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.     

Yellow mealworm,Tenebrio molitor (Col: Tenebrionidae), larvae powder as dietary protein sources for broiler chickens: Effects on growth performance,carcass traits,selected intestinal microbiota and blood parameters

This study was conducted to evaluate the effects of dietary Tenebrio molitor larvae meal (TM meal) inclusion on the performance, carcass traits, caecum microbiota and blood parameters of broiler chickens. A total of 180-day-old chickens were allocated to three dietary groups with five replicate pens (12 birds/pen). Experimental diets were included a corn-soybean basal diet, and two diets with 2.5% and 5% TM. TM-included diets have been administered for periods of starter (1–10 days) and grower (11–25 days). A regular finisher diet was given to all groups during the 26–42 days of age. Diets containing TM meal did not influence feed intake and mortality percentage. Feed conversion ratio was lower with the 2.5% TM meal diet compared with the control group at starter period (p < .05). Besides, broiler chickens fed TM meal included diet had a higher body weight gain than the control group during 1–10 days of age (p < .05). The carcass characteristics and length and weight of different parts of small and large intestines were not influenced by diets. TM meal inclusion decreased the albumin-to-globulin ratio compared with the control group (p < .05). Additionally, lower total count of aerobic and Escherichia coli bacteria was observed in response to diet inclusion of 5% TM meal. Overall, the inclusion of TM meal in broilers' diet improved body weight gain and feed conversion ratio in the starting period, decreased albumin-to-globulin ratio and E. coli bacteria content.     

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。     

Effect of herbal feed additives on performance parameters,intestinal microbiota,intestinal morphology and meat lipid oxidation of broiler chickens

ABSTRACT

1. This feeding trial investigated the effects of herbal feed additives on performance of broiler chickens, jejunal and caecal microbiota, jejunal morphology, meat chemical composition and oxidative stability during refrigerated storage.

2. In a 42 days trial, 320 one-day-old broiler chickens were randomly allocated to 4 groups with 4 replicate pens each containing 20 chicks. The control group was fed maize–soybean-based diets. The diets of the other three groups were supplemented with herbal feed additives: HRB1 with Stresomix^TM (0.5 g/kg feed); HRB2 with Ayucee^TM (1.0 g/kg feed); HRB3 with Salcochek Pro^TM (1.0 g/kg feed). The GC/MS analysis of the feed additives showed that the major components of HRB1 were β-caryophyllene (14.4%) and menthol (9.8%); HRB2 were n-hexadecanoic acid (14.22%) and β-caryophyllene (14.4%); and HRB3 were menthol (69.6%) and clavicol methyl ether (13.9%).

3. Intestinal samples were taken at 42 day to determine bacterial populations (total aerobe counts, Lactobacilli, and Escherichia coli) and perform gut morphology analysis. Meat samples were analysed for chemical composition and oxidative stability under storage.

4. The HRB1 group had improved (P < 0.05) body weight gain and tended to have improved (0.05 ≤ P < 0.10) feed conversion ratio, compared to the control group. Jejunum lactic acid bacteria counts were increased (P < 0.001) in groups HRB1 and HRB3, compared to the control group, whereas caecal lactic acid bacteria counts tended to increase (0.05 ≤ P < 0.10) in group HRB1, compared to the control group. Breast meat fat content tended to be lower (0.05 ≤ P < 0.10) in group HRB1. Meat oxidative stability was improved (P < 0.001), and jejunum villus height, crypt depth and goblet cells numbers were increased (P < 0.001) in all three herbal supplemented groups, compared to the control.

5. In conclusion, herbal feed additives may be able to improve both growth performance and antioxidant activity of broiler chickens, based on their phenolic compound content.     

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.     

The effects of benzoic acid supplementation on the performance of broiler chickens

To determine the efficiency of benzoic acid, a feeding experiment was carried out with 240 one‐day old ROSS 308 cockerels. Birds were divided into three dietary treatments: I (C) – no additives, II (B1) – 0.1% inclusion of benzoic acid, III (B2) – the inclusion of 0.2% benzoic acid. The performance results were similar in birds fed the control diet and the diet with 0.1% of benzoic acid. The dietary inclusion of benzoic acid at 0.2% depressed the growth of broiler chickens (p < 0.05). The dry matter of the digesta increased in the crop and caeca after benzoic acid supplementation. The pH of the caecal contents decreased following benzoic acid supplementation and was the lowest in the B2 group. No differences were found in the pH of the crop, ileal, gizzard digesta and rectum content. Lactic acid bacteria populations were the lowest in the caeca of the B1 group (p < 0.05). Coliform bacteria decreased in the caeca contents following increased benzoic acid supplementation.     

Performance and physiological responses of broiler chickens to supplemental guanidinoacetic acid in arginine-deficient diets

1. The present experiment was designed to examine arginine (Arg)-sparing effects of guanidinoacetic acid (GAA) on production performance, intestinal morphology and certain blood parameters in broiler chickens.

2. A total of 300 male broiler chicks (Ross 308) were randomly allotted to 5 dietary treatments in a completely randomised design during 1–15 and 15–35 d rearing periods. Experimental treatments consisted of a basal diet with a reduction in Arg content (CON?; starter: 12.1 g/kg and grower: 11.3 g/kg Arg), a CON? diet supplemented with synthetic Arg equal to the required Arg level (CON+; starter: 13.80 g/kg; grower: 12.37 g/kg) and CON? diet supplemented with three levels of GAA (GAA0.6; GAA1.2; and GAA1.8; g/kg).

3. Supplemental GAA at 0.6 and 1.2 g/kg reduced the adverse effects of the CON? diet on weight gain and feed conversion ratio in the starter period, equal to the effect of CON+ diet. During the entire rearing period, such an effect was only seen for broilers fed CON? diet supplemented with 1.2 and 1.8 g/kg GAA (P < 0.05). Proportional weights of liver and abdominal fat were decreased in birds fed diets containing supplemental Arg and 1.8 g/kg GAA compared to the CON? broilers (P < 0.05). Jejunal villus height and villus height to crypt depth ratio were increased in broilers receiving the CON? diet (P < 0.05). Serum concentration of nitric oxide increased when the CON? diet was supplemented with either Arg or 1.8 g/kg GAA (P < 0.05).

4. Dietary inclusion of GAA had an Arg-sparing effect, whereby 1.2 and 1.8 g/kg of supplemental GAA resulted in greater growth performance during the starter and entire rearing periods, respectively. Supplementation with 1.8 g/kg GAA had significant effects on some carcass and physiological parameters.     

Effects of dietary oregano essential oil supplementation on growth performance,intestinal antioxidative capacity,immunity, and intestinal microbiota in yellow-feathered chickens

Essential oils are plant-derived aromatic volatile oils, and they contain bioactive compounds that have been shown to improve poultry nutrition. In this study, we investigated the effects of oregano essential oil (OEO) on intestinal antioxidative capacity, immunity, and gut microbiota of young yellow-feathered chickens. A total of nine hundred and sixty 1-d-old female Qingyuan partridge chickens were randomly allocated to four treatment groups with six replicates of 40 birds each, and the feeding trial was lasted for 30 d. The controls were fed on a basal diet without in-feed antibiotics; the birds in the antibiotic group were fed the basal diet supplemented with 20 mg/kg virginiamycin; the remaining birds were fed the basal diet containing 150 or 300 mg/kg OEO, respectively. Dietary supplementation with 150 or 300 mg/kg OEO increased average daily feed intake (P = 0.057) and average daily gain (P < 0.05). The activities of glutathione peroxidase and total antioxidative capacity in plasma, jejuna, and ileal mucosa were increased by OEO supplementation (P < 0.05), with a trend of lower jejunal content of malonaldehyde (P = 0.062). Moreover, dietary OEO increased the content of secretory immunoglobulin A (P = 0.078) and the relative expression of Claudin 1, Mucin 2, and Avain beta-defensin 1 in ileum (P < 0.05). Sequencing data of 16S rRNA indicated that dietary OEO increased the relative abundance of Firmicutes phylum, and Clostridium and Lactobacillus genera, and decreasing that of Romboutsia. Functional analyses indicated that microbial amino sugar and nucleotide sugar metabolism, replication, and repair systems were higher in OEO groups than those of controls and antibiotic treatment. In conclusion, dietary supplementation with OEO enhanced growth performance, alleviated local oxidative stress in intestine, improved production of natural antibodies, and favorably modulated intestinal microbiota composition.     

Plantago ovata in broiler chicken nutrition: Performance,carcass criteria,intestinal morphology,immunity, and intestinal bacterial population

In this experiment, the effect of dietary Plantago ovata (PO) on performance, carcass criteria, intestinal morphology, immunity, and intestinal bacterial population of broiler chickens was evaluated. A total of 250 one‐day‐old male broiler chicks (Ross 308) were randomly assigned to five treatments containing 0, 5, 10, 15, or 20 g/kg of PO with five replicate pens and 10 birds in each replicate. Dietary PO increased body weight gain and decreased feed conversion ratio in the finisher period, improving the performance index (p < .05). Dietary treatments had no effects on carcass criteria, but breast meat percentage showed an increasing trend with incremental levels of PO in the diet (p = .069). The length of small intestine, especially jejunum section, as well as the villus height, villus width, villus area, and goblet cell numbers were significantly increased with supplemental PO (p < .05). Humoral and cellular immunity parameters, and oxidation stability of meat were improved due to use of dietary PO (p < .05). Dietary PO decreased the CFU of Escherichia coli, whereas the Lactobacilli population was increased (p = .001). Broken‐line regression revealed that dietary PO at the rate of 10 g/kg may results in the best performance in broiler chickens. This study showed that PO at the level of 10 g/kg could be considered as a beneficial feed additive in broiler diet.     

Performance and intestinal microbiota of chickens receiving probiotic in the feed and submitted to antibiotic therapy

The purpose of this study was to verify the ability of a probiotic in the feed to maintain the stability of the gut microbiota in chickens after antibiotic therapy and its association with growth performance. One thousand six hundred twenty 1‐day‐old Cobb male were housed in floor pens (36 pens, 45 birds/pen) and were fed corn‐/soya bean meal‐based diets supplemented with or without probiotic (Bacillus subtilis) during the entire rearing phase. From 21 to 24 days of age (three consecutive days), the chickens were submitted to antibiotic therapy via drinking water (bacitracin and neomycin) in order to mimic a field treatment and induce dysbiosis. Growth performance was monitored until 42 days of age. At 2, 4 and 6 days after antibiotic therapy, three chickens from each pen were euthanized and the contents of the small intestine and caeca were collected and pooled. The trial was conducted with four treatments and nine replicates in a 2 × 2 factorial arrangement for performance characteristics (with and without probiotic × with and without antibiotic therapy); for the intestinal microbiota, it was in a 2 × 2 × 3 factorial arrangement (with and without probiotic × with and without antibiotic therapy × 2, 4 and 6 days after the antibiotic therapy) with three replicates per treatment. Terminal restriction length polymorphism (T‐RFLP) analysis showed that the structure of gut bacterial community was shaped by the intestinal segment and by the time after the antibiotic therapy. The number of 16S rDNAs copies in caecum contents decreased with time after the therapeutic treatment. The antibiotic therapy and dietary probiotic supplementation decreased richness and diversity indexes in the caecal contents. The improved performance observed in birds supplemented with probiotic may be related to changes promoted by the feed additive in the structure of the intestinal bacterial communities and phylogenetic groups. Antibiotic therapy modified the bacterial structure, but did not cause loss of broiler performance.     

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.     

Evaluation of the chicory inulin efficacy on ameliorating the intestinal morphology and modulating the intestinal electrophysiological properties in broiler chickens

Chicory (Cichorium intybus) belongs to plants of the Compositae family accumulating energy in the form of inulin fructan. Chicory, a prebiotic, is a fermentable oligosaccharide and oligofructose that may affect the intestinal mucosal architecture and the electrophysiological parameters. Therefore, this study was conducted to evaluate the effectiveness of adding chicory fructans in feed on the intestinal morphology and electrogenic transport of glucose in broilers. Four hundred, 1 day old broiler chicks were randomly divided into two groups (200 bird per group) for 5 weeks. The dietary treatments were (i) control, (ii) basal diets supplemented with the dried, grinded ground chicory pulp containing inulin (1 kg of chicory/ton of the starter and grower diets). In duodenum, dietary chicory increased the villus height and villus width and villus height to crypt depth ratio (p < 0.05), but the duodenal crypt depth remained unaffected (p > 0.05). However, in jejunum, the villus height, crypt depth and villus height to crypt depth ratio were decreased by dietary chicory compared with control birds (p < 0.05). In ileum, the villus height and villus crypt depth was decreased by dietary chicory supplementation compared with control (p < 0.05), but, the villus height to crypt depth ratio was increased (p < 0.05). Moreover, dietary chicory relatively affected the electrophysiological parameters of the intestine but did not reach significance. The amount of ΔIsc after d ‐glucose addition to the jejunal mucosa was numerically higher for chicory fed birds (19 μA/cm²) than control birds (10 μA/cm²). The percentage of increase in the Isc after d ‐glucose addition (ΔIsc %) was higher for chicory group upto (90%) of the control group. In colon, the actual Isc value and Isc after d ‐glucose addition was numerically higher for chicory fed birds than control birds (p > 0.05). Moreover, the conductance of jejunal and colonic tissues after d ‐glucose addition remained unaffected by the dietary chicory. In conclusion, addition of chicory to broilers diet increased the duodenal villus height, villus width and villus height to crypt depth ratio and decreased the villus height and crypt depth in both jejenum and ileum. Furthermore, dietary chicory relatively modified the small intestinal electrogenic transport of glucose in broilers.     

Effects of soybean oligosaccharide,stachyose, and raffinose on growth performance and cecal microbiota in broiler chickens

This study was conducted to examine the effects of soybean oligosaccharide, stachyose, and raffinose on growth performance and cecal microbiota in broiler chickens. Three-hundred 1-day-old Arbor Acres broiler chickens were randomly assigned to five dietary treatments: a basal diet (control diet); diets supplemented with soybean oligosaccharide, stachyose, or raffinose at levels of 0.6% total sugar content; and diet with soybean meals (positive control). Results showed that feed-to-gain ratio (F/G) was improved (p < 0.05) in broiler chickens that received stachyose, but not raffinose, while addition of soybean oligosaccharide to the diet significantly (p < 0.05) resulted in decreases in the average daily feed intake (ADFI) and average daily gain (ADG). Supplementation with soybean oligosaccharide, stachyose, or raffinose had a positive effect (p < 0.05) on the nutrient digestibility, but not on the blood immune parameters (p > 0.05). The results of the gene sequencing indicated that, at the family level, Ruminococcaceae, Lachnospiraceae, and Lactobacillaceae were comparatively present in all the treatments but the cecal microbial composition was changed after dietary addition of different oligosaccharides. Our study had led to a greater understanding of prebiotic effects of these oligosaccharides on growth and intestinal health.     

A deficient or an excess of dietary threonine level affects intestinal mucosal integrity and barrier function in broiler chickens

The aim of this study was to investigate the effects of deficient or excess of dietary threonine (Thr) levels on intestinal integrity and barrier function of broilers. A total of 432 1‐day‐old commercial broilers (Arbor Acre) were assigned to four experiment groups consisting of six replicates of 18 birds. The treatments were designed as follows: 85%, 100%, 125% and 150% of NRC (Nutrient requirements of poultry (9th edn). Washington, DC: The National Academies Press, 1994) recommendations. The results indicated that expressions of jejunal and ileal secretory immunoglobulin A (sIgA) mRNA were increased linearly or quadratically by increasing Thr (p < .05), and the highest sIgA mRNA abundance was obtained in 125% Thr level. Likewise, the intestinal sIgA content showed similar increasing trend with the intestinal sIgA gene expression in this instance. The high level of Thr inclusion upregulated mucin 2 (MUC2) mRNA expression in the jejunum and ileum (p < .05). In addition, on day 21, the expression levels of jejunal zonula occludens‐2 (ZO‐2) and ileal zonula occludens‐1 (ZO‐1) decreased then increased with increasing Thr level (p < .05), whereas, the mRNA expressions of occludin in the jejunum and ileum had no significant difference amongst groups (p >.05). On day 42, Thr treatments did not affect the mRNA abundance of measured genes in the jejunum and ileum (p > .05). These findings suggested that Thr might be a nutrient immunomodulator that affects intestinal barrier function, moreover, 125% of the NRC (1994) recommendations Thr level was optimum.     

Effects of oxidative stress induced by high dosage of dietary iron ingested on intestinal damage and caecal microbiota in Chinese Yellow broilers

The objective of this trial was to test the effects of oxidative stress induced by a high dosage of dietary iron on intestinal lesion and the microbiological compositions in caecum in Chinese Yellow broilers. A total of 450 1‐day‐old male chicks were randomly allotted into three groups. Supplemental iron (0, 700 and 1,400 mg/kg) was added to the basal diet resulting in three treatments containing 245, 908 and 1,651 mg/kg Fe (measured value) in diet respectively. Each treatment consisted of six replicate pens with 25 birds per pen. Jejunal enterocyte ultrastructure was observed by transmission electron microscopy. The results showed that a high dosage of dietary iron induced oxidative stress in broilers. Dilated endoplasmic reticulum (ER), autophagosome formation of jejunal enterocytes and decreased villi were caused by this oxidative stress. Compared to the control, concentration of the malondialdehyde (MDA) in jejunal mucosa in the 908 and 1,651 mg/kg Fe groups increased by 180% (p < .01) and 155% respectively (p < .01); activity of copper‐zinc superoxide dismutase (Cu/ZnSOD) increased in jejunum (p < .01); and the concentration of plasma reduced glutathione (GSH) decreased by 34.9% (p < .01) in birds fed 1,651 mg/kg Fe. Gene expression of nuclear factor, erythroid‐derived 2‐like 2 (Nrf2) and zonula occludens‐1 (ZO‐1), in the higher dietary Fe groups was enhanced (p < .05). Species of microbial flora in caecum increased caused by oxidative stress. The PCR‐DGGE (denaturing gradient gel electrophoresis) dendrograms revealed different microbiota (65% similarity coefficient) between the control and iron‐supplemented groups (p < .05). These data suggest high dosage of iron supplement in feed diet can induce oxidative stress in Chinese Yellow broilers, and composition of microbiota in the caecum changed. It implied there should be no addition of excess iron when formulating diets in Chinese Yellow broilers.