河蟹早期溞状幼体病害防治研究

在河蟹育苗中常见早期溞状幼体（ＺⅠ、ＺⅡ）被聚缩虫（Ｚｏｏｔｈａｍｎｉｕｍｓｐ）寄生引起细菌性或病毒性感染，造成突发性死亡。作者用孔雀绿、制霉素、氯霉素、高锰酸钾等药物对聚缩虫进行杀灭试验，结果显示以孔雀绿和制霉素的杀虫效果最好。但从早期　状幼体的病害防治效果来看，由于越冬亲蟹是病原体的携带者，因此作者强调防治工作必须从亲蟹暂养时做起，立足于早防早治；且以多种药物综合性措施为妥。     

Effect of poly-β-hydroxybutyrate on Chinese mitten crab, Eriocheir sinensis, larvae challenged with pathogenic Vibrio anguillarum

This study investigated the protective effect of poly-β-hydroxybutyrate (PHB) on Chinese mitten crab, Eriocheir sinensis, zoea larvae challenged with pathogenic Vibrio anguillarum. PHB was delivered to the crab larvae through rotifer and Artemia bioencapsulation. Zoea 3 larvae were challenged with V. anguillarum at a final concentration of 10(5) CFU mL(-1). PHB-enriched rotifers and Artemia nauplii were added to the culture water 24 h prior to, upon and 24 h after challenge. The results confirmed that PHB could enhance the survival and growth of unexposed E. sinensis larvae. Moreover, PHB protected larvae from the pathogen as the larvae fed PHB-enriched live food showed the highest survival and development rate in all challenged groups (P < 0.05). Furthermore, larval performance was the best when PHB was delivered to the larvae 24 h before challenge (P < 0.05). In conclusion, our results indicate that PHB can be used as part of an effective strategy to protect E. sinensis larvae from V. anguillarum resulting in higher survival and better growth, especially when applied before the challenge.     

文蛤病原菌——需钠弧菌的鉴定和生物学特性分析

从患病文蛤Meretrix meretrix体内分离出一优势菌株WT01,回归感染试验证明是文蛤的致病菌。对该菌形态、生理生化特征、盐度、温度和PH值生长条件以及16S rRNA基因序列进行了研究。结果表明,该菌为革兰氏阴性菌,发酵葡萄糖产酸不产气,氧化酶阳性,生长需NaCl,无色素,不发光,在TCBS平板上形成圆形黄色菌落,对弧菌抑制剂O／129敏感,具有弧菌属的典型特征。16S rRNA基因序列分析表明,该菌与需钠弧菌Vibrio natriegen的亲缘关系最为接近,其同源性达999／6以上。因此,将该菌鉴定为需钠弧菌。实验显示其半数致死量为5．5×10^6CFU／g。19种抗菌药物药敏试验结果表明,该菌对氟哌酸、复方新诺明、菌必治、先锋必、链霉素、氯霉素和庆大霉素等抗生素较为敏感。并对其胞外产物的生物学特性进行了分析。     

Bacteria associated with early life stages of halibut, Hippoglossus hippoglossus L., inhibit growth of a pathogenic Vibrio sp.

Abstract Bacteria capable of inhibiting growth of a pathogenic Vibrio sp. were isolated from the gastrointestinal tract of cultured halibut larvae during the first feeding and weaning stages. No such bacteria were found among isolates from the surface of eggs or the gastrointestinal tract of yolk sac larvae. The fraction of pathogen-inhibitors among the total number of isolates ranged between 0–100% (first feeding) and 0–66% (weaning). All pathogen-inhibitors were Gram-negative rods, and 95% were oxidase and catalase positive fermentative isolates, capable of producing acid aerobically from a varying range of carbohydrates. These isolates possessed the characteristics of the Vibrio/Aeromonas -group, but only 19% were sensitive to the vibriostatic agent 0/129. Isolates from eggs and yolk sac larvae were dominated by bacteria belonging to the Cytophaga/Flexibacter/Flavobacterium - group. The high fraction of isolates with the ability to inhibit growth of the pathogenic Vibrio sp. among the total number of isolates indicates that pathogen inhibition may be an important mutualistic role of the intestinal flora of early life stages of halibut.     

养殖大菱鲆黑瘦症的病原菌鉴定及杀菌中草药筛选

从患黑痩症的大菱鲆幼体内分离得到一株优势菌株,人工感染实验证实其为大菱鲆致病菌,并经过生理生化及16SrDNA序列分析鉴定为灿烂弧菌Vibrio splendidus。组织病理学显示,病鱼的肝脏、肾脏、脾脏、肠道及脑均呈现不同程度的组织破坏。用煎煮法提取11种常用中草药成分,测定该致病菌对中草药的杀菌效果。实验表明,五倍子和白头翁的杀菌效果最好,黄柏、穿心莲次之,当归、党参、杜仲、秦皮、金银花、乳香和没药的杀菌效果最差。以五倍子、白头翁为主要成分的初步治疗试验证明,药浴和口服给药可有效治愈大菱鲆苗期的黑瘦症。     

福建海区网箱养殖刺参“腐皮综合症”病原分析与鉴定

2011年冬季,福建省宁德市霞浦县海区网箱养殖刺参发生"腐皮综合症",并伴有死亡现象出现。刺参的发病症状表现为:厌食、排脏、身体萎缩、体表局部溃烂乃至大面积溃烂。从患病刺参病灶处分离得到两种优势细菌CS1和CS2。经人工回接感染实验证明两种菌对健康刺参都具有较强的感染性,且感染病参的症状与自然发病刺参的症状相同。通过生理生化试验、16S rDNA序列分析及系统进化树分析,结果表明两株菌分别与灿烂弧菌(Vibrio splendidus)和假交替单胞菌(Pseudoalteromonas sp.)相似。菌株CS1与灿烂弧菌的亲缘关系最近,相似率达到99%,菌株CS2与假交替单胞菌的亲缘关系最近,相似率达到95%。菌株CS1可鉴定为灿烂弧菌,菌株CS2可鉴定为假交替单胞菌。另外,在患病刺参呼吸树膜和腔体内发现大量后口虫(Boveria sp.)。所以导致本次刺参"腐皮综合症"的原因可能是致病性细菌和寄生虫共同作用的结果。本文首次揭示了该地区"腐皮综合症"导致养成刺参大规模死亡的致病原因,对刺参病害防治和健康养殖具有重要的指导意义。     

Diet particle size preference and optimal ration for mud crab, Scylla serrata, larvae fed microbound diets

This paper reports on experiments to determine particle size preference and optimal ration for the various larval stages of the mud crab, Scylla serrata, fed microbound diets (MBD). All experiments used ¹⁴C-labelled rotifers as components of MBD, and ingestion was determined by assessing the ¹⁴C content of S. serrata larvae after feeding on the MBD. Five size ranges of MBD (<150, 150–250, 250–400, 400–600 and 600–800 μm) were used to assess ingestion by Zoea I, Zoea III, Zoea V and Megalopa. All were fed a ration equivalent to twice the equivalent dry weight of live foods provided to each larval stage under standard rearing conditions (standard ration). MBD particle size preference increased with increasing larval development. Highest ingestion rates by Zoea I, Zoea III, Zoea V and Megalopa were found for particle size ranges of <150, 150–250, 250–400 and 400–600 μm, respectively. MBD within these particle size ranges were used in subsequent experiments to assessed optimum rations for each of the four larval stages. Larvae were fed rations based on multiples or fractions of the standard ration (100%): 12.5%, 25%, 50%, 100%, 200% and 300%. There were no significant increases in ingestion, for any of the four larval stages, when ration was increased above 100%. For Zoea I, Zoea III and Zoea V larvae, there was no significant difference in the rate of MBD ingestion when ration was reduced to 50%; however, a 25% ration brought about a significant decrease in ingestion. There was no significant decrease in the rate of ingestion by Megalopa when ration was reduced from 100% to either 50%, 25% or 12.5%. The results suggest that rations currently used for larval rearing of S. serrata may be excessive. Little is currently known of the feeding behaviour and nutritional requirements of S. serrata larvae, and the results of this study are a significant development in this field.     

养殖牙鲆弧菌病病原菌初步研究

1998年 7月自荣成市寻山水产集团总公司养鱼场养殖牙鲆病死鱼体内分离到 1株细菌 ,经人工感染试验证明是牙鲆弧菌病病原菌。该菌主要特征为 :革兰氏阴性 ,弧状 ,以极生单鞭毛运动。氧化酶阳性 ,发酵葡萄糖产酸不产气 ,不产生 H2 S,不产生色素。生长温度范围 10～ 4 2℃ ,p H范围 6～10 ,盐度范围 0 .5～ 6。经 Biolog Microstation System鉴定为河川弧菌 - 。该菌株对环丙沙星、氟哌酸、利福平、呋喃唑酮、呋喃妥因、红霉素、链霉素等药物敏感。     

网箱养殖大黄鱼诺卡氏菌病的初步研究

首次报道了大黄鱼诺卡氏菌病的发生情况。病鱼以体表和心、脾、肾等内脏出现白色结节为主要症状，平均死亡率15％。对病鱼进行细菌分离培养和光镜、电镜检查，均发现长或短的丝状分枝状杆菌。用分离菌株作回归感染，证实为该大黄鱼结节病的病原菌。对病原菌的基本生物学特性进行了研究，确定病原为诺卡氏菌。     

Isolation of a highly pathogenic Vibrio pelagius strain associated with mass mortalities of turbot,Scophthalmus maximus (L.), larvae

A bacterial strain, characterized as Vibrio pelagius (Hq 222), was isolated from a turbot, Scophthalmus maximus (L.), larvae mass mortality in a commercial fish farm in Spain. Turbot larvae, post-larvae (0.2 g) and juveniles (5 and 15 g) were experimentally infected. The bacterium appeared to be very virulent for larvae and post-larvae, LD50 being < 5 bacteria mL(-1) for larvae 1 week post-infection and 3.9 x 10(5) bacteria mL(-1) in post-larvae at day 12 post-infection. The bacterial strain was recovered in pure culture from the internal organs of infected fish. Histological lesions in post-larvae exhibited swelling and necrosis of gill secondary lamellae, sloughing of intestinal mucosa and necrosis of haematopoietic tissue in the kidney. Vibrio pelagius (Hq 222) was able to grow in sterile sea water when incubated at room temperature or at 15 degrees C. Vibrio pelagius (Hq 222) was more adherent to the turbot cell lines TV-1 and TF than Escherichia coli. In both cell lines, the number of adhered bacteria increased with incubation time.     

基于InDel标记的斑节对虾早期性别鉴定方法的建立

为分辨斑节对虾(Penaeus monodon)发育早期的性别,利用改良的插入/缺失(InDel)标记追踪了从受精卵、无节幼体、溞状幼体、糠虾幼体、仔虾后1 d(PL1)、仔虾后30 d(PL30)及亚成虾个体的性别,建立了测序和荧光定量PCR熔解曲线的检测方法。结果表明,InDel标记可扩增获得雌雄特异性序列,用测序法判定亚成虾性别的结果与外部观察结果一致,准确率达100%,对受精卵、无节幼体、溞状幼体及糠虾幼体均可得到雌雄特异性序列;进而建立了基于SYBR Green实时荧光定量PCR(Quantitative real-time polymerase chain reaction,q RT-PCR)熔解曲线的快速鉴定斑节对虾性别的方法,其中雄性特异序列的熔解温度(Melting temperature,Tm)为(79.10±0.10)℃,雌性为(78.45±0.20)℃,通过特异的熔解曲线可准确区分PL30、亚成虾和无节幼体Ⅵ期个体,准确率可达96.3%以上。     

Successful rearing of whiteleg shrimp Litopenaeus vannamei larvae fed a desiccation‐tolerant nematode to replace Artemia

The nematode Panagrolaimus sp. was tested as live feed to replace Artemia nauplii during first larval stages of whiteleg shrimp Litopenaeus vannamei. In Trial 1, shrimp larvae were fed one of four diets from Zoea 2 to Postlarva 1 (PL1): (A) Artemia nauplii, control treatment; (NC) nematodes enriched in docosahexaenoic acid (DHA) provided by the dinoflagellate Crypthecodinium cohnii; (N) non‐enriched nematodes; and (Algae) a mixture of microalgae supplemented in C. cohnii cells. In Trial 2, shrimp were fed (A), (NC) and a different treatment (NS) with nematodes enriched in polyunsaturated fatty acids (PUFAs) provided by the commercial product S.presso^®, until Postlarva 6 (PL6). Mysis 1 larvae fed nematodes of the three dietary treatments were 300 μm longer (3.2 ± 0.3 mm) than control larvae. At PL1, control shrimp were 300 μm longer (4.5 ± 0.3 mm) than those fed DHA‐enriched or PUFAs‐enriched nematodes. No differences were observed in length and survival at PL6 between control larvae and those fed DHA‐enriched nematodes (5.1 ± 0.5 mm; 33.1%–44.4%). Shrimp fed microalgae showed a delay in development at PL1. This work is the first demonstration of Panagrolaimus sp. suitability as a complete substitute for Artemia in rearing shrimp from Zoea 2 to PL6.     

中国对虾糠虾幼体病原菌（非01群霍乱弧菌）的研究

本文对中国对虾糠虾幼体的一种病原菌———非０１群霍乱弧菌作了研究报道。这种病的症状是病虾运动能力差、趋光性弱、镜检发现肠道肿胀。从垂死病虾中分离到５株细菌，经感染健康糠虾幼体得到与病虾相同症状。５株细菌的４７项形态及生理生化特性与霍乱弧菌相同，但不被０１群霍乱弧菌多价血清凝集。血清学分型结果均为不同的ＶＢＯ血清型ＶＢＯ５，１４，２６，４７，５６。用ＤＮＡ霍乱ＣＴ基因探针菌落原位杂交及小鼠肠结扎法测肠毒素，表明菌株有强毒力。分离菌株对小鼠的ＬＤ５０为１５８×１０８个／只。     

Does Ralstonia eutropha,rich in poly‐β hydroxybutyrate (PHB), protect blue mussel larvae against pathogenic vibrios?

The natural amorphous polymer poly‐β‐hydroxybutyrate (PHB‐A: lyophilized Ralstonia eutropha containing 75% PHB) was used as a biological agent to control bacterial pathogens of blue mussel (Mytilus edulis) larvae. The larvae were supplied with PHB‐A at a concentration of 1 or 10 mg/L for 6 or 24 hr, followed by exposure to either the rifampicin‐resistant pathogen Vibrio splendidus or Vibrio coralliilyticus at a concentration of 10⁵ CFU/ml. Larvae pretreated 6 hr with PHB‐A (1 mg/L) survived a Vibrio challenge better relative to 24 hr pretreatment. After 96 hr of pathogen exposure, the survival of PHB‐A‐treated mussel larvae was 1.41‐ and 1.76‐fold higher than the non‐treated larvae when challenged with V. splendidus and V. coralliilyticus, respectively. Growth inhibition of the two pathogens at four concentrations of the monomer β‐HB (1, 5, 25 and 125 mM) was tested in vitro in LB₃₅ medium, buffered at two different pH values (pH 7 and pH 8). The highest concentration of 125 mM significantly inhibited the pathogen growth in comparison to the lower levels. The effect of β‐HB on the production of virulence factors in the tested pathogenic Vibrios revealed a variable pattern of responses.     

基于toxR基因病原哈氏弧菌PCR检测方法的建立

基于toxR基因序列设计检测哈氏弧菌的1对特异性引物,通过PCR反应条件优化、PCR反应特异性及敏感性检测,建立一种哈氏弧菌的特异性分子检测方法.试验结果表明,该引物可使哈氏弧菌扩增出大小为390 bp的toxR基因片段,3种水产动物病原弧菌未扩增出任何条带,敏感性检测结果为该PCR反应最低能检测出0.375 ng/μ...     

仿刺参幼体烂胃病及其致病原鉴定

山东、辽宁两省在仿刺参（Apostichopus japonicus）苗种培育期间的5-7月，耳状幼体易发生一种以胃壁增厚、萎缩、溃烂为主要特征的“烂胃病”。从患烂胃病的耳状幼体中分离到1株细菌LW-1，人工回接感染实验证实其具有较强的致病性，并与自然发病症状相同。通过API半自动化鉴定和常规的生理生化试验的结果表明，LW-1具有弧菌属的特征，其表型特征与灿烂弧菌（Vibrio splendidus）相似。对该菌株16S rRNA基因序列分析和构建系统发生树。结果显示，菌株LW-1与灿烂弧菌的亲缘关系最近，相似率达到99．8％。菌株LW-1可鉴定为灿烂弧菌，并视为耳状幼体烂胃病的致病原之一。通过对多起仿刺参苗期烂胃病的调查研究表明，烂胃病病原也具有多样性和复杂性，并与投喂老化腐败的单胞藻饵料有关。     

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。     

Susceptibility of Three Penaeus Species to a Vibrio campbellii-like Bacterium

The susceptibility of larvae and post-larvae of Penaeus indicus, P. monodon , and P. semisulcatlls to a Vibrio campbellii-like bacterium was studied by immersion challenge. Exposure to bacteria caused up to 100% mortality, depending upon bacterial dose and time of exposure. Mortality rate was higher for nauplius and protozoeal stages than for mysis and post-larval stages. Based upon LC50 values at different time intervals, P. monodon was found to be the most sensitive of the three shrimp species to this pathogen.     

Characterization by whole-cell hybridization of bacterial populations associated with shrimp hatchery biofilms

The impact of shrimp larvae development, as well as water and food inputs upon the increase of bacterial populations within the bacterial community of hatchery tank biofilms, was studied. For this study, a total of 68 biofilm samples were collected from concrete tanks at three larvae production times in a commercial shrimp hatchery. Seventeen samples were taken at each larval development stage (Zoea I, Mysis I, postlarvae 1 and postlarvae 16), as well as 37 samples from water, shrimp nauplii and food, introduced into the shrimp hatchery tanks. Culturable and direct bacterial counts were performed and 16S‐rRNA‐targeted oligonucleotide probes were used to quantify the presence of specific bacterial groups. An average of 27–70% of DAPI total cell counts were detected with the EUB338 probe, while the GAM42a probe signal ranged from 1% to 11%. Vibrio‐like bacteria (VLB) counts in TCBS agar ranged from <10 to 10¹ VLB/cm⁻², with a tendency to increase at the last postlarvae stage. The most significant external source of bacteria registered with GAM42a probe and TCBS agar were found in live Artemia nauplii, used as food; nevertheless, biofilms remain with low counts of these groups.     

虎纹蛙烂趾病病原菌分离鉴定与药敏试验

从息典型烂趾病的虎纹蛙病灶组织分离到1株细菌.暂命名为WD-01,经人工感染证实该细菌为虎纹蛙烂趾病的病原菌.通过形态学、生理生化特性对其进行鉴定,并用纸片法进行药敏试验.鉴定结果表明,细菌WD-01为嗜水气单胞菌.药敏试验结果表明,新霉素、利福平、先锋霉紊、诺氟沙星、庆大霉素、强力霉素、链霉素、卡那霉素、土霉素和金霉素等12种药物对菌株WD-01具有明显的抑制作用.