1株军曹鱼病原弧菌的鉴定及其系统发育树分析

从养殖的患病军曹鱼身上分离出1株病原菌JT2，革兰氏阴性，短杆状，有极生鞭毛，能运动，菌落半透明。经回归感染实验，证明该菌为军曹鱼病原菌。进行了常规生理生化实验，并用API-ID32E系统鉴定该菌。再经Biolog-GN细菌鉴定系统鉴定，结果JT2与鲨鱼弧菌（Vibrio carchariae）相似度最高。为了进一步确定该菌的分类学地位，测定了其16SrDNA序列，分析了相关细菌的同源性，构建其系统发育树。结果表明，该菌株与V．carchariae的亲缘关系最近。综合上述几种方法的鉴定结果，最后鉴定该株菌为鲨鱼弧菌V．carchariae。     

九孔鲍消化道及养殖水体中弧菌胞外毒力因子的分析

从汕尾健生鲍鱼养殖场成鲍养殖水体和消化道中分离筛选到22株弧菌,其中14株来自成鲍消化道,8株来自养殖水体。本文对这两种不同来源的菌株进行了致病因子(胞外酶及溶血毒素)的分析比较,同时采用API条带法对其进行了种类鉴定。结果发现,消化道中除1株溶藻弧菌和3株最小弧菌外,其余均为河流弧菌;而水体中除副溶血弧菌和创伤弧菌各1株外,其他也均为河流弧菌。实验还发现,在5株胞外酶分泌能力最强的弧菌中,只有Sh031株是副溶血弧菌,另外4株(Bh14、Sh02、Sh08、Sh05)均为的河流弧菌。结果显示,无论是养殖水体还是消化道,河流弧菌都应视为一种海水养殖贝类(鲍)的主要条件致病菌,是能力较强的胞外致病因子生产者。     

28株水产动物致病菌的编码鉴定

采用国产革兰氏阴性杆菌编码鉴定系列培养基对28株细菌进行鉴定。结果显示：除2株细菌未能得到鉴定外，其余26株细菌均鉴定到属或种水平，可鉴定率为92．86％．但各菌株的鉴定概率和模式频率有所不同。26株可鉴定菌分布于气单胞菌属(Aeromonas)、假单胞菌属(Pseudomonas)、不动杆菌属(Acinetobacter)、摩根氏菌属(Morganella)、弧菌属(Vibrio)、变形杆菌属(Proteus)、黄单胞菌属(Xanthomonas)、邻单胞菌属(Plesiomonas)和埃希氏菌属(Escherichia)9个菌属，其中13株为气单胞菌(Aeromonas)；3株为恶臭假单胞菌(P．putide)；2株为摩氏摩根氏菌(M.morganella)；2株为不动杆菌(Acinetobacter)；2株为类志贺邻单胞菌(P．shigelloides)；拟态弧菌(V．mimicus)、普通变形杆菌(P．vulgaris)、嗜麦芽黄单胞菌(X．maltophilia)和大肠埃希氏菌(E．coli)各l株。基本上摸清了安徽省水产养殖动物常见病原菌的种类。     

养殖大菱鲆(Scophthalmus maximus)腹水病的病原多样性及其耐药性分析

为了解引起养殖大菱鲆(Scophthalmus maximus)腹水病的病原多样性及其耐药性情况,针对2002–2010年由不同地区病样分离的27株细菌性病原进行了16S r DNA鉴定,并采用K-B法测定了27株细菌对22种抗生素的耐药性,分析了病原菌的耐药谱及耐药率变化。结果显示,大菱鲆腹水病病原菌主要有大菱鲆弧菌(Vibrio scophthalmi)、迟钝爱德华氏菌(Edwardsiella tarda)、鳗弧菌(Vibrio anguillarum)、哈维氏弧菌(Vibrio harveyi)、假交替单胞菌(Pseudoalteromonas espejiana)。山东青岛地区以大菱鲆弧菌为主,威海地区以迟钝爱德华氏菌为主,烟台地区菌株种类分布平均。5类细菌对青霉素类、头孢菌素类、大环内酯类、复方新诺明耐药率高于50%。只有1株迟钝爱德华氏菌对氟苯尼考产生了耐药,其余菌株对其均没有耐药性,且在长期使用中不易产生耐药性,证实氟苯尼考为当前防治腹水病的一种良好抗菌药物。27株病原菌的耐药谱数量为27个,每个菌株具备自己独特的耐药谱,74.1%的菌株对10种以上的抗菌药物产生了耐药性,均有多重耐药性。     

养殖大菱鲆(Scophthalmus maximus)腹水病的病原多样性及其耐药性分析

为了解引起养殖大菱鲆(Scophthalmus maximus)腹水病的病原多样性及其耐药性情况,针对2002-2010年由不同地区病样分离的27株细菌性病原进行了16S rDNA鉴定,并采用K-B法测定了27株细菌对22种抗生素的耐药性,分析了病原菌的耐药谱及耐药率变化.结果显示,大菱鲆腹水病病原菌主要有大菱鲆弧菌(Vibrio scophthalmi)、迟钝爱德华氏菌(Edwardsiella tarda)、鳗弧菌(Vibrio anguillarum)、哈维氏弧菌(Vibrio harveyi)、假交替单胞菌(Pseudoalteromonas espejiana).山东青岛地区以大菱鲆弧菌为主,威海地区以迟钝爱德华氏菌为主,烟台地区菌株种类分布平均.5类细菌对青霉素类、头孢菌素类、大环内酯类、复方新诺明耐药率高于50％.只有1株迟钝爱德华氏菌对氟苯尼考产生了耐药,其余菌株对其均没有耐药性,且在长期使用中不易产生耐药性,证实氟苯尼考为当前防治腹水病的一种良好抗菌药物.27株病原菌的耐药谱数量为27个,每个菌株具备自己独特的耐药谱,74.1％的菌株对10种以上的抗菌药物产生了耐药性,均有多重耐药性.     

南移仿刺参体表溃疡症病原菌的分离鉴定及药敏分析

从福建省霞浦县网箱吊笼养殖的患病仿刺参(Apostichopus japonicus)体壁中分离得到1株优势菌。人工感染实验证实该菌株为致病菌,经形态学、生理生化指标、16S rRNA序列分析和Biolog自动微生物鉴定系统鉴定,确定该病原菌为哈维氏弧菌(Vibrio harveyi)。药敏试验表明,该病原菌对阿奇霉素、大观霉素、呋喃妥因、氯霉素、头孢吡肟、头孢呋辛、头孢噻肟、头孢他啶、头孢哌酮和复方新诺明等11种抗生素类药物敏感。     

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.     

发病鲆鲽类分离菌株的16S rRNA基因测序分析

细菌性疾病是中国海水养殖鲆鲽类的主要病害,为全面了解病原菌种类,本研究对1999~2012年从山东、江苏、河北、天津等沿海地区养殖场发病鲆鲽鱼类中分离得到的124株优势菌株进行了16S rRNA基因测序和系统发育学分析。将基因序列与GenBank核酸序列数据库进行相似度比对分析,结果显示,有83株与弧菌属(Vibriosp.)细菌相似度最高,11株与气单胞菌属(Aeromonas sp.)细菌相似度最高,4株与爱德华氏菌属(Edwardsiella sp.)细菌相似度最高,26株为其他15种属的细菌。根据系统发育学分析结果,进一步将66株菌鉴定为16个种,优势种为溶藻弧菌(V.alginolyticus)、哈氏弧菌(V.harveyi)、鳗弧菌(V.anguillarum)、杀鲑气单胞菌(A.salmonicida)和迟缓爱德华氏菌(E.tarda)。选择其中的9株鳗弧菌和4株迟缓爱德华氏菌进行人工感染实验,结果显示,其中7株鳗弧菌和3株迟缓爱德华氏菌对大菱鲆(Scophthalmus maximus)有较强的致病性。研究结果可为阐明中国海水养殖鲆鲽类的流行病发生历史、病原种类、病原监测及疾病控制提供重要参考。     

池塘养殖刺参病原菌塔式弧菌的分离与鉴定

2013年春季,福建漳州地区池塘养殖刺参(Apostichopus japonicus)感染不明病原而患病。病参表现出排脏、身体萎缩、体表溃烂等症状,俗称为"腐皮综合征"。为确定引起该病的病原菌,从患病刺参病灶部位分离得到1株优势菌FJY001,经回接感染试验,证实所分离的细菌为刺参的病原菌。经形态、生理生化、16S rRNA序列分析和Biolog微生物自动鉴定系统等多项指标鉴定,确定该病原菌为塔式弧菌(Vibrio tubiashii)。     

致病性拟态弧菌的敏感药物和拮抗菌的筛选

采用纸片扩散法,系统测试了3株致病性拟态弧菌对8类23种抗菌药物的敏感性与联合用药的敏感性。结果显示:氨基糖甙类、头孢类、喹诺酮类和磺胺类等多种抗菌药物对病原弧菌均有抑制或杀死作用。庆大霉素与四环素类、头孢类、喹诺酮类、红霉素或氯霉素联用以及多粘菌素与头孢类、喹诺酮类、红霉素或氯霉素联用均表现出一定程度的累加或协同作用;同时,采用琼脂平板扩散法从9株非致病菌中筛选对致病性拟态弧菌具有拮抗作用的益生菌并对其抑菌活性进行测定。结果显示:HX-01和HX-05两株细菌的菌液和无细胞上清液对3株致病性拟态弧菌均具有拮抗作用。在拮抗菌—病原菌混合体系中,病原菌经24 h培养,菌数由原来的105CFU/mL增至2ⅹ108CFU/mL,但从第2天开始,随着拮抗菌产生的抗性产物浓度的增加,菌数快速下降,至第6天病原菌完全被杀死。而拮抗菌培养至第6天,菌数仍保持较高水平(2ⅹ107CFU/mL)。表明这两株拮抗菌对致病性拟态弧菌具有较强的拮抗作用。经数值鉴定法确定两株拮抗菌均为产碱杆菌。     

Enzyme producing bacterial flora isolated from fish digestive tracts

Isolationand enumeration of aerobic bacterial flora in the gastrointestinal tract of nineculturable freshwater teleosts, namely catla, rohu, mrigal, silver carp, grasscarp, common carp, tilapia, walking catfish and murrel have been carried out.Amylolytic, cellulolytic, lipolytic and proteolytic microflora were identifiedfrom the culture plate using selective media. The isolates were qualitativelyscreened on the basis of their extracellular enzyme producing ability. Theselected strains were further quantitatively assayed for amylase, cellulase,lipase and protease activities. Protease activity was exhibited by almost allthe bacterial isolates, while strains isolated from tilapia, grass carp andcommon carp showed considerable amylolytic and cellulolytic activities. Maximumactivity of lipase was exhibited by a strain isolated from silver carp. Thestudy indicates that there is a distinct microbial source of the digestiveenzymes – amylase, cellulase, lipase and protease, apart from endogenoussources in fish gut. The information generated from the present investigationmight contribute towards better feed formulations for carp at low cost,incorporating the enzyme producing bacterial isolates as probiotics.     

人工雌核发育鲢近交F_2微卫星DNA变异分析

采用17对鲢微卫星引物,以野生鲢、养殖鲢、雄鲤作对照对人工雌核发育鲢近交F2及其亲本进行了微卫星分析。结果表明:人工雌核发育鲢近交F2、养殖鲢和野生鲢群体的平均等位基因数范围为2.1～4.0;平均观测杂合度范围为0.2762～0.9588;期望杂合度范围为0.2774～0.7360;遗传多样性指数范围为0.4500～1.2258,人工雌核鲢近交F2为0.4500,显著低于养殖鲢(1.0273)和野生鲢(1.2258),揭示人工雌核发育鲢近交F2遗传多样性水平较低,纯合度较高。从遗传距离来看,人工雌核发育鲢近交F2与对照群体之间的遗传距离都要大于野生鲢与养殖鲢群体之间遗传距离,表明人工雌核发育鲢近交F2发生了一定程度的遗传分化。     

鳜鱼、青鱼、草鱼、鲤、鲫、鲢消化酶活性的研究

鳜鱼、青鱼、草鱼、鲤、鲫、鲢肝脏蛋白酶活性低于肠蛋白酶。肠蛋白酶活性，鳜鱼最高，其余依次为青鱼、鲤、鲢、草鱼、鲫。鳜鱼、鲢的肠蛋白酶活性由前肠向后肠递减；而青鱼、鲤、草鱼的则由前肠向后肠递增；鲫则中肠活性最低。六种鱼不同组织的脂肪酶活性因鱼而异。青鱼、鳜鱼、鲤、鲢肝脏中脂肪酶活性低于肠脂肪酶活性（Ｐ＜０．０１）；而草鱼和鲫肝脏中脂肪酶活性与肠脂肪酶活性差异不明显（Ｐ＞０．０５）。六种鱼肝脏中脂肪酶活性由高到低依次为鳜鱼＞鲢＞鲫＞草鱼＞青鱼＞鲤；肠脂肪酶活性顺序为鳜鱼＞鲢＞青鱼＞鲤＞草鱼＞鲫。脂肪酶活性与鱼类食性无明显相关性。不同组织间淀粉酶活性存在差异。鳜鱼的肝脏和肠均有较高淀粉酶活性，青鱼和鲫肝脏中淀粉酶活性低于肠中的；草鱼、鲤和鲢肝脏中淀粉酶活性高于肠中的，但差异不显著。六种鱼中鳜鱼的肝脏淀粉酶活性明显高于其它五种无胃鱼，它们的淀粉酶活性顺序依次为鳜鱼＞鲢＞鲤＞草鱼＞青鱼＞鲫；肠淀粉酶活性顺序为鳜鱼＞鲫＞鲤＞青鱼＞鲢＞草鱼。     

Food habits and growth of silver and bighead carp in cages and ponds

Analysis of intestinal contents of silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) showed that silver carp consumed primarily phytoplankton while bighead carp consumed large quantities of zooplankton and detritus in addition to phytoplankton. The size of particles filtered by the bighead carp was larger (17–3,000 μm) than that filtered by silver carp (8–100 μm). Artificial feed was readily consumed by bighead carp but not by silver carp. No growth difference was indicated for silver carp in fertilized ponds and ponds receiving artificial feed. Growth of bighead carp increased substantially with the addition of artificial feed. Silver carp grew more rapidly in cages than bighead carp.     

松浦镜鲤与长丰鲢食用鱼混养试验

2012年5-10月,在面积为0.19hm2的试验池1中放养体质量160g的松浦镜鲤（Cyprinus carpio Songpu）117,700尾·hm^-2,混养体质量160g的长丰鲢（Hypophthalmichthys molitrix）春片、乌子头和鳙（Aristichthys no-bilis）夏花鱼种。在面积为0.19hm2的试验池2中放养体质量149.5g的松浦镜鲤3,450尾·hm^-2,只混养鲢和鳙夏花,采用常规饲养方法。2012年10月2日,试验池1平均每hm2产鱼21,025.5kg,其中松浦镜鲤平均全长34.3cm,体质量1425g,产量18,294.0kg;长丰鲢春片平均体质量674g,平均产量1,816.5kg,长丰鲢夏花平均全长18.6cm,体质量112.4g,平均产量592.5kg;鳙夏花平均全长达12.1cm,体质量39.9g,平均产量322.5kg。试验池2平均每hm2产鱼3,069.0kg,其中松浦镜鲤平均全长35.0cm,体质量1225g,平均产量2,766.0kg;鲢夏花平均全长达11.3cm,体质量24.9g,平均产量130.5kg;鳙夏花平均全长达11.1cm,体质量35.1g,平均产量172.5kg。试验表明,高密度养殖的松浦镜鲤产量显著高于密度低时,长丰鲢夏花的出池体质量是普通鲢的4.5倍,特殊生长率（6.27%·d^-1）是普通鲢（3.5%·d^-1）的1.79倍。文中还讨论了松浦镜鲤养殖池的水质和技术特点。     

Improved survival of rohu, Labeo rohita (Hamilton-Buchanan) and silver carp, Hypophthalmichthys molitrix (Valenciennes) fingerlings using low-dose quinaldine and benzocaine during transport

The effects of fingerlings immersion in low‐dose benzocaine (15 and 30 mg L⁻¹, silver carp and rohu) and quinaldine (100 μL L⁻¹ silver crap and 250 μL L⁻¹ rohu) for 1, 3 and 6 h on stress responses and survival of rohu, Labeo rohita and silver carp, Hypophthalmichthys molitrix fingerlings were evaluated in a transport simulation experiment. Both quinaldine and benzocaine showed low mortalities (0–2%). The total mortality in control (with no anaesthesia) was 30% for rohu and 14% for silver carp. Quinaldine and benzocaine‐treated fingerlings had significantly higher plasma chloride levels than the control in both species. Benzocaine, quinaldine, as well as the control, had an initial elevation of plasma cortisol levels. Benzocaine lost its effectiveness after 3 h exposure while quinaldine persisted throughout the 6 h experimental period. Both sedatives reduced bacterial build‐up compared with the control. No post‐exposure mortality was observed for any of the transport methods assessed 48 h after the treatment. This study suggests that the use of low‐dose benzocaine or quinaldine during transport has positive effect on the survival and health of rohu and silver carp fingerlings.     

太湖梅梁湾生物控藻围栏内鲢、鳙比肠长和比肝重的动态变化

为了解滤食性鱼类的消化生理指标及其生态适应性,2005年1-10月,调查了太湖梅梁湾生物控藻围栏中鲢(Hypophthalmichthys molitrix)、鳙(Aristichthys nobilis)在不同生长阶段的比肠长和比肝重指数的动态变化,分析了比肠长和比肝重与温度、体长、体重以及饵料组成的关系。结果表明,鲢、鳙比肠长和比肝重的季节变化都非常明显,比肠长均值分别为9.66和5.73,夏季的比肠长均明显增大。总的来看,鲢的比肠长和季节变化幅度均大于鳙,表明在滤食和消化浮游植物上,鲢具有更强的环境适应能力;鲢、鳙的比肠长与水温显著正相关(鲢P<0.01;鳙P<0.05),而与体长、体重以及饵料组成的相关性不显著;温度导致的摄食强度变化应是鲢、鳙比肠长季节变化的主要原因。鲢、鳙的比肝重均值分别为1.57%和1.91%,鳙的比肝重明显大于鲢,这可能与鳙摄食更多的浮游动物有关;鲢、鳙的比肝重与温度、体长、体重以及饵料组成均有显著的相关性(P<0.05),表明生物个体的形态特征与其所利用的资源环境有非常紧密的联系。     

The effect of fish density and species combination on growth and utilization of natural food in ponds

Abstract. Interactions between bottom-feeding fish (common carp, Cyprinus carpio L., and hybrid tilapia) and a filter feeder (silver carp, Hypophthalmichthys molitrix (Valenciennes)) in polyculture were studied in fertilized ponds with no supplementary feeding. The silver carp were stocked at two densities: 1300 and 2600/ha. Growth rate and yield of each of the species were also compared with those in polyculture of bottom feeders alone, and in a monoculture of silver carp alone, at the same densities. The consumption of natural feed as melabolizable energy (ME) was calculated by the energy balance (requirement for maintenance and growth divided by the utilization efficiency).
Increasing the density of silver carp reduced proportionally their own growth rate, due to the limited amount of food. Above a density of about 1000/ha, silver carp inhibit the growth rate of common carp and tilapia. This inhibition was small at 1300 silver carp/ha, but considerable at 2600/ha. However, the presence of bottom feeders increased the growth rate of silver carp at both densities. Calculated natural food consumption showed that growth interactions between species were brought about through the availability of food. The synergistic effect of the bottom-feeding fish on the silver carp is assumed to be due to the upwelling of bottom nutrients to the upper layers of water by the burrowing of the fish in the mud. Combined yield of all species in the polyculture was highest at the density of 1300 silver carp/ha (2116 kg/ha in 156 days). At 2600 silver carp/ha the inhibition of growth rate of silver carp itself, reduced total yield as compared with the lower density.     

Effect of three commercial microbial products on bacterial community in a freshwater fish polyculture system

A 30‐day experiment was conducted to evaluate the effect of three commercial microbial products, Novozymes Pond Plus, Zhongshui BIO‐AQUA and Effective Microorganisms on bacterial community in polyculture tanks stocked with grass carp (Ctenopharyngodon idellus), gibel carp (Carassius auratus) and silver carp (Hypophthalmichthys molitrix). Four treatments were tested. One treatment with no supplementation of the microbial products served as control. In the other three treatments, the microbial products were added at the intervals of 10 days respectively. During the experiment, grass carp and gibel carp were fed with a commercial formulated feed daily. Bacterial count and bacterial composition in water column of the tanks were monitored at the intervals of 3 days, and bacterial composition in sediment was determined at the end of the experiment. Bacterial composition in water column varied with progress of the experiment. Some bacteria from Novozymes Pond Plus and Effective Microorganisms could colonize in the tanks but did not dominate in bacterial community. This study reveals that the competition between the exogenous bacteria and native bacteria might be a factor determining the efficacy of the microbial products in improving water quality.