Changes in the gut microbiome of the Chinese mitten crab (Eriocheir sinensis) in response to White spot syndrome virus (WSSV) infection

Intestinal microorganisms play important roles in maintaining host health, but their functions in aquatic animal hosts have yet to be fully elucidated. The Chinese mitten crab, Eriocheir sinensis, is one such example. We attempted to identify the shift of gut microbiota that occurred in response to infection of white spot syndrome virus (WSSV), an emerging viral pathogen in the crab aquaculture industry. The microbiota may exert some control over aspects of the viral pathogenesis. We investigated the changes in composition and structure of the crab gut microbiome during various WSSV infection stages of 6 h post‐infection (hpi) and 48 hpi, using a 16S rRNA approach on the MiSeq Illumina sequencing platform. Four phyla (Firmicutes, Proteobacteria, Tenericutes and Bacteroidetes) were most dominant in the gut of E. sinensis regardless of the WSSV infection stages. However, further analysis revealed that over 12 bacterial phyla, 44 orders and 68 families were significantly different in abundance at various states of WSSV infection. Several intriguing aspects of E. sinensis gut bacteria that had not been previously reported were also uncovered, such as class Mollicutes was dominant here, but absent in crabs from Yangtze River estuary and Chongming Islands. Overall, this study provided the first evidence that changes in gut microbiome were closely associated with the severity of WSSV infection and that indicator taxa could be used to evaluate the crab health status.     

Effect of dietary GroBiotic®‐A supplementation as a prebiotic on the intestinal microflora,growth performance,haemato‐serological parameters,survival rate and body composition in juvenile beluga (Huso huso Linnaeus, 1754)

The objectives of the present study were to investigate of GroBiotic^®‐A (GBA) on growth, autochthonous intestinal microbiota and haemato‐serological parameters of beluga juvenile. A total of 180 fish (40.82 ± 5.81 g) were fed diets containing graded levels of GBA (0, 5, 10 and 20 g kg^?1 diet) for 8 weeks. No significantly differences in body composition, total viable aerobic bacteria, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration, neutrophils, eosinophils, monocytes, albumin, glucose, triglyceride, cholesterol, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase were detected between treatment groups. The fish fed on 10 and 20 g kg^?1 GBA significantly showed higher lactic acid bacteria, final weight, body weight increases, weight gain and lower feed conversion ratio compared with the control and 5 g kg^?1 groups. The group fed 20 g kg^?1 GBA showed a highly significant difference in condition factor, survival rate, final length, total red and white blood cells, lymphocytes, haematocrit, haemoglobin, total protein, total immunoglobulin and lysozyme activity. The specific growth rate of the treatment groups was significantly elevated compared with the control groups. These results indicated that GBA at level 20 g kg^?1 improved growth, welfare and survival of beluga juvenile.     

In vitro and in vivo selection of probiotic purple nonsulphur bacteria with an ability to inhibit shrimp pathogens: acute hepatopancreatic necrosis disease‐causing Vibrio parahaemolyticus and other vibrios

Shrimp cultivation has been faced with huge losses in productivity caused by infectious shrimp pathogenic vibrios, especially Vibrio parahaemolyticus that causes acute hepatopancreatic necrosis disease (AHPND). Hence, purple nonsulphur bacteria (PNSB) were isolated from shrimp ponds for investigating their abilities to control shrimp pathogenic Vibrio spp. and their use as probiotics for sustainable shrimp cultivation. Based on their probiotic properties, strains S3W10 and SS15 were selected because of their strong abilities to produce amylase, gelatinase and vitamin B12. However, only three PNSB strains (SS15, TKW17 and STW181) strongly inhibited V. harveyi_KSAAHRC and V. vulnificus_KSAAHRC including V. parahaemolyticusAHPND strains by secreting antivibrio compounds. Four selected PNSB also grew in the presence of pancreatic enzymes, and they were identified as Rhodobacter sphaeroides for strains S3W10, SS15 and TKW17 and Afifella marina for strain STW181. The effects of a mixed culture were also investigated as follows: T1 (S3W10 + SS15), T2 (S3W10 + TKW17) and T3 (S3W10 + STW181) on postlarval white shrimp (Litopenaeus vannamei) for 60 days by comparison with a control. All three probiotic PNSB sets significantly improved the digestive enzyme activities and shrimp growth with their proliferation in shrimp gastrointestinal tract although the shrimp survival was not significantly different. They also significantly reduced the cumulative mortality of shrimp exposed to a virulent AHPND strain (V. parahaemolyticusSR2). This is the first to conclude that selected probiotic PNSB strains have great potential to be used for shrimp cultivation to control vibrios including AHPND strains.     

抑霉乳酸菌的筛分及其抑菌特性研究

试验旨在获取生物抑霉的优良菌株以及为质量安全青贮饲料生产提供高效菌剂。从青贮饲料、酸奶以及奶豆腐中分离纯化出乳酸菌和霉菌,并且通过双层平板法对具有抑制霉菌活性的乳酸菌菌株进行筛选,测定其上清液抑霉效果;采用蛋白酶处理和高温处理法分析乳酸菌抑霉的有效成分。结果表明:分离筛选的乳酸菌和霉菌经鉴定为植物乳杆菌和黄曲霉菌,且植物乳杆菌对黄曲霉菌的生长有明显抑制作用。蛋白酶处理对乳酸菌无细胞上清液的抑菌活性有不同影响,而加热处理并不改变其抑菌效果。研究表明,植物乳杆菌对黄曲霉菌生长有明显的抑制作用,推测其抑菌活性物质为蛋白质、肽类等,并且这些抑菌活性物质的热稳定性较高。     

乳酸菌代谢研究进展

乳酸菌代谢研究已进入一个新的时代，本文主要阐述目前乳酸菌在糖代谢、柠檬酸代谢及蛋白质代谢等方面的研究，以及基于下一代测序技术的基因组学、比较基因组学和宏基因组学等技术手段在乳酸菌代谢研究中的应用，旨在为现代乳酸菌代谢研究提供理论支持。     

Clostridium butyricum as probiotic for promoting growth performance,feed utilization,gut health and microbiota community of tilapia (Oreochromis niloticus × O. aureus)

The feeding trial was conducted to evaluate the potentials of Clostridium butyricum in the diet of tilapia. Fish (~14 g) were fed with basal diet supplemented with 0 (Control), 0.5 (C‐1), 1 (C‐2), 2 (C‐3), 4 (C‐4) and 8 (C‐5) g/kg commercial probiotic‐containing C. butyricum (1.5 × 10⁸ CFU/g) for 8 weeks. The results showed that weight gain significantly increased, and feed conversion ratio decreased in the C‐2, C‐3 and C‐4 groups (p < .05). The protein retention (except C‐1 group), lipid retention and apparent digestibility coefficient (ADC) of dry matter in probiotic supplementation groups were significantly enhanced, and ADC of protein in the C‐4 group was also improved (p < .05). The supplementation of probiotic significantly increased villus height in anterior intestines and reduced the numbers of intestinal Escherichia coli (p < .05). High‐throughput sequencing showed that top three phyla namely Planctomycetes in all probiotic‐containing groups, Proteobacteria in the C‐1 and C‐2 groups and Chloroflexi in the C‐3 group had higher level than the NC group. The cumulative mortality was reduced by dietary probiotic after challenging with Aeromonas hydrophila (p < .05). In conclusion, C. butyricum can be supplemented at 1–2 g/kg feed for promoting the growth, feed utilization, gut health and microbiota of tilapia.     

A bacterium with close genetic identity to Pseudomonas mandelii associated with spring fish kills in wild bluegill Lepomis macrochirus Rafinesque and pumpkinseed sunfish Lepomis gibbosus (Linnaeus)

Pseudomonas fluorescens are known bacterial pathogens in fish. The P. fluorescens group contains at least nine different bacterial species, although species from fish have rarely been differentiated. Two isolated fish kills affecting wild bluegills, Lepomis macrochirus Rafinesque, and pumpkinseed sunfish, Lepomis gibbosus (Linnaeus), occurred in the spring of 2015 during cool water temperatures (12.5°C–15.5°C). Disease signs included severe bacteraemia with rare gross external signs. Pure bacterial cultures isolated from kidneys of all affected fish were identified as P. fluorescens using the API 20NE system, while no bacteria were isolated from asymptomatic fish. To further identify the species of bacterium within the P. fluorescens complex, genetic analysis of the 16S rRNA, rpoD and gyrB genes was conducted. DNA sequences of bacterial isolates from both mortality events were identical and had close identity (≥99.7%) to Pseudomonas mandelii. Although likely widespread in the aquatic environment, this is the first report of a bacterium closely resembling P. mandelii infecting and causing disease in fish. The bacterium grew at temperatures between 5°C and 30°C, but not at 37°C. It is possible that infections in fish were a result of immunosuppression associated with spring conditions combined with the psychrotrophic nature of the bacterium.     

外源溶磷菌对不同土壤条件下大豆生长特性的影响

为了明确溶磷菌在多种土壤中的应用效果,探讨了两株溶磷菌对黑土及盐碱土种植大豆的促生效应。采用盆栽试验分析了不同处理对大豆光合作用、渗透调节物质含量、生物量、氮磷吸收及产量的影响。结果表明:加菌处理可增加植株Pn,显著降低R6、R8两时期叶片可溶性蛋白、可溶性糖及游离氨基酸含量,增加R2~R8时期大豆株高和根干重,混合土壤中加菌处理也可提高植株地上部分氮、磷及植株全磷、全氮含量。添加wj1和混合菌处理的植株干重优于添加wj3的处理。黑土加混合菌可显著提高大豆R2期叶片WUE,增加V3、R2和R8这3个时期大豆地上、地下部分氮、磷及植株全氮、全磷的含量。黑土加wj1与混合土壤加菌处理的大豆株荚数、总粒数、单株粒重及百粒重均高于未加菌处理。添加外源溶磷菌,对黑土及盐碱土栽培大豆均有不同程度的促生效果,能明显增强大豆在混合盐碱土壤中的适应性,增加植株生物量、氮磷吸收量和产量,且混合菌处理对大豆的促生效应优于wj1及wj3单一菌株处理。     

生防细菌复配对苹果主要病原真菌抑菌活性筛选及其稳定性

采用生长速率法测定4种生防细菌及其复配对5种苹果主要病原菌的抑制效果，并将筛选出的最佳复配生防细菌发酵液进行稳定性测定。结果表明，单一生防细菌发酵液中多粘类芽孢杆菌FS-1206对苹果树腐烂病菌的抑菌效果最好，为78.19%；当多粘类芽孢杆菌FS-1206与解淀粉芽孢杆菌TS-1203复配比例为 2∶1时，其对苹果树腐烂病菌的抑菌率高达86.64%，抑菌效果显著优于单一生防细菌发酵液。稳定性测定结果表明，复配生防细菌（FS-1206∶TS-1203= 2∶1）发酵液在紫外照射、高温和强酸强碱环境条件下，其抑菌活性均具有良好的稳定性。