口服急性毒性试验中Cry1C蛋白对鼠肠道菌群的影响

利用变性梯度凝胶电泳(DGGE)分析不同ICR(institute of cancer researcch)小鼠在口服急性毒性试验中摄入Cry1C蛋白前后其肠道菌群的变化,进而对该抗虫蛋白的食用安全性进行较为深入的研究.DGGE图谱经UPGAMA聚类分析显示:小鼠间菌群结构存在较明显的个体差异;实验期间,对照组小鼠灌胃前后其菌群基本保持稳定;实验组小鼠试验前后肠道菌群变化差异较对照组明显.随着灌胃的停止其变化差异在经历过高峰后会随着停喂时间的延长而减小,且菌相有逐渐恢复灌胃前结构的趋势.从肠道菌相的角度分析可知:该抗虫Cry1C蛋白对小鼠是基本安全的.     

指纹图谱技术在动物肠道微生物多样性研究中的应用

随着分子生物技术的发展,不可培养微生物多样性研究的难题得到了解决。肠道微生物处于特殊的生态环境条件下,分子生物学技术的应用使得肠道微生物多样性的研究进入了一个崭新的阶段。本文主要介绍了基于16S rRNA基因片段的一些肠道微生物研究工作中常用的分子生物学分析方法,主要包括变性梯度凝胶电泳(DGGE),温度梯度凝胶电泳(TGGE),单链构象多态性(SSCP),限制性片段长度多态性(RFLP),放大片断长度多态性(AFLP)和随机扩增多态性DNA(RAPD)等指纹图谱技术。     

The Application of Microbiome Culturomics in Veterinary Medicine

Under laboratory conditions, the number of cultured microorganisms accounts for about 1% of the total number of microorganisms in nature, which limits people's understanding and utilization of 99% of the unknown microorganisms. However, relevant researches show that those "uncultured microorganisms" can be developed and utilized, and the uncultured microorganisms are the main body of the unknown microorganisms. The microbial culturomics explored the application of multiple culture conditions and long-term culture, it was combined with Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) and 16S ribosomal RNA (rRNA) sequencing to identify all kinds of microorganisms on a large scale. At the same time, whole-genome sequencing (WGS) and Metagenomics sequencing technology were used to analyze unknown microorganisms in depth. In this paper, the latest progress of culturomics in the ruminant gastrointestinal tract, poultry cecum, and livestock nasal microflora in recent years was reviewed, and the feasibility of applying the method of microflora culturomics in animal disease prevention and control was discussed. As a new research idea, culturomics has some immature aspects, but its development prospect is very broad. The complementary of microflora culturomics and other research methods have gradually become a breakthrough in the development of veterinary microbiology.     

High Throughput Sequencing to Study the Effect of Chronic Renal Failure on the Diversity and Gene Function Prediction of Gut Microbiota in Dogs

This study aimed to investigate the effect of chronic renal failure(CRF) on the gut microbiota diversity and predict the gene function of the flora. A total of 30 2-year-old dogs were selected and randomly divided into the chronic renal failure model group (CRF, established by renal artery ligation), sham operation control group (Sham) and healthy control group (HCG). All animals were fed normally during the 56 days of test period. The serum creatinine (Scr), blood urea nitrogen (BUN) and urine protein / creatinine ratio (UP/C) were detected regularly during the experiment. The effects of chronic renal failure on the structure, diversity and function of gut microbiota were analyzed according to the result of bacterial 16S rDNA sequencing from fresh feces collected without contamination. The flora markers index (FMI) was constructed based on the principal component Logistic regression analysis of different microbiota in CRF group to predict the development of chronic renal failure. The results showed that: 1) The levels of Scr, BUN and UP/C in CRF group from the start of the 28th day of the experiment were significantly higher than those of HCG group and Sham group (P<0.05), and significantly higher than that of the first day of CRF group (P<0.05). 2) Compared with those before chronic renal failure (CRF group at day 0 and 28), HCG group and Sham group, the Chao 1 diversity and Shannon diversity of gut microbiota in CRF group at day 56 were significantly lower (P<0.05), while the relative abundance of Bacteroidetes, Proteobacteria and Actinobacteria significantly increased and the number of Firmicutes significantly decreased (P<0.05). 3) LEfSe analysis showed that 20 species were enriched in CRF group, mainly including Pseudomonas, Escherichia, Proteus and so on, and most of them had negative correlation with other intestinal bacteria. Functional prediction revealed that genes of those different species were mainly enriched in amino acid metabolism, sugar biosynthesis and metabolism and carbohydrate metabolism in CRF group. The area under ROC curve (AUC) of the FMI constructed with those species enriched was 0.788, which could be used as the intestinal microbial marker for CRF in dogs. In summary, chronic renal failure can reduce the diversity of intestinal microbiota, lead to the imbalance of bacterial structure and change of bacterial function. Moreover, the enriched gut microbiota in CRF group can be used as the intestinal microbial marker of CRF in dogs, and the best prediction effect can be obtained by FMI.     

Research Progress on the Effects of Gut Microbiota and Its Functional Metabolites on Fat Metabolism and Intramuscular Fat Deposition

The meat quality is influenced by many factors, among which intramuscular fat (IMF) is one of the most prominent factors. IMF content is closely related to the tenderness and flavor of meat. Numbers of studies suggested that gut microbiota and its functional metabolites (such as short fatty acids, bile acids, lipopolysaccharides, trimethylamines, tryptophan and their derivatives) played an important role in host fat metabolism. In this review, we present the role of gut microbiota and its functional metabolites in regulating fat metabolism and IMF deposition. This paper would provide new insights and feasible ways through nutrition regulation to increase IMF deposition and improve meat quality.     

Acquired antimicrobial resistance in the intestinal microbiota of diverse cat populations

The aim of this study was to investigate the prevalence of acquired antimicrobial resistance in the resident intestinal microbiota of cats and to identify significant differences between various cat populations. Escherichia coli, Enterococcus faecalis, E. faecium and Streptococcus canis were isolated as faecal indicator bacteria from rectal swabs of 47 individually owned cats, 47 cattery cats and 18 hospitalised cats, and submitted through antimicrobial sensitivity tests. The results revealed that bacteria isolated from hospitalised and/or cattery cats were more frequently resistant than those from individually owned cats. E. coli isolates from hospitalised cats were particularly resistant to ampicillin, tetracycline and sulfonamide. Both enterococci and streptococci showed high resistance to tetracycline and in somewhat lesser extent to erythromycin and tylosin. Most E. faecium isolates were resistant to lincomycin and penicillin. One E. faecalis as well as one E. faecium isolate from hospitalised cats showed 'high-level resistance' (MIC > 500 microg/ml) against gentamicin, a commonly used antimicrobial agent in case of human enterococcal infections. The results of this research demonstrate that the extent of acquired antimicrobial resistance in the intestinal microbiota of cats depends on the social environment of the investigated population. It is obvious that the flora of healthy cats may act as a reservoir of resistance genes.     

溶葡萄球菌素对小鼠肠道菌群结构的影响

【目的】研究溶葡萄球菌素对小鼠肠道菌群结构的影响,为溶葡萄球菌素的食用安全性评估提供参考。【方法】将24只7周龄ICR小鼠随机分为3个试验组和对照组,3个试验组分别提供溶葡萄球菌素含量为1 μg/mL（低剂量）、5 μg/mL（中剂量）和15 μg/mL（高剂量）的饮水10 mL/(只·d),于饮用第0(饮用前),1,3,7,14,21天以及停饮第3,7,14天采集小鼠新鲜粪便样品,采用PCRDGGE技术检测小鼠肠道菌群结构变化。【结果】DGGE图谱显示,各组小鼠试验前后不同时间肠道内的优势菌种相似;经UPGMA聚类分析,与对照组相比,低、中剂量溶葡萄球菌素对小鼠肠道菌群结构影响较小,高剂量溶葡萄球菌素对小鼠肠道菌群结构影响较大;差异性分析结果显示,试验组饮用各阶段带谱与试验前的相似性与对照组相应阶段的相似性差异均不显著（P＞0.05）,溶葡萄球菌素低、中和高剂量组在停饮后第7天带谱与饮用前的相似性分别达到了70.8%,66.9%和63.8%;对图谱中主要条带进行测序和比对分析,结果表明,高剂量溶葡萄球菌素对小鼠肠道中Lactobacillus taiwanensis和1株不可培养细菌有促进作用。【结论】溶葡萄球菌素对小鼠肠道菌群结构的改变具有浓度依赖性,且停饮后小鼠肠道菌群结构有一定程度的恢复。     

Constraints using the liquid fraction from roadside grass as a bio-based fertilizer

Background

Roadside grass cuttings are currently considered a waste product due to their association with road sweepings as contaminated waste, therefore, their potential as a biofertilizer is understudied.

Aim

This study aimed to determine whether grass liquid fraction (GLF) collected from a roadside verge in Maldegem, Belgium, and pressed using a screw press was suitable as a biofertilizer.

Methods

The characterization of the heavy metal content of the GLF was conducted using an ICP-OES. From May to September 2019, a pot experiment was set up using a randomized block design to compare tomato plant growth, yield, and nutrition for GLF-treated plants to two commercial fertilizers and tap water as a control.

Results

The heavy metal content of the GLF was below the maximum permissible concentrations (MPCs) for organic fertilizers as set out by the European Comission fertilizer regulation 1069/2009 and 1107/2009 (European Comission, 2019). However, despite having a fairly well-balanced nutrient content (0.1% N, 0.04% P₂O₅, and 0.2% K₂O), GLF had a negative effect on the growth, root weight, and yield of the tomato plants, killing six out of ten plants. GLF also promoted mold growth in the soil of some plants. Since the GLF was uncontaminated, heavy metal toxicity did not cause the negative effect.

Conclusions

Previous research showed that liquid fractions from some plants negatively affect the growth of others due to allelopathic chemicals; this, together with the stimulation of fungal growth, could have caused the negative effects observed. Future experiments will investigate the herbicidal property of GLF and possible treatments to potentially recover the nutrients contained within the GLF for application as a biofertilizer.