Correlations between changes in intestinal microbiota composition and performance parameters in broiler chickens

Growing male Cobb broiler chickens were fed on diets supplemented with additives reported as able to influence intestinal microbiota composition. The diets used were a balanced commercial diet (no additive), inulin (20 g/kg), fructose caramel (FC, 20 g/kg) and the garlic derivative PTS‐O (propyl propane thiosulfonate, 45 and 90 mg/kg diet). The composition of the intestinal microbiota was analysed by qPCR at different points of the intestinal tract, and a number of nutritional parameters were also determined. The relative amounts of bacteroides (bacteroides/total bacteria) in the ileal contents correlated (p < 0.05) positively with faecal NDF, ADF, hemicellulose and cellulose digestibility. The relative amounts of Escherichia–Shigella (Escherichia–Shigella/total bacteria) in the crop contents correlated (p = 0.05) negatively with weight gain of broilers. Faecal N digestibility correlated (p < 0.05) negatively with total bacteria in the ileal contents of chickens. The relative amounts of Escherichia–Shigella (Escherichia–Shigella/total bacteria) in the caecal contents correlated (p = 0.05) negatively with faecal fat digestibility of broilers. Total bacteria in ileal or caecal contents of growing chickens correlated (p < 0.05) negatively with ileal N digestibility. The results here reported suggest that positive or negative correlations can be found between performance parameters and changes in intestinal microbiota composition of growing broiler chickens.     

The efficacy of selected probiotic and prebiotic combinations in reducing Campylobacter colonization in broiler chickens

Campylobacter is one of the leading causes of foodborne illness worldwide and is associated with consumption of poultry products. One potential strategy to reduce Campylobacter colonization in poultry is the use of probiotics. In an attempt to reduce Campylobacter in poultry, we isolated 117 GRAS (generally recognized as safe) bacterial isolates from the ceca of healthy birds and evaluated their efficacy against Campylobacter, using a soft agar overlay technique. Twenty-six isolates (Bacillus and Lactobacillus spp.) with greatest ability to inhibit Campylobacter in vitro, were tested in birds. In these studies, bacterial isolates were administered orally on day-of-hatch and, chicks were challenged with a four-strain mixture of wild type C. jejuni (∼10⁶ CFU/mL) on day 7. Cecal samples were collected on day 14 for Campylobacter enumeration. Out of 26 isolates, 3 isolates demonstrated a reduction in Campylobacter counts (1 to 2 logs). In an attempt to improve the efficacy of these 3 isolates, follow-up studies were conducted with various doses of 2 prebiotics (fructooligosaccharide [FOS] or mannan oligosaccharide [MOS]) in the feed. Campylobacter counts were not reduced for any of the FOS treatments (0.125%, 0.25%, or 0.5%) or the MOS treatments (0.04%, 0.08%, or 0.16%), but were reduced for the combination of isolate 3 with 0.04% MOS in feed. The results of this study indicate that the selection and administration of bacterial isolates in combination with selected prebiotics may reduce enteric Campylobacter counts in preharvest poultry.     

The effect of dietary lysozyme with EDTA on growth performance and intestinal microbiota of broiler chickens in each period of the growth cycle

Lysozyme is a potential replacement for antibiotics in the poultry industry. Two trials were conducted using new or used litter to determine the effect of 100 ppm lysozyme in feed on growth performance and intestinal microbiota of broiler chickens in each period of the growth cycle. The new litter trial and used litter trial were each analyzed as a one-way analysis of variance with length of time in which 100 ppm lysozyme was fed to the birds as the main factor (no lysozyme d zero to 35; positive lysozyme d zero to 4; positive lysozyme d 5 to 14; positive lysozyme d 15 to 24; positive lysozyme d 25 to 35; positive lysozyme d zero to 35; positive virginiamycin d zero to 35). Effects of lysozyme were not detected under clean conditions (P > 0.05). When used litter was provided, growth performance was not improved by lysozyme (P > 0.05). However, feeding lysozyme to birds from d 5 to 14 and throughout the trial reduced the number of E. coli in the ileum compared with feeding virginiamycin (positive lysozyme d 5 to 14 and zero to 35, and virginiamycin d zero to 35: 1.7 × 10⁴ cfu g⁻¹, 1.9 × 10⁴ cfu g⁻¹ and 9.3 × 10⁴ cfu g⁻¹, respectively) to birds (P < 0.05). Dietary lysozyme at 100 ppm can change intestinal microbiota of broiler chickens.     

Effect of a synbiotic on the intestinal microflora of chickens

The current experiment was conducted to evaluate the effect of a synbiotic (Biomin Imbo) on intestinal microflora of Ross broiler chickens. A total of 200 male chickens were randomly divided into 20 groups of 10 birds each, kept in 20 pens (1.5 × 1 m each), and fed for 42 d at different synbiotic levels. Treatments included (1) a basal diet without synbiotics (control), (2) a basal diet with synbiotic levels proposed by the manufacturer, (3) a basal diet with synbiotic levels 25% lower than those proposed by the manufacturer, (4) a basal diet with synbiotic levels 50% higher than those proposed by the manufacturer, and (5) a basal diet with synbiotic levels 25% higher than those proposed by the manufacturer. At the end of the 42-d feeding period, 1 bird was randomly selected from each experimental unit, humanely euthanized, and the cecum was removed to measure the microbial population. The cecal contents were collected in discharge containers for microbial cultures, and counts were conducted after microbial culture. The addition of the synbiotic reduced Escherichia coli and total coliform populations in the intestines of broiler chickens. Conversely, different levels of probiotic increased the numbers of lactobacilli in the intestine of broiler chickens. Concentrations of the synbiotic higher than the recommended levels in the diet increased the lactic acid bacteria population in the gut of broiler chickens.     

Evaluating two multistrain probiotics on growth performance,intestinal morphology,lipid oxidation and ileal microflora in chickens

An experiment was conducted to investigate the supplementation of two commercially available multistrain probiotics as an alternative to antibiotics on growth performance, intestinal morphology, lipid oxidation and ileal microflora in broiler chickens. A total of 280‐day‐old ROSS 308 mixed‐sex broiler chickens with an average initial body weight of 42 ± 0.5 g were randomly divided into four treatments with five replicate cages of 14 birds each cage in a completely randomized design and fed with the following diets for 42 day: (a) control (CON) (antibiotic‐free diet), (b) antibiotic (ANT) (CON + Avilamycin 150 g/ton feed), (c) probiotic A (CON + Protexin^® 150 g/ton feed) and (d) probiotic B (CON + Bio‐Poul^® 200 g/ton feed). The results showed the broilers fed the ANT diet had greater average daily gain than broilers fed the CON diet during day 1–14 (p < 0.05). At day 42, two birds were randomly selected per replicate for evaluation intestinal morphology, lipid oxidation and ileal microflora. birds fed diet supplemented with probiotic A and probiotic B increased villus height and goblet cells numbers in the jejunum and villus height to crypt depth ratio and villus height in the ileum as compared to birds fed CON diet (p < 0.05). The malondialdehyde value was reduced (p < 0.05) in the ANT, probiotic B and probiotic A groups compared with the CON group. The Lactobacillus population was increased and Clostridium spp. population decreased in the ileum of broilers fed diets containing the probiotic B and probiotic A compared with those fed CON diet (p < 0.05). The results from this study indicate that the probiotic A (Protexin^®) and probiotic B (Bio‐Poul^®) used in this trial may serve as alternatives to ANT.     

黑麦作为能量原料对肉鸡细菌移位、肠道黏度及微生物组成的影响

本研究开展了两批试验,旨在评估黑麦作为能量饲料原料对肉鸡细菌移位、小肠黏度和完整性、肠道微生物含量及胫骨钙化的影响。每个试验分别选择40只1日龄肉鸡,随机分为两组,对照组饲喂玉米型日粮,试验组饲喂黑麦型日粮,在10d时,两批试验各组分别选择10只鸡口服2.0mg/mL葡萄糖荧光素,2.5h后检查血清葡萄糖荧光素含量,收集肝脏用于分析细菌移位,十二指肠、回肠和盲肠用于分析微生物含量及肠道黏度,胫骨用于测定强度和分析骨成分。试验结果:两批试验中,黑麦日粮组肉鸡体重均显著低于玉米日粮组(P<0.05),但黑麦日粮组较玉米日粮组显著提高了小肠黏度(P<0.05),同时伴随高水平的血清葡萄糖荧光素和肝脏细菌移位(P<0.05)。试验1和试验2黑麦日粮组较玉米日粮组均显著提高了十二指肠、回肠和盲肠乳酸杆菌含量(P<0.05)。同时,黑麦组日粮肉鸡十二指肠和回肠大肠杆菌含量显著升高(P<0.05),而总厌氧菌含量仅在十二指肠表现为显著升高(P<0.05)。与黑麦日粮组相比,两批试验中,玉米日粮组肉鸡胫骨强度以及灰分、钙和磷含量较均显著提高(P<0.05)。结论:黑麦作为肉鸡日粮能量原料来源增加了鸡肠道黏度和体重,降低了血清葡萄糖荧光素,改变了鸡的微生物群组成和胫骨矿化。     

Microstructure of the small intestine in broiler chickens fed a diet with probiotic or synbiotic supplementation

The aim of the study was to determine the effect of dietary supplementation of a probiotic and a synbiotic on the morphometric parameters of the small intestine of broiler chickens. The experiment was conducted on three hundred sixty, one‐day‐old female Ross 308 chicks, which were randomly selected from 20,000 birds and divided into three treatment groups (n = 120) with ten replicates per treatment. The control group (C) was fed a commercial diet, the probiotic group (PRO) was fed the same diet with an added 1% of the probiotic Lavipan® (Lactococcus lactis, Carnobacterium divergens, Lactobacillus casei, Lactobacillus plantarum and Saccharomyces cerevisiae), and the synbiotic group (SYN) was fed the commercial diet with an added synbiotic: 0.8% of the prebiotic RFO (extracted from lupin seeds) and 1% Lavipan®. According to the manufacturer's data, apart from the typical probiotic action,microorganisms contained in the preparation release anti‐bacterial substances (hydrogen peroxide and bacteriocins) and, therefore, are antagonistic towards pathogenic bacteria present in the gut of animals. Supplementation took place in the first seven days of rearing, and all birds had ad libitum access to water and feed during the 42 days of the experiment. On the last day, all birds were slaughtered and samples from three segments of the small intestine were taken. Villi area, height, width and crypt depth ratios were read using Multiscan software. Synbiotic supplementation increased the BWG of broilers from first to tenth day of rearing, compared to the control group. The PRO group had improved villi morphometric parameters of the duodenum. In the jejunum and ileum, both bioactive substances improved villus width and villus surface area. Crypts were deeper in the small intestine of birds supplemented with bioactive substances, which allows greater renewal of the villi. As expected, the intestinal morphometric parameters of broiler chickens benefited from bioactive substance supplementation.     

Supplemental magnolol improves the antioxidant capacity and intestinal health of broiler chickens

Magnolol is a multifunctional polyphenol rich in Magnolia officinalis. The objective of this study was to investigate the effects of magnolol on growth performance, carcass traits, antioxidant capacity, and gut health of broiler chickens. A total of 240 1-day-old broilers were randomly allocated into five dietary treatments: control (Ctrl); control diet supplemented with 100, 200, or 300 mg/kg of magnolol (M100, M200, and M300); and control diet supplemented with 200 mg/kg of bacitracin zinc (PC). The results showed that magnolol linearly decreased the feed conversion ratio between d 0 and d 14, linearly decreased the amount of malondialdehyde and increased the activity of total superoxide dismutase (T-SOD) in both serum and ileal mucosa on d 42 with increasing magnolol levels (p < 0.05). Moreover, the ileal villus height, the ileal villus height to crypt depth ratio, and the jejunal gene expressions of SOD1, glutathione peroxidase, and Claudin1 were linearly up-regulated with increasing magnolol levels (p < 0.05). The supplementation of magnolol had no effect on carcass traits or cecal short chain fatty acids (p > 0.05). The results indicated that magnolol could be applied in the diet of broiler chickens to benefit their antioxidant capacity and intestinal health.     

Linkage between the intestinal microbiota and residual feed intake in broiler chickens

Background: Intestinal microbiota plays a key role in nutrient digestion and utilization with a profound impact on feed efficiency of livestock animals. However, the intestinal microbes that are critically involved in feed efficiency remain elusive.Methods: To identify intestinal bacteria associated with residual feed intake(RFI) in chickens, male Cobb broiler chicks were individually housed from day 14 to day 35. Individual RFI values were calculated for 56 chickens.Luminal contents were collected from the ileum, cecum, and cloaca of each animal on day 35. Bacterial DNA was isolated and subjected to 16 S rRNA gene sequencing. Intestinal microbiota was classified to the feature level using Deblur and QIIME 2. High and low RFI groups were formed by selecting 15 and 17 chickens with the most extreme RFI values for subsequent LEfSe comparison of the difference in the microbiota. Spearman correlation analysis was further performed to identify correlations between the intestinal microbiota composition and RFI.Results: No significant difference in evenness, richness, and overall diversity of the microbiota in the ileum, cecum,or cloaca was observed between high and low RFI chickens. However, LEfSe analysis revealed a number of bacterial features being differentially enriched in either high or low RFI chickens. Spearman correlation analysis further identified many differentially enriched bacterial features to be significantly correlated with RFI(P 0.05). Importantly,not all short-chain fatty acid(SCFA) producers showed a positive association with RFI. While two novel members of Oscillibacter and Butyricicoccus were more abundant in low-RFI, high-efficiency chickens, several other SCFA producers such as Subdoligranulum variabile and two related Peptostreptococcaceae members were negatively associated with feed efficiency. Moreover, a few closely-related Lachnospiraceae family members showed a positive correlation with feed efficiency, while others of the same family displayed an opposite relationship.Conclusions: Our results highlight the complexity of the intestinal microbiota and a need to differentiate the bacteria to the species, subspecies, and even strain levels in order to reveal their true association with feed efficiency. Identification of RFI-associated bacteria provides important leads to manipulate the intestinal microbiota for improving production efficiency, profitability, and sustainability of poultry production.     

Potential effect of two Bacillus probiotic strains on performance and fecal microbiota of breeding sows and their piglets

The effect of long-term administration of two Bacillus strains was tested on 98 breeding sows and their litters allotted into three treatments: a control group (CON); supplemented with 5 × 10⁸ cfu/kg B. subtilis − 541 (BSU); or with 5 × 10⁸ cfu/kg B. amyloliquefaciens − 516 (BAM). Reproductive and performance variables were recorded over three cycles with 56 dams remaining through the third lactation. Blood and fecal samples were taken longitudinally from 12 sows per treatment on days 8 and 21 of the third lactation and milk samples were taken on day 21. Feces from one piglet per litter was sampled on days 21 and 33 and jejunal gene expression was assessed in two piglets on day 21. Changes in fecal microbiota were assessed by 16S rRNA gene sequencing (Illumina MiSeq) and gene expression by Open-Array technology. Metabolomic responses were analyzed in milk by NMR and Ig-G and Ig-A specific antibodies were determined by ELISA. No significant differences were observed on feed intake, body weight, or fat mobilization of the sows. However, a significant increase in the total number of piglets born was observed in supplemented sows. Although the increase was seen from the first cycle with BAM, improvements were not seen with BSU until the third cycle. BAM also increased the number of born-alive and weaned piglets. NMR analysis showed an impact of BAM on milk composition. No differences were found in milk or blood immunoglobulins. A different structure of the fecal microbiota was found in supplemented sows, with changes across phylum, family, and genus. These changes were greater at day 8, suggesting a relevant role of probiotics establishing a new intestinal balance after labor. Shifts in the microbiota were also seen in the piglets, with a clearer impact post-weaning than in suckling. In this regard, correlations between microbial groups of sows and piglets showed a higher link with weaned (d33) than with suckling pigs (d21), reinforcing the idea of an early maternal carry-over. No changes due to treatment in jejunal gene expression were detected; however, piglet size had a clear impact on different genes. In summary, the addition of both probiotics, and particularly Bacillus amyloliquefaciens, demonstrated potential benefits on the prolificacy of sows. Daily feeding of Bacillus amyloliquefaciens resulted in an increase in the number of weaned piglets. The high correlations between the compositions of the microbiota of sows and their piglets are evidence of maternal imprinting, with effects lasting beyond weaning.     

益生菌制剂调控畜禽肠道健康与消化道微生物互作的免疫机制

益生菌制剂作为饲料添加剂,在维护畜禽机体肠道健康、菌群平衡以及营养生理方面发挥着极其重要的作用。一方面,益生菌制剂能够替代部分抗生素,为实现畜禽养殖过程中的"减抗"目标,开辟了一条优质、安全、绿色的道路;另一方面,益生菌制剂能够调控肠道生理结构和改善肠道屏障完整性,进而缓解畜禽养殖中的"亚健康"状况。本文围绕益生菌制剂的种类与作用,益生菌制剂对畜禽肠道免疫、应激、疾病的调控机制,益生菌制剂的发展前景进行阐述,为开发更多益生菌制剂的产品提供理论支撑。     

Administration of Lactobacillus johnsonii FI9785 to chickens affects colonisation by Campylobacter jejuni and the intestinal microbiota

1. Campylobacter jejuni is the most common bacterial cause of human food-borne gastroenteritis in the world. A major source of human infection is the consumption of contaminated meat, particularly poultry. New control measures to reduce or eliminate this pathogen from the animal gastrointestinal tract are urgently required, and the use of probiotics as competitive exclusion agents is a promising biocontrol measure to reduce C. jejuni in the food chain.

2. In this study, we assessed the potential of Lactobacillus johnsonii FI9785, which has shown efficacy against Clostridium perfringens, to combat C. jejuni. The effect of prophylactic administration of L. johnsonii on the ability of C. jejuni to colonise chickens was determined.

3. Two doses of L. johnsonii given a week apart led to a reduction in C. jejuni colonisation in the caecal contents, but this biocontrol seemed reliant upon a high level of initial colonisation by the probiotic.

4. The microbial composition in the chicken gut was significantly altered by the probiotic treatment, as shown by denaturing gradient gel electrophoresis of 16S rRNA gene amplicons.

5. Together these results demonstrate the potential of this probiotic strain to be tested further as a competitive exclusion agent in poultry against C. jejuni.     

Use of blends of organic acids and oregano extracts in feed and water of broiler chickens to controlSalmonella Enteritidis persistence in the crop and ceca of experimentally infected birds

The aim of the present evaluation was to assess the efficacy of blends of organic acids and oregano extracts supplied from d 1 to 21 and from d 35 to 42 in feed, drinking water, or both, in controllingSalmonella Enteritidis persistence in the crop and ceca of broiler chicken. A total of 105 one-day-old male broiler breeder chicks were randomly distributed into 5 different treatments according to the supply of referred blends andSalmonella Enteritidis challenge.Salmonella Enteritidis was inoculated directly in the crop of birds at the 15 d of age. Treatments were unsupplemented unchallenged, unsupplemented challenged, supplemented through water and challenged, supplemented through feed and challenged, and supplemented through feed and water and challenged. Use of the additives in feed and water and in water alone efficiently controlledSalmonella shedding and reduced cecal persistence. Immune mechanisms involved are proposed.     

Evaluation of bee venom as a novel feed additive in fast-growing broilers

1. The present study was designed to evaluate purified bee venom (BV) as an alternative to antibiotics in broiler chickens. The experimental treatment diets were formulated by adding BV into a maize-soybean meal-based diet to give 0, 10, 50, 100, and 500 μg BV per kg of diet.

2. Dietary BV quadratically improved (P < 0.05) feed conversion ratio and increased body weight gain at 1–21 d as level in diet increased. Higher BV levels lowered relative weight of spleen (linear and quadratic, P < 0.05), bursa of Fabricius (quadratic, P < 0.05), and liver (linear and quadratic, P < 0.05) at 21 d of age. Relative breast meat yields were increased quadratically at 21 d and linearly at 35 d with supplementation levels. Dietary BV increased (linear and quadratic, P < 0.05) lightness (L*) value for meat at 21 d, decreased (linear, P < 0.05) ileal villus height and narrowed (quadratic, P < 0.05) width.

3. Dietary BV inclusion linearly increased the concentration of secretory immunoglobulin A (sIgA) on ileal mucosa at 21 d and decreased (quadratic, P < 0.05) nitric oxide contents in serum samples at 21 d and 35 d. Total short-chain fatty acids (SCFA) in caecal digesta were reduced with increasing venom in diets at 21 d of age. None of the serum parameters except for creatinine was affected by dietary BV.

4. It was concluded that dietary BV exhibited wide range of in vivo biological properties in broiler chickens and could be incorporated into feed to promote growth and animal health.