Probiotics-induced Changes in Intestinal Structure and Gut Microbiota Are Associated with Reduced Rate of Pimpled Eggs in the Late Laying Period of Hens.

Production of pimpled or sandpaper-shelled eggs (SE) is a major problem in aged hens. Probiotics can improve eggshell quality; however, the relationship between SE production and gut bacteria remains unclear. Here, 1200 450-d-old Hy-line hens were assigned to four groups (300 hens each), with the control group fed basal diet and treatment groups fed basal diet plus 500, 1000, and 1500 mg/kg of Clostridium butyricum and Bacillus subtilis, respectively. After 4 weeks, probiotics significantly decreased the SE rate from 42.51% to 28.02%. To address why probiotics reduced SE rate, the hens that only produced normal eggs (NE) or SE based on a 2-week assessment were assigned to three groups (NE, SE, and SEP groups; 10 hens each), with the NE and SE groups fed a basal diet and SEP group fed a basal diet plus 1000 mg/kg probiotics. After 4 weeks, ileal tissues from eight birds/group were collected for histomorphological and gene expression analyses, and the ileal content was collected from five birds/group for 16S rDNA sequencing analysis. The data showed that probiotics significantly increased the villus length and ratio of villus length to crypt depth. Quantitative PCR analysis indicated that there were no significant differences in the expression of genes related to tight junctions, nutrient transport, and calcium absorption among the groups (except TRPV6, P<0.001). The 16S rDNA sequencing analysis indicated that the alpha-diversity of gut bacteria in the SEP group was the highest among the groups. The Firmicutes phylum was dominant in the NE and SEP groups, whereas the Proteobacteria phylum was dominant in the SE group. Together, these results suggest that probiotics can significantly influence the intestinal structure and composition of the intestinal microbiota, which may lead to a reduction in the SE rate in aged hens.

Characterization and functional analysis of a myeloid differentiation factor 88 in Ostrinia furnacalis Guenée larvae infected by Bacillus thuringiensis.

Myeloid differentiation factor 88 (MyD88) is a pivotal adapter protein involved in activating nuclear factor NF-κB of the Toll pathway in insect innate immunity. MyD88 has been extensively studied in vertebrates and Drosophila. However, the information ascribed to MyD88 in Lepidoptera is scarce. In the present study, an Ostrinia furnacalis MyD88 (OfMyD88) cDNA was cloned and functionally characterized (GenBank accession no. MN906311). The complete cDNA sequence of OfMyD88 is 804 bp, and contains a 630 bp open reading frame encoding 209 amino acid residues. OfMyD88 has the death domain (DD), an intermediate domain, and the Toll/interleukin 1 receptor (TIR) domain. OfMyD88 was widely expressed in immune-related tissues such as hemocytes, fat body, midgut, and integument, with the highest expression level in hemocytes, and the lowest expression level in integument. To clarify the immune function of MyD88, O. furnacalis larvae were challenged with Bacillus thuringiensis (Bt) through feeding. Bt oral infection had significantly up-regulated the expression of OfMyD88 and immune genes, including PPO2 (prophenoloxidase 2), Attacin, Gloverin, Cecropin, Moricin, GRP3 (ß-1, 3-Glucan recognition protein 3), and Lysozyme, and increased the activities of PO and lysozyme in hemolymph of O. furnacalis larvae. Knockdown of OfMyD88 by RNA interference suppressed the expression levels of immune related genes, but not PPO2 in the larvae orally infected with Bt, suggesting that OfMyD88 is involved in defending against Bt invasion through the Toll signaling pathway, but does not affect the PPO expression in O. furnacalis larvae.

Subacute ruminal acidosis in dairy herds: Microbiological and nutritional causes, consequences, and prevention strategies.

Dairy cattle are frequently fed high-concentrate (HC) diets in modern intensive feeding systems, especially in the transition period. During this period, cows face many alterations that include hormonal changes and shifting to a lactating state. Switching to a HC diet that may disrupt the ruminal microbiota balance can lead to subacute ruminal acidosis (SARA). Moreover, the main factor shaping the rumen microbiota is dietary composition, especially the ratio of starch to fibrous carbohydrates. Feeding highly fermentable carbohydrate diets after adaptation to forage diets leads to a rumen fermentation rate that exceeds rumen absorption and buffering rates, resulting in a reduction in ruminal pH. As a result of Gram-negative bacterial cell lysis, an increase in harmful ruminal bacterial metabolites, including lipopolysaccharide, lactic acid, and histamine, is observed. The interactions between the host immune system and the ruminal microbiota play an essential role in many physiological processes and the development of the disorder. Progress in DNA sequencing and bioinformatics platforms provides new opportunities to investigate the composition of ruminal microbes and yields unique advances in understanding ecology of the rumen. Subacute ruminal acidosis is linked with a change in the ruminal microbiota structure and richness and with other metabolic disorders; such as rumenitis, milk fat depression, laminitis, and liver abscesses. Therefore, this review aims to explore a better understanding of the crosstalk between diet and microbiota in the prevalence of rumen acidosis and its consequences, which is crucial for control strategies such as feeding management, and supplementation with thiamine, prebiotics, and probiotics.

Screening of MicroRNAs with Potential Systemic Effects Released from Goose Fatty Liver.

Communication between tissues and organs plays an important role in the maintenance of normal physiological functions as well as the occurrence and development of diseases. Communication molecules act as a bridge for interactions between tissues and organs, playing not only a local role in the tissues and organs where they are secreted but also in exerting systemic effects on the whole body via circulation. In this study, blood microRNA-omics analysis of overfed vs. normally fed (control) Landes geese revealed that the content of each of the 21 microRNAs (miRNAs) in the blood of overfed geese was significantly higher than that in the blood of control geese. These miRNAs may have systematic effects in the development of goose fatty liver as well as being candidate markers for the diagnosis of goose fatty liver. We determined the expression of miR-143, miR-455-5p, miR-222a-5p, miR-184, miR-1662, and miR-129-5p using quantitative PCR in goose fatty liver vs. that in normal liver. The expression of these miRNAs, except miR-129-5p, in goose fatty liver was also significantly higher than that in normal liver (P<0.05), suggesting that these blood miRNAs are released from goose fatty liver. In addition, we found that expression of IGFBP5, the predicted target gene of miR-143, was significantly decreased in goose fatty liver vs. the normal liver (P<0.05), indicating that miR-143 may exert both local and systematic effects by inhibiting the expression of IGFBP5, thus promoting the development of goose fatty liver. In conclusion, we identified several miRNAs, including those we validated (i.e., miR-143, miR-455-5p, miR-222a-5p, miR-184, miR-1662, and miR-129-5p) that may serve as candidate markers in the diagnosis of goose fatty liver as well as local and global regulators contributing to the development of goose fatty liver.

Thiamine modulates intestinal morphological structure and microbiota under subacute ruminal acidosis induced by a high-concentrate diet in Saanen goats.

Ruminant animals are generally fed with starch-rich grain as the main energy source, and the incidence of metabolic diseases such as subacute ruminal acidosis (SARA) is high due to the intensive farming. Thiamin has been reported to alleviate SARA caused by high-concentrate diets, but the exact mechanism is not well understood. The goal of this study was to examine the role of thiamine in intestinal inflammation and microbiota caused by high-concentrate diets. The SARA model was induced by low neutral detergent fibre/starch ration to study the effects of thiamine on intestinal tissue structure and microbiota. 18 mid-lactation (148 ± 3 d in milk; milk yield = 0.71 ± 0.0300 kg/d) Saanen goats (BW = 36.5 ± 1.99 kg; body condition score = 2.73 ± 0.16, where 1 = emaciated and 6 = obese) in parities 1 or 2 were selected. The goats were randomly divided into three groups with six replicates: (1) control diet (C; concentrate:forage 30:70), (2) high-concentrate diet (H; concentrate:forage 70:30), and (3) high-concentrate diet with 200 mg of thiamine/kg of DM intake (H + T;concentrate:forage 70:30). The experimental period was lasted for 56 d. The small and large intestine, expression of inflammatory factor genes, tight junction protein genes, total antioxidant capacity, and intestinal microbiota were measured. The results showed that SARA was observed in treatment H, whereas rumen fluid pH was improved in treatment H + T. Treatment H + T also significantly repaired the intestinal tissue structure damaged by SARA, improved the total antioxidant capacity of the small intestinal mucosa, reduced mRNA expression of inflammatory factors in the small intestine tissue, and increased the mRNA expression of tight junction genes in small intestine tissue. The high-concentrate diet reduced the diversity of intestinal microbiota. When thiamine is added to the high-concentrate diet, the relative abundance of intestinal Firmicutes and beneficial bacteria represented by Lactobacilli were upregulated, and the relative abundance of Proteus, a marker of intestinal dysbacteriosis, returned to normal. In conclusion, thiamine supplementation could alleviate the damage to the intestinal tissue structure and microbial environment caused by SARA condition in dairy goats fed a high-concentrate diet.

Uterine structure and function contributes to the formation of the sandpaper-shelled eggs in laying hens.

The avian eggshell is formed in the uterus, and eggshell quality usually decreases markedly in the late phase of hen laying cycles. Production of sandpaper-shelled eggs (SE), a category of eggs with relatively less eggshell quality, causes a great economic loss. Underlying mechanisms of SE formation, however, remain unclear. For the present study, it was hypothesized that alterations in uterine structure and function contribute to SE formation. To test this hypothesis, uterine samples were collected from 450-day-old hens that produced normal eggs (NE) and SE (based on 2-week-long assessments, n = 10) for histomorphological and transcriptome analyses. Compared with the NE group, uteri of the SE group were apparently atrophied. Furthermore, a total of 211 differentially expressed genes (DEGs) were identified in the uteri of hens of the two groups. These DEGs were clustered into 145 gene ontology terms (FDR < 0.05) and enriched in 12 KEGG pathways (P < 0.10), which are primarily related to organ morphogenesis and development, cell growth, differentiation and death, ion transport, endocrine and cell communication, immune response, and corticotropin-releasing hormones. In particular, corticotropin may be an important factor in SE formation because of effects on ion transport. Furthermore, as indicated by lesser abundances of relevant mRNA transcripts, the lesser expression of genes related to ion transport and matrix proteins also contribute to SE production because of effects on eggshell formation. In conclusion, results from this study revealed there were structural and functional differences in the hen uterus in NE and SE groups.