Ethoxyquin attenuates enteric oxidative stress and inflammation by promoting cytokine expressions and symbiotic microbiota in heat-stressed broilers.

Intestinal oxidative stress in broilers is produced by chronic heat stress (HS) and has a negative impact on poultry performance as it induces intestinal inflammation and promotes the invasion of gram-negative bacteria, such as bacterial lipopolysaccharide (LPS). Therefore, dietary inclusion of the antioxidant compound, ethoxyquin (EQ), could improve enteric antioxidant capacity, immune responses, and the epithelial barrier, and maintain the symbiotic gut microbiota community. To investigate the effects of EQ supplementation on alleviating enteric oxidative stress in heat-stressed broilers, 200 one-day-old male Ross 308 broilers were randomly assigned to 4 groups (n = 50 chicks/group; n = 10 chicks/replicate) and fed a basal diet supplemented with 0 (CT), 50 (EQ-50), 100 (EQ-100), and 200 (EQ-200) mg EQ/ kg-1 for 5 wk. The chicks were raised in floor pens inside the broiler farm at a temperature and humidity index (THI) of 29 from d 21 to d 35. Growth performance traits, relative organ index, hepatic antioxidant enzymes, serum immunity, total adenylate, and cytokine activities were improved in the EQ-50 group (linear or quadratic P < 0.05), promoting the relative mRNA expression of cytokine gene-related anti-inflammatory and growth factors. A distinct microbial community colonised the gut microbiota in the EQ-50 group, with a high relative abundance of Lactobacillus, Ligilactobacillus, Limosilactobacillus, Pediococcus, Blautia, and Faecalibacterium compared to the other groups. Dietary supplementation with 50 mg EQ/ kg-1 for 5 wk attenuates enteric oxidative stress and intestinal inflammation by enhancing serum immune and cytokine content (IgG, IL-6, and TGF-ß,) and symbiotic microbiota in heat-stressed broilers. EQ promotes the expression of Hsp70, SOD2, GPx 4, IL-6, and IGF-1 cytokine gene-related anti-inflammatory and growth factors in heat-stressed hepatic broilers. Collectively, EQ-50 could be a suitable feed supplement for attenuating enteric oxidative stress and intestinal inflammation, thereby promoting the productivity of heat-stressed broilers.

Effects of exposing Japanese quail eggs to a low dose of gamma radiation and in ovo feeding by two sources of trace elements on embryonic development activities.

The present study investigated the influence of exposing quail eggs to low-dose gamma radiation (GR) and in ovo feeding with 2 sources of a mixture of trace elements (Zn, Fe, and Cu), including sulfate (TES) and loaded with montmorillonite (TEM), on embryonic development activities and prehatch quality. A total of 960 eggs on the seventh day of incubation were randomly divided into 6 groups (160 eggs/group) with 4 replicate of 40 eggs in each. A 3 × 2 factorial arrangement experiment was performed and included 3 sources in ovo feeding with a mixture of trace elements (Zn, Fe, and Cu), including 0 mg/egg, 50 mg TES/egg, and 50 mg TEM/egg with egg irradiation using 0 and 0.2 Gy from GR. Eggs injected with 50 mg TEM/egg and exposed to 0.2 Gy from GR (TEM/GR) was significantly (P ≤ 0.05 and 0.01) higher in hatchability, hatch body weight, and relative organ weight (liver, gizzard, proventriculus, heart, and intestine). The obtained results indicated significant (P ≤ 0.05) decreased in the serum concentration of malondialdehyde (MDA) in TEM/GR group. There was significant (P ≤ 0.05) increased of catalase (CAT) activity and the concentrations of growth hormone (GH) and insulin-like growth factor-1 (IGF-1) in TEM/GR group; however; total antioxidant capacity (T-AOC) was significant (P ≤ 0.05) increased in CT/GR group. Serum concentrations of immunoglobulin M (IgM) (P ≤ 0.05) and tumor necrosis factor-alpha (TNF-α) were increased in the TEM/CR group; the concentration of transforming growth factor beta (TGF-ß) significant (P ≤ 0.05) increased in the TEM/GR group; and interleukins (IL6 and IL10) showed no significant differences among the groups. Our results showed increase in thyroxine and myostatin concentrations with TES/CR and CT/GR of our study groups, respectively. The relative mRNA expression levels of the GH, IGF-1, and Fas cell surface death receptor (FAS) genes were significantly (P ≤ 0.05 and 0.01) upregulated in the liver tissue of the TEM/GR group compared with the other groups. In conclusion, TEM/GR was the best treatment for improving prehatch quality, increasing serum antioxidant enzyme activities, and promoting the expression of growth and immune genes in fertilized quail eggs.

Effect of Zinc Oxide Nanoparticles as Feed Additive on Blood Indices, Physiological, Immunological Responses, and Histological Changes in Broiler Chicks.

A feeding trial of 5-week duration was performed to assess the response of broiler chicks to dietary supplementation with different doses of myco-fabricated zinc oxide nanoparticles (ZONPs) on blood indices, physiological, immunological response, antioxidant status, intestinal microbial count, and histological changes in immune organs. A total of 162 3-day-old Ross 308 broiler chicks were weighed individually and distributed equally into 3 dietary treatments with 6 replicate of 9 chicks in each in a completely randomized design. Chicks were fed ad libitum a basal ration prepared as starter, grower, and finisher supplemented with 0 (T1, control), 40 (T2), and 60 (T3) mg zinc oxide nanoparticles (ZONPs)/kg feed. Results showed that supplementing with ZONPs at both studied levels increased the relative weights of the spleen, bursa, thymus, and liver and decreased the relative weight of the kidney, gizzard, and intestine. A significant increase in the concentrations of hemoglobin (Hb), hematocrit (PCV%), red and white blood cell counts, total protein (TP), globulin (GLOB), aspartate transferase (AST), alanine transferase (ALT), alkaline phosphatase (ALP), superoxide dismutase (SOD), glutathione peroxidase (GPx), and total antioxidant capacity (TAC) and a significant decrease in malonaldehyde (MDA), uric acid, and creatinine concentration were observed. Furthermore, all immunological organs showed histological alteration and increased both types of immunity in ZONPs groups with more pronounced effects in the T2 group.

Gut Microbiota, Intestinal Morphometric Characteristics, and Gene Expression in Relation to the Growth Performance of Chickens.

this study aimed to investigate the growth mechanism in a local breed of chickens by comparing the highest weight (HW) and the lowest weight in their microbiota, histological characteristics, and gene expression. Golden Montazah chickens, an Egyptian breed, were reared until they were 49 days old. All of the birds were fed ad libitum by a starter diet from day 1 until day 21, followed by a grower diet from day 21 to the end of the study. At 49 days old, the forty-eight birds with the heaviest body weight (HW) and the lightest body weight (LW) were chosen. Blood biochemical and histological morphometric parameters, electron microscopy, and intestinal nutrient transporter gene expression were studied in the sampled jejunum. The microbial composition and functions of the content and mucosa in HW and LW chickens were studied using 16S rRNA gene sequencing. The histological morphometric parameters were all more significantly (p < 0.05) increased in the HW chickens than in the LW chickens. Total protein, albumin, and triglycerides in serum were significantly higher (p < 0.05) in the HW chickens than in the LW chickens. The microbiome profile in the gut showed that Microbacterium and Sphingomonas were positively correlated with the body weights. In the local breed, there were significant differences in the intestinal microstructure which could enhance the growth mechanism and body weight. Our findings showed that some microbial components were significantly associated with body weight and their interactions with the host could be inferred to explain why these interactions might alter the host's metabolic responses. Further investigation into combining bioinformatics with lab experiments in chickens will help us to understand how gut bacteria can change the host's metabolism by special metabolic features in the gastrointestinal system.

Early life microbiota transplantation from highly feed-efficient broiler improved weight gain by reshaping the gut microbiota in laying chicken.

Starting phase of laying chicken life is the building stone for rearing and production stages. Since, fecal microbial transplantation (FMT) regulates the gut microbial diversity and affects the productive performance of the bird. The aim of this study is to evaluate the effect of FMT from feed-efficient broiler chicken could program the diversity of gut microbiota and growth of recipient native slow growing egg-laying chicks. For this, a total of 150 (one-day-old) Jing Hong chicks were randomly assigned into two groups, each group consisted of 5 replicates (n = 15 bird/ replicate). The control group (CON) and FMT recipient birds (FMT) fed on basal diet, the FMT group received an oral daily dose of FMT prepared from Cobb-500 chickens. The FMT performed from the 1d to 28d of age, through the experimental period, feed intake and body weight were recorded weekly. At the end of a 28-day trial, carcass traits were assessed and cecal samples were collected for microbiome assessment via 16S rRNA-based metagenomic analysis to characterize the diversity and functions of microbial communities. The data were statistically analyzed using R software. Body weight and body weight gain increased, and FCR decreased (p = 0.01) in FMT group. The relative abundance of Firmicutes and the Firmicutes/Bacteroidetes (F/B) ratio were increased due to FMT administration (p = 0.01). A higher relative abundance of Lactobacillus, Lactococcus, and Bifidobacterium were presented in the FMT group. Meanwhile, Enterococcus, Helicobacter, and Bacteroides were more abundant in the CON group (p < 0.01). Kyoto encyclopedia of genes and genomes (KEGG) pathways for microbial functions regarding amino acid metabolism, secondary metabolites biosynthesis, carbohydrate metabolism, energy metabolism, and enzyme families, cofactors, and vitamins were significantly annotated in the FMT group. Overall, FMT administration from the donor of highly feed-efficient broilers improved weight gain by reshaping a distinct gut microbiome, which may be related to the metabolism and health in the recipients laying chicks, providing new insight on the application of the FMT technique for early life programming of laying chickens.

Immunopathological comparison of in ovo and post-hatch vaccination techniques for infectious bursal disease vaccine in layer chicks.

This study was designed to compare immunopathological effects of in ovo vaccination with post-hatch vaccination against IBD in White Leghorn chicks. A total of 189 embryonated eggs were divided into six groups. At day 18 of incubation, groups A-C were administered in ovo with 228E, Winterfield 2512:10/3 and 2512/90:10/2.7, respectively, group D (post-hatch vaccination) and group E as shamed control (for quality evaluation of in ovo vaccination technique), and group F as control. The results showed that antibody titers against IBD detected by ELISA on days 2, 17, and 28 were significantly higher in all in ovo groups as compared to control groups E and F. On day 17, all vaccinated groups (in ovo and post-hatch vaccinated) showed no significant differences in antibody titers among themselves; however, at day 28, only the post-hatch group showed significantly higher antibody titers followed by in ovo vaccinated groups. The cell-mediated immunity determined by PHA-P assay was significantly higher in all vaccinated groups than the non-vaccinated groups. No clinical signs of IBD infection were observed in any of the vaccinated groups. There was only increase in bursa size of groups vaccinated with intermediate plus strains (groups A, C, and D) at day 28. The histopathology showed that all the treatment groups had mild lesions induced by IBD virus in bursa. This study concluded that in ovo vaccination with live IBD vaccines provides protective immunity to the chickens even in the presence of IBD-specific MDA; therefore, the onset of immunity was much earlier than the post-hatch vaccination and in ovo groups also maintained protective immunity against IBD for longer time.

Computational Insights into the Structural and Functional Impacts of nsSNPs of Bone Morphogenetic Proteins.

BMPs (bone morphogenetic proteins) are multipurpose (transforming growth factor)TGF-superfamily released cytokines. These glycoproteins, acting as disulfide-linked homo- or heterodimers, are highly potent regulators of bone and cartilage production and repair, cell proliferation throughout embryonic development, and bone homeostasis in the adults. Due to the fact that genetic variation might influence structural functions, this study is aimed to determine the pathogenic effect of nonsynonymous single-nucleotide polymorphisms (nsSNPs) in BMP genes. The implications of these variations, investigated using computational analysis and molecular models of the mature TGF-ß domain, revealed the impact of modifications on the function of BMP protein. The three-dimensional (3D) structure analysis was performed on the nsSNP Y316S, V386G, E387G, C389G, and C391G nsSNP in the TGF-ß domain of chicken BMP2 and H344P, S347P, V357A nsSNP in the TGF-ß domain of chicken BMP4 protein that was anticipated to be harmful and of high risk. The ability of the proteins to perform variety of tasks interact with other molecules depends on their tertiary structural composition. The current analysis revealed the four most damaging variants (Y316S, V386G, E387G, C389G, and C391G), highly conserved and functional and are located in the TGF-beta domain of BMP2 and BMP4. The amino acid substitutions E387G, C389G, and C391G are discovered in the binding region. It was observed that the mutations in the TGF-beta domain caused significant changes in its structural organization including the substrate binding sites. Current findings will assist future research focused on the role of these variants in BMP function loss and their role in skeletal disorders, and this will possibly help to develop practical strategies for treating bone-related conditions.

Effect of Dietary Rosemary and Ginger Essential Oils on the Growth Performance, Feed Utilization, Meat Nutritive Value, Blood Biochemicals, and Redox Status of Growing NZW Rabbits.

This study was conducted to assess the impacts of using two essential oils, rosemary and ginger, on growing rabbits' performance, carcass traits, meat composition, blood biochemicals, and the redox status of growing New Zealand White (NZW) rabbits. A total of 120 unsexed NZW rabbits, 42-days-old, were assigned randomly to five experimental groups (n = 24, 6 replicates with 4 rabbits each). The first group received a basal diet (control), the second to fifth groups were dietary supplemented daily with rosemary essential oil (REO) and ginger essential oil (GEO) at doses of 0.25 and 0.5% for each supplementation (REO-0.25, REO-0.5, GEO-0.25, and GEO-0.5), respectively. The growth traits were studied for 7 weeks, from the 7th to the 13th week of the rabbits' age. The results revealed that final body weight, weight gain, and average daily gain increased significantly (p < 0.01) in the REO-0.5 and GEO-0.5 treatments compared to the control group. Daily feed intake decreased (p = 0.005) in essential oil treatments. Meanwhile, the feed conversion ratio improved significantly (p = 0.001) in REO and GEO at the high doses compared to the control group. The weight percentages of liver and giblets increased (p < 0.001) with both treatments of REO and GEO compared to the control group. The dietary supplementation with REO and GEO did not affect (p > 0.05) the meat composition of Longissimus dorsi and hind leg muscles. Meanwhile, REO and GEO supplementation significantly decreased cholesterol levels in the rabbit meat. Thiobarbituric acid reactive substance concentrations decreased by 10 and 15% in the meat of REO-0.5 and GEO-0.5 treatments, respectively, compared to the other groups. In the same trend, REO and GEO treatments induced a significant (p = 0.001) reduction in the plasma cholesterol concentrations and triglycerides compared to the control. The total antioxidant capacity increased by 7.60% and the malondialdehyde decreased by 11.64% in the plasma of GEO-0.5 treatment than the control. Thus, the dietary supplementation of REO and GEO have a beneficial effect in improving the productivity and meat quality of growing rabbits.

Caecal microbiota could effectively increase chicken growth performance by regulating fat metabolism.

It has been established that gut microbiota influences chicken growth performance and fat metabolism. However, whether gut microbiota affects chicken growth performance by regulating fat metabolism remains unclear. Therefore, seven-week-old chickens with high or low body weight were used in the present study. There were significant differences in body weight, breast and leg muscle indices, and cross-sectional area of muscle cells, suggesting different growth performance. The relative abundance of gut microbiota in the caecal contents at the genus level was compared by 16S rRNA gene sequencing. The results of LEfSe indicated that high body weight chickens contained Microbacterium and Sphingomonas more abundantly (P < 0.05). In contrast, low body weight chickens contained Slackia more abundantly (P < 0.05). The results of H & E, qPCR, IHC, WB and blood analysis suggested significantly different fat metabolism level in serum, liver, abdominal adipose, breast and leg muscles between high and low body weight chickens. Spearman correlation analysis revealed that fat metabolism positively correlated with the relative abundance of Microbacterium and Sphingomonas while negatively correlated with the abundance of Slackia. Furthermore, faecal microbiota transplantation was performed, which verified that transferring faecal microbiota from adult chickens with high body weight into one-day-old chickens improved growth performance and fat metabolism in liver by remodelling the gut microbiota. Overall, these results suggested that gut microbiota could affect chicken growth performance by regulating fat metabolism.

The therapeutic effect of bromocriptine as mesylate and estradiol valerate on serum and blood biochemistry of common quails.

Hematology and serum biochemistry study may provide antique knowledge about the physical status of individuals, making them a valuable tool to differentiate healthy animals from affected animals. We aimed to investigate Steroid safety levels in birds through ex-situ studies at regular therapeutic doses. A total of 100 birds were used for hematology and serum biochemistry. This study was designed into 2 trials over the summer and winter, each comprised 5, 10, 15, and 20 d. Each study group was based on 5 control group birds and 20 experimental group birds. A sum of 2 groups representing 2 different steroids trial groups was treated with therapeutic doses to the stretch of 5, 10, 15, and 20 d each season. A therapeutic dose of each of the steroids was given at the rate of 3 drops 2 times a day to each bird. Analysis of data reveals that steroids had severe effects on bird's (Coturnix coturnix) hematological parameters. In most trials, the hematological effects of bromocriptine as mesylate showed an increase in red blood cell count and white blood cell count. On the other hand, steroid estradiol valerate showed a decrease in these parameters. Effect of steroids on serum biochemistry profile indicate acute damage to vital organs, especially to liver and kidney, indicating an increase in cholesterol, total protein, albumin, urea, and uric acid. The overall effect of steroids on the bird's serum and biochemistry of quails were nearly similar but different only in their intensity.

Role of clay in detoxification of aflatoxin B1 in growing Japanese quail with reference to gender.

The present study investigated the influence of the quail diet polluted with aflatoxin B1 (AFB1) and its detoxification by using clay as a feed additive on the growth performance and some blood biochemical components of growing Japanese quail with reference to sex. A total number of 120 Japanese quail chicks (1 week old), was randomly divided into 10 groups (24 chicks/ group). A 5 × 2 factorial arrangement experiment was performed and included five levels of AFB1 (0 ppm, 1 mg/kg AFB1, 1 mg/kg AFB1 + 1% clay, 2 mg/kg AFB1 and 2 mg/kg AFB1 + 1% clay) and two sexes. Birds fed with aflatoxin free diet had significantly (P ≤ 0.05 and 0.01) higher final live body weight, weight gain and lower mortality rate than the other groups. Addition of 1% clay significantly (P ≤ 0.05 and 0.01) improved the growth performance traits and diminished aflatoxin effect when compared to groups without the addition of clay. Obtained results indicated significant (P ≤ 0.05) differences between the two sexes in their response to aflatoxicosis in the final live body weight and weight gain. Our results showed significant (P ≤ 0.01) changes in all blood biochemicals (total protein, albumin, globulin, total cholesterol, creatinine, uric acid) and activities of serum enzymes studied due to the toxicity of AFB1. Conclusively, the consumption of polluted diets with AFB1 caused deleterious effects on the growth performance and blood biochemicals components of Japanese quail, while dietary addition of natural clay to the diet of growing Japanese quail caused beneficial effects.

Feeding Bacillus subtilis ATCC19659 to Broiler Chickens Enhances Growth Performance and Immune Function by Modulating Intestinal Morphology and Cecum Microbiota.

This study investigated dietary supplementation with Bacillus subtilis (BS) ATCC19659 on growth performance, biochemical indices, intestinal morphology, and cecum microflora in broiler chicks. A total of 600 Arbor 1-day Acres broilers of either sex were allotted to 5 treatments: chicks were fed a corn- and soybean-based diet (CON); chicks were fed basal diet containing 500 mg ZnB/kg (ZnB); chicks were fed basal diet containing 1 × 108 CFU/g feed of BS-ATCC19659 (BS-1); chicks were fed basal diet containing 3 × 108 CFU/g feed of BS-ATCC19659 (BS-3); and chicks were fed basal diet containing 5 × 108 CFU/g feed of BS-ATCC19659 (BS-5). Each treatment comprised 6 replicates with 20 birds for each replicate pen. Chicks in the BS-5 and BS-3 groups had higher body weight at the 21st and 42nd days and average daily gain from 1 to 21 days than that in the CON group (p < 0.05). Chicks in the BS-5 and ZnB groups had higher serum antioxidant activities and immunity response than those in the CON group (p < 0.05). Compared with the CON group, the liver mRNA abundance of GHR, TGF-ß, IGF-1, IFN-γ, SOD, CAT, and GPX of chicks in three BS groups and the ileum villus length (µm) of chicks in BS-3 and ZnB groups was increased (p < 0.05). Compared with the CON group, the villus height-to-crypt depth ratio of the ileum of chicks in the BS-5 and BS-3 groups and the crypt depth and villus height-to-crypt depth ratio of the jejunum in the BS-5 and ZnB groups were increased (p < 0.05). The abundance of the Cyanobacteria phyla in the cecum decreased in response to treatment with both BS-ATCC19659 and ZnB groups (p < 0.05). Compared with the CON group, the cecum abundance of genera GCA-900066575 (Lachnospiraceae), Anaerofustis, and Papillibacter (Firmicutes phylum) in three BS groups were increased (p < 0.05); The abundance of genus Escherichia-Shigella reduced in the BS-3 group (p < 0.05). Compared with the CON group, the cecum abundance of genus Clostridia_unclassified in ZnB and BS-5 groups was decreased (p < 0.05) of broilers. Generally, Bacillus subtilis ATCC19659 as feed additive positively affected growth performance, immunity response, and cecal microflora of broilers.

Study the Effect of Microinjection of Zn, Fe, and Cu Loaded in Montmorillonite on Development Activities in Fertilized Chicken Eggs.

This study aims to reduce embryonic mortality, increase body weight, and improve immune system in chicken. A total of 240 eggs were assigned to three treatments (n = 60) and injected with cooper (Cu), zinc (Zn), and iron (Fe) loaded by montmorillonite (Mnt), and one untreated group (n = 60). Some hormones and enzymes related with growth were measured in terms of serum, and expression of some genes related to growth, immune, and programmed cell deaths that were determined in the liver and spleen of chicken by RT-qPCR. The embryonic death on the fifth and seventh days after injecting eggs with Fe-Mnt was less obvious than in other groups. The heaviest body weight was recorded for Fe-Mnt and Cu-Mnt treatment. Fe-Mnt treatment had higher serum GSH, SOD, GH, and Myostatin contents and lower MDA than those in the other treatments. Cu-Mnt treatment included the highest contents of CAT enzyme and IGF-1 hormone in serum. The highest expression of IGF-1, GH, BCL6, and SYK genes in liver tissue were recorded by Zn-Mnt, IGFBP2, FGF8, and IFNW1 genes by Cu-Mnt, and TC1RG1 and IFNW1 genes by Fe-Mnt in spleen tissue. In conclusion, Fe-Mnt was the best treatment for reducing embryonic mortality, and increasing body weight of chickens and expression of growth and immune genes, followed by Cu-Mnt treatment.

Sex differences in growth performance are related to cecal microbiota in chicken.

In poultry industry, male chickens have a better growth performance than female ones under the same genetic background and diet. Emerging evidences proposed an important role of intestinal microbiota in chicken's growth performance. This study aimed to determine gut microbiota related gender based differences in the growth performance of chickens. Therefore, male and female chickens (n = 20) at 7-week age were used to carry out histomorphological, molecular, gene expression analysis with their liver, chest and leg muscle, as well as 16S rRNA sequencing analysis for gut microbiota. The results revealed that Bacteroides and Megamonas genera were more prominently colonized in the cecum of male chickens. The male chicken's cecal microbiota indicated a closer relation with glycan metabolism, while in the female chickens it was more related with lipid metabolism. Gene expression levels associated with glycan and lipid metabolism were different between male and female chickens. Further, using Spearman correlation analysis, we found a positive correlation between glycan and lipid metabolism, and the relative abundance of Bacteroides, Megamona and Lactobacillus in male chickens. Similarly, we also found a positive correlation between the lipid metabolism and the relative abundance of Ruminococcaceae and Enterococcus in female chickens. These findings revealed the association of chicken growth performance with cecal microbiota that contributed to the metabolism of glycan and lipid in a sex-dependent manner.

A genome-wide single nucleotide polymorphism scan reveals genetic markers associated with fertility rate in Chinese Jing Hong chicken.

The function of the sperm storage tubules is directly correlated with the fertility of laying hens. However, little is known about the molecular mechanisms regulating the fertility traits in chicken. To identify genetic markers associated with reproductive traits, we calculated fertility rate at 61 to 69 wk (51 D) of Jing Hong chickens parent generation as the phenotype and the genotype were detected by the chicken 600K Affymetrix Axiom High Density single nucleotide polymorphisms (SNP)-array. The genome-wide association study using 190 Jing Hong hens showed that the 20 SNP in chromosomes 3 and 13 were significantly associated with fertility rate. To verify these results, a total of 1900 Jing Hong laying hens from 2 populations (P1 and P2) were further genotyped by polymerase chain reaction-restriction fragments length polymorphisms method. The association analysis results revealed that 12 polymorphisms (AX-75769978, AX-76582632, AX-75730546, AX-75730496, AX-75730588, AX-76530282, AX-76530329, AX-76529310, AX-75769906, AX-75755394, AX-80813697 and AX-76582809) out of 20 showed highly significant effects (P < 0.0001) on fertility rate in P1, P2 and P1+P2. Six haplotypes (TTAA, TTGG, TTAG, CTAA, CTGG, and CTAG) were inferred based on significant loci (AX-75730546 and AX-76530282) also showed significant association with fertility rate, where haplotype CTAG was shown to be markedly associated with the significantly highest (P < 0.0001) fertility rate (in P1, 86.42 ± 0.59; P2, 85.98 ± 0.59 and P1+P2, 86.16 ± 0.42) followed by other haplotypes for the irrespective of population studied. Collectively, we report for the first time that 12 SNP in the chromosomes 3 and 13 were significantly associated with fertility rate during the later stage of egg production, which could be used as the potential genetic markers that would be able to facilitate in the selection and improvement of fertility rate through chicken breeding.

Investigation of the Impacts of Antibiotic Exposure on the Diversity of the Gut Microbiota in Chicks.

The dynamic microbiota in chickens can be affected by exposure to antibiotics, which may alter the composition and substrate availability of functional pathways. Here, 120 Jing Hong chicks at 30 days of age were randomly divided into four treatments totaling seven experimental groups: control chicks not exposed to antibiotics; and chicks exposed to enrofloxacin, diclazuril, and their mixture at 1:1 for 14 days and then not exposed for a withdrawal period of 15 days. Fecal samples were collected from the 7 groups at 8 time-points (exposure to 4 antibiotics and 4 withdrawal periods) to perform in-depth 16S rRNA sequencing of the gut microbiota. Taxon-independent analysis showed that the groups had significantly distinct microbial compositions (p < 0.01). Based on the microbial composition, as compared with the control group, the abundances of the phyla Firmicutes, Actinobacteria, Thermi, and Verrucomicrobia, as well as the families Lactobacillus, Lactococcus, S24-7, and Corynebacterium, were decreased in the antibiotic-exposed chicks (p < 0.01). Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) analyses revealed significant differences in microbiota metabolite pathways due to the genera of the antibiotic-responsive microbes (p < 0.01), especially the pathways relating to cell growth and death, immune system diseases, carbohydrate metabolism, and nucleotide metabolism. Oral treatment with enrofloxacin, diclazuril, and their mixture modified the gut microbiota composition and the microbial metabolic profiles in chickens, with persistent effects (during the withdrawal period) that prevented the return to the original community and led to the formation of a new community.

Adaptive selection in the evolution of programmed cell death-1 and its ligands in vertebrates.

Programmed cell death-1 (PD-1) and its ligands, particularly PD-L1 and PD-L2, are the most important proteins responsible for signaling T-cell inhibition and arbitrating immune homeostasis and tolerance mechanisms. However, the adaptive evolution of these genes is poorly understood. In this study, we aligned protein-coding genes from vertebrate species to evaluate positive selection constraints and evolution in the PD1, PD-L1 and PD-L2 genes conserved across up to 166 vertebrate species, with an average of 55 species per gene. We determined that although the positive selection was obvious, an average of 5.3% of codons underwent positive selection in the three genes across vertebrate lineages, and increased positive selection pressure was detected in both the Ig-like domains and transmembrane domains of the proteins. Moreover, the PD1, PD-L1 and PD-L2 genes were highly expressed in almost all tissues of the selected species indicating a distinct expression pattern in different tissues among most species. Our study reveals that adaptive selection plays a key role in the evolution of PD1 and its ligands in the majority of vertebrate species, which is in agreement with the contribution of these residues to the mechanisms of pathogen identification and coevolution in the complexity and novelties of vertebrate immune systems.

Investigation of the impact of gut microbiotas on fertility of stored sperm by types of hens.

Owing to the practical interest in understanding duration of fertility (DF) to reduce the cost of producing hatching eggs by decreasing the frequency of artificial insemination, as well to uncover the mechanism of the estrogen-gut microbiome axis, elucidating the interaction between the maternal microbiome and the function of sperm storage tubules (SST) has become important for revealing the DF in laying hens. In this study, we investigated the compositional, structural, and functional differences in gut microbiomes between hens with high (HSST, n = 8) and low SST activity (LSST, n = 10) by performing phenotypic selection from approximately 400 individual hens based on their DFs. Their cecal microbial communities were analyzed by sequencing the V4 region of the 16S rRNA gene. The microbiome abundance estimators from the ceca of HSST and LSST hens were not significantly different at the phylum and genus taxonomic levels, although the relative abundances for the phylum Firmicutes and the genus Lactobacillus were higher in the HSST group. Furthermore, some taxonomic levels of bacteria expressing the components of several metabolic pathways differed between the HSST and LSST groups. Moreover, predicting functional microbiomes by Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed that certain pathways, such as the metabolism of carbohydrates and protein, cellular processes, and organismal systems, of the HSST group exhibited higher expression of genes associated with bioactivity and energy biosynthesis than those in the LSST group. Our results may provide insights into hen-microbe interactions with respect to DF and will be useful in establishing a strategy for new research to uncover the functional regulation of SST in laying hens.

Characterisation of stem and proliferating cells on the retina and lens of loach Misgurnus anguillicaudatus.

The eye of the fish has a lifelong persistent neurogenesis unlike eye of mammals, so it's highly interesting to study retinal neurogenesis and its genetic control to give complete knowledge about the cause of this property in fish in comparison to mammals. We performed fluorescent in situ hybridisation for loach Misgurnus anguillicaudatus bmi1, msi1 and sox2 genes, which are used as an indicator of the sites of multipotent stem cells. Proliferating cell nuclear antigen (PCNA), bromodeoxyuridine (BRDU) and KI67 markers were used as indicators of proliferating cells and glial fibrillary acidic protein (GFAP) immunofluorescence was used for detection of the glial property of cells, as well as, immunohistochemistry detected the role of peroxisome proliferator-activated receptor (PPAR)α and γ in retinal neurogenesis. Our results determined that the lens and the retina of loach M. anguillicaudatus contain proliferative and pluripotent stem cells that have both glial and neuroepithelial properties, which add new cells continuously throughout life even without injury-induced proliferation. The PPARα has an essential function in providing energy supply for retinal neurogenesis more than PPARγ.

Zinc and Copper with New Triazine Hydrazone Ligand: Two Novel Organic Complexes Enhanced Expression of Peptide Growth Factors and Cytokine Genes in Weaned V-Line Rabbit.

Two novel transitional organic Zn/Cu complexes based on a new biocompatible bidentate triazine-hydrazone ligand (Thz) was designed, synthesized, and evaluated in this study. This study evaluated the effects of injecting 60 mg of Zn and 40 mg of Cu in three different forms, twice per week, for eight weeks on growth performance, expression of growth factors and cytokine genes, carcass yield, blood biochemicals, and intestinal morphology in weaned rabbits. The tested complexes were sulfate (Cu/ZnSO4), montmorillonite (Cu/Zn-Mnt), and triazine hydrazone (Cu/Zn-Thz). A total of 60 V-line weaned rabbits at four weeks of age were assigned to four treatments (n = 15), which were intramuscularly injected with 0.5 mL of either (1) saline (control) or saline containing (2) Cu/ZnSO4, (3) Cu/Zn-Mnt, or (4) Cu/Zn-Thz. Compared to the controls, the rabbits injected with Cu/Zn-Thz showed a higher (p < 0.01) growth rate, carcass yield (p < 0.05), and liver expression of insulin like growth factor-1 (IGF-1), growth hormone receptor (GHR), fibroblast growth factor-1 (FGF1), and transforming growth factor beta-1 (TGFB1) (p < 0.05), as well as better jejunum morphometric variables (p < 0.05). On the other hand, mRNA of FGF1, TGF1, TCIRG1, and adenosine deaminase (ADA) were higher expressed (p < 0.05) in the spleen tissues of Cu/Zn-Mnt group. Collectively, the results indicated that our novel synthesized organic complexes of Zn/Cu-Thz proved to be a suitable feed supplement, as it increased rabbit productive performance through enhancing expression of peptide growth factors and cytokine genes.