells/microm of villus length; P=0.09) as dietary Thr increased in experiment 2. For experiment 4, intestinal MUC2 mRNA abundance increased (P=0.03) as dietary Thr increased for the 14-d feeding trial but not for the 7-d feeding trial. The data establish a link between dietary Thr and intestinal crude mucin dynamics in chicks for experiment 1 and ducklings for both experiments.

Effect of aflatoxin culture on intestinal function and nutrient loss in laying hens

The negative effects of aflatoxins (AFLA) on hepatic necrosis and total tract digestibility of energy, N, and amino acids have been well documented. However, the question remains if this is an effect on nutrient metabolizability alone or an effect on the intestine, or both, resulting in increased endogenous nutrient loss or reduced nutrient retention, or both. Therefore, a 2-wk feeding study with a crude AFLA culture was conducted with laying hens to measure endogenous losses and digestive functionality of the intestine. Hy-Line W36 hens were fed 1 of 4 experimental diets containing a crude AFLA culture from 20 to 22 wk of age. Diets were analyzed to contain 0, 0.6, 1.2, or 2.5 mg/kg of AFLA B(1). Dietary AFLA concentration had no effect on BW, egg production, or feed intake. Intestinal crypt depth (but not villus length) increased linearly with increasing AFLA concentration. Similarly, specific activity of the intestinal disaccharidase, maltase, increased quadratically by feeding up to 1.2 mg/kg of AFLA and declined at 2.5 mg/kg of AFLA (P <or= 0.022). Although there was no effect of AFLA on goblet cell number, density, or crude mucin excretion (P > 0.05), sialic acid excretion increased quadradically such that it was increased 12% when 0.6 and 1.2 mg/kg of AFLA was fed versus the control (P <or= 0.025). Digestibility of DM and N per hen per day were unaffected by AFLA. Feeding of 0.6 and 1.2 mg/kg reduced the apparent digestible and AME(n) of the hen by 10 and 4%, respectively (P <or= 0.025). Because intestinal characteristics (intestinal morphology, sialic acid production, and apparent digestible energy) were altered by AFLA feeding, it can be surmised that AFLA can have a direct or indirect effect, or both, on functionality of the gastrointestinal tract.

Factors affecting endogenous amino acid flow in chickens and the need for consistency in methodology

Accurate estimation of ileal endogenous amino acid (EAA) losses is important when formulating diets on a standardized ileal digestible amino acid basis. In addition to the undigested and unabsorbed amino acids of dietary origin, amino acids of endogenous origin, which can be basal or diet specific, are found in digesta. The improvement in the techniques used in amino acid analysis as well as a shift from sampling excreta to ileal digesta has resulted in more accurate amino acid digestibility coefficients. Despite this, however, it is important to determine the amino acids in the digesta that are of endogenous origin. Although the need for standardization and its associated advantages is still subject to debate, it is important to evaluate how values from various methodologies compare. Several methods have been used to estimate ileal EAA flow. The classical methods, including the regression method, the use of nitrogen-free diet (NFD), and the fasted cecectomized rooster method, are the most widely used. The criticisms with the last 2 methods are that birds are not in a normal physiological state and the ileal EAA flow is, therefore, underestimated. Different methods have resulted in different endogenous flow estimates, with the NFD method having the lowest values when compared with flows from the regression and highly digestible protein methods. In addition to the influence of methods on ileal EAA flows, the influence of the age of the birds on flow is important. Data on EAA losses are copious in the literature; however, variation in data across and within laboratories calls for investigation of factors contributing to the variation. This review compares results from different methods and examines the issue of repeatability and consistency of EAA losses data from different laboratories. Finally, composition of an NFD for estimating EAA losses is proposed.

Dietary L-arginine supplementation attenuates lipopolysaccharide-induced inflammatory response in broiler chickens.

In the present study, two experiments were conducted to investigate the effect of dietary L-arginine (Arg) supplementation on the inflammatory response and innate immunity of broiler chickens. Expt 1 was designed as a 2 × 3 factorial arrangement (n 8 cages/treatment; n 6 birds/cage) with three dietary Arg concentrations (1.05, 1.42 and 1.90%) and two immune treatments (injection of lipopolysaccharide (LPS) or saline) given at an interval of 48 h between 14 and 21 d of age. In Expt 2, correlation between dietary Arg concentration (0.99, 1.39, 1.76, 2.13 or 2.53%) and percentage of circulating B cells (percentage of circulating lymphocytes) was determined. In Expt 1, LPS injection decreased body-weight gain and feed intake and increased feed conversion ratio of the challenged broilers (14-21 d; P< 0.05). LPS injection suppressed (P< 0.05) the percentages of splenic CD11+ and B cells (percentages of splenic lymphocytes) and phagocytic activity of splenic heterophils and macrophages; Arg supplementation linearly decreased the percentages of CD11+, CD14+ and B cells in the spleen (P< 0.10). LPS injection increased (P< 0.05) the expression of IL-1β and IL-6 mRNA in the spleen and caecal tonsils. Arginine supplementation decreased (P< 0.05) the expression of IL-1β, Toll-like receptor 4 (TLR4) and PPAR-γ mRNA in the spleen and IL-1β, IL-10, TLR4 and NF-κB mRNA in the caecal tonsils. In Expt 2, increasing dietary Arg concentrations linearly and quadratically reduced the percentage of circulating B cells (P< 0.01). Collectively, Arg supplementation attenuated the overexpression of pro-inflammatory cytokines probably through the suppression of the TLR4 pathway and CD14+ cell percentage. Furthermore, excessive Arg supplementation (1.76%) suppressed the percentages of circulating and splenic B cells.

The nutritional value of high-protein corn distillers dried grains for broiler chickens and its effect on nutrient excretion

Two experiments were conducted with a co-product of corn endosperm fermentation. The first experiment determined nutrient digestibility of high-protein corn distillers dried grains (HP-DDG; 54% CP) after feeding semipurified diets from 15 to 22 d of age. The AMEn of HP-DDG was 2,526 kcal/kg, whereas standardized ileal Lys, Met, and Thr digestibilities were 73.0, 84.9, and 73.0%, respectively. In a second experiment, an industry control diet (I) regimen was compared with that of either an approximate 25 or 50% replacement for the level of 48% CP soybean meal (SBM) inclusion in the diet utilizing the amino acid digestibility and AMEn determined from the first experiment. From 0 to 14, 14 to 28, and 28 to 42 d of age, the HP-DDG in the 50% SBM replacement diet was added at 25, 23.5, and 21% of the diet, respectively. To meet digestible amino acid needs, the diet containing 50% SBM replacement with HP-DDG contained 3.2, 3.6, and 4.4% units more CP than the I diet regimen from 1 to 14, 14 to 28, and 28 to 42 d of age, respectively. Dietary replacement of up to 50% of SBM inclusion with HP-DDG had no effect on bird performance at 14 or 42 d of age or breast fillet yield at 42 d of age; however, it decreased BW gain and increased feed:gain ratio from 14 to 28 d of age. Birds consuming a diet with 50% replacement of SBM with HP-DDG consumed 17.1% more N compared with those consuming I diets. This additional N and fiber consumed resulted in birds being fed the 50% replacement for SBM diet excreting 21.9 and 31.8% more manure DM and N, respectively. Due in large part to the amino acid profile and digestibility of HP-DDG, use of this feedstuff as a large proportion of the diet is feasible but results in more manure and manure N from broilers.

Prevalence and effects of mycotoxins on poultry health and performance, and recent development in mycotoxin counteracting strategies

Extensive research over the last couple of decades has made it obvious that mycotoxins are commonly prevalent in majority of feed ingredients. A worldwide mycotoxin survey in 2013 revealed 81% of around 3,000 grain and feed samples analyzed had at least 1 mycotoxin, which was higher than the 10-year average (from 2004 to 2013) of 76% in a total of 25,944 samples. The considerable increase in the number of positive samples in 2013 may be due to the improvements in detection methods and their sensitivity. The recently developed liquid chromatography coupled to (tandem) mass spectrometry allows the inclusion of a high number of analytes and is the most selective, sensitive, and accurate of all the mycotoxin analytical methods. Mycotoxins can affect the animals either individually or additively in the presence of more than 1 mycotoxin, and may affect various organs such as gastrointestinal tract, liver, and immune system, essentially resulting in reduced productivity of the birds and mortality in extreme cases. While the use of mycotoxin binding agents has been a commonly used counteracting strategy, considering the great diversity in the chemical structures of mycotoxins, it is very obvious that there is no single method that can be used to deactivate mycotoxins in feed. Therefore, different strategies have to be combined in order to specifically target individual mycotoxins without impacting the quality of feed. Enzymatic or microbial detoxification, referred to as “biotransformation” or “biodetoxification,” utilizes microorganisms or purified enzymes thereof to catabolize the entire mycotoxin or transform or cleave it to less or non-toxic compounds. However, the awareness on the prevalence of mycotoxins, available modern techniques to analyze them, the effects of mycotoxicoses, and the recent developments in the ways to safely eliminate the mycotoxins from the feed are very minimal among the producers. This symposium review paper comprehensively discusses the above mentioned aspects.

The use of distillers dried grains plus solubles as a feed ingredient on air emissions and performance from laying hens

The objectives of the current study were to evaluate the effect of feeding diets containing 0, 10, or 20% distillers dried grains plus solubles (DDGS) to laying hens (21 to 26 wk of age) on emissions of NH3 and H2S. Hy-Line W-36 hens (n = 640) were allocated randomly to 8 environmental rooms for a 5-wk period (hens in 3 rooms were offered the 10% and 20% DDGS diets each; hens in 2 rooms were offered the 0% DDGS diet). Diets were formulated to contain similar CP levels (18.3%), nonphytate P (0.46%), and Ca (4.2%). On an analyzed basis, the 0, 10, and 20% DDGS diets contained 0.22, 0.27, and 0.42% S. Egg weight (50.9 g), egg production (85%), and feed intake (87.9 g/hen per d) were unaffected by diet (P > 0.05) over the study period. Daily NH3 emissions from hens fed diets containing 0, 10, and 20% DDGS were 105.4, 91.7, and 80.2 mg/g of N consumed, respectively (P < 0.05). Daily H2S emissions from hens fed commercial diets containing 0, 10, and 20% DDGS were 2.6, 2.4, and 1.1 mg/g of S consumed, respectively. Overall, feeding 21- to 26-wk-old laying hens diets containing 20% DDGS decreased daily NH3 emissions by 24% and H2S emissions by 58%. Each hen emitted approximately 280 mg of NH3 and 0.5 mg of H2S daily when fed a control diet containing 18% CP and 0.2% S. The results of this study demonstrate that 20% DDGS derived from ethanol production can be fed to laying hens, resulting in lower emissions of NH3 and H2S with no apparent adverse effects on hen performance.