Mycobacteriosis caused by Mycobacterium marinum in reared mullets: first evidence from Sardinia (Italy)

Mycobacterium marinum is a slow‐growing non‐tuberculous mycobacterium, and it is considered the most common aetiologic agent of mycobacteriosis in wild and cultured fish. The diagnosis is principally made by histology when positive Ziehl–Neelsen stain granulomas are detected. The aim of this study was to investigate the occurrence of mycobacteriosis in extensively cultured Mugilidae of two lagoons (Cabras and San Teodoro) from Sardinia by the use of histology, microbiology, PCR and DNA sequencing. Nine of 106 mullets examined were affected by mycobacteriosis, and the spleen was the most affected organ. The histology detected higher rate (100%) of infection in spleen than the culture and PCR (75% and 62.5%, respectively). The sequencing of hsp65 gene identified M. marinum as the primary cause of mycobacteriosis in the mullets examined. Mullets affected by mycobacteriosis were mainly fished in the San Teodoro lagoon characterized by critical environmental conditions. Histology remains the most common method in detecting fish affected by mycobacteriosis, and PCR‐based methods are essential for species identification. Our finding are worthy of attention because mycobacteriosis caused by M. marinum in reared mullets was evidenced for the first time in Sardinia, suggesting that this disease may be underestimated also in other cultured fish species.     

基于高通量测序技术分析石蝉草内生细菌多样性

以石蝉草根、茎、叶为材料,采用Illumina Mi Seq高通量测序技术研究石蝉草内生细菌多样性。结果表明,根、茎、叶样品共测得167个OTUs,归属于11个门,分别为变形菌门、Ignavibacteriae、芽单胞菌门、梭杆菌门、厚壁菌门、异常球菌-栖热菌门、绿弯菌门、拟杆菌门、放线菌门和酸杆菌门,各样品的菌群组成和多样性虽有差异,但以变形菌门为优势菌群,约占62.48%~93.24%。在各样品菌群丰度最高的10个OTUs中,叶部优势群落芽孢杆菌属、根部特有群落粘球菌目与石蝉草中具有抗炎、抗肿瘤活性的物质存在潜在相关性。     

Soil sterilization effects on root growth and formation of rhizosheaths in wheat seedlings

Sterilized soils are frequently used in experiments related to soil biology. Soil sterilization is known to alter physicochemical characteristics of soil, plant growth and community structure of the newly developed bacterial population. However, little information exists regarding soil sterilization effects on belowground processes mediated through root–microbe–soil interactions, e.g., development of rhizosheaths which significantly promote the plant growth under stress environments. The present study was conducted to elucidate effects of soil sterilization on wheat root growth and formation of rhizosheaths in relation to chemical changes caused by soil sterilization and the proportion of expolysaccharide (EPS)-producers in bacterial population recolonizing the sterilized soils. Wheat plants were grown for two weeks under greenhouse conditions either in the unsterilized soil or in soils sterilized by autoclaving (121 °C, 1 h) or by gamma (γ)-irradiation (50 kGy). While soil sterilization had no effect on the release of macronutrients, both sterilization procedures significantly increased the electrical conductivity, water-soluble carbon and DTPA-extractable Mn. Seedlings grown in sterilized soils produced higher root biomass and the rhizosheath soil (RS) mass as compared to those grown in the unsterilized soil. Soil sterilization also increased the root length, surface area, volume and number of tips. In bulk soil, RS and on roots, the proportion of EPS-producers in the total bacterial population was higher in sterilized treatments than in the unsterilized. Amending the unsterilized soil with glucose-C increased the root biomass, whereas adding Mn II increased the RS mass. The results showed that soil sterilization by autoclaving or γ-irradiation increases the root growth and RS mass of wheat seedlings. The water-soluble C and DTPA-extractable Mn released upon sterilization, and the increased proportion of EPS-producers in the bacterial population recolonizing the sterilized soils were involved in the observed effects. The results may have implications in studies using autoclaved or γ-irradiated soils to investigate soil–plant–microbe interactions and signify the need to account for intrinsic stimulatory effects of soil sterilization.     

Rate-specific responses of prokaryotic diversity and structure to nitrogen deposition in the Leymus chinensis steppe

Serious nitrogen (N) deposition in terrestrial ecosystems causes soil acidification and changes the structure and function of the microbial community. However, it is unclear how these changes are dependent on N deposition rates, other factors induced by N (e.g., pH), and their interactions. In this study, we investigated the responses of soil prokaryotic community structure and stability after a 13-year N addition in the semi-arid Leymus chinensis steppe in Inner Mongolia, China. Our results demonstrated that the prokaryotic community structure changed at the low N addition rate of 1.75 g N m⁻² yr⁻¹; however, dramatic changes in microbial abundance, respiratory quotient, and prokaryotic diversity occurred at N addition rates of more than 5.25 g N m⁻² yr⁻¹ when the soil pH dropped below 6.0. The two patterns indicated the difference in driving forces for different microbial properties. The N-driven and pH-driven processes are likely the most important mechanisms determining the responses of bacterial community to N. Some copiotrophic/oligotrophic bacteria, e.g., Proteobacteria and Acidobacteria, changed their relative abundances with the N addition continuously even at a low rate, indicating that they were more sensitive to N directly. Some bacterial groups significantly changed their relative abundance at a high N addition rate when pH dropped below 6.0, e.g., Verrucomicrobia and Armatimonadetes, indicating that they were more sensitive to pH below 6.0. N addition altered the prokaryotic community structure through enrichment of copiotrophic bacteria (species adjustment) at low N addition rates and through enrichment of nitrophilous taxa and significant loss of diversity at high N rates. The results also demonstrated that a high N addition diminished the stability of the prokaryotic community structure and activity through reduction in species diversity and bacterial interaction. Overall, this study supported the hypothesis that the responses of prokaryota to N were dependent on deposition rates, and N-driven and pH-driven processes were the important mechanisms to control the shift of the prokaryotic community.     

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。     

固态热带假丝酵母菌菌剂的发酵工艺研究

试验在单因素优化试验的基础上,采用正交试验进一步对热带假丝酵母菌固态发酵工艺进行研究,包括发酵时间、接种量、料水比、碳源、氮源、无机盐等因素,最终确定热带假丝酵母菌的固态发酵工艺。发酵条件:在自然pH值条件下,发酵温度30℃、发酵时间48 h、接种量11%、料水比1∶0.75。培养基成分:麸皮与甘油8.5∶1.5 (g/g)、尿素0.5%、磷酸二氢钾0。依照此工艺进行发酵,热带假丝酵母菌活菌数最高可达32.94×10⁸个/g。