miRNA-16-5p inhibits the apoptosis of high glucose-induced pancreatic β cells via targeting of CXCL10: potential biomarkers in type 1 diabetes mellitus

Introduction: We aimed to elucidate the relationship between CXC chemokine ligand 10 (CXCL10) and miR-16-5p, and their functions on the biological behaviour of type 1 diabetes mellitus (T1DM). Material and methods: The GSE72492 dataset from the GEO database was used to analyse gene expression. We discovered that CXCL10 was highly expressed in T1DM patients. The up-stream miRNA was predicted by Targetscan website. Low glucose (2.8 mmol/L) and high glucose (HG, 16.7 mmol/L) were utilised to treat β-TC-tet (pancreatic β cell) cells to form the model. The direct interaction between miR-16-5p and CXCL10 was verified by a dual-luciferase reporter assay. Real-time quantitative PCR (qRT-PCR) and western blotting analyses were used to detect RNA and protein expression. CCK8 and flow cytometry were used to detect cell proliferation and apoptosis. Results: We discovered that CXCL10 was highly expressed in T1DM patients. MiR-16-5p, which was lowly expressed in T1DM patients, was verified the upstream regulatory miRNA of CXCL10. The facilitating influence of miR-16-5p up-regulation on the proliferation of HG-induced β-TC-tet cells was reversed by CXCL10 over-expression, while the knockdown results were opposite. More importantly, the restraining impact of miR-16-5p high expression on the apoptosis of HG-induced β-TC-tet cells was accelerated by CXCL10 over-expression. Correspondingly, the level of Bcl-2 was enhanced while the levels of Bax and Cleaved Caspase-3 were lowered by miR-16-5p mimic, which were reversed by CXCL10 over-expression in HG-treated β-TC-tet cells. Conclusions: Our data offered evidence that miR-16-5p implicated in T1DM cell proliferation and apoptosis through targeting CXCL10, which might provide novel therapeutic information for T1DM.

Control of Streptomyces alfalfae XY25T Over Clubroot Disease and Its Effect on Rhizosphere Microbial Community in Chinese Cabbage Field Trials

Clubroot caused by Plasmodiophora brassicae is one of the most destructive diseases in cruciferous crops. Streptomyces alfalfae XY25T, a biological control agent, exhibited great ability to relieve clubroot disease, regulate rhizosphere bacterial and fungal communities in Chinese cabbage, and promote its growth in greenhouse. Therefore, field experiments were carried out to investigate the effects of S. alfalfae XY25T on clubroot and rhizosphere microbial community in Chinese cabbage. Results showed that the control efficiency of clubroot by S. alfalfae XY25T was 69.4%. Applying the agent can alleviate soil acidification; increase the contents of soil organic matter, available nitrogen, available phosphorus, and available potassium; and enhance activities of invertase, urease, catalase, and alkaline phosphatase. During Chinese cabbage growth, bacterial diversity decreased first and then increased, and fungal diversity decreased gradually after inoculation with S. alfalfae XY25T. High-throughput sequencing analysis showed that the main bacterial phyla were Proteobacteria, Bacteroidetes, Acidobacteria, and Planctomycetes, and the major fungal phyla were Ascomycota and Basidiomycota in rhizosphere soil. The dominant bacterial genera were Flavobacterium, Candidatus, Pseudomonas, Stenotrophomonas, Sphingomonas, Flavisolibacter, and Gemmatimonbacteria with no significant difference in abundance, and the major fungal genera were Monographella, Aspergillus, Hypocreales, Chytridiaceae, Fusarium, Pleosporales, Agaricales, Mortierella, and Pleosporales. The significant differences were observed among Pleosporales, Basidiomycota, Colletotrichum, two strains attributed to Agaricales, and another two unidentified fungi by using S. alfalfae XY25T. Moreover, quantitative real-time PCR results indicated that P. brassicae content was significantly decreased after the agent inoculation. In conclusion, S. alfalfae XY25T can affect rhizosphere microbial communities; therefore, applying the agent is an effective approach to reduce the damage caused by clubroot

The Influence of Typical Forest Types on Soil Erosion Resistance in the Water Source Areas of Central Yunnan

In order to clarify the influence of different forest types on soil erosion resistance in water source area of Central Yunnan, with the soils under three different kinds of typical forest in Yizhe watershed as the research object, this paper uses field simulation method and principal component analysis to analyze the soil erosion resistance of three kinds of soils. The results show that there is a significant difference in the shear strength of soil among three types of typical forest, and the size of soil shear strength is in the order of Pinus yunnanensis forest land>mixed broadleaf-conifer forest land>eucalyptus forest land. The difference in the soil erosion coefficient among different forests is not significant, and the soil erosion resistance is highest in mixed broadleaf-conifer forest land (39.0%), followed by eucalyptus woodland (37.0%) and Pinus yunnanensis forest land (24.0%). Under heavy rain intensity and long duration of rainfall, the ability of soil under Eucalyptus ×Pinus yunnanensis mixed forests to resist disintegration is more obvious. Using principal component analysis to analyze soil erosion resistance of soils under three different forests, we get the comprehensive evaluation model for soil erosion resistance: Y=0.763Y1+0.236Y2. The soil erosion resistance is in the order of mixed broadleaf-conifer forest land (0.150)> eucalyptus forest land (0.127)> Pinus yunnanensis forest land (-0.079), indicating that the mixed forests have better water loss and soil erosion control effect than pure forests

Advances and Limitations of Next Generation Sequencing in Animal Diet Analysis

Diet analysis is a critical content of animal ecology and the diet analysis methods have been constantly improving and updating. Contrary to traditional methods of high labor intensity and low resolution, the next generation sequencing (NGS) approach has been suggested as a promising tool for dietary studies, which greatly improves the efficiency and broadens the application range. Here we present a framework of adopting NGS and DNA metabarcoding into diet analysis, and discuss the application in aspects of prey taxa composition and structure, intra-specific and inter-specific trophic links, and the effects of animal feeding on environmental changes. Yet, the generation of NGS-based diet data and subsequent analyses and interpretations are still challenging with several factors, making it possible still not as widely used as might be expected. We suggest that NGS-based diet methods must be furthered, analytical pipelines should be developed. More application perspectives, including nutrient geometry, metagenomics and nutrigenomics, need to be incorporated to encourage more ecologists to infer novel insights on they work

Effects of KN-42 on Growth Performance, Diarrhea and Faecal Bacterial Flora of Weaned Piglets

This research focused on the effects of different doses of Bacillus subtilis KN-42 on the growth performance, diarrhea incidence, faecal bacterial flora, and the relative number of Lactobacillus and Escherichia coli in faeces of weaned piglets to determine whether the strain can serve as a candidate antimicrobial growth promoter. A total of 360 piglets (initial body weight 7.14±0.63 kg) weaned at 26±2 days of age were randomly allotted to 5 treatment groups (4 pens per treatment with 18 pigs per pen) for a 28-day trial. Dietary treatments were basal diet without any antimicrobial (negative control; NC), basal diet supplemented with 120 mg/kg feed of neomycin sulfate (positive control; PC) and basal diet supplemented with 2×109 (L), 4×109 (M) and 20×109 (H) CFU/kg feed of B. subtilis KN-42. During the overall period, average daily gain and feed efficiency of piglets were higher in groups PC, M, and H than those in group NC (p<0.05), and all probiotics and antibiotics groups had a lower diarrhea index than group NC (p<0.05). The 16S rDNA gene-based methods were used to analyze faecal bacterial flora on day 28 of experiment. The result of denaturing gradient gel electrophoresis analysis showed that supplementation of B. subtilis KN-42 to the diet changed the bacterial communities, with a higher bacterial diversity and band number in group M than in the other four groups. Real-time polymerase chain reaction analysis showed that the relative number of Lactobacillus were higher in groups PC and H than in group NC (p<0.05), and the supplemented B. subtilis KN-42 to the diet also reduced the relative number of E. coli (p<0.05). These results suggest that dietary addition of B. subtilis KN-42 can improve the growth performance and gastrointestinal health of piglets

Metagenome-assembled genome reveals species and functional composition of Jianghan chicken gut microbiota and isolation of Pediococcus acidilactic with probiotic properties

Abstract Background Chickens are one of the most widely farmed animals worldwide and play a crucial role in meat and egg production. Gut microbiota is essential for chickens’ health, disease, growth, and egg production. However, native chickens such as Jianghan chickens have better meat and egg production quality than centralized chickens, their intestinal microbial diversity is richer, and the potential gut microbial resources may bring health benefits to the host. Results The bacterial species composition in the gut microbiota of Jianghan chickens is similar to that of other chicken breeds, with Phocaeicola and Bacteroides being the most abundant bacterial genera. The LEfSe analysis revealed significant differences in species composition and functional profiles between samples from Jingzhou and the other three groups. Functional annotation indicated that the gut microbiota of Jianghan chickens were dominated by metabolic genes, with the highest number of genes related to carbohydrate metabolism. Several antibiotic resistance genes (ARGs) were found, and the composition of ARGs was similar to that of factory-farmed chickens, suggesting that antibiotics were widely present in the gut microbiota of Jianghan chickens. The resistance genes of Jianghan chickens are mainly carried by microorganisms of the Bacteroidota and Bacillota phylum. In addition, more than 829 isolates were selected from the microbiota of Jianghan chickens. Following three rounds of acid and bile tolerance experiments performed on all the isolated strains, it was determined that six strains of Pediococcus acidilactici exhibited consistent tolerance. Further experiments confirmed that three of these strains (A4, B9, and C2) held substantial probiotic potential, with P. acidilactici B9 displaying the highest probiotic potential. Conclusions This study elucidates the composition of the intestinal microbiota and functional gene repertoire in Jianghan chickens. Despite the absence of antibiotic supplementation, the intestinal microbial community of Jianghan chickens still demonstrates a profile of antibiotic resistance genes similar to that of intensively reared chickens, suggesting resistance genes are prevalent in free-ranging poultry. Moreover, Jianghan and intensively reared chickens host major resistance genes differently, an aspect seldom explored between free-range and pastured chickens. Furthermore, among the 829 isolates, three strains of P. acidilatici exhibited strong probiotic potential. These findings provide insights into the unique gut microbiota of Jianghan chickens and highlight potential probiotic strains offering benefits to the host. Video Abstrac

Bacterial and Archaeal Water and Sediment Communities of Two Hot Spring Streams in Tengchong, Yunnan Province, China

In Tengchong County, springs with wide physicochemical diversity provide a multitude of niches for extremophilic microorganisms. In this study, eight middle-low temperature spring sites along two continuous small streams with low water flow and slow speed in the fourth geothermal experience area of Rehai scenic spot were chosen, and geochemical characteristics and HTS of the 16S rRNA V4 region were used to analyze the prokaryotic community structure and diversity in the water and sediment of these sites. The effect of environmental factors on the microbial communities was explored via redundancy analysis (RDA). All sediment samples had higher alpha diversity values than the corresponding water samples. Twenty-five phyla were annotated; Euryarchaeota, Crenarchaeota, Aquificae, Thermotogae and Proteobacteria were the dominant phyla, accounting for 95.31% of all prokaryotes, with relative abundances above 5%. Aquificae dominated in water samples, while Euryarchaeota dominated in sediment samples. RDA indicated that temperature was the main factor influencing the microbial communities in the two streams. The study expands the current understanding of the microbiology of Tengchong hot springs and provides a basis for further mining of hot spring microbial and functional gene resources