Kitasamycin (KM), a broad-spectrum macrolide antibiotic, has implications for growth performance and residue in animals and humans. This study aimed to explore the effects of different KM doses on intramuscular fat accumulation, cecal microflora, and short-chain fatty acids (SCFAs) using a growing-finishing pig model. Forty-two pigs were divided into three groups: control, subtherapeutic KM (50 mg/kg, KM50), and therapeutic KM (200 mg/kg, KM200) diets over 8 weeks. KM50 led to increased back fat thickness, fat content in the longissimus dorsi muscle (LM), and elevated plasma total cholesterol (TC) levels (p < 0.05), supported by upregulated lipid synthesis gene expression (Acc1, Fas, Scd1) (p < 0.05) in the LM. KM50 altered cecal microflora, reducing Lactobacillus spp. and Bifidobacterium spp. abundance, while increasing SCFA concentrations (acetic acid, propionic acid, total SCFAs) (p < 0.05). KM200 had minimal effects on intestinal weight and density, with increased apparent digestibility of nutrients. These findings highlight the dose-dependent impact of KM on intramuscular fat deposition. Subtherapeutic KM induced ectopic fat deposition, emphasizing potential risks in disease treatment for humans and animals.

This trial aimed to determine the effects of tryptophan (Trp) on the rectal temperature, hormone, humoral immunity, and cecal microflora composition in broiler chickens under heat stress (HS). One hundred and eighty 18 days-old female Arbor Acres broilers were randomly divided into three treatment groups, with six replicates of ten birds in each replicate. The broilers were either raised under thermoneutral conditions (TN, 23 ± 1°C) or subjected to heat stress (34 ± 1°C for 8 h daily). The TN group received a basal diet, and another two heat-stressed groups were fed the basal diet (HS) or the basal diet supplemented with 0.18% Trp (HS + 0.18% Trp) for 21 consecutive days. The basal diet contained 0.18% Trp. Results revealed that HS increased the rectal temperature, serum epinephrine (EPI), and corticotropin-releasing hormone (CRH) concentrations (p < 0.05), reduced the bursal index, the levels of serum immunoglobulin A (IgA), IgG, IgM, and serotonin (5-HT) as well as the relative abundance of Actinobacteria in cecum (p < 0.05) compared with the TN group. Dietary supplementation of Trp decreased the rectal temperature, serum dopamine (DA), EPI, and the levels of CRH and L-kynurenine (p < 0.05), increased the bursal index, the levels of serum IgA, IgM, and 5-HT as well as the relative abundance of Ruminococcus torques group in cecum of heat-stressed broilers (p < 0.05) compared to HS group. In conclusion, dietary Trp supplementation decreased rectal temperature, improved cecal microbiota community and Trp metabolism, and enhanced humoral immunity of heat-stressed broilers.

The primary goal of this research was to elucidate the novel influence of Brachiaria decumbens supplementation on broiler chicken growth performance, nutritional digestibility, cecal microbiota, intestinal histomorphology, carcass characteristics, and meat quality. A total of 300 male day-old Ross 308 broiler chickens were randomly subjected to six different treatment groups having five replicates per treatment with 10 birds in each replicate. In treatment 1, broiler chickens were fed commercial diets with no added additives; in treatment 2, broiler chickens were offered commercial diets containing 100 mg/kg of the antibiotic oxytetracycline. However, in treatments 3, 4, 5, and 6, broiler chickens received similar commercial diets supplemented with 25 mg/kg, 50 mg/kg, 75 mg/kg, and 100 mg/kg of B. decumbens ground leaf powder, respectively, without antibiotics. Throughout the 42-day trial, the body weight gain and total feed intake for each replicate were recorded every week to determine the growth performance. Then, on 21th and 42nd day, ten broilers from each treatment (two in each replicate) were randomly selected and slaughtered to assess the digestibility of nutrients, histomorphology of the small intestine, the population of the cecal microbiota, carcass characteristics, as well as quality of both breast and drumstick muscle. There were differences (p < 0.05) in the growth performance, apparent ileal nutrient digestibility, intestinal histomorphology, carcass characteristics, and meat quality. Animals supplemented with 25 mg/kg of B. decumbens had higher growth performance and better ileal nutrient digestibility of protein, fiber, and ether extract, as well as higher villi height and carcass percentage with superior meat quality. Besides, the growth of E. faecalis during the starter phase was inhibited. In summary, B. decumbens supplementation at 25 mg/kg may be suggested as an optimal dose of as a prophylactic as well as an alternative for antibiotic growth promoter in stimulating the productivity of commercial broilers. This unconventional phytobiotic supplementation could be the key to replacing unnecessary antibiotics used in poultry feed.

Lipopolysaccharides (LPS) induces liver inflammatory response by activating the TLR4/NF-κB signaling pathway. Antrodia cinnamomea polysaccharide (ACP) is a medicinal mushroom that can protect from intoxication, liver injury, and inflammation. Nevertheless, the effect of ACP on the liver antioxidant, anti-inflammatory capacity and cecal flora structure of LPS-challenged broilers remains unclear. The aim of this experiment was to investigate the effects of ACP on the anti-oxidative and anti-inflammatory capacities of the liver, and cecal microbiota in slow-growing broilers stimulated by LPS. A total of 750 slow-growing broilers (9-day-old) were assigned to five treatments with 6 replicates of 25 chicks per replicate: a control diet, the chicks were fed a control diet and challenged with LPS. Dietary treatments 3 to 5 were the control diet supplemented with 100, 200, 400 mg/kg ACP challenged with LPS, respectively. The groups of 100 mg/kg ACP supplementation significantly increased liver index, pancreas index, and bursa of Fabricius index (P < 0.05). The GSH-Px content of LPS-challenged broilers was lower than that of the control group (P < 0.001), but the content of MDA increased (P < 0.001). Feeding with 100 mg/kg ACP resulted in increased the activity of T-AOC, GSH-Px, and T-SOD, and decreased MDA content (P < 0.05). The activity of TNF-α, IL-1β, and IL-6 of the LPS group increased, but these indicators were decreased with supplemental 100 mg/kg ACP (P < 0.05). Dietary application of ACP up to 100 mg/kg down-regulated (P < 0.05) the expression of TLR4/NF-κB pathway in the liver induced by LPS. The results of 16S rRNA demonstrated that feeding with 100 mg/kg ACP can change the diversity and composition of the gut microbiota, and restrained the decline of beneficial cecal microbiota (typically Lactobacillus, Faecalibacterium, and Christensenellaceae R-7 group) in the challenged LPS group (P < 0.05). Conclusively, feeding a diet with 100 mg/kg ACP may have beneficial effects on liver damage and the bacterial microbiota diversity and composition in the ceca of LPS-stressed slow-growing broiler breeds, probably because of its combined favorable effects on antioxidants and cytokines contents, and restoration the decline of beneficial cecal microbiota.

Improving the nutritional quality of unconventional feed ingredients such as fava bean by-products can enhance their utilization by broiler chickens. Hence, the quality of fermented fava bean by-products (FFB), in addition to growth, nutrient digestibility, digestive enzyme, and intestinal barrier-related gene expression, and serum biochemical and immunological parameters were evaluated in response to different levels of FFB. A total of 500 1-day-old broiler chicks (46.00 ± 0.388 g) were allocated to five groups with 10 replicates each (100 chicks per treatment). The first group was fed a corn-soybean diet (control diet), and the other four groups were fed a diet containing 5, 15, 25, and 35% FFB for 38 days. Birds fed 25% FFB exhibited maximum body weight gain (increase by 12.5%, compared with the control group) and the most improved feed conversion ratio. Additionally, birds fed FFB at 15, 25, and 35% showed improved dry matter and crude protein digestibility. Moreover, birds fed FFB at 25 and 35% exhibited a decrease in ileal pH and an increase in fiber digestibility (p < 0.05). Upregulation of digestive enzyme genes (AMY2A, PNLIP, and CCK) was observed in groups fed with FFB. The most prominent upregulation of genes encoding tight junction proteins (claudin-1, occludin, and junctional adhesion molecules) in the duodenum was observed in chicks fed 25 and 35% FFB (increase of 0.66-, 0.31-, and 1.06-fold and 0.74-, 0.44-, and 0.92-fold, respectively). Additionally, the highest expression level of enterocyte protective genes [glucagon-like peptide (GLP-2), mucin-2 (MUC-2), and fatty acid-binding protein (FABP-6)] was detected in duodenum of chicks fed high levels of FFB. Substitution of corn-soybean diet with FFB had an inhibitory effect on cecal pathogenic microbes (Escherichia coli and Clostridium perfringens) and increased beneficial microflora (Lactobacilli and Bifidobacterium), especially at high levels. Additionally, an increase was observed in IgM and lysozyme activity, with no effect on IgA in all groups fed FFB. All levels of FFB decreased cholesterol levels. Based on our results, we concluded that substitution of corn-soybean diet with FFB can improve the growth rate and nutrient digestibility of broiler chickens, enhance their intestinal barrier functions, and increase the number of beneficial microorganisms. Using FFB at 25% had a positive effect on the growth performance of broiler chickens, and it could be utilized in poultry farms.

Grape seed is rich in vitamin E, flavonoids, and proanthocyanidins and has the potential to be used as an antibiotic substitute in broilers. We investigated the effects of grape seed proanthocyanidin extract (GSPE) on growth performance, immune responses, cecal microflora, and serum metabolism in early stage broilers. Data indicated that GSPE improved broiler growth performance by strengthening antioxidant capacity, enhancing immune responses, and increasing cecal short chain fatty acids. 16S rRNA sequencing indicated that GSPE changed the predominant cecal microflora and induced the metabolism of amino acids, lipids, and carbohydrates. An UPLC-Q-TOF/MS-based metabolomics analysis identified 23 serum metabolites (mainly related to lipid, amino acid, and alkaloid) were extremely changed by GSPE treatment. The correlations between the changes of cecal microflora and serum metabolites in birds fed with GSPE were analyzed. Hence, GSPE potentially provides active ingredients that may be used as antibiotic substitute and reduces environmental pollution by grape by-products.

The objective of this study was to examine the effects of chronic cyclic heat stress (HS) on the intestinal morphology, oxidative stress and cecal bacterial communities of broilers. One-day-old Arbor Acres (AA) male broilers (n = 100) were acclimated for 3 weeks and then randomly allocated into two groups, normal control (NC) group (22 ± 1 °C, 24 h/day) and HS group (32 ± 1 °C, 10 h/day lasted for 2 weeks). At 35 d of age, intestinal segments (duodenum, jejunum and ileum) and cecal digesta were collected for detection. HS affected intestinal morphology, inducing epithelial cell abscission, inflammatory cell infiltration, and lamina propria edema. Compared with the NC group, HS significantly decreased (P < 0.01) villus height (VH) and the VH-to-crypt depth (CD) ratio (VCR), increased (P < 0.05) CD in the duodenum and ileum, but had no effect on the VH in the jejunum. Moreover, HS induced oxidative stress with antioxidant enzymes activity decreasing (P < 0.05) while malondialdehyde (MDA) content increasing (P < 0.05) in small intestine. Pearson\'s correlation analysis indicated that MDA content was negatively correlated with VH (P < 0.05). The result of 16S rRNA sequencing showed that HS exposure impacted cecal microbiota alpha diversity (phylogenetic diversity whole-tree index) and beta diversity. Based on principal coordinate analysis (PCoA) plots for weighted UniFrac metrics and unweighted pair group method with arithmetic mean (UPGMA), there were 8 discriminative features at the genus level (linear discriminant analysis score > 2). Parabacteroides, Saccharimonas, Romboutsia and Weissella were reduced, while Anaerofustis, Pseudonocardia, Rikenella and Tyzzerella were enriched in heat-stressed broilers. Collectively, these results indicated that chronic cyclic HS induced oxidative stress that caused damage to intestinal villus-crypt structures, and then altered the cecal microflora profile.

The aim of this study was to investigate the effects of tea tree oil (TTO) supplementation on the growth performance, cecal microflora composition, immunity, and antioxidant status of Partridge Shank chickens. A total of 144 one-day-old chicks were allocated into three treatments with six replicates of eight chicks each and fed with a basal diet supplemented with 0 (Control group), 500, and 1000 mg/kg TTO for 50 days. Compared with the control group, the broilers fed with the basal diet supplemented with 1000 mg/kg TTO exhibited an increase in average daily gain from 22 to 50 days (P=0.035) and in both relative thymus weight (P<0.001) and Lactobacillus colonies in the cecal contents (P=0.045) at 50 days of age, but a reduction in the feed/gain ratio during 1 to 50 days (P=0.048). Additionally, dietary TTO supplementation, irrespective of dosage, increased the relative spleen weight (P=0.003) and total antioxidant capacity in the jejunum (P=0.049) and ileum (P=0.001) at 21 days, but decreased the malondialdehyde content in the ileum at both 21 (P=0.003) and 50 days (P<0.001) and in the jejunum at 50 days (P=0.012). The results suggested that TTO supplementation could improve the growth performance, cecal microflora composition, immunity, and antioxidant capacity of Partridge Shank chickens.

Specific pathogen-free (SPF) experimental animals are recognized as standard laboratory animals in the fields of biomedical, animal husbandry and veterinary research and production. Intestinal flora plays a critical role in nutrient absorption, improving health and protecting the host from pathogens. We therefore explored the variation and maintenance of intestinal flora in SPF chicks in order to better understand the composition of intestinal microflorain SPF chickens, and provide reference for the study of intestinal flora of SPF experimental animals. Five chicks were randomly selected at each of 14, 28, and 42 days, and ceca were removed for DNA extraction. The Illumina Miseq platform was used for microbiome analysis of the V3-V4 region of the 16S rRNA gene. During the course of chick gut microbiome development, we observed major changes in diversity, especially between day 14 and day 28. Firmicutes, Proteobacteria, and Bacteroidetes were the main bacterial taxa, and Firmicutes increased significantly with age. The genus with the highest relative abundance was Lactobacillus, followed by Faecalibacterium. In addition, while abundance of Ruminococcaceae spp., Ruminococcus, and Blautia increased with age, Lactobacillus, Enterobacteriaceae spp., and Oscillospira decreased with age. Interestingly, the abundance of Faecalibacterium first increased and then decreased over time. The characteristics of SPF chicken gut flora at different ages establish a basis for the regulation of intestinal flora in the early stage of brooding, and also provide a theoretical foundation for controlling and preventing infections and poultry diseases in newborn chickens.

The effects of dietary supplementation with phytogenic blend (PB) of Aerva lanata, Piper betle, Cynodon dactylon, and Piper nigrum on growth performance, ileal nutrient digestibility, intestinal morphology, and cecal microflora were determined in a 42-day broiler feeding trial. A total of 192 broilers were assigned to 4 dietary treatments (6 replicates and 8 birds/replicate): basal diet, basal diet supplemented with antibiotic (chlortetracycline), 1% and 2% PB, respectively. The body weight gain (BWG) of starter chicks increased linearly (P = 0.023) as dietary supplementation levels of PB increased. At grower phase, broilers fed diet supplemented with 1% PB had similar BWG with the antibiotic group, but other treatments had reduced (P = 0.0001) BWG. Dietary supplementation with 1% PB resulted in the highest (P < 0.0001) BWG during the study. Feed intake was not affected by the treatments during the starter, finisher, and overall rearing periods. Broilers fed diet supplemented with 1% PB had the best (P < 0.0001) feed conversion ratio during the study. Overall, broilers fed only basal diet had the highest (P = 0.0450) mortality. Ileal organic matter (OM) digestibility increased linearly (P = 0.044) with broilers fed diet supplemented with PB, but reduced with antibiotic group. Dietary supplementation with 1% PB had the highest (P = 0.0402) ileal digestibility of tryptophan. In the duodenum, broilers fed diet supplemented with PB had longer (P = 0.0006) villi heights than the birds fed only basal diet, but similar with antibiotic group. Broilers fed diet supplemented with PB had longer (P = 0.0064) villi height in the jejunum than the antibiotic group. Bifidobacterium concentration of the cecum content showed a slight increase (P = 0.053) with increasing supplementation levels of PB. In conclusion, the current study shows that dietary supplementation with PB improves growth performance, intestinal morphology, and apparent ileal digestibility of OM and tryptophan in a dose-dependent manner with the best response at 1% inclusion level.