鲢,鳙溶血性腹水病研究

报道了链，鳙，鱼溶血性腹水病的病原和防治方法。研究结果表明，该病无病毒性病原存在，病原为嗜水气单胞菌嗜水亚种。采用作者研制的鱼康乐进行治疗，治愈率达８６．５％。     

工厂化养殖暗纹东方鲀致病菌的分离和鉴定

从不同养殖场患病暗纹东方鲀（Takifagu obscurus）的鳃、肠、肝、肌肉及溃疡部位分离到9株菌株，经人工感染致病性试验，其T-22、T-11、J-2、H-3、H-5表现出较强的致病性，出现症状与自然发病鱼相同。通过对菌体形态特征、培养特性和API20E细菌生化快速鉴定试条的测定，分别鉴定为温和气单胞菌（Aeromonas sobria）、豚鼠气单胞菌（Aeromonas calyioe）、嗜水气单胞菌（Aeromonas hydrophila）、类志贺邻单胞菌（Plesiomonas shigelloides）、霍乱弧菌（Vrbrio cholerae）。证实这5株菌为致病菌。药敏试验结果表明9种药物对温和气单胞菌、豚鼠气单胞菌、霍乱弧菌均有较好的抑菌效果；嗜水气单胞菌表现出明显的耐药性。氟嗪酸、环丙沙星、复合磺胺对类志贺邻单胞菌有一定抑菌效果。     

铌（Ⅴ）的极谱催化波研究及其应用

在ＨＮＯ＿３·ＥＤＴＡ─ＦｅｚＯ＿３·Ｈ＿２ＳＯ＿４介质中，铌（Ⅴ）─氟化铵─酒石酸─偶氮胂（Ⅲ）体系形成灵敏的导数极谱催化波，峰电位在─０．８６Ｖ（ＶＳ·ＳＣＥ），Ｎｂ＿２Ｏ＿５量在０─２．５μｇ／２５ｍｌ范围内与峰电位成线性关系，拟定了一个不经分离，适用于硅酸盐，碳酸盐及化探样品中微量铌的测定方法。     

化探样品中微量铀的示波极谱测定

在醋酸－醋酸钠－铜铁试剂一二苯胍体系中对微量铀进行极谱测定。在峰电位为－１．１７Ｖ，铀浓度在０．０５μｇ·ｍＬ￣（－１）－２μｇ·ｍＬ￣（－１）之间时铀的浓度与峰电流成线性关系。本方法选用碳酸铵小体积沉淀分离干扰元素，直接取其清液测定微量铀。最低检出限为０．０２μｇ·ｍＬ￣（－１）。     

应用塑性图对陕西特殊土的判别

塑性图是由卡萨格兰德（Ａ·Ｃａｓａｇｒａｎｄｅ）于１９４２年首次提出的，该图将阿太量（Ａ·Ａｅｔｅｒｂｅｒｇ）界限（液限、塑限）应用于土质分类．本文根据塑性图判别理论并应用塑性图，对陕西省特殊土（黄土、三趾马红土、膨胀土等）进行了判别，结果为黄土位于ＣＬＹ区，三趾马红土和膨胀土位于ＣＨＥ区，其效果较好，达到了分类的目的．     

6300ZC型柴油机加中冷节能研究

在６３００ＺＣ型柴油机上加装中冷器以实现节能，并对其技术关键的热阻与风阻损失问题进行了研究，设计研制了二代样机。经实船试验，节油率在３％～８％；经台架对比试验，耗油率从２１５ｇ／（ｋＷ·ｈ）降到２０９／（ｋＷ·ｈ）。     

多鳞鱚Sillago sihama Forskal外周血细胞的显微结构观察

对多鳞鱚（Sillago sihama Forskal）Wright染色的血涂片进行显微观察。结果表明多鳞鱚外周血细胞可区分为红细胞、血栓细胞、淋巴细胞、嗜中性粒细胞、嗜酸性粒细胞和单核细胞等6种细胞,未发现嗜碱性粒细胞。血栓细胞、淋巴细胞、嗜中性粒细胞和单核细胞在白细胞中所占比例分别为47．5％、32．5％、12．5％和7．5％,嗜酸性粒细胞在整个实验中只发现7个。并发现较多未成熟的红细胞以及少量正在分裂的红细胞和晚幼嗜中性粒细胞。     

黄鳍鲷细菌性溃疡病致病菌研究

研究了黄鳍鲷细菌性溃疡病的病原。从患病濒死的黄鳍鲷体内分离到两株细菌,经人工感染试验确定为致病菌,通过测定其形态及生理生化特性得出：致病菌为革兰氏阴性杆菌,极生单鞭毛,发酵葡萄糖产酸不产气,氧化酶阳性、过氧化氢酶阴性,发酵乳糖,甘露醇发酵阳性,明胶酶阳性,在70g／LNaCl培养基中生长良好,对O／129不敏感;由于两株菌对山梨醇、水扬苷、七叶苷及麦芽糖等氧化发酵的结果不一致,分别被鉴定为嗜水气单胞菌不产气亚种（Aeromonashydropilaanaerogenes）和温和气单胞菌（A．sobria）变异亚种。药敏试验显示,两个菌株对氯霉素高度敏感而对氟哌酸中度敏感。两株菌对黄鳍鲷的半致死剂量（按每g鱼计算）分别为2．3×106个菌体和2．6×105个菌体。     

鳖甲溃疡和穿孔病的病原及其防治

对鳖甲溃疡和穿孔病的病原进行了研究,提出了勒氏假单胞菌（Pseudomonaslemoignei）、产碱菌（Alcaligenessp）和枯草芽孢杆菌（Bacillussubtilis）3种病原菌;同时在病原药敏试验的基础上筛选治疗药物（鳖康乐1号）,并进行了生产性防治试验。     

九孔鲍外套膜表皮细胞的超微结构

利用透射电镜观察九孔鲍（Haliotidaediversicoloraquatilis）的外套膜表皮细胞,结果表明细胞可分为4大类,即普通柱状表皮细胞、嗜碱性粒细胞、嗜酸性粒细胞和感觉细胞。它们在不同区域的分布、形态和数量变化与外套膜的功能分化密切相关。     

Protective efficacy evaluation of immunogenic protein AHA_3793 of Aeromonas hydrophila as vaccine candidate for largemouth bass Micropterus salmoides

Journal of Oceanology and Limnology - Aeromonas hydrophila is a Gram-negative pathogen that can infect various fish, including largemouth bass (Micropterus salmoides), which have caused huge...     

Isolation and characterization of AHL-degrading bacteria from fish and pond sediment

Quorum sensing(QS)disruption is considered as a potential alternative strategy to combat bacterial diseases in aquaculture.In this study,we isolated and identified bacteria degrading QS molecules from pond sediment and fish intestine.A total of 132 strains were obtained in the enrichment culture,of which two strains were identified as Enterobacter sp.f003 and Staphylococcus sp.sw120,being isolated from the fish intestine and pond sediment,respectively.We found that strains f003 and sw120 could degrade acyl-homoserine lactones(AHLs)and cause no hemolysis of sheep red blood cells.The AHL lactonase(aiiA)homologous gene in the two strains was detected in PCR amplification and the high-degrading activity to N-hexanoyl-L-homoserine lactone(C6-HSL)and AHLs secreted from pathogenic Aeromonas hydrophila was assessed.Meanwhile,the artificial infection of cyprinid Carassius auratus gibelio with intraperitoneal injection showed that the two strains were avirulent.Therefore,the obtained indigenous bacteria are candidate probiotics against pathogenic A.hydrophila in aquaculture.     

Non-specific immune response of bullfrog Rana catesbeiana to intraperitoneal injection of bacterium Aeromonas hydrophila

Non-specific immune response of bullfrog Rana catesbeiana to pathogenic Aeromonas hydrophila was studied to 60 individuals in two groups.Each bullfrog in bacterium-injected group was injected intraperitoneally(i.p.) with 0.2 ml bacterial suspension at a density of 5.2 × 106 CFU/ml,while each one in control group injected i.p.with 0.2 ml sterile saline solution(0.85%,w/v).Three bullfrogs in both groups were sampled at 0,1,3,7,11,15 and 20 days post-injection(dpi) for the evaluation of non-specific immune parameters.It was observed that intraperitoneal injection of A.hydrophila significantly increased the number of leucocytes and that of NBT-positive cells in peripheral blood.Significant increases in serum bactericidal activity and serum acid phosphatase activity were also observed in the bacterium-injected frogs when compared with those in the control group.However,a significant reduction was detected in vitro in phagocytosis activity of peripheral blood phagocytes.No significant difference in changes in the number of peripheral erythrocytes,serum superoxide dismutase(SOD) activity,and lysozyme activity was detected between the two groups.It is suggested that bullfrogs may produce a series of non-specific immune reactions in response to the A.hydrophila infection.     

Rapid,sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification

Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming.The rapid detection and identification of V.anguillarum,and other pathogens that infect marine organisms,is crucial to effective disease management.In this study,we developed a loop-mediated amplification (LAMP) assay to detect V.anguillarum in an hour in a single tube without the need for thermal cycling.Conserved regions of the metalloproteinase (empA) gene of V.anguillarum served as the targets for primer design.A fragment of the empA gene was amplified at 65°C in the presence of the primer mixture and Bst DNA polymerase.In the optimized LAMP assay,6.7 pg of V.anguillarum DNA could be detected.Six strains of V.anguillarum and 17 strains of non-V.anguillarum bacteria were used in this study to evaluate the species specificity of the primers.The six V.anguillarum strains gave a positive result in the LAMP assay.This method was also validated in V.anguillarum-infected fish.This LAMP method is more sensitive than PCR in the detection of V.anguillarum and shows good species specificity.The LAMP assay is therefore an effective method for the quick detection of V.anguillarum both in the laboratory and in the field.     

免疫胶体金检测中华鳖抗毒素抗体和嗜水气单胞菌外毒素

以中华鳖的嗜水气单胞菌纯化其外毒素,用胶体金建立斑点免疫检测方法,测定中华鳖对嗜水气单胸菌外毒素抗体效价。结果表明：该方法有高的特异性,与血清中的其它成分交叉反应小;检测灵敏度达4ng。     

工厂化养殖暗纹东方鱼屯致病菌的分离和鉴定

从不同养殖场患病暗纹东方鱼屯(Takifaguobscurus)的鳃、肠、肝、肌肉及溃疡部位分离到9株菌株,经人工感染致病性试验,其T-22、T-11、J-2、H-3、H-5表现出较强的致病性,出现症状与自然发病鱼相同。通过对菌体形态特征、培养特性和API20E细菌生化快速鉴定试条的测定,分别鉴定为温和气单胞菌(Aeromonassobria)、豚鼠气单胞菌(Aeromonascaviae)、嗜水气单胞菌(Aeromonashydrophila)、类志贺邻单胞菌(Plesiomonasshigelloides)、霍乱弧菌(Vibriocholerae)。证实这5株菌为致病菌。药敏试验结果表明9种药物对温和气单胞菌、豚鼠气单胞菌、霍乱弧菌均有较好的抑菌效果;嗜水气单胞菌表现出明显的耐药性。氟嗪酸、环丙沙星、复合磺胺对类志贺邻单胞菌有一定抑菌效果。     

Optimization of protectant,salinity and freezing condition for freeze-drying preservation of <Emphasis Type="Italic">Edwardsiella tarda</Emphasis>

Novel preservation condition without ultra-low temperature is needed for the study of pathogen in marine fishes. Freeze-drying is such a method usually used for preservation of terrigenous bacteria. However, studies using freeze-drying method to preserving marine microorganisms remain very limited. In this study, we optimized the composition of protectants during the freeze-drying of Edwardsiella tarda, a fish pathogen that causes systemic infection in marine fishes. We found that the optimal composition of protectant mixture contained trehalose (8.0%), skim milk (12.0%), sodium citrate (2.0%), serum (12.0%) and PVP (2.0%). Orthogonal and interaction analyses demonstrated the interaction between serum and skim milk or sodium citrate. The highest survival rate of E. tarda was observed when the concentration of NaCl was 10.0, 30.0 and between 5.0 and 10.0 g L?1 for preparing TSB medium, E. tarda suspension and protectant mixture, respectively. When E. tarda was frozen at ?80°C or ?40°C for 6 h, its survival rate was higher than that under other tested conditions. Under the optimized conditions, when the protectant mixture was used during freeze-drying process, the survival rate (79.63%–82.30%) of E. tarda was significantly higher than that obtained using single protectant. Scanning electron microscopy (SEM) image indicated that E. tarda was embedded in thick matrix with detectable aggregation. In sum, the protectant mixture may be used as a novel cryoprotective additive for E. tarda.     

16S rRNA Gene Phylogenesis of Culturable Predominant Bacteria from Diseased Apostichopus japonicus(Holothuroidea,Echinodermata)

Cultured Apostichopusjaponicus in China suffers from a kind of skin ulceration disease that has caused severe economic loss in recent years. The disease, pathogens of which are supposed to be bacteria by most researchers, is highly infectious and can often cause all individuals in the same culture pool to die in a very short time. The 16S rRNA gene phylogenesis of the culturable bacteria from the lesions of diseased individuals was conducted to study the biodiversity of the bacterial communities in the lesions and to identify probable pathogen(s) associated with this kind of disease. S. japonica samples were selected from a hatchery located in the eastern part of Qingdao, China. Bacterial universal primers GM5F and DS907R were used to amplify the 16S rRNA gene of bacteria colonies, and touchdown PCR was performed to amplify the target sequences. The results suggest that γ- proteobacteria (Alteromonadales and Vibrionales) of CFB group, many strains of which have been also determined as pathogens in other marine species, are the predominant bacterial genera of the diseased Apostichopusjaponicus individuals.     

壳寡糖对吉富罗非鱼幼鱼生长性能、非特异性免疫及血液学指标的影响

用添加质量分数0.00(对照组)、0.10%、0.30%、0.50%和0.70%壳寡糖的饲料饲喂初始体重(3.81±0.23)g的吉富罗非鱼幼鱼(Oreochromis niloticus)10周,研究不同浓度壳寡糖的添加对吉富罗非鱼幼鱼的生长、非特异性免疫功能以及血脂指标的影响。结果表明:与对照组相比,在4个不同浓度添加组中,添加质量分数0.50%壳寡糖能显著提高幼鱼的增重率(P<0.05)、特定生长率(P<0.05),并降低饲料系数(P<0.05);添加质量分数0.3%和0.5%壳寡糖能显著提高幼鱼抗嗜水气单胞菌感染的能力(P<0.01);添加质量分数0.10%、0.30%、0.50%壳寡糖能显著提高幼鱼血清中碱性磷酸酶活性(P<0.05)。同时,各浓度的壳寡糖均能明显提高幼鱼血清中溶菌酶和超氧化物歧化酶活性以及抗嗜水气单胞菌感染的能力(P<0.05),并降低幼鱼血清中总胆固醇和低密度脂蛋白胆固醇水平(P<0.05)。在本实验条件下,添加壳寡糖可提高吉富罗非鱼幼鱼生长性能、饲料利用率、非特异性免疫功能和调节血脂水平,添加量以质量分数0.30%~0.50%为宜。     

广东省罗非鱼主养区无乳链球菌的分离、鉴定与致病性

从广东省5个罗非鱼主养区患暴发性的罗非鱼中分离到15株菌,挑选其中的5株(不同地域、不同组织分离的菌株)进行人工感染试验,表现出自然发病的症状,确定此5株分离菌为导致罗非鱼暴发病的主要病原。通过形态学观察和生理生化实验,并结合16SrRNA基因序列分析,5株分离菌株均为无乳链球菌(Streptococcus agalactiae)。5株分离菌在形态学和生理生化实验结果上保持一致,16SrRNA基因序列同源性达98.6%～99.8%,但在致病性和对药物的敏感性上却呈现一定的差异。另外10株分离菌株通过生理生化实验鉴定也为无乳链球菌。研究表明,广东省罗非鱼流行性暴发病的病原菌主要为无乳链球菌。