团头鲂病原嗜水气单胞菌的分离鉴定及药敏试验

从患病的团头鲂肝脏中分离到1株优势菌WJ-8。将菌株WJ-8进行人工回归感染试验,其患病症状同自然发病症状,证明其对团头鲂有致病性。该菌株的形态特征、主要理化特性、16SrRNA序列测定结果及系统发育树的构建表明该菌株为嗜水气单胞菌。药敏试验结果显示,该菌株对氟苯尼考、甲哌利福霉素、链霉素、卡那霉素等22种药物敏感,对青霉素、氨苄西林、羧苄青霉素、苯唑西林等12种药物表现出耐药性。     

副猪嗜血杆菌的分离鉴定及药敏试验

该研究从广东中山某猪场疑似患有副猪嗜血杆菌(HPS)病的病死猪心脏和肺脏中分离到一株细菌。经形态学观察、培养特性鉴定、生化试验、PCR检测和致病性试验,显示该菌为HPS。药敏试验结果表明该菌株对恩诺沙星、诺氟沙星、红霉素、庆大霉素和新霉素等表现高敏感性;对阿米卡星、土霉素、青霉素、强力霉素和四环素等则耐药。     

七彩神仙鱼肠炎病原菌的分离鉴定及药敏试验

为确定七彩神仙鱼(Symphysodon aequifasciatus)肠炎的致病菌并筛选敏感药物,利用传统培养法,从该鱼的肠道组织中分离出优势菌,通过生理生化试验及16S r DNA序列测序比对确定菌种信息,并使用药敏纸片法筛选致病菌的敏感药物。试验结果表明:优势菌株QC3为肠型点状气单胞菌(Aeromonas punctata),是七彩神仙鱼肠炎的致病菌株,此菌对氟苯尼考、强力霉素、庆大霉素、诺氟沙星高度敏感,对链霉素中度敏感,对土霉素、红霉素、青霉素、头孢唑林、氨苄西林表现出耐药性。根据研究结果,建议使用氟苯尼考、诺氟沙星、强力霉素、庆大霉素等药物对七彩神仙鱼肠炎病进行治疗。     

中华倒刺鲃迟缓爱德华菌的分离鉴定

从患病中华倒刺鲃(Spinibarbus sinensis Bleeker)肝脏中分离得到1株优势菌,命名为SS01。将分离菌株感染健康中华倒刺鲃,人工试验发现其患病症状与自然发病症状一致,半致死剂量(LD50)为2.88×106CFU。根据分离菌株的形态特征、生理生化特性、16S rRNA序列测定结果及系统发育树分析表明菌株SS01为迟缓爱德华菌(Edwadsiella tarda)。药敏试验表明菌株SS01对氟哌酸、恩诺沙星、氟苯尼考、克拉霉素等9种药物敏感,对青霉素、利福平、强力霉素等13种药物表现出耐药性。     

养殖斜带髭鲷源鳗利斯顿氏菌分离鉴定、系统发育分析及药敏试验

斜带髭鲷(Hapalogenys nitens)作为我国东南沿海主要海水网箱养殖鱼类之一,常年受到以细菌性病害为主的侵袭,但相关病原检疫和诊断的研究较少。本文以福建罗源湾养殖濒死斜带髭鲷为研究对象,无菌解剖后对其肝、肾和脾脏进行细菌分离,对纯培养菌使用常规方法和API进行形态和生理生化鉴定,使用16S rDNA、gyrB和rpoD基因进行分子生物学鉴定和系统发育树构建及分析,并对该菌进行药敏试验。菌株鉴定结果为鳗利斯顿氏菌(Listonella anguillarum),但在系统发育树中与多株鳗弧菌(Vibrio anguillarum)聚为一支,药敏试验表明该菌对新生霉素、左氟沙星、罗红霉素、氟苯尼考等7种药物敏感,对磺胺甲基异恶唑、利福平、四环素、强力霉素、氨苄西林等8种药物耐药。     

虹鳟源嗜水气单胞菌(Aeromonas hydrophila)分离及16SrRNA基因鉴定

从发病虹鳟(Oncorhynchus mykiss)肾脏组织中分离到了1株优势菌HZS-01,将分离纯化的菌株进行形态、生理生化特征研究和16S rRNA基因鉴定分析。结果显示,HZS-01株菌的形态和生理生化反应结果符合嗜水气单胞菌(Aeromonas hydrophila)的特征。16S rRNA基因序列分析结果显示,分离菌株在系统发育树上与嗜水气单胞菌(A.hydrophila)属同一分支,同源性达99%以上。综合菌株HZS-01的形态学特征和16S rRNA基因分析结果,将其鉴定为嗜水气单胞菌。药敏试验结果表明,分离菌株对强力霉素、诺氟沙星、左氧氟沙星、新生霉素、阿奇霉素敏感度高,对生产用药预防和治疗有实际指导意义。     

鲟源迟钝爱德华氏菌的分离鉴定与药敏特性研究

从患病鲟鱼的肾脏组织中分离到菌株K9,通过菌株的形态特征、生理生化鉴定及16S rDNA序列分析等,鉴定菌株K9为迟钝爱德华氏菌。人工感染试验证明该菌株为引起鲟鱼疾病及死亡的病原菌;药敏特性研究表明,K9对链霉素、阿米卡星、左氧氟沙星、阿齐霉素、新霉素、氯霉素(禁药)等6种抗生素敏感,而对青霉素G、妥布霉素、麦迪霉素等9种抗生素具有耐药性。本研究结果为有效防控鲟鱼迟钝爱德华氏菌病害提供参考。     

紫贻贝(Mytilus edulis Linnaeu)大量死亡原因的初步研究

从发病紫贻贝（Mytilus deulis）的内脏团内分离出一株菌SS1，通过人工感染试验，证实其为致病菌。经形态、生理生化等几项指标鉴定，该菌株性状与溶藻弧菌基本相符。药物敏感试验结果表明羧苄青霉、万古霉素、克林霉素、氨苄青霉素、阿莫西林等9种药物对该菌株均有明显的抑制作用。     

克氏原螯虾源异常嗜糖气单胞菌的分离鉴定

为了给克氏原螯虾源异常嗜糖气单胞菌的防治提供科学依据,从湖北省某养殖场濒死的克氏原螯虾中分离得到1株优势菌,命名为X1。经生理生化试验分析,初步鉴定该菌为异常嗜糖气单胞菌(Aeromonas allosaccharophila)。进一步的序列分析显示,该菌的16S r DNA与异常嗜糖气单胞菌的16S r DNA一致性达90%,证明其为异常嗜糖气单胞菌。药敏试验结果表明,分离菌株X1对强力霉素、头孢噻肟、诺氟沙星、庆大霉素等敏感。通过动物回归试验,可分离到与自然发病的克氏原螯虾中相同的细菌,且患病症状一致。     

乌龟鲍曼不动杆菌的分离鉴定及药敏试验

自患病乌龟肝脏中分离出1株优势菌DL01221,对其进行形态学观察、生化试验、16SrRNA基因序列分析,同时采用Kriby-Bauer纸片扩散法进行药物敏感性试验。试验结果表明,该菌为革兰氏阴性球杆菌,将该菌株16SrRNA序列进行同源性和系统进化树分析,发现它与鲍曼不动杆菌处于同一群,同源性超过98%,结合菌株形态和生化指标,确定该菌株为鲍曼不动杆菌。致病性试验结果显示,其对乌龟具有较强的致病作用;耐药性结果显示其对头孢他啶、卡那霉素及氟罗沙星敏感,对其他多种抗生素耐药。     

Isolation and characterization of pathogenic Vibrio alginolyticus from diseased postlarval abalone, Haliotis diversicolor supertexta (Lischke)

Outbreaks of serious mortality among cultured abalone postlarvae have occurred across Southern China since July 2002. Five motile bacterial strains were isolated from diseased abalone postlarvae on tryptic soy agar supplemented with 1% NaCl (TSA1) and/or thiosulphate citrate bile salt (TCBS) sucrose agar plates during an outbreak in August 2003 in Shanwei, Guangdong province. All isolates were characterized and identified as Vibrio alginolyticus on the basis of biochemical characteristics and comparisons with those of the reference strain V. alginolyticus ATCC 17749. Strain 19 (a representative of five similar isolates) was virulent to abalone postlarvae with an LD₅₀ value of1.00 × 10⁴ colony‐forming units mL^?1. All abalone postlarvae exhibited the same signs as in natural outbreaks. The same bacterium could be re‐isolated from abalone postlarvae after bacterial challenge using TSA1 and TCBS plates. The results reveal that V. alginolyticus is an infectious agent of abalone postlarvae.     

杂色鲍健康苗种与患病苗种异养细菌致病毒力因子的比较研究

2002年以来,南方杂色鲍苗出现大规模死亡的问题,严重影响到南方鲍产业的发展。为寻找病因,于2004年自广东深圳下沙鲍鱼场培苗过程中变白掉板的杂色鲍苗以及官湖养殖场健康鲍苗各分离到19株和15株异养细菌,在此基础上研究了它们蛋白酶、淀粉酶、明胶酶、磷脂酶的产酶能力及溶血现象,并对它们的产酶能力及溶血能力进行了比较分析。结果表明,虽然来自下沙病鲍苗的菌株比来自官湖的健康鲍苗的溶血能力差;但它们大多数具有较强的分泌蛋白酶、明胶酶、淀粉酶、磷脂酶的能力。考虑到胞外酶是细菌的毒力因子之一,据此推测,下沙鲍鱼场鲍苗发生脱板死亡的原因可能与这些菌株能分泌胞外酶有关。     

Characterization of Pseudomonas aeruginosa associated with diseased postlarval abalone in Shenzhen, China

Outbreaks of mass mortality of postlarval abalone, Haliotis diversicolor supertexta, have occurred in south China since 2002 and have forced many abalone farms to close. About 30 representative bacterial strains were isolated from a sample of five diseased postlarval abalone, taken 25 days post-fertilization during an outbreak of postlarval disease in Shenzhen, China, in October 2006. Bacterial challenge tests showed that the predominant strain, designated as strain 22, was highly virulent to postlarvae with an LD₅₀ value of 7.8 × 10⁴ colony forming units (CFU) ml⁻¹, while four of the other isolates were weakly virulent with LD₅₀ values ranging from 1 × 10⁶ to 1 × 10⁷ CFU ml⁻¹, and the remaining 25 isolates were classified as avirulent with LD₅₀ values greater than 1 × 10⁸ CFU ml⁻¹. By means of API 20NE and 16S rDNA and ITS sequencing analyses, strain 22 was identified as Pseudomonas aeruginosa. Antibiotic susceptibility tests showed that strain 22 exhibited around 75% of susceptibility to 16 various antibiotics tested. The results of this study show P. aeruginosa as one of the bacteria involved in the mortality of abalone postlarvae in Shenzhen, China.     

Growth of Red Abalone,Haliotis rufescens,Fed Cold‐stored Benthic Diatoms

We measured the proximal composition of four benthic diatoms that were stored for 24 mo in the dark at low temperature (4 C by refrigeration) and examined their potential as feed for abalone, Haliotis rufescens, postlarvae. The proximal composition of the four diatoms was modified by species‐specific responses as a function of time in storage. The cultures of all stored diatoms contained low or undetectable concentrations of Vibrio‐like bacteria (<0.01 VLB/mL). As feed for abalone postlarvae, the four diatoms promoted growth under all experimental conditions. Greater shell lengths were measured on Day 14 when Navicula sp. and Navicula incerta were used as feed. Postlarvae that were fed N. incerta and Navicula sp. had higher growth rates. In contrast, lower growth rates were observed on Day 7 with fresh and stored Nitzschia thermalis as food. Survival was high in postlarvae that were fed the four stored diatoms (100%). This report demonstrates that cultures of benthic diatoms that have been stored by refrigeration for 24 mo can be used to feed abalone postlarvae and have an effect on improving growth and survival.     

不同单胞藻饵料培养九孔鲍早期稚贝的研究

以单种培养的6种底栖硅藻和具有底栖习性的亚心形扁藻(Platymoras sp.)为饵料,培养九孔鲍(Haliotis diversi-color supertexta)受精后12 d的早期稚贝并观察其生长和存活,培养期为28 d,目的是筛选能维持早期稚贝高存活率和生长率的藻种。实验结果表明,培养稚贝成活率最高的3个藻种由大到小依次是爪哇曲壳藻亚缢变种(Achnanthes javanica var.)、流水双眉藻(Amphora fluninensis)和亚历山大菱形藻(Nitzschia alexandrina),使稚贝生长最快的3个藻种由大到小依次是流水双眉藻、爪哇曲壳藻亚缢变种和亚历山大菱形藻。研究结果揭示了适宜的底栖硅藻种类对鲍早期稚贝阶段生长和存活的重要性。个体小或者胞外产物量大的藻类很可能对九孔鲍早期稚贝的培养是有利的。     

Performance of a new artificial abalone hatchery culture system in terms of settlement of larvae and growth and survival of postlarval Haliotis discus discus (Reeve)

ABSTRACT:   A new abalone postlarval culture system, Stott's abalone postlarval production system (SAPPS), is being developed as an alternative method for settling larvae and on-growing postlarval abalone. SAPPS was tested in trial 1 using two commercially available artificial diets, Adam and Amos (Adam; supplied by Adam and Amos, Mt Barker, SA, Australia) and Cosmo (Cos; supplied by Cosmo, Matsuyama, Japan), against the diatom biofilm method (Diatom). In trial 2, SAPPS was tested against the previous production system (PPS, used by Stott et al. 2002). Larvae were induced to metamorphose and the resulting postlarvae were on-grown in 10-L flow-through tanks for 28 days in their respective treatments. The final survival of postlarvae was significantly higher ( P  < 0.05) in the SAPPS-Cos group (56.7 ± 11.15%) than in the Diatom (9.4 ± 2.7%) and SAPPS-Adam groups (8.5 ± 1.1%). The final length of postlarvae in SAPPS-Cos (1065 ± 73 µm) was significantly higher ( P  < 0.05) than those in the other two treatments (average of 841–883 µm). In trial 2 the metamorphosis rate was improved by using Spirulina platensis. The final length of postlarvae in SAPPS (1449–1471 µm) was significantly higher ( P  < 0.05) than PPS (1065–1075 µm) and Diatom (993 µm). There is potential for SAPPS to be used as an alternative to the current diatom method in the culture of postlarval abalone.     

Effect of three bacterial isolates from a commercial hatchery on early red abalone (Haliotis rufescens) postlarvae

Three bacterial strains, GHrC11, GHrC13 and GHrC15, were isolated from an abalone postlarval culture system in a commercial farm at Baja California, México. The strains were phenotypically characterized and sequenced (16S rDNA). Strain GHrC11 was a Gram-positive coccobacillum, while strains GHrC13 and GHrC15 were Gram-negative bacilli. Strain GHrC11 was identified as Exiguobacterium sp. The strains GHrC13 and GHrC15 were identified as Vibrio splendidus. The effects of these strains for the development of early abalone postlarvae (2 days old) were evaluated following a completely randomized design with three replicates using 5-mL-volume Petri dishes as experimental units. The experiment considered two different bacterial concentrations of each strain (10³ and 10⁵?cells?ml^?1) and two controls (with and without the benthic diatom Navicula incerta). After 10?days of experimentation, the highest mortality (90?±?5.8?%) and the lowest growth rate (4.1?±?0.1?μm?day^?1) were recorded for the strain GHrC11. In contrast, the lower mortality (16.7?±?3.3?%) and the highest growth rate (11.2?±?0.9?μm?day^?1) corresponded to the control fed N. incerta. Our results suggest that pathogenic effects of these bacterial strains were stronger than any potential benefits derived from the ingestion of bacteria by early abalone postlarvae. In conclusion, the most pathogenic strain was GHrC11, and the intensity of pathogenicity could be ordered as Exiguobacterium sp.?>?V. splendidus (C13)?>?V. splendidus (C15).     

Histological evidence of accumulation of iron in postlarvae of red abalone,Haliotis rufescens

The effect of iron on abalone postlarvae (Haliotis rufescens) was investigated in a controlled‐culturing system. Three iron concentrations (0.15, 1.5 and 15 mg L^?1 of Fe) and a control (no iron added) were used to culture H. rufescens postlarvae while being fed the diatom Navicula inserta over 10 days. Results indicate that H. rufescens postlarvae accumulate iron granules in the stomach, digestive gland and mantle, but not in the gills or other tissues. The number and diameter of iron granules in tissues increased with increasing iron concentration in the culturing environment. The iron accumulation is assumed to have been acquired in the digestive system through the iron‐enriched diatom feed and in the mantle through subcutaneous iron transfer. The lack of iron granules in the gills suggests that iron is not absorbed through the respiratory system, as is the case for many filter feeding bivalves. Exposure to the highest iron concentration (15 mg L^?1) resulted in tissue abnormalities where granules accumulated, and may have significantly affected the health of H. rufescens postlarvae. These findings provide valuable information for the regulation of appropriate iron levels within aquaculture settings and highlights the importance of monitoring iron levels within abalone larval culturing environments.     

Vibrio parahaemolyticus Associated with Mass Mortality of Postlarval Abalone,Haliotis diversicolor supertexta (L.), in Sanya,China

Outbreaks of mass mortality in postlarval abalone, Haliotis diversicolor supertexta (L.), have swept across south China since 2002 and in turn have resulted in many abalone farms closing. Twenty‐five representative bacterial isolates were isolated from a sample of five diseased postlarval abalone, taken 15 d postfertilization during an outbreak of postlarval disease in Sanya, Hainan Province, China in October 2004. A dominant isolate, referred to as Strain 6, was found to be highly virulent to postlarvae in an experimental challenge test, with a 50% lethal dose (LD₅₀) value of 3.2 × 10⁴ colony forming units (CFU)/mL, while six of the other isolates were weakly virulent with LD₅₀ values ranging from 1 × 10⁶ to 1 × 10⁷ CFU/mL, and the remaining 18 isolates were classified as avirulent with LD₅₀ values greater than 1 × 10⁸ CFU/mL. Using both an API 20E kit and 16S recombinant DNA sequence analysis, Strain 6 was shown to be Vibrio parahaemolyticus. It was sensitive to 4 and intermediately sensitive to 5 of the 16 antibiotics used when screening the antibiotic sensitivities of the bacterium. Extracellular products (ECPs) prepared from the bacterium were lethal to postlarvae when used in a toxicity test at a concentration of 3.77 mg protein/mL, and complete liquefaction of postlarvae tissues occurred within 24 h postexposure. Results from this study implicate V. parahaemolyticus as the pathogen involved in the disease outbreaks in postlarval abalone in Sanya and show that the ECPs may be involved in the pathogenesis of the disease.     

Poly‐beta‐hydroxybutyrate‐enriched Artemia sp. for giant tiger prawn Penaeus monodon larviculture

The beneficial effects of PHB as supplement for giant tiger prawn Penaeus monodon postlarvae using a short‐term enrichment strategy via Artemia were examined. The effects of co‐supplementing with a lipid emulsion were also evaluated to determine whether it yielded an additional benefit. Results on the average weight and larval development were not significantly different among postlarvae fed the different dietary treatments, indicating that PHB supplementation could not be used to stimulate growth in P. monodon postlarvae while such positive results have been reported in other aquaculture species. Nonetheless, significantly higher survival was obtained in postlarvae fed PHB‐enriched Artemia irrespective of lipid enrichment. In addition, PHB increased the survival of the postlarvae after exposure to a lethal dose of ammonia. Lipid supplementation nullified this effect. The cumulative mortality of postlarvae subjected to a sublethal concentration of ammonia for 24 h and subsequent exposure to pathogenic Vibrio campbelli showed that PHB but not lipids could effectively enhance the resistance of the postlarvae. Co‐supplementing lipids even significantly decreased this outcome. Our study indicates that PHB supplementation increases the quality of larval P. monodon and their chance of surviving under adverse environmental conditions. The short‐term co‐supplementation with lipid emulsion did not add to these effects.