Effect of dietary canola oil on long-chain omega-3 fatty acid content in broiler hearts.

Young and healthy broilers are susceptible to sudden death syndrome (SDS), which is caused by cardiac arrhythmia. The long-chain 'fish-type' omega-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), have cardioprotective anti-arrhythmic effects in animals and humans. Raising the cardiac level of EPA and DHA in chickens may protect against SDS. However, fish oil as a source of EPA and DHA in poultry feed is costly and introduces undesirable properties to the meat. Whilst omega-3 vegetable oils, such as canola oil, are cheaper and do not have a strong odour, they contain the short-chain fatty acid α-linolenic acid, which requires conversion to EPA and DHA after ingestion. We investigated the capacity for dietary canola oil to elevate cardiac EPA and DHA in broilers. Broilers were fed with diets containing either 3% canola oil or tallow, which is currently used in some commercial feeds. Upon completion of a 42 day feeding trial, canola oil significantly increased EPA and EPA + DHA in heart phospholipids relative to tallow. The elevation in cardiac EPA and EPA + DHA may provide anti-arrhythmic effects and protect against SDS in poultry. This proof-of-concept biochemical study suggests that a larger study to assess the clinical outcome of SDS may be warranted.

Dietary alpha-linolenic acid enhances omega-3 long chain polyunsaturated fatty acid levels in chicken tissues.

The effects of enriching broiler chicken diets with a vegetable source of n-3 fat in the form of alpha-linolenic acid (ALA, 18:3n-3) on the accumulation of n-3 long chain polyunsaturated fatty acids (LCPUFA) in chicken meat were investigated. Sixty unsexed one-day-old broiler chickens (Cobb 500) were randomly allocated to one of six diets (n=10 birds/diet) for 4 weeks. The ALA levels varied from 1 to 8% energy (%en) while the level of the n-6 fatty acid linoleic acid (LA, 18:2n-6) was held to less than 5%en in all diets. At harvest (day 28) the levels of n-3 LCPUFA including eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA) in breast and thigh meat increased in a curvilinear manner as dietary ALA increased, reaching 4- to 9-fold above the levels seen in control birds. In contrast, arachidonic acid (AA) was reduced in response to increasing dietary ALA.

Quantitative analyses of genes associated with mucin synthesis of broiler chickens with induced necrotic enteritis.

Clostridial infection of the intestine can result in necrotic enteritis (NE), compromising production and health of poultry. Mucins play a major role in protecting the intestinal epithelium from infection. The relative roles of different mucins in gut pathology following bacterial challenge are unclear. This study was designed to quantify the expression of mucin and mucin-related genes, using intestinal samples from an NE challenge trial where birds were fed diets with or without in-feed antimicrobials. A method for quantifying mucin gene expression was established using a suite of reference genes to normalize expression data. This method was then used to quantify the expression of 11 candidate genes involved in mucin, inflammatory cytokine, or growth factor biosynthesis (IL-18, KGF, TLR4, TFF2, TNF-α, MUC2, MUC4, MUC5ac, MUC5b, MUC13, and MUC16). The only genes that were differentially expressed in the intestine among treatment groups were MUC2, MUC13, and MUC5ac. Expression of MUC2 and MUC13 was depressed by co-challenge with Eimeria spp. and Clostridium perfringens. Antimicrobial treatment prevented an NE-induced decrease in MUC2 expression but did not affect MUC13. The expression of MUC5ac was elevated in birds challenged with Eimeria spp./C. perfringens compared with unchallenged controls and antimicrobial treatment. Changes to MUC gene expression in challenged birds is most likely a consequence of severe necrosis of the jejunal mucosa.

The effects of lactoferrin on the intestinal environment of broiler chickens.

The influence of in-feed lactoferrin (Lf) on bird production, intestinal microbiota, mucosal immune system and gut microarchitecture was assessed in male Cobb 500 broiler chickens. Birds were given one of four diets from day of hatch: Control (basal diet with no additives), ZnB (basal diet + 50 mg/kg zinc bacitracin), Lf 250 mg/kg (basal diet + 250 mg/kg Lf) and Lf 500 mg/kg (basal diet + 500 mg/kg Lf); n = 24 birds/treatment. An apparent metabolisable energy study was performed between d 25-32. Lf did not affect growth rate or feed conversion in the period 0-21 d of age, nor performance or energy metabolism during the 7 d metabolism experiment which commenced at 25 d of age.The profiles of caecal microbial communities were significantly different in birds given ZnB compared with birds given a diet with no additives, or supplemented with 250 mg/kg Lf. Birds given 250 mg/kg Lf also had a different microbial profile compared with birds given 500 mg/kg Lf. In comparison to control birds, Lf treated birds showed some differences in the T cell proportions in caecal tonsil and spleen. No differences in ileal villus height, crypt depth or goblet cell proportions were observed amongst dietary treatments. Whilst Lf had little effect on the measured parameters, the use of an integrated approach to study the influence of novel feed additives may facilitate a greater understanding of the relationships between nutrition, gut health and bird performance.

Effects of necrotic enteritis challenge on intestinal micro-architecture and mucin profile.

1. This study investigated the effect of Eimeria spp./Clostridium perfringens induced necrotic enteritis and traditional antibiotic preventatives on intestinal micro-architecture and mucin profile. 2. A total of 600 Cobb 500 broiler chickens were randomly assigned to the following three groups: (i) unchallenged, (ii) challenged, and (iii) zinc bacitracin/monensin (ZnB/monensin) (n = 25 chickens/pen, 8 pens/group). The challenged and ZnB/monensin chickens were individually inoculated with Eimeria acervulina, E. maxima and E. tenella and C. perfringens type A (EHE-NE18) at 9 and 15 d post-hatch respectively, to induce necrotic enteritis. 3. The challenge procedure significantly decreased villus height, increased villus width and increased crypt depth in the challenged compared to the unchallenged chickens. Zinc bacitracin and monensin maintained villus-crypt structure similar to that of the unchallenged chickens. 4. Mucin profile was not affected by Eimeria spp./C. perfringens challenge as demonstrated by periodic acid-Schiff and high iron diamine-alcian blue pH 2 x 5 staining. Zinc bacitracin and monensin decreased the number of intestinal mucin-containing goblet cells. 5. Lectin histochemistry showed a trend towards greater Arachis hypogea (PNA) reactivity in unchallenged chickens. 6. In summary, Eimeria spp./C. perfringens challenge disrupted intestinal micro-architecture; however, challenge did not appear to affect intestinal mucin profile. Traditional antibiotics, zinc bacitracin and monensin maintained micro-architecture.

Comparison of alternatives to in-feed antimicrobials for the prevention of clinical necrotic enteritis.

AIMS: The capacity for Lactobacillus johnsonii and an organic acid (OA) blend to prevent Clostridium perfringens-induced clinical necrotic enteritis (NE) in chickens was studied. METHODS AND RESULTS: Cobb 500 birds were allocated into six groups (n = 25 birds/pen, eight pens/treatment); Unchallenged, Challenged, Antimicrobial (zinc bacitracin (ZnB)/monensin), OA, probiotic Lact. johnsonii and probiotic sham (Phosphate-buffered saline). All birds were challenged with Eimeria spp. and Cl. perfringens except for unchallenged controls. Birds fed antimicrobials were protected from NE development as indicated by maintenance of body weight, low mortality and clostridium levels, and decreased intestinal macroscopic lesion scores compared to challenged controls (P < 0.05). Lactobacillus johnsonii-fed birds had reduced lesion scores, whilst OA-fed birds had decreased Cl. perfringens levels. Both Lact. johnsonii and OA-fed birds had improved feed efficiency between days 0 and 28 compared to challenged controls; however, mortality and body weights were not improved by either treatment. Microbial profiling indicated that the challenge procedure significantly altered the jejunal microbiota. The microbiota of antimicrobial-fed birds was significantly different from all other groups. CONCLUSIONS: Whilst Lact. johnsonii and OA altered specific intestinal parameters, significant protection against NE was not observed. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactobacillus johnsonii and OA did not prevent NE; however, some improvements were evident. Other related treatments, or combinations of these two treatments, may provide greater protection.

Dietary omega-3 polyunsaturated fatty acid does not influence the intestinal microbial communities of broiler chickens.

The capacity for n-3 polyunsaturated fatty acids (PUFA) to improve broiler chicken growth, influence the intestinal microbial communities, and modify the PUFA content of meat was studied. Male Cobb 500 chickens were fed 1 of 4 diets from hatch: control (standard diet with no additives), ZnB (standard diet with added antibiotics), 2% SALmate (standard diet with 2% SALmate, which is composed of 42% fish oil and 58% starch), and 5% SALmate (standard diet with 5% SALmate). A 7-d energy metabolism study was conducted between d 15 and 22 posthatch. Birds were killed at d 25 and intestinal samples were collected to assess microbial communities by terminal restriction fragment length polymorphism and Lactobacillus PCR-denaturing gradient gel electrophoresis. Diet did not affect BW, feed intake, feed conversion, or ileal digestible energy (P > 0.05). Apparent ME was greater in ZnB-fed birds compared with all other diets (P < 0.05). Breast tissue levels of eicosapentaenoic acid, docosahexaenoic acid, docosapentaenoic acid, and total n-3 PUFA were elevated significantly in 2% SALmate- and 5% SALmate-fed chickens compared with control and ZnB diets (P < 0.05). No significant differences in overall microbial communities were observed in the ileum or cecum as assessed by terminal RFLP (P > 0.05). Birds fed 2% SALmate had a significantly different cecal Lactobacillus species profile compared with birds fed the control diet (P < 0.05); however, no differences were observed in birds fed 5% SALmate compared with birds fed all other diets. In addition to the expected increase in breast tissue n-3 fatty acid levels, a low level of dietary n-3 PUFA also altered the intestinal Lactobacillus species profiles. However, n-3 PUFA supplementation did not alter the overall microbial communities or broiler performance.

Indigestible carbohydrates alter the intestinal microbiota but do not influence the performance of broiler chickens.

AIMS: Prebiotics are a potential alternative to in-feed antimicrobials to improve performance of chickens. We investigated the effects of mannanoligosaccharide (MOS) and fructooligosaccharide (FOS) on growth, performance and the intestinal microbiota. METHODS AND RESULTS: Cobb 500 birds were fed either: Control, starter diet without antimicrobials; ZnB, Control + 50 ppm zinc bacitracin; MOS, Control + 5 g kg(-1) MOS; or FOS, Control + 5 g kg(-1) FOS. An energy metabolism study was conducted and intestinal microbial communities assessed by T-RFLP and Lac PCR-DGGE. Diet did not influence performance. Ileal microbial communities were significantly different in ZnB-fed birds compared to all diets, and FOS-fed chickens compared to Control. MOS-fed chickens had a different caecal profile to ZnB and FOS-fed birds. Consensus Lac PCR-DGGE profiles indicated Lactobacillus communities clustered according to diet with Lactobacillus johnsonii characteristic of ZnB diet. Control and MOS-fed chickens displayed significantly different jejunal Lactobacillus profiles to each other whilst ileal profiles were different between MOS and FOS-fed birds. CONCLUSION: Prebiotics influenced the intestinal microbiota, but did not affect performance. SIGNIFICANCE AND IMPACT OF THE STUDY: In light of pressure for in-feed antimicrobial withdrawal, the impact of alternative compounds on the intestinal microbiota and bird performance is critical to the poultry industry.