Transcriptome analysis of the uterovaginal junction containing sperm storage tubules in heat-stressed breeder hens.

Sperm storage tubules (SSTs) in the uterovaginal junction (UVJ) of the oviduct are major sites of sperm storage after artificial insemination or mating. Female birds may regulate sperm motility in the UVJ. Heat stress can decrease the reproductive ability of broiler breeder hens. However, its effects on UVJ remain unclear. Changes in gene expression aid in understanding heat stress-affected molecular mechanisms. Herein, we wanted to conduct a comparative transcriptomic analysis to identify the differentially expressed genes (DEGs) in the UVJ of breeder hens under thermoneutral (23°C) and heat stress (36°C for 6 h) conditions. The results indicated that cloacal temperatures and respiratory rates were significantly increased in heat-stressed breeder hens (P < 0.05). Total RNA was extracted from the hen UVJ tissues containing SSTs after heat exposure. Transcriptome analysis identified 561 DEGs, including 181 upregulated DEGs containing heat shock protein (HSP) transcripts and 380 downregulated DEGs containing immune-related genes, such as interleukin 4-induced 1, radical S-adenosyl methionine domain containing 2, and 2'-5'-oligoadenylate synthetase like, in heat-stressed hens. Gene Ontology analysis revealed the significantly enriched terms involving HSPs. Kyoto Encyclopedia of Genes and Genomes analysis identified 9 significant pathways, including the protein processing in endoplasmic reticulum (11 genes including HSPs), neuroactive ligand-receptor interaction (13 genes including luteinizing hormone/choriogonadotropin receptor), biosynthesis of amino acids (4 genes including tyrosine aminotransferase), ferroptosis (3 genes including heme oxygenase 1), and nitrogen metabolism (carbonic anhydrase [CA]-12 and CA6) pathways. Protein-protein interaction network analysis of DEGs revealed 2 large networks, one containing upregulated HSPs and the other containing downregulated interferon-stimulating genes. Overall, heat stress inhibits innate immunity in the UVJ tissues of broiler chickens, and heat-stressed chickens protect their cells by increasing the expression levels of HSPs. The identified genes are potential candidates for further exploration of the UVJ in heat-stressed hens. The identified molecular pathways and networks increase our understanding of the sperm storage reservoirs (UVJ containing SSTs) within the reproductive tract and may be used to prevent heat stress-induced fertility loss in breeder hens.

Assessment of lactic acid bacteria isolated from the chicken digestive tract for potential use as poultry probiotics.

OBJECTIVE: The use of probiotics as an alternative to antibiotics in animal feed has received considerable attention in recent decades. Lactic acid bacteria (LAB) have remarkable functional properties promoting host health and are major microorganisms for probiotic purposes. The aim of this study was to characterize LAB strains of the chicken digestive tract and to determine their functional properties for further use as potential probiotics in poultry. METHODS: A total of 2,000 colonies were isolated from the ileum and cecal contents of the chickens based on their phenotypic profiles and followed by a preliminary detection for acid and bile tolerance. The selected 200 LAB isolates with exhibited well-tolerance in acid and bile conditions were then identified by sequencing the 16S rDNA gene, followed by acid and bile tolerance, antimicrobial activity, adhesion to epithelial cells and additional characteristics on the removal of cholesterol. Then, the two probiotic strains (L. ingluviei and L. salivarious) which showed the greatest advantage in vitro testing were selected to assess their efficacy in broiler chickens. RESULTS: It was found that 200 LAB isolates that complied with all measurement criteria belonged to five strains, including L. acidophilus (63 colonies), L. ingluviei (2 colonies), L. reuteri (58 colonies), L. salivarius (72 colonies), and L. saerimneri (5 colonies). We found that the L. ingluviei and L. salivarius can increase the population of LAB and Bifidobacterium spp. while reducing Enterobacteria spp. and Escherichia coli in the cecal content of chickens. Additionally, increased concentrations of valeric acid and short chain fatty acids were also observed. CONCLUSION: This study indicates that all five Lactobacillus strains isolated from gut contents of chickens are safe and possess probiotic properties, especially L. ingluviei and L. salivarius. Future studies should evaluate the potential for growth improvement in broilers.

Extraction of dietary fibers from cassava pulp and cassava distiller's dried grains and assessment of their components using Fourier transform infrared spectroscopy to determine their further use as a functional feed in animal diets.

OBJECTIVE: The present study was to investigate the extraction conditions of dietary fiber from dried cassava pulp (DCP) and cassava distiller's dried grains (CDG) under different NaOH concentrations, and the Fourier transform infrared (FTIR) was used to determine the dietary fiber components. METHODS: The dried samples (DCP and CDG) were treated with various concentrations of NaOH at levels of 2%, 4%, 6%, and 8% using a completely randomized design with 4 replications of each. After extraction, the residual DCP and CDG dietary fiber were dried in a hot air oven at 55°C to 60°C. Finally, the oven dried extracted dietary fiber was powdered to a particle size of 1 mm. Both extracted dietary fibers were analyzed for their chemical composition and determined by FTIR. RESULTS: The DCP and CDG treated with NaOH linearly or quadratically or cubically (p< 0.05) increased the total dietary fiber (TDF) and insoluble fiber (IDF). The optimal conditions for extracting dietary fiber from DCP and CDG were under treatment with 6% and 4% NaOH, respectively, as these conditions yielded the highest TDF and IDF contents. These results were associated with the FTIR spectra integration for a semi-quantitative analysis, which obtained the highest cellulose content in dietary fiber extracted from DCP and CDG with 6% and 4% NaOH solution, respectively. The principal component analysis illustrated clear separation of spectral distribution in cassava pulp extracted dietary fiber (DFCP) and cassava distiller's dried grains extracted dietary fiber (DFCDG) when treated with 6% and 4% NaOH, respectively. CONCLUSION: The optimal conditions for the extraction of dietary fiber from DCP and CDG were treatment with 6% and 4% NaOH solution, respectively. In addition, FTIR spectroscopy proved itself to be a powerful tool for fiber identification.

Mix of cassava pulp, napier grass, and enzymes can be used as low cost alternative feed ingredient for laying hens.

Two experiments were carried out to evaluate the effect of mixed cassava pulp and napier grass (MCN) supplemented with multi-component enzymes on the hen performance and egg quality as well as nutrient digestibility. In experiment 1, forty laying hens (Isa Brown) of 34 weeks in age were randomly allotted to 5 groups: control and 5, 10, 15, and 20% MCN for 10 days (one bird per cage, eight cage replicates). The results indicated that dry matter, and organic matter digestibilities, or nitrogen retention (P > 0.05) showed no significant alteration when used MCN up to 20%. In experiment 2, one hundred and eighty Isa Brown laying hens of 67 weeks in age were randomly assigned to 5 groups (control and 4 MCN: 5, 10, 15, and 20%) and raised for 8 weeks. It revealed that the inclusion rate of MCN up to 20% did not affect the productive performance, plasma cholesterol concentration, cecal microbial populations, or ammonia production (P > 0.05). In addition, egg yolk cholesterol was found to be lower in laying hens fed 20% MCN compared to 5% MCN (P < 0.05), but no significant differences compared to the control. In conclusion, it is indicated that MCN can be used as a partial part of an energy source in laying hen diets with no adverse effects on productive performance, nutrient digestibility or egg quality.

Free Faecal Water: Analysis of Horse Faecal Microbiota and the Impact of Faecal Microbial Transplantation on Symptom Severity.

Free faecal water (FFW) in equines results in pollution of the hindquarters and tail and can also involve clinical signs. Though the cause of FFW is unknown, it was hypothesized that it may involve the gut microbiota. This hypothesis was addressed as follows. First, the faecal prokaryotic community composition of horses suffering from FFW relative to healthy controls (n = 10) was compared. Second, FFW horses were treated with a standardised faecal microbiota transplantation (FMT) protocol (n = 10), followed by assessment of FFW symptom severity and faecal prokaryotic community composition over a follow-up period of 168 days. No significant differences were found in the faecal microbiota composition of FFW horses compared to healthy controls (p > 0.05). Relative to before FMT, FFW symptom severity decreased in affected horses 14 days after FMT (p = 0.02) and remained decreased for the remainder of the study (p < 0.02). However, individual animal responses to FMT varied. FMT had no effect on FFW horse faecal prokaryotic community composition in terms of alpha or beta diversity. Alpha diversity of the donor inocula used in the FMT was always lower than that of the faecal microbiota of the FFW treated horses (p < 0.001). In conclusion, whilst findings indicate FFW horses do not have an altered hindgut microbiota, some horses that received FMT had a temporary alleviation of FFW symptom severity without causing changes in the faecal microbiota. Future studies using controls are now needed to confirm the effectiveness of FMT to treat FFW.

Lactobacillus ingluviei C37 from chicken inhibits inflammation in LPS-stimulated mouse macrophages.

Probiotics are growing alternatives to antibiotics, and can contribute to the prevention and treatment of diseases and enhance livestock production. Lactobacillus (L.) ingluviei is a novel probiotic species with growth-enhancement effects; however, this species remains poorly understood, and there have been (to our knowledge) no studies focusing on its immunological effects. Here, we isolated L. ingluviei C37 (LIC37) from chicken and evaluated the bacterium's immunomodulatory properties to explore its probiotic potential. Real-time quantitative PCR and ELISA showed that in vitro exposure of inflammation-stimulated mouse peritoneal macrophages to heat-killed LIC37 led to decreases in tumor necrosis factor-α and interleukin (IL)-6 levels and an increase in IL-10. These findings suggested that LIC37 exerts anti-inflammatory effects by modulating cytokine profiles. This species may be an attractive probiotic bacterial strain for use in animal production.

Digestibility, productive performance, and egg quality of laying hens as affected by dried cassava pulp replacement with corn and enzyme supplementation.

Two experiments were conducted to investigate the potential use of dried cassava pulp (DCP) supplemented with enzymes as an alternative feed ingredient in laying hen diets. In experiment 1, 45 laying hens (Isa Brown) aged 45 weeks were placed in individual cages to measure nutrient digestibility for 10 days. Nine dietary treatments were control and DCP as a replacement for corn at 20, 25, 30, and 35% supplemented with mixed enzymes (cellulase, glucanase, and xylanase) at 0.10 and 0.15%. Results showed that the use of DCP at 20-35% added with mixed enzymes had no negative effects on dry matter digestibility, while organic matter digestibility and nitrogen retention decreased with increased DCP up to 30-35% in diets. Both enzyme levels (0.10 and 0.15%) showed similar results on nutrient digestibility and retention. In experiment 2, a total of 336 laying hens aged 32 weeks were randomly allocated to seven dietary treatments (control and DCP-substituted diets at 20, 25, and 30%) supplemented with mixed enzymes (0.10 and 0.15%). Diets incorporated with 20-30% of DCP and supplemented with mixed enzymes at both levels had no significant effects on egg production, egg weight, feed intake, egg mass, feed conversion ratio, protein efficiency ratio, or egg quality, except for egg yolk color being decreased with an increase of DCP in diets (P < 0.05). In conclusion, it is suggested that DCP supplemented with enzymes can be used as an energy source in laying hen diets up to 30% without showing negative effects on nutrient digestibility, productive performance, or egg quality.