方斑东风螺“脱壳病”病原及病理初步研究

为明晰方斑东风螺"脱壳病"的病原或病理,对东风螺"脱壳病"进行了病原分离纯化、病原菌形态观察、组织病理观察、人工感染试验,并对病原菌16S rDNA序列进行PCR扩增和测序,以及系统发育树分析。试验中分离纯化了3种纯菌W、B、Y。致病菌W杆状,端生单鞭毛。致病菌Y杆状,两端略尖,周生菌毛。致病菌B短杆状,无鞭毛。结果显示,患病个体和人工感染个体的腹足及连壳肌肉组织均出现明显病变,肌纤维受损,出现空腔,并存在细菌;肝胰腺组织无明显病变,但血细胞显著减少。腹足肌注射人工感染试验结果表明,致病菌W在3种细菌中毒力最强。混合菌感染的毒力在混合比例接近原始比例时达到最大。致病菌B与报道的鲍希瓦氏菌相似度为99%;致病菌Y与报道的哈维弧菌相似度为98%;致病菌W与报道的变形假单胞菌相似度为99%。推测致病菌W、B、Y可能是此次方斑东风螺脱壳病的重要病原菌。     

鸢乌贼(Sthenoteuthis oualaniensis)减菌处理及其对方斑东风螺(Babylonia areolata)摄食率影响的研究

为了探讨不同灭菌方法对鸢乌贼(Sthenoteuthis oualaniensis)减菌处理效果及其对方斑东风螺(Babylonia areolata)摄食率的影响,本研究通过H₂O₂溶液处理、酒精处理、臭氧处理及紫外线处理等4种灭菌方法对鸢乌贼进行减菌处理,并将预处理后的鸢乌贼作为食物投喂体质量为(0.53±0.14)g的方斑东风螺稚螺5 d。结果显示,不同灭菌方法处理后的样品,其异养细菌和大肠埃希氏菌(Escherichia coli)数量显著减少(P<0.05);通过H₂O₂溶液和酒精处理后的鸢乌贼对方斑东风螺摄食率的影响显著低于对照组(P<0.05),而经过紫外线处理和臭氧处理后,其摄食率与对照组差异不显著(P>0.05)。综上,在实际生产作业中,使用臭氧处理方法对鸢乌贼进行20 min减菌化处理为最优方案,不仅可以除去鸢乌贼中大量有害细菌,而且不会影响方斑东风螺的摄食率。[中国渔业质量与标准,2022,12(4):42-46]     

"翻背症"对方斑东风螺主要消化酶及免疫相关酶的影响

为探索方斑东风螺消化及免疫系统对"翻背症"的应答机制,采集了健康(对照组)和"翻背症"早期、中期和晚期的方斑东风螺,采用TCBS平板计数法检测螺体内弧菌含量,试剂盒法检测螺体内组织超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、溶菌酶(LZM)等免疫相关酶和胃蛋白酶、脂肪酶、淀粉酶等消化酶活性。结果显示,方斑东风螺体内弧菌含量随病情的发展而显著增加,晚期达到最大值。发病早期,CAT、SOD、POD、ACP和胃蛋白酶等酶活性均显著上升。发病中期,POD、AKP、LZM活性均上升,且显著高于对照组和发病早期;与发病早期相比,CAT、SOD、ACP、胃蛋白酶、脂肪酶活性有所下降;与对照组相比,CAT、ACP活性仍显著升高,胃蛋白酶、脂肪酶活性显著降低,而SOD、淀粉酶活性无显著变化。发病晚期,各消化酶与免疫相关酶活性均较发病中期有所下降,但POD、ACP活性仍显著高于对照组,CAT、SOD、AKP、LZM活性与对照组无显著差异,而胃蛋白酶、脂肪酶、淀粉酶活性显著低于对照组。研究表明,"翻背症"方斑东风螺体内弧菌的暴发过程中,机体的免疫系统与消化系统均参与了机体病害免疫应答反应,且可以采用CAT、POD和ACP活性作为方斑东风螺"翻背症"的检测评价指标。     

温度对方斑东风螺生长与摄食影响的初步试验

本文是在小水体、自动控温的条件下,研究不同水温方斑东风螺(B.areolata)的生长、摄食和存活.2种不同时间的对比试验,分别为约1个月和3个月,不同时间的试验结果相近.在22～32℃的水温范围,水温22℃和32℃方斑东风螺的生长速度略有降低.但在此范围内,不同水温方斑东风螺的生长差异不明显.根据试验结果和养殖生产的实际情况,可以认为,方斑东风螺人工养殖适宜水温22℃～28℃,最适水温24℃～28℃.另外,方斑东风螺耐受的高温是34℃,35.5℃出现死亡.能耐受7～8℃的低温.     

海南养殖方斑东风螺暴发性疾病病原分离鉴定及药敏分析

2011年4月,从海南工厂化养殖的患病方斑东风螺体内分离得到3株优势菌,经感染实验确定菌株DFL11-01为该暴发性疾病的致病菌,其对方斑东风螺注射感染的LD50为2.6×106CFU/g。采用常规形态和生理生化特征,对菌株DFL11-01进行鉴定,并以16SrRNA基因序列同源性分析法对所有3株分离菌进行同源性分析,结果表明,3株细菌均为哈氏弧菌Vibrio harveyi。药敏试验结果表明,菌株DFL11-01对阿洛西林、利福平等15种药物耐药;对头孢他啶、四环素等3种药物中介敏感;对氨苄西林、恩诺沙星、头孢三嗪、哌拉西林、左旋氧氟沙星等5种药物敏感。     

方斑东风螺“翻背症”的病原及初步治疗研究

对海南发生"翻背症"的方斑东风螺进行了病原分离、鉴定,药敏分析及药物治疗试验。结果显示,病原为哈维弧菌,该菌对喹诺酮类、磺胺类药物敏感,对青霉素、阿莫西林等耐药;药物治疗结果显示,试验组诺氟沙星、复方磺胺甲恶唑、苯扎溴铵及对照组的最终存活率分别为39.05%、32.38%、29.52%和14.29%,诺氟沙星的治疗效果较好,可用于治疗方斑东风螺"翻背症"。     

水泥池养殖方斑东风螺Babylonia areolata的生长特性

对室外水泥池养殖方斑东风螺的生长特性进行了研究。对1龄内不同生长发育阶段的1400余个方斑东风螺的壳高、体重生长参数进行测量和分析,结果表明:(1)水泥池养殖方斑东风螺壳高和体重呈幂函数关系,W=0.2349H~(2.89)(R~2=0.9822,P<0.01);(2)1龄内养殖方斑东风螺的壳高生长曲线和体重生长速度曲线呈现出不断升高趋势,壳高日均增长和体重日均增重为0.11 mm·d~(-1)和0.03 g·d~(-1);壳高特定增长率 IGR_H 和体重特定增重率 IGR_W 平均为0.69%和1.96%;养殖方斑东风螺在1龄以内为快速生长期,远没有达到生长的拐点;同时,方斑东风螺的生长明显受水温影响,当水温降到20.2℃时,其日均增长和日均增重降到最低,分别为0.09 mm·d~(-1)和0.01 g·d~(-1);特定增长率和特定增重率分别降到0.42%和0.64%。     

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<正>水霉净本品具有强氧化作用,可以破坏或分解真菌的细胞壁中的甘露聚糖和甘露聚糖-蛋白质复合物,迅速扩散进入细胞内,氧化细胞内的酶或RNA、DNA,从而致死菌原体。此外,本品还可以同细菌的细胞壁中的脂蛋白或细胞膜中的磷脂质、蛋白质发生化学反应,从而使细菌的细胞壁和细胞受到破坏(即所谓的溶菌作用),细胞膜的通透性增加,细胞内物质外流,使其失去活性。     

东风螺生物学及养殖生态学研究进展

东风螺(Babyloniasp.)属软体动物的腹足纲、新腹足目、蛾螺科、东风螺属[1-3]。我国的主要经济种类有方斑东风螺(B.areolata)、泥东风螺(B.lutosa)和台湾东风螺(B.form osae)3种[4-5],分布于我国东南沿海、东南亚及日本。东风螺肉质鲜美、酥脆爽口、经济价值高,是近年国内外市场十分畅销的优质海产贝类,具有广阔的发展前景。本研究就东风螺的生物学及其养殖生态学研究进展作了概述,为东风螺的人工增养殖提供参考。1生物学特性1.1栖息环境东风螺一般分布于潮下带数米至数十米水深的海区,不同种类对底质的要求不同。方斑东风螺和台湾东风螺…     

方斑东风螺吻管水肿病病原菌的初步研究

从患病方斑东风螺(Babylonia areolata)分离到一株致病菌Balo001,运用生理生化方法并结合分子生物学方法对病原菌进行分类鉴定,并对病原菌进行药敏分析及半致死剂量(LD50)测定.结果表明:致病菌Balo001为哈氏弧菌(Vibrio harveyi),它对东风螺的LD50值为6.8×105 CFU/g;该菌对氟哌酸、氟苯尼考、复方新诺明等抗菌药有较高的敏感性.     

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.