硝化毛球和底沙对硝化细菌脱氮效果的影响

本实验模拟工厂化养殖模式建立养殖水体净化装置,研究硝化毛球和底沙对硝化细菌净化效果的影响,结果表明:装载硝化毛球、铺设底沙和只投加硝化细菌制剂的三个实验组对养殖水体水质具有一定的净化效果,氨氮、亚硝氮等指标均低于空白组。其中装载硝化毛球的实验组氨氧化细菌、亚硝酸盐氧化细菌可在短时间大量生长繁殖,形成优势,使养殖池氨氮、亚硝酸盐浓度维持在较低水平;铺设底沙的实验组对硝化细菌净化水质效果影响不大。装载硝化毛球的实验组,水质最清澈,无异味,养殖池底部无残渣碎屑,青虾生长状况良好,增重最多。     

四联活菌制剂对养殖水体中氨氮及亚硝酸盐的降解

利用四联活菌制剂,在室内进行了对养殖池塘水体中氨氮及亚硝酸盐的降解试验.结果表明,光合细菌、纳豆芽孢杆菌、乳酸菌、硝化细菌具有较好的氨氮、亚硝酸盐降解性能,随着添加质量浓度的增加,氨氮、亚硝酸盐的去除率增加;各菌株氨氮降解能力依次为:乳酸菌＞光合细菌＞硝化细菌＞纳豆芽孢杆菌;亚硝酸盐降解能力依次为:硝化细菌＞纳豆芽孢杆菌＞光合细菌＞乳酸菌.四联活菌制剂对养殖水体中氨氮及亚硝酸盐降解试验结果表明,乳酸菌、光合细菌、硝化细菌、纳豆芽孢杆菌的协同作用对氨氮、亚硝酸盐的降解效果更显著、快速.当制剂添加量分别为1.5、3.0、4.5 kg/hm~2时,5 d氨氮的去除率分别为52%、80%、74%,亚硝酸盐的去除率接近100%,结果均显著高于添加同剂量单一菌株时的氨氮、亚硝酸盐的去除率.     

电气石对硝化菌生长和生物膜形成、熟化的影响

在培养基中添加电气石培养硝化菌,研究电气石对硝化菌生长的影响,并在此基础上将电气石添加到普通陶粒(CM)原料中制备了功能性陶粒(FCM),通过测定水中氨氮、亚硝酸盐氮和硝酸盐氮的含量变化,比较功能性陶粒和普通陶粒两种载体上生物膜的生长状况。结果表明:添加电气石的培养基中的亚硝化细菌和硝化细菌数量明显高于未添加电气石的对照组;FCM上的生物膜熟化过程对氨氮的去除率在第14 d趋于稳定,硝酸盐氮含量从第12 d逐渐升高,分别比CM早7 d和6 d,能较早发挥生物硝化功能。     

微生物对水产养殖水质调控作用研究

正本实验研究了光合细菌、放线菌、枯草芽孢杆菌三种细菌优化配比成的复合功能菌去除养殖水体有机氮效果。结果表明,光合细菌、放线菌、枯草芽孢杆菌混合培养生态制剂能有效去除养殖水体中的有机氮,对高浓度的氨氮、亚硝酸盐氮的去除率可达99.6%和94%。     

两种斑节对虾养殖模式水质净化效果与细菌群落结构的比较分析

为了提高对虾养殖系统水质净化能力,改善对虾养殖水环境,利用红糖+枯草芽孢杆菌和聚氨酯填料+硝化细菌分别构建生物絮团和内循环斑节对虾(Penaeus monodon)养殖系统,比较两个系统在对虾标苗和养殖阶段的水质净化效果和细菌群落结构,从微生物学层面探究其水质净化机理。将体长(0.5±0.1)cm的5日龄仔虾养殖在1 m³水体的帆布池中,标苗期密度为5 000 ind.·m^-3,养殖期降为400 ind.·m^-3。标苗结束时,各帆布池底铺约3 cm厚的砂子。生物絮团系统中加活化后的枯草芽孢杆菌,每天一次性加50%日投饲量的红糖,每隔7 d加0.5 L活化后的芽孢杆菌,每隔14 d换水20%。内循环系统中悬挂已挂膜的聚氨酯填料包,其上表面浸没在水中,内部放一个气石。每个系统设2组平行,各阶段养殖周期分别为20 d和40 d。实验结束时,采集生物絮团系统水样、底砂和内循环系统生物膜、水样及底砂的细菌样品,提取DNA,利用Illumina Mi Seq平台对DNA样品进行高通量测序。结果显示,生物絮团系统氨氮和亚硝酸盐氮浓度波...     

循环水养殖系统生物滤池细菌群落的PCR-DGGE分析

通过模拟实验对循环水养殖系统中不同初始NH ₄N浓度的生物滤池中生物膜上和水中的细菌数量及群落种类组成进行了研究。对成熟生物膜及水体样品中的异养菌、氨氧化菌、亚硝酸盐氧化菌的培养计数结果表明,随着生物滤池初始氨氮浓度增大,除异养细菌数量逐渐下降外,生物膜上的氨氧化菌和亚硝酸盐氧化菌数量呈逐渐增加趋势,且均高出水样3~4个数量级;同时对上述样品的16S rRNA基因片段的PCR扩增产物进行变性梯度凝胶电泳(DGGE)分析及其序列同源性分析的结果表明,生物膜和水中都有较高的细菌多样性,同一初始氨氮浓度的滤池中生物膜上的细菌多样性高于水中的。生物滤池中的细菌主要由拟杆菌门的黄杆菌纲和变形菌门的α-、β-、γ-变形菌纲的15种细菌组成。生物膜上的优势菌包括奥雷氏菌属、湖饲养者菌属、泥滩杆菌属、沉积杆菌属、雷辛格氏菌属、冷蛇形菌属和亚硝化单胞菌属等;水体中的优势菌则有明显差异,主要有蛋黄色杆菌属、Nautella,玫瑰杆菌属和一种硫氧化菌等。初始氨氮越高的滤池中,亚硝化单胞菌属的细菌在生物膜上所占比例越高,逐渐成为优势菌之一。实验证实,挂膜初期,提高水体中初始氨氮浓度,有利于硝化细菌的富集和固着,提高生物滤池的除氮效率。     

枯草芽孢杆菌改善水质提高凡纳滨对虾幼体抗逆性的研究

投放不同浓度梯度的枯草芽孢杆菌于凡纳滨对虾幼体养殖环境中,监测不同枯草芽孢杆菌使用浓度下与养殖环境相关的水质因子（氨氮、亚硝酸盐氮、化学需氧量）的变化;观察不同枯草芽孢杆菌使用浓度下凡纳滨对虾幼体对人工制造的胁迫环境的抗逆性;观测不同枯草芽孢杆菌使用浓度下凡纳滨对虾幼体的成活率与体重增长率。研究枯草芽孢杆菌对水质改善和凡纳滨对虾幼体抗逆性的影响。结果表明,枯草芽孢杆菌能显著（P〈0.05）降低化学需氧量、氨氮含量,抑制亚硝酸盐氮的产生;当枯草芽孢杆菌投放浓度为1.25×10^4cfu/ml时,水体中化学需氧量、氨态氮、亚硝酸盐氮含量均值比对照组分别降低了65.30%、59.70%、88.64%,成活率比对照组提高了10.00%,体重增长率是对照组的2.44倍;当枯草芽孢杆菌使用浓度为1.25×10^4～1.25×10^6cfu/ml时,对虾抗逆性显著（P〈0.05）增强,对虾在氨氮、亚硝酸盐氮、高锰酸钾、甲醛胁迫下24小时的成活率均值分别比对照组提高了16.94%、24.44%、44.17%、18.61%。     

生物絮凝技术处理水产养殖用水效果的初步研究

在水产养殖中应用生物絮凝技术(BFT),可以将养殖过程中产生的残饵和粪便转化为可被部分养殖对象重新摄食的絮体饵料,而且在絮凝体形成过程中对水体氨氮等物质的去除有明显作用。试验在自建的生物絮凝反应器中分别接种活性污泥和枯草芽胞杆菌(Bacillus subtilis),研究絮凝体形成过程中主要水质指标的变化情况。结果表明:接种活性污泥的装置中,氨氮、亚硝酸盐氮的去除率分别为72.25%、94.04%;接种枯草芽孢杆菌的装置中,氨氮、亚硝酸盐氮的去除率分别为81.53%、97.89%;同时接种活性污泥和枯草芽孢杆菌的装置中,氨氮、亚硝酸盐氮的去除率分别为40.85%、63.19%;对照装置中不接种任何物质,氨氮、亚硝酸盐氮的去除率分别为11.41%、70.56%。分别接种活性污泥和枯草芽孢杆菌的装置在去除氨氮、亚硝酸盐氮方面较好。试验装置中所形成的絮体颗粒直径在0.1~1.0 mm,粒径大小适合作为部分养殖鱼类稚鱼期的开口饵料。     

不同基质构建海水养殖系统硝化功能的比较分析

为比较不同基质构建海水养殖系统硝化功能的强弱,选取纤维毛球、陶粒、螺旋式生物绳等7种基质,其中陶粒、流化床填料、纤维毛球采取不同放置方式,共建立14个模拟海水养殖系统,比较不同基质硝化功能建立过程以及同种基质不同放置方式对氨氮和亚硝氮的去除效果。结果表明,单位体积珊瑚骨的氨氧化活性和亚硝酸盐氧化活性高于其他载体,在氨氮初始浓度20 mg/L条件下,氨氮和亚硝氮降解至检测不出分别需要3 d和11 d,而纤维毛球、陶粒、螺旋式生物绳、流化床填料、丝带内芯悬浮球、海绵内芯悬浮球硝化系统的建立分别需要18、26、30、25、27和22 d。纤维毛球100目筛绢悬挂、陶粒网兜悬挂、流化床填料100目筛绢悬挂优于其他放置方式,其中纤维毛球100目筛绢悬挂硝化功能建立时间为18 d,效果最优,流化床填料100目筛绢悬挂、陶粒网兜悬挂硝化系统建立分别需要21、24 d。     

墨瑞鳕循环水养殖系统中不同生物膜反应器水处理效率及微生物群落对比分析

固定床生物膜反应器(fixed-bed biofilm bioreactor, FBBR)和移动床生物膜反应器(moving- bed biofilm reactor, MBBR)在养殖水体氨氮(NH4+-N)和亚硝酸氮(NO2–-N)污染控制中已有较为广泛的研究,然而相关研究大多是在实验室完成的,目前尚缺乏实际生产的循环水养殖系统(recirculating aquaculture system, RAS)中FBBR和MBBR水体净化效能的对比研究。因此,本研究将FBBR (弹性毛刷滤料)和MBBR (PVC多孔环滤料)并联接入实际生产的墨瑞鳕(Macculochella peeli) RAS中,实现二者的同步连续运行(35 d),考察了其出水水质变化和微生物群落结构。出水水质变化表明,FBBR和MBBR中氨氧化能力的形成快于亚硝氮氧化能力,硝化能力渐趋成熟,可以有效控制养殖水体中的NH4+-N和NO2–-N浓度,但会导致养殖水体中硝酸氮(NO3–-N)积累和pH下降;单因素方差分析表明,FBBR出水中NH4+-N、NO2–-N、NO3–-N浓度和pH与MBBR出水无显著差异,两反应器的硝化效率相似。FBBR和MBBR在微生物群落上的相同点在于：优势菌门为变形菌门(Proteobacteria) (相对丰度分别为69.42%和86.92%),优势菌纲为γ-变形菌纲(γ-Proteobacteria) (40.71%和63.36%)和α-变形菌纲(α-Proteobacteria) (26.58%和21.74%),优势菌属为不动杆菌属(Acinetobacter) (27.50%和53.29%);硝化菌由亚硝化单胞菌属(Nitrosomonas)和硝化螺菌属(Nitrospira)构成;硝化螺菌属的相对丰度远高于亚硝化单胞菌属,两反应器中可能存在完全氨氧化菌。两反应器在微生物群落上的不同点在于FBBR微生物群落的丰富度和多样性以及硝化菌的相对丰度均高于MBBR。本研究可以为RAS养殖水体净化提供技术支撑,助推循环水养殖模式的推广应用。     

Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China

Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.     

Growth hormone (GH) and reproduction: a review

Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL^?1 (1.25 ng well^?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

Evaluation of a Fast Method Based on the Presence of Two Restriction Sites in the Mitochondrial ND5 (mt ND5) Gene for the Identification of Scomber Species

The purpose of this work was to evaluate the suitability of a method based on the presence of two restriction sites (for Hae III and Hindf I) in the mitochondrial NADH dehydrogenase subunit 5 (mt ND5) gene to identify Scomber species. The evaluation was performed on 144 reference and market samples by sequencing of the entire 505-bp fragment of the mt ND5 gene and of a 464-bp fragment of the Kocher fragment of the cytochrome b gene (mt Cytb). Sequence analysis of any of the two fragments allows the identification of each of the four Scomber species, but S. japonicus and S. colias had the same restriction sites at the ND5 amplicon and would not have been differentiated by this analysis. Similarly, loss of the Hae III site in some S. scombrus individuals would have misidentified them as not being Scomber. All the market products were correctly labeled except one acquired in Spain labeled as originating in the Atlantic and containing S. japonicus.     

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.     

Effect of iodophor disinfection of non‐hardened Salmo trutta eggs on their bacterial and fungus load

This study investigated the efficiency of iodophor disinfection (135 ppm active iodine for 15–30 min) of non‐hardened Salmo trutta eggs against different groups of bacteria and against fungus. Egg samples were taken from non‐disinfected and from disinfected eggs, microorganisms were cultured on specific nutrient media and their mass was measured by turbidimetric methods. Bacteria and fungus mass of non‐hardened eggs could be reduced but not eliminated by iodophor disinfection with 135 ppm active iodine for 15 min. The extent of reduction was 47–65% (Experiment 1). The efficiency of disinfection increased with disinfection time as the reduction in bacteria and fungus mass was 40–55% after 15 min and 58–74% after 30 min (Experiment 2). Disinfection efficiency of iodophor solution diluted in water (reduction 49–57%) and of iodophor solution diluted in sodium chloride solution iso‐osmolar to the oocytes (reduction 52–61%) was similar (Experiment 3). The reduction in bacteria and fungus mass was persistent as it was 39–72% lower in embryos deriving from disinfected eggs than in embryos deriving from non‐disinfected ones (Experiment 4). In conclusion, the tested disinfection method is inadequate to eliminate pathogens completely but it could positively influence immune defence of eggs and embryos.     

Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.