Utilisation de souches bacteriennes selectionees dans l'alimentation des larves de Mytilus galloprovincialis Lmk (Mollusque Bivalve) en elevages experimentauxUse of selected bacterial strains for feeding larvae of Mytilus galloprovincialis Lmk in experimental rearing

The authors studied the possibilities of using several bacterial strains, alive or heat-killed, alone or associated with an algal diet, as food for mussel larvae. Living bacteria offered with an algal culture improved larval growth, whereas the addition of heat-killed bacteria seemed to favour the appearance of an exogenous bacterial infection. Utilization of bacteria without algae sets the problems of their food value under these conditions and the concentration at which they should be used. Therefore, further complementary research will be necessary.     

养殖水体添加嗜酸小球菌对鳜发育过程肠道微生物构成的影响

从鳜(Siniperca chuatsi)出膜仔鱼阶段开始向养殖水体添加5×10~7CFU/L的嗜酸小球菌(Pediococcus acidilactici),利用PCR-DGGE技术对鳜受精卵、出膜仔鱼、仔稚鱼和其开口饵料白鲢仔稚鱼的肠道微生物群落结构组成和多样性进行了分析。DGGE图谱的条带克隆测序结果显示,组成鳜鱼苗各阶段的细菌主要隶属于α-变形菌纲(Alphaproteobacteria)、β-变形菌纲(Betaproteobacteria)、γ-变形菌纲(Gammaproteobacteria)、异常球菌纲(Deinococci)、黄杆菌纲(Flavobacteria)。其中黄杆菌属(Flavobacterium)为鳜受精卵特有细菌;异常球菌属(Deinococcus)、红杆菌属(Rhodobacter)和菊苣假单胞菌(Pseudomonas cichorii)是受精卵和仔鱼共有优势细菌,但在仔稚鱼中没有或只有极少分布;肠杆菌属(Enterobacter)是鳜仔稚鱼和开口饵料白鲢仔稚鱼的共有优势细菌;类志贺邻单胞菌(Plesiomonas shigelloides)和豚鼠气单胞菌(Aeromonas caviae)在对照组仔稚鱼的分布明显比试验组丰富。     

Evaluation of membrane filtration and UV irradiation to control bacterial loads in recirculation aquaculture systems

Ultraviolet (UV) irradiation is commonly used to control pathogen loads in recirculation aquaculture systems (RAS), although these micro-organisms can be shielded by particles in the water, and some species tolerate very high UV doses. The objective of this study was to evaluate membrane filtration (MF) as an alternative, or complimentary, treatment to UV irradiation for pathogen control in RAS, as well as examine the operation and cost of each treatment. In a pilot-scale RAS, both MF and UV were used to treat wastewater for 30 days and water samples were collected biweekly and analysed for culturable bacteria, suspended solids, UV transmittance and other parameters. Bacterial control efficiencies were similar between both MF and UV treatments, which removed 99% of total bacteria and 98% of heterotrophic bacteria, respectively. Surface fouling was negligible for the UV while MF required biweekly cleaning to maintain operation. However, MF had the additional benefit of removing 96% of suspended solids, which resulted in increased UV transmittance. Capital and operating costs of MF were similar to UV, but only when MF treated a fraction of the wastewater compared with UV. We conclude that MF represents a potential complimentary technology to enhance UV irradiation, especially to minimise pathogens in RAS that are shielded by particles or tolerate UV.     

Antibiotic resistance in potentially bacteriocinogenic probiotic bacteria in aquaculture environments

Bacteriocin‐producing bacteria in aquaculture may prevent spreading of potentially pathogenic microorganisms, and could be used as an alternative to the empirical use of antimicrobial drugs, especially for prophylaxis. Bacteriocinogenic bacteria inoculated as probiotics should not carry undesirable traits, such as antibiotic resistance. This study aimed to find potentially bacteriocinogenic bacteria in an aquaculture system and evaluate their antimicrobial susceptibility patterns. Selective cultures for enterobacteria, non‐fermenting Gram‐negative rods and Gram‐positive cocci were obtained from water samples before bacterial isolation and biochemical identification. Overall, 160 representative strains were recovered and for 57 antagonism was observed against selected strains such as Staphylococcus aureus, with antagonism being expressed better on Brain Heart Infusion medium. After exclusion of interfering factors, bacteriocin or bacteriocin‐like substances were suggested to be related to the antagonism observed. Higher drug‐resistant rates were observed among potentially bacteriocinogenic bacteria for different antimicrobials of clinical relevance. Although antibiotic resistance is a global health problem and bacteriocins are attractive alternatives to classical antibiotic even to multiresistant bacteria, the data obtained suggest that bacteriocin‐producing bacteria may harbour resistance genes available for transference in different environments. From the ecological and biotechnological perspective, antimicrobial susceptibility tests must be always performed when prospecting potentially bacteriocinogenic bacteria as probiotic candidates in the environment.     

Drug resistance of bacteria isolated from the intestine of yellowtail cultured with feeds containing various drugs

ABSTRACT: In order to ascertain whether the use of drugs as feed additive for fish may cause drug resistance of bacteria in the intestinal tract, viable counts and bacterial flora in the intestine of yellowtail cultivated with the feeds in the presence or absence of various drugs, drug resistance of the flora were determined. Viable counts in the intestine were lowered remarkably by the administration of each drug, suggesting that bacterial growth in the intestinal tract was inhibited by various drugs. Bacterial flora in the intestine consisted mainly of coryneform bacteria, Staphylococcus and Bacillus regardless of whether drugs were administered or not. In these floras, significant differences could not be observed for their resistance to respective drugs among the isolates from fish cultivated with feeds containing or not containing each drug. In addition, these floras mainly showed resistance to various combinations comprising from one to seven drugs; moreover, the majority of resistant strains were multidrug-resistant.     

两种疾病发生对养殖半滑舌鳎肠道菌群结构的影响分析

为研究疾病发生情况下鱼类肠道菌群的结构变化,运用高通量测序方法分别对感染腹水病和皮肤溃疡病的工厂化养殖半滑舌鳎病鱼及同一养殖系统中健康鱼的肠道菌群结构进行了对比分析。结果发现,养殖半滑舌鳎肠道中的细菌种类要明显少于环境中的细菌种类,仅为养殖池进水和颗粒饲料中细菌种类总数的14.3%~44.7%。在疾病发生后,病鱼肠道中的细菌种类数量仅是同一养殖系统中健康鱼肠道细菌种类数量的45.8%和64.5%,病鱼肠道中细菌多样性下降非常明显。进一步分析肠道菌群结构的组成发现,虽然发病鱼和健康鱼肠道中超过60%的细菌种类不相同,但优势菌群的结构组成差异不明显,丰度最高的前10种细菌种类基本相同,其比例超过肠道相对总菌量的80%。不同疾病的发生对半滑舌鳎肠道菌群结构的影响也不同,腹水病会造成肠道中优势菌株的种类和相对丰度都发生比较明显的改变,而皮肤溃疡病发生前后肠道菌群结构组成几乎没有变化。研究表明,养殖半滑舌鳎的肠道菌群结构相对比较保守,而且并不是所有的疾病种类都会引起肠道菌群结构发生改变。     

Imported ornamental fish are colonized with antibiotic‐resistant bacteria

There has been growing concern about the overuse of antibiotics in the ornamental fish industry and its possible effect on the increasing drug resistance in both commensal and pathogenic organisms in these fish. The aim of this study was to carry out an assessment of the diversity of bacteria, including pathogens, in ornamental fish species imported into North America and to assess their antibiotic resistance. Kidney samples were collected from 32 freshwater ornamental fish of various species, which arrived to an importing facility in Portland, Oregon from Colombia, Singapore and Florida. Sixty‐four unique bacterial colonies were isolated and identified by PCR using bacterial 16S primers and DNA sequencing. Multiple isolates were identified as bacteria with potential to cause disease in both fish and humans. The antibiotic resistance profile of each isolate was performed for nine different antibiotics. Among them, cefotaxime (16% resistance among isolates) was the antibiotic associated with more activity, while the least active was tetracycline (77% resistant). Knowing information about the diversity of bacteria in imported ornamental fish, as well as the resistance profiles for the bacteria will be useful in more effectively treating clinical infected fish, and also potential zoonoses in the future.     

鱼类肠道微生物总DNA的提取

尝试提取鲫(Carassius auratus)肠道菌群总DNA,为研究鱼类肠道菌群结构提供依据。以鲫肠道内容物为样本,用PBS多次洗涤,离心样品,沉淀菌体。使用试剂盒法提取肠道菌群总DNA,电泳结果显示,样品DNA条带明亮,无降解现象,可用于后续分子生物学试验研究;通过设计的细菌通用引物,对其16S rDNA基因进行PCR扩增,得到较清晰的图谱,条带整齐;表明采用该方法提取鱼类肠道微生物群落的DNA较为简单、准确、可行。     

Bacteriological and physico-chemical analysis in a bivalve hatchery: Techniques and preliminary results

The bacteriological and physico-chemical methods used in the survey of a bivalve hatchery are presented and the preliminary results discussed. It was concluded that food, through its contribution of nutrient salts and bacteria, had a major influence on larval culture.The physico-chemical characteristics of sea water were not modified by heat or filtration treatments. The bacteria were not always eliminated from sea water by filtration and were sometimes more numerous after treatment.In effect, the larvae were raised in an artificial environment subject to marked fluctuations which had mechanical and biological origins. This survey must be complemented by a qualitative bacteriological study and the establishment of critical values for the physico-chemical parameters involved in the culture of these bivalve larvae.     

Use of probiotics to control furunculosis in rainbow trout, Oncorhynchus mykiss (Walbaum)

Aerobic heterotrophic bacteria were isolated from the intestinal contents of Atlantic salmon, Salmo salar , rainbow trout, Oncorhynchus mykiss , and turbot, Scophthalmus maximus , on tryptone soya agar and De Man Rogosa and Sharpe agar, of which 11 of 177 (6% of the total) of the isolates were antagonistic to Aeromonas salmonicida . Four of these cultures, which were identified tentatively as A. hydrophila , Vibrio fluvialis , Carnobacterium sp. and an unidentified Gram-positive coccus, were beneficial to fish when fed singly or as an equi-mixture. Feed supplemented with the putative probiotics indicated survival of the organisms in the gastrointestinal tract for 7 days. Feeding with the probiotics for 7 and 14 days led to better survival following challenge with A. salmonicida . There was no indication of serum or mucus antibodies to A. salmonicida , but there was an increased number of erythrocytes, macrophages, lymphocytes and leucocytes, and enhanced lysozyme activity in the fish.     

近江牡蛎养殖水体中细菌产酶能力的研究

为了解近江牡蛎养殖水体中细菌的产酶特性 ,筛选出有应用潜力的有益菌 ,本实验从近江牡蛎养殖水体中分离到 32株菌。革兰氏染色表明 ,10株为革兰氏阳性菌 ,2 2株革兰氏阴性菌。在此基础上研究了它们蛋白酶、脂肪酶、淀粉酶和纤维素酶的产酶能力。结果表明 ,6 1.9% ( 17株 )的菌株能分泌蛋白酶 ,71.9% ( 2 3株 )菌株能分泌脂肪酶或淀粉酶 ,34.4 % ( 11株 )的菌株产纤维素酶。这些产酶菌株均以革兰氏阴性菌为主。在这 32株菌中 ,产四种酶的有 5株菌 ,产三种酶的有 9株菌 ,产两种酶的有 9株菌 ,产一种酶的有 9株菌 ,也就是说 ,所有的菌株都能产酶。由此表明近江牡蛎养殖水体中细菌在该环境物质循环中的重要地位。     

剑尾鱼在检测细菌毒力方面的应用

用１８株从鱼类所分离的细菌攻毒剑尾鱼,结果显示细菌对剑尾鱼的毒力与回归感染或攻毒敏感鱼的毒力结果较为一致,另外,用剑尾鱼感染病原菌的适宜途径为背肌注射,与原代比较,近交高代剑尾鱼个体之间对病原菌的反应较为一致,感染后死亡时间相对集中。水生实验动物剑尾鱼在检测鱼类细菌的毒力方面有较好的应用前景。     

杭州湾南岸滩涂贝类养殖环境中微生物数量分布及其类群

研究了杭州湾南岸慈溪滩涂贝类养殖区的微生物在水平、垂直及季节变化的分布规律。结果表明:3个采样区的异养细菌的数量变动在1.62×103~6.62×105cfu.g-1之间,平均值为4.28×104cfu.g-1,细菌数量在春季和夏季的数量偏低,10月份异养细菌的数量开始上升,并于11月份达到最高,表层高于底层;其中以革兰氏阳性菌比例最高,为主要的优势种,具有陆源性特点。各站位反硝化细菌、氨化细菌和硫酸还原菌的检出率均为100%,都维持在一个较高的水平,整体上数量具有表层高于底层的特点,3种微生物在数量上的季节演替与异养细菌具有类似的结果,均为10-11月达到最大。     

Antibiotic sensitivity of bacteria isolated from the canned eggs of the Californian brine shrimp (Artemia salina)

Bacteria presum ptively classified as Bacillus spp. were isolated from the surfaces of canned Californian brine shrimp eggs. These bacteria were screened for their sensitivity to antibiotics used in the process of hatching marine fish. It was found that where antibiotics were used together they were effective in preventing the growth of these bacteria both at the concentrations applied to the fish hatching process itself and also at lower concentrations. The methods employed to screen the bacteria are discussed.     

獭兔巴氏杆菌病病原的分离与药敏试验

通过对海晏县某兔场病死獭兔无菌采取肝、脾、肺、肾等8份病料,应用常规分离鉴定技术进行病原的分离与鉴定,同时以多杀性巴氏杆菌C47-8株为质控菌株,结果显示,分离菌株与C47-8株间有一定的生化差异,但符合多杀性巴氏杆菌的生化特性。最后结合流行病学、临床症状、病理变化,确诊为獭兔巴氏杆菌病,并对分离菌株进行了药物筛选,选取敏感药物进行交替治疗,取得了较好的效果。     

河口水体中的菌群分析及其脱氮能力的研究

从天津渤海湾三个河口六个站位采集水样,用Zobell2216E培养基分离共得到92株异养细菌,其中G+52株,G-40株。对其形态和生理生化性状进行分析,初步研究结果表明,他们主要分属葡萄球菌、微球菌、不动杆菌、莫拉氏菌、芽孢杆菌、弧菌、产碱杆菌、肠杆菌、假单胞菌。选择假单胞菌属、葡萄球菌属、产碱杆菌属、芽孢杆菌属,研究其在含氮量较高的所采河口水样中(无机氮含量>0.3 mg/L)的脱氮能力,于0、3、6、9、12、15 h不同时间测定水样中的亚硝酸盐氮、硝酸盐氮、氨氮含量。实验结果表明经过9 h的培养测定,水样中的无机氮含量明显降低,但9 h以后除假单胞菌的水样无机氮含量继续降低之外,其他三个菌种水样无机氮含量开始上升。确定四个菌种的最佳脱氮时间为9 h,且芽孢杆菌的脱氮能力最强。     

Bacterial diversity studied by next‐generation sequencing in a mature phototrophic Navicula sp‐based biofilm promoted into a shrimp culture system

The bacterial diversity of a phototrophic biofilm used as nutrient‐recycler into a shrimp aquaculture system and as direct food‐source for shrimp was studied by next‐generation sequencing, considering the 16S rDNA and metagenomics sequence classification using exact alignments. Biofilm was promoted by the modification of cabon:nitrogen ratio and by the addition of the diatom Navicula sp. as promoter. Results revealed a wide diversity of bacteria thriving into the biofilm; most of the bacteria detected in the biofilm belonged to the Proteobacteria (47%), Chlamydiae/Verrucomicrobia (11%), Bacteriodetes (8%), Planctomycetes (5%) Phylum. Species involved in the decomposition of organic matter, nitrogenous‐ and sulfurous metabolites were detected; moreover, filamentous species known as biomass‐bulking enhancers and producers of adhesin‐like compounds were also detected. Surprising results were also obtained by detecting both, strictly anaerobic and aerobic bacteria involved into the metabolism of nitrogenous compounds. Other species not belonging to the marine environment were also detected, but their role is unclear. Finally, the detection of most of these species may constitute a first case report for a phototrophic biofilm. The results suggest an important role of bacteria in this type of biofilm and a complex microbial‐interaction network.     

健康和患病稚黑鲍中微生物产胞外酶的分析比较

2005年初自福建漳浦鲍鱼场患病以及健康稚黑鲍中各分离到27和56株异养细菌,在此基础上研究了它们产胞外蛋白酶、淀粉酶、明胶酶、磷脂酶及溶血能力。结果表明,虽然来自健康鲍的菌株产胞外酶的数量明显大于来自病鲍的菌株,但来自病鲍的菌株具有强分泌胞外酶能力的比例明显大于来自于健康鲍的菌株。综合考虑每一种胞外酶的潜在致病作用,以及菌株分泌多种胞外酶的能力,最后确认10株菌为潜在的致病菌。     

Clinical and bacteriologic studies of the frequency of mastitis during and after parturition in heifers lactating for the first time

The udders of 100 heifers were examined for clinical changes during parturition. Of each quarter colostral samples were taken and analyzed bacteriologically. 35 heifers (13.5% of the quarters) showed clinical changes of the quarters and/or of colostral samples. 14 of these animals (4.75% of the quarters) suffered from cellulitis-like mastitis. 13 (5.25% of the quarters) had acute catarrhalic mastitis and eight heifers (3.5% of the quarters) showed acute galactophoritis. In all milk samples of animals with pathological changes of the udder or of colostrum the causing bacteria could be identified. Besides the group of 35 heifers with clinical lesions udder-pathogenic bacteria were found in samples of 24 apparently healthy animals (20.5% of the quarters) during parturition. However, these animals failed to show clinical signs of inflammation at least until the end of this study (10 days post partum). Isolated bacteria included facultative and obligate pathogenic species, 70% of which were gram-positive cocci (i.e. Streptococcus [Sc.] uberis, Sc. dysgalactiae, Sc. agalactiae, Staphylococcus aureus and other staphylococci) versus 30% of gram-negative bacteria (i.e. Escherichia [E.] coli, Klebsiella and other coliform species. In-vitro-susceptibility tests showed best effectiveness of Penicillin against Sc., Cloxacillin against staphylococci and Gentamycin and Polymyxin B against coliform bacteria. Possible pathways, leading to bacterial infection of first lactating heifers during parturition, are discussed. Pathways of infection seem to be of lesser importance for the development of apparent, sometimes severe mastitis with coliform bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)     

Purification and characterization of a 630 kDa bacterial killing metalloprotease (KilC) isolated from plaice Pleuronectes platessa (L.), epidermal mucus

Antibacterial chemicals in the mucus of fish such as lysozyme, lectins, peptides and proteases provide an efficient first line of defence against pathogens. This study shows that there are at least three antibacterial proteins in plaice skin mucus in addition to lysozyme. One of these proteins is responsible for approximately 74% of the antibacterial activity and is a 630 kDa protease complex designated KilC (bacterial killing metalloprotease C). Purified KilC kills the bacteria Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Pseudomonas aeruginosa efficiently. The protease activity of KilC is dependent upon the divalent cation Mg(2+) and shows pH dual optima of 5.0 and 8.0. The enzyme has a temperature optimum of 25 degrees C and is made up of at least five different sized peptides. Studies with protease inhibitors show that the catalytic site of KilC may be cysteine- or serine protease-like. KilC may kill bacterial cells by acting directly upon the bacteria or by producing low molecular weight bioactive compounds such as peptides.