冬季菲律宾蛤仔育苗系统几类主要细菌数量变化

试验结果表明:水中异养细菌为2.3×103-1.1×105cfu/m l,其变化趋势与温度变化一致;弧菌为5.0×103-6.6×103cfu/m l,其变化趋势与温度变化不一致,规律性不强。沉淀池泥中异养细菌为7.6×104-6.7×105cfu/g,弧菌为1.6×103-5.4×103cfu/g,反硝化细菌为2.5×102-3.2×104MPN/g,反硫化细菌为35-4.9×105MPN/g;温度高时,沉淀池泥中反硫化细菌占总异养菌比例最高,反硝化细菌次之,弧菌最小。从异养细菌的数量来看,沉淀池水质达到富营养化水平。因此,温度一旦升高,水中细菌会急剧增加,造成水质恶化,产生有毒的H2S、NO2-等物质,不利于菲律宾蛤仔的生长。     

流沙湾海区异养菌和弧菌动态变化规律研究

对流沙湾海区异养菌、弧菌动态变化规律及其优势菌群进行了初步探讨。采用平板稀释法对异养菌和弧菌数量进行了周年检测;对优势菌株进行分离纯化,并对代表性月份样品优势菌中的弧菌进行菌种鉴定。结果显示,流沙湾海区的异养菌和弧菌密度分别为8.8×103~9.60×105cfu/mL和1.0×103~2.45×104cfu/mL,其中异养菌数量高峰出现在6月,弧菌数量高峰出现在12月,异养菌、弧菌数量与水温变化的相关性不明显;流沙湾海区优势菌群中存在着溶藻弧菌等多种致病性弧菌。     

跑道式南美白对虾养殖池中弧菌的数量动态

采用TCBS平板计数法对南美白对虾跑道式养殖池和土塘养殖塘水样弧菌数量进行了一个养殖周期的连续监测。结果表明:跑道式养殖池水体中弧菌数在放苗后的前70 d均较低,随后迅速增加并维持在一个较高水平,变化幅度为15.0~1.42×104cfu/ml,平均3.96×103cfu/ml,平均数比土塘高2倍。跑道式养殖对虾始终未发病,跑道式养虾病害少,产量高。     

军曹鱼肠道及水体异养菌和弧菌的周年变化

采用总活异养菌2216E平板计数法和弧菌TCBS平板计数法对广东深圳大鹏养殖军曹鱼(Rachycentron canadum Linnaeus)养殖水体及鱼肠道细菌进行了周年监测。结果显示,育苗初期养殖水体异养菌和弧菌密度分别为0.63×104～6.2×104CFU/mL和0.30×102～1.03×104CFU/mL,鱼肠道异养菌和弧菌密度分别为0.80×106～7.5×107CFU/g和0～1.30×107CFU/g;网箱养殖监测期间,养殖水体异养菌和弧菌密度分别为4.20×103～5.40×105CFU/mL和0.70×102～1.14×105CFU/mL,鱼肠道异养菌和弧菌密度分别为1.50×107～8.78×108CFU/g和1.00×107～3.50×108CFU/g。周年监测结果显示,水体异养菌、弧菌的数量高峰均出现于8-9月份;肠道异养菌及弧菌的数量高峰均出现于6-7月份,分别又在12月份和2月份出现次高峰,弧菌变化趋势始终与异养菌一致。对分离得到的407株菌鉴定到属并进行多样性分析。结果显示,育苗期水体以假单胞菌(Pseudomonas)、黄单胞菌(Xanthomonas)和芽孢杆菌(Bacillus)为优势菌,三者约占水体异养菌总数的70%;养殖后期,弧菌(Vibrio)和发光杆菌(Photobacterium)的数量逐渐上升并占据一定优势,其中假单胞菌、黄单胞菌、弧菌和芽孢杆菌常年出现。鱼肠道异养菌以假单胞菌和芽孢杆菌为优势菌,其次是肠杆菌科(Enterbacteriaceae)细菌,也多次检测到弧菌、发光杆菌、气单胞菌(Aeromonas)和小球菌(Microccus)等。总之,养殖微生态系统的可持续稳定平衡与系统的种群多样性、群落组成及水质因子等因素相关,而整个环境生态系统的平衡直接影响到鱼类肠道的细菌生态系统。     

凡纳滨对虾(Litopenaeus vannamei)低盐度围隔调控养殖环境微生物研究

在广东省珠海市斗门一个大小为0.3hm2的空白虾池内,设立10个大小为5×5×1.6m(l×b×d)的陆基池塘围隔,对凡纳滨对虾低盐度调控养殖环境微生物进行实验研究,结果低盐度围隔调控养殖系统,水体异养细菌平均为1.33~9.27×104cfu.ml-1,致病性弧菌平均为2.83~7.91×102cfu.ml-1,养殖水体致病性弧菌的数量比异养细菌平均低2个数量级。各围隔水体异养菌在养殖早期都有一个高峰期,中后期低而稳定;围隔异养细菌的数量并未因定期施菌而增高,反而呈下降的趋势。对虾养殖健康的围隔,中后期水体致病性弧菌保持较低的数量,而对虾发病的围隔,致病性弧菌数量高。围隔调控养殖系统,异养细菌的数量相对比较稳定,而致病性弧菌数量呈较大波动。     

凡纳滨对虾工厂化养殖水环境分析

2018年6—10月对4口凡纳滨对虾工厂化养殖池水质理化因子和微生物环境的基本变化特征进行研究。结果表明:虾池水体中,NO~-_3是DIN(NH~+_4、NO~-_2和NO~-_3)主要的存在形式,占比达55%～85%;NH~+_4在0.005～0.060 mg/L低浓度范围内波动;5#池中NO~-_2出现累积现象,其变化趋势与NO~-_3高度一致。虾池中PO_4~(3-)含量为0.012～0.400 mg/L,随时间延长呈上升趋势。各池中Chl-a含量呈先上升后下降的趋势,最高值均出现在9月份。水体中总异养菌数量为2.80×10~4～2.40×10~(5 )CFU/mL,弧菌数量为8.00×10~2～1.00×10~(4 )CFU/mL,弧菌在总异养菌中所占比例在10%以下。     

高位虾塘细菌数量变化特点及其与理化因子的关系

本试验选取两口凡纳滨对虾高位池塘,通过连续地周期性采样和测定分析,研究了虾池异养细菌数量和弧菌数量变动规律及其与理化因子的相关关系。结果表明：两口试验虾池异养细菌数量变化范围在9.07×10^3～2.13×10^5 cfu/mL之间,平均为6.45×10^4 cfu/mL,呈现持续缓慢上升的趋势;两口试验虾池的弧菌数量变化趋势较为一致,变化范围为2.12×10^2～2.91×10^3 cfu/mL,养殖生产前期较低,中期高,后期随着换水量的加大而有所下降。相关性分析表明：虾池水体的异养细菌和弧菌数量均与透明度、 DO、 pH 呈负相关,与COD、 TSS含量呈正相关,其中弧菌数量与TSS含量呈显著正相关（ P〈0.05）。     

葡萄糖对对虾养殖水体水质的影响

研究葡萄糖的不同添加量(1.25~5×10-3 g/L)对对虾养殖水体水质指标(氨氮、活性磷)和微生物数量(总异养菌、弧菌)的影响。结果显示,与对照组相比,水体中添加葡萄糖能明显提高异养菌、弧菌密度(P<0.05),显著降低养殖水体中氨氮、活性磷浓度(P<0.05)。且在一定浓度范围内,葡萄糖浓度越高,氨氮、活性磷浓度越低,异养菌、弧菌密度越高。     

甘蔗渣载体填料在海水曝气生物滤池中的应用

为研究甘蔗渣作为载体填料用于海水曝气生物滤池中的可行性,在海水曝气生物滤池中培养生物膜,并以此为基础构建海水养殖排放水处理系统。通过监测水体总氨氮(TAN)、亚硝酸盐氮(NO2--N)等水质指标浓度变化,水体游离细菌与载体附着细菌密度变化,评价甘蔗渣载体生物滤池的降解效果。结果显示,以甘蔗渣为载体的生物滤池挂膜所需时间为26 d,挂膜完成后甘蔗渣附着可培养总菌和芽孢杆菌密度分别为3×108cfu/g和7.8×107cfu/g。在处理养殖水体时,生物滤池中水体氨氮和亚硝酸盐氮浓度分别控制在0.2 mg/L和0.05 mg/L以下,同时,水体中芽孢杆菌数量由3.3×103cfu/L增加至7×104cfu/L,弧菌数量由4.9×103cfu/L下降至3.1×101cfu/L。研究表明,以甘蔗渣为载体的海水曝气生物滤池能快速有效地完成挂膜,并在海水养殖排放水处理中取得较好效果。     

九孔鲍养殖水体及消化道细菌学的研究

2002年对汕尾健生鲍鱼养殖场养殖水体和鲍消化道中异养细菌及弧菌的数量和类群组成进行了研究。研究结果表明:养殖水体中异养细菌数量的四季变化从2 4×104～1 3×105cfu ml,平均为7 6×104cfu ml;消化道中异养细菌数量四季变化则从1 6×107～5 4×107cfu g(湿重),平均为3 3×107cfu g。水体中弧菌数量四季变化从1 2×104～5 1×104cfu ml,平均为2 5×104cfu ml;而消化道中弧菌数量四季变化则从2 8×105～3 8×105cfu g,平均为3 2×105cfu g。水体中细菌类群由Sphingomonas,Vibrio,Pasteurella,Moraxella,Pseudomonas,Aeromonas,Flavobacterium,Klebsiella,Weeksella和Alcall组成,消化道中细菌类群由Sphingomonas,Vibrio,Aeromonas,Flavobacterium,Shewanella,Sphingobacterium,Pseudomonas组成,水体和消化道弧菌种类主要为Vibriofluvialis,Vibrioparahaemolyticus,Vibriovulnificus,Vibriocholerae,Vibriominicus,Vibrioalginolyticus。并对不同养殖场水源弧菌数量也进行了比较。     

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).     

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.     

Influence of the antibacterial herbs, Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia on the survival, growth and bacterial load of Penaeus monodon post larvae

The indiscriminate use of antibiotics and chemicals in shrimp hatcheries has led to biomagnification and that in turn could lead to rejection of a whole consignment. The application of the bioencapsulation technique as a tool for curative treatment in shrimp larvae was investigated. Herbs having antibacterial properties such as Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia (methanolic extracts) were bioencapsulated in Artemia and fed to Penaeus monodon post larvae PL 1–25. The post larvae were reared in a medium inoculated with pathogenic bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Vibrio sp. Post larvae reared in the non-inoculated water and fed with non-enriched Artemia exhibited 90% survival, highest specific growth rate (12.43%) and reduced bacterial load. P. monodon reared in the bacterial inoculated water and fed with the non-enriched Artemia exhibited the lowest survival (10–30%), specific growth rate (8.42–9.1%) and increased bacterial load (2.86 × 10³ to 3.76 × 10⁵ cfu/g). The methanolic extracts of the herbs helped to increase survival and specific growth rate and reduced bacterial load in the P. monodon culture system. Among the three herbal extracts, P. corylifolia enriched Artemia fed post larvae showed the tendency to higher survival (>50%), growth rate (11.5 averaged) and low bacterial load (1.12 × 10⁵ cfu/g). This revised version was published online in August 2006 with corrections to the Cover Date.