中华鳖出血性肠炎病免疫防治的初步试验

将中华鳖出血笥肠炎病原病菌－－嗜水气单胞菌制成灭活菌苗家兔,６０天家兔血液中产生抗体,能与抗原凝集反应,凝集效价为６４０～１２８０。用嗜水气单胞菌苗免疫２０～６９ｇ的鳖,３周抗攻击保护率为７３．３３％,５周为８６．６７％;土法灭活轩免疫ｇ的鳖,３周抗攻击保护率为６６．６７％,５周为８０％。中华鳖出血性肠炎病免疫防治是可行的。     

3种免疫途径下迟缓爱德华菌菌蜕疫苗对欧洲鳗的免疫保护效果

为了探讨迟缓爱德华菌菌蜕疫苗预防鳗鲡爱德华菌病的可行性,实验采用腹腔注射、口服、浸泡3种途径探讨了迟缓爱德华菌菌蜕疫苗对欧洲鳗的免疫保护效果。结果表明,免疫后欧洲鳗血清抗体效价均明显升高,但菌蜕疫苗(ETG)组与福尔马林灭活疫苗(FKC)组间血清抗体效价无显著性差异。欧洲鳗注射、口服、浸泡ETG组的相对免疫保护率分别为75.0%、52.5%、37.5%,分别高于FKC组的55.0%、40.0%、32.5%,且ETG组欧洲鳗死亡时间明显推后,表明菌蜕疫苗的免疫保护效果优于福尔马林灭活疫苗。易操作、对鱼体应激少的ETG疫苗口服免疫欧洲鳗获得了52.5%的免疫保护率,在预防鳗鲡爱德华菌病中具有良好的应用前景。     

嗜水气单胞菌灭活苗对林蛙免疫效果的初步研究

用分离于患红腿病的长白山中国林蛙的嗜水气单胞菌,采用热灭活的方法制成嗜水气单胞菌灭活苗,通过不同的途径免疫接种林蛙,通过测定受免林蛙血清中凝集抗体效价,探讨林蛙对热灭活的嗜水气单胞菌菌苗的免疫应答状况.结果表明,受免林蛙血清中凝集抗体效价可达到27,明显高于对照组.通过检测免疫林蛙的免疫保护率发现受免林蛙中口服免疫组、浸泡免疫组、腹腔注射组、肌肉注射组的免疫保护力可达71%、76%、59%、47%.     

罗非鱼海豚链球菌疫苗及其免疫效果的研究

用筛选到的海豚链球菌(Streptococcus iniae)临床分离菌株CMS005,甲醛灭活制备成疫苗并对其进行了注射、浸泡和口服免疫奥尼罗非鱼的效果研究。结果显示:实验室水族箱试验的注射、浸泡和口服免疫的最佳免疫保护率分别为90.5%、61.9%和14.3%,水泥池小网箱试验的最佳免疫保护率分别为100%、86.1%和66.7%。免疫剂量对浸泡免疫效果的影响大于注射和口服免疫,加强免疫可显著提高浸泡和口服免疫效果,超声波处理可以提高浸泡免疫效果。注射和口服免疫7 d和15 d后均可控制罗非鱼海豚链球菌病的继续发生;未免疫组罗非鱼月累计死亡率为4%~16%,而免疫组罗非鱼月累计死亡率低于0.5%;疫苗产生的免疫保护力可持续2~3个月。     

中华鳖爱德华菌病病原菌的分离鉴定及致病因子研究

采用API 20E系列生化鉴定及16S rDNA和gyrB基因序列同源性分析方法,对从患病中华鳖(Trionyx sinen-sis)肝脏中分离到的一株细菌TL5m进行了鉴定,并通过人工感染试验,对该菌株进行了毒力检测;此外分别提取该TL5m株的主要致病因子外膜蛋白、脂多糖和胞外产物,对中华鳖进行毒力和免疫保护率试验。结果显示:菌株TL5m的API 20E鉴定编码为4544000,99.9%为迟钝爱德华氏菌(Edwardsiella tarda);其16SrDNA序列和gyrB基因序列(GenBank登录号分别:EF121756和GU563803)与迟钝爱德华氏菌的同源性最高(分别为94%和98%);菌株TL5m对中华鳖的半数致死量LD50为2.45×106 CFU/ind。药敏感结果显示菌株对磷霉素、菌必治、头孢孟多、头孢噻吩、壮观霉素高度敏感。外膜蛋白攻毒剂量60μg/ind和脂多糖攻毒剂量400μg/ind时,对中华鳖的致死率都为33.3%,胞外产物对中华鳖的LD50为31.73μg/ind;全菌灭活苗、胞外产物、外膜蛋白和脂多糖的免疫保护率分别为75%、62.5%、25%和87.5%。结果表明,发病中华鳖的病原菌为迟钝爱德华氏菌,其分泌的胞外产物对中华鳖具有较高毒力;提取的脂多糖对中华鳖遭受迟钝爱德华氏菌攻击具有较高免疫保护率。     

嗜水气单胞菌疫苗免疫效果研究

选取对鲤鱼毒力强的嗜水气单胞菌菌株制成灭活疫苗,研究疫苗的免疫保护效应.采用甲醛灭活制备疫苗,分别对鲤鱼进行注射和浸泡试验,测定白细胞吞噬活性、淋巴细胞转化能力、血清、皮肤黏液的抗体效价及活菌攻毒后的免疫保护率.结果表明,受免鱼各项指标均明显高于对照组,注射组免疫保护率达90%,其中浸泡组各项略低于注射组.嗜水气单胞菌灭活疫苗对鲤鱼有显著免疫保护效应,可作为预防细菌性败血症感染的疫苗.     

鳜浸泡嗜水气单胞菌全菌疫苗后皮肤黏液抗体的变化

应用间接ELISA方法,研究经嗜水气单胞菌(Aeromonas hydrophila)疫苗浸泡免疫后鳜(Siniperca chuatsi)皮肤黏液中抗体消长规律,以揭示鳜局部黏膜免疫在免疫保护中的作用及浸泡免疫的保护效果。结果表明,鳜皮肤黏液中抗体滴度在免疫后第7天达到峰值211,抗体从开始形成到消失持续21 d;添加佐剂(IMS1312、葡聚糖、莨菪碱、食盐)可以提高鳜皮肤黏液及血清中抗体滴度和相对免疫保护率,其中添加IMS1312组免疫保护率最高,达77.8%,免疫保护与抗体滴度成正相关(R2=0.79,P<0.05);以溶菌酶为代表的非特异性免疫在免疫后1周内即抗体未形成时,对鱼体起主要保护作用。此外,通过比较鳜血清与皮肤黏液中的抗体消长规律,发现血清中抗体滴度峰值出现时间较迟(第14天),抗体持续时间较长(42 d),初步推断鳜皮肤黏膜中可能存在相对独立于全身免疫系统的局部黏膜免疫系统。     

鲖爱德华菌口服疫苗对斑点鲖的免疫效果

为评价鲖爱德华菌口服微球疫苗对斑点鲖的免疫效果,实验以天然高分子聚合物海藻酸钠和鲖爱德华菌灭活疫苗为材料,制备鲖爱德华菌口服微球疫苗。将实验动物随机分为鲖爱德华菌微球疫苗组、鲖爱德华菌灭活疫苗组、空微球组和对照组,以拌料口服方式进行免疫,通过检测血清中溶菌酶活力、总超氧化物歧化酶(T-SOD)活力、补体替代途径(ACH50)活性等非特异性免疫指标,抗体效价以及相对免疫保护率评价疫苗免疫效果,采用荧光定量PCR检测口服疫苗对斑点鲖免疫相关基因表达量的影响。结果显示,鲖爱德华菌口服微球疫苗能够较长时间增强斑点鲖非特异性免疫功能;血清凝集效价于第5周达到峰值,为1∶16,免疫后第7周仍可检测到特异性抗体;口服鲖爱德华菌微球疫苗的斑点鲖获得的抗鲖爱德华菌相对免疫保护率为60.7%,远高于灭活疫苗组(14.3%)及空微球组(10.7%);荧光定量分析结果显示,攻毒后48 h相比攻毒前各免疫基因表达量均有上调,鲖爱德华菌微球疫苗对受免鱼肾脏、脾脏中免疫基因的表达影响尤为明显。结果表明,鲖爱德华菌口服微球疫苗能增强斑点鲖非特异性免疫功能,对鲖爱德华菌病起到一定的预防作用。     

海豚链球菌灭活疫苗对罗非鱼免疫效果的初步研究

将海豚链球菌(Streptococcus iniae)淡水分离株TBY-1菌株灭活后制备成无佐剂和添加白油佐剂的疫苗,利用浸泡和腹腔注射复合免疫方式对罗非鱼(Oreochromis niloticus)进行免疫,比较其相对保护率及血清中抗体凝集效价,以评价佐剂的存在与否及免疫次数对海豚链球菌灭活疫苗的免疫效果的影响,同...     

鸡蛋抗体治疗病鳖的初探

从病鳖分离得到的不同气单胞菌,制备四联菌苗,免疫母鸡,取鸡血清抗体效价为１：３２０时的鸡蛋,制马鸡蛋抗体制剂,治疗病鳖,治愈率为３６．６％     

草鱼出血病的免疫学研究进展

由于草鱼病毒性出血病发病率高,且尚未找到有效防治的药物,至今一直成为草鱼养殖发展中"瓶颈"问题的根本原因。因此,采取免疫学途径防治草鱼出血病是解决此问题的关键之所在。综述了采用草鱼出血病疫苗、干扰素及免疫增强剂进行草鱼出血病防治的研究现状;提出口服型免疫增强剂是生产上防治草鱼出血病最为有效的途径之一。     

精制禽流感高免卵黄抗体的研究

用禽流感油乳剂灭活苗按一定的免疫程序免疫SPF蛋鸡,制备高免卵黄抗体,用改良的水稀释法对IgY进行纯化,经过抗体效价检测、无菌检验、安全试验、动物试验,该卵黄抗体可以用于田间试验。保存期试验证明:精制禽流感高免卵黄抗体室温条件下可保存10个月,4℃条件下保存15个月,-20℃条件下保存18个月。     

金枪鱼延绳钓钓具的最适浸泡时间

根据2010年10月—2011年1月金枪鱼延绳钓海上调查数据,分两种起绳方式,建立每次作业每一根支绳的浸泡时间计算模型。将钓具的浸泡时间以1 h为间隔分别统计每个区间的支绳数量及大眼金枪鱼(Thunnus obesus)、黄鳍金枪鱼(Thunnus albacores)的渔获尾数,并计算其钓获率(CPUE)。结果表明:1)大眼金枪鱼和黄鳍金枪鱼的CPUE都随浸泡时间的增加呈现先增后减的趋势,这是由于饵料的诱引效果变化及渔获的丢失引起的;2)二次曲线可拟合浸泡时间与大眼金枪鱼和黄鳍金枪鱼CPUE的关系;3)大眼金枪鱼和黄鳍金枪鱼CPUE最高的浸泡时间分别为9.9 h和10.1 h。建议:1)今后在金枪鱼延绳钓作业中,保证每一根支绳在水中的浸泡时间为9.5~10.5 h,以提高捕捞效率并减少副渔获物;2)可把延绳钓钓具的浸泡时间作为有效捕捞努力量,并用于CPUE的标准化。研究结果可用于提高捕捞效率并减少副渔获物的技术方案制订,并为渔业生产和CPUE的标准化提供科学参考。     

无乳链球菌不同途径感染吉富罗非鱼后病原菌在体内的分布规律研究

为探讨无乳链球菌(Streptococcus agalactiae)感染罗非鱼(Oreochromis spp)的致病途径。采用腹腔注射、灌胃和浸泡三种方式对吉富罗非鱼进行无乳链球菌(HN016菌株)胁迫感染,利用平板活菌计数法统计三种方式感染后病原菌在体内组织的分布。注射和灌胃两种方式感染后均出现典型的链球菌感染发病症状,其中注射组在感染24 h后出现死亡高峰,死亡率为92.5%;灌胃组感染48 h后出现死亡高峰,死亡率为90%;而浸泡组,感染后均没有出现明显的发病症状,也没有出现死鱼。注射组和灌胃组在感染后2 h,其脾脏、肝脏、前肾、胃、腮、皮肤和肌肉组织中均可分离出病原菌,5 h后在脑组织中均可分离出病原菌,8 h后各组织分离出的病原菌数达到峰值;而浸泡组在感染8 h后才从各组织中分离出病原菌,且它们的数量均低于同时期的注射组和灌胃组。注射和灌胃两种方式可使吉富罗非鱼快速感染无乳链球菌而发病,而浸泡方式感染后病原菌虽可以侵入机体,但不表现出症状。由此,我们推测在自然条件下养殖的罗非鱼是通过口腔采食携带无乳,链球菌的食物而被感染。     

蛭弧菌的生物学特性及其在刺参养殖中的应用前景

蛭弧菌是一种以捕食细菌为生的寄生性细菌,对诸多水产动物病原菌具有吞噬和裂解作用。蛭弧菌能够预防水产动物的细菌性疾病、改善养殖环境以及增强水产动物机体免疫力。养殖刺参的疾病以细菌性病害为主,利用蛭弧菌可以预防刺参疾病的发生。关键词蛭弧菌水产养殖刺参应用前景     

Protective properties of egg yolk IgY containing anti-Edwardsiella tarda antibody against paracolo disease in the Japanese eel, Anguilla japonica Temminck & Schlegel

Abstract. Immunized hens are known to contain a high level of immunoglobulin Y (IgY) in their egg yolk. In this study, the present authors obtained anti- Edwardsiella tarda IgY (containing 20% specific IgY, agglutination titre, 1:128) from hens vaccinated by injection with formalin-killed bacterin. The IgY was stable against eel digestive factors, and therefore, was orally administered with viable E. tarda to the Japanese eels and the efficacy of protection against E. tarda infection was evaluated. Orally administered IgY at a dose of 400mg fish^-1 cleared E. tarda inoculated simultaneously at 10^5–6 CFU fish^-1 from the intestine within 24h. Moreover, orally administered IgY at doses of 200 and 400mg fish^-1 inhibited the penetration of E. tarda inoculated simultaneously at 10^4–6 CFU fish^-1 into the liver and kidney via the damaged intestine. The fish orally administered with IgY showed reduced mortality. These results suggest that egg yolk containing anti- E. tarda IgY is effective in preventing edwardsiellosis.     

Rainbow trout silage as immune stimulant and feed ingredient in diets for Mozambique tilapia (Oreochromis mossambicus)

Peptides and free amino acids resulting from protein hydrolysis could act as stimulants of fish non‐specific immunity. The study aimed to determine the potential of rainbow trout viscera silage as immune stimulant and feed ingredient for Mozambique tilapia, and to establish whether formic acid used during silage preparation contributed to any effects. Four diets were evaluated: a reference diet (R), one containing 6.5 g kg^?1 formic acid (FA) and two silage diets containing 160 g kg^?1 (SL, low inclusion) and 285 g kg^?1 (SH, high inclusion) silage. Low silage inclusion improved phagocytic activity of leucocytes compared with the reference, while high inclusion showed no improvement. No other non‐specific immunity parameters or haematology were affected by any treatments. High silage inclusion significantly decreased growth and led to higher mortality, while formic acid had no effect on growth. It is concluded that rainbow trout viscera silage can stimulate the cellular non‐specific immunity of Oreochromis mossambicus, and that protein hydrolysis products (and not formic acid) is responsible for the stimulation. The silage can also serve as source of dietary protein and essential amino acids in tilapia diets. However, both fish growth performance and improvement in cellular immunity are dependent on silage inclusion level.     

Influence of high dietary α-tocopherol intakes on specific immune response, nonspecific resistance factors and disease resistance of healthy and aflatoxin B1-induced immunocompromised Indian major carp, Labeo rohita (Hamilton)

The effect of aflatoxin treatment and/or feeding of a high level of α‐tocopherol on immune response and disease resistance was investigated in Indian major carp, Labeo rohita. Group A served as a healthy control, group B was treated with aflatoxin, group C was fed a high level of α‐tocopherol whereas group D was exposed both to aflatoxin and a high level of dietary α‐tocopherol for 60 days. Aflatoxin B₁ (AFB₁) was injected once intraperitoneally into fish on the first day of the experiment (groups B & D). High levels of DL‐α‐tocopherol (1000 mg kg^–1 feed) were provided to healthy as well as AFB₁‐treated immunocompromised fish for 60 days (groups C & D). At the end of the experiment blood samples were assayed for changes in nonspecific immunity and humoral protein levels. Disease resistance against two common bacterial pathogens viz., Aeromonas hydrophila and Edwardsiella tarda were evaluated in all groups. Significant (P < 0.05) suppression of specific immunity as measured through haemagglutination (HA) titre against sheep red blood cells (SRBCs) as well as bacterial (formalin‐killed E. tarda) agglutination titre; nonspecific resistance factors viz., globulin level, serum bactericidal and lysozyme activities, neutrophil activities, and disease resistance against two bacterial pathogens only in aflatoxin‐treated fish with respect to the control group, clearly indicated the immunosuppressive nature of aflatoxin. Feeding of a high level of α‐tocopherol to AFB₁‐treated immunocompromised fish significantly (P < 0.05) raised specific immunity, nonspecific resistance factors and disease resistance capacity when compared with aflatoxin‐exposed fish. Disease resistance and enhancement of immune status through feeding of high levels of α‐tocopherol to healthy as well as AFB₁‐treated immunocompromised fish confirmed the potential of α‐tocopherol in carp feed for prevention of disease and for combating natural/environmental immunosuppressants.     

以可消化氨基酸为基础研制产蛋鸡无鱼粉饲粮试验

选用272只罗曼褐商品蛋鸡，采用2×3重复交叉试验设计进行了以可消化氨基酸为基础研制产蛋鸡无鱼粉饲粮的研究。结果表明，根据饲料可消化氨基酸含量。采用全植物性蛋白质饲料研制的产蛋鸡无鱼粉饲粮，试鸡的产蛋性能、饲料转化率均达到相同营养水平的鱼粉型饲粮的效果，料蛋比为2．4％。每吨无鱼粉型饲粮可降低原料成本74．96元，平均每只试鸡每天可多获毛利0．0168元，提高11．28％，经济效果良好。     

Seasonal variation in the prevalence of 'red spot' disease in estuarine fish with particular reference to the sea mullet, Mugil cephalus L.

Abstract. In March 1972 an epizootic of a non-specific fish disease (red spot), was reported in estuarine fish stocks of the Burnett River in central Queensland, Australia. The disease, characterized by the presence of external lesions, infected a wide range offish species. Red spot has now reportedly spread to the river systems of New Guinea in the north and southwards to central New South Wales. The sea mullet, Mugil cephalus , was selected as a target species for the monitoring of epizootics within the Noosa River system of southern Queensland. Disease epizootics appeared to be related to the occurrence of certain environmental changes such as low or rapidly changing temperatures and rapid or prolonged depressions of salinity in the estuarine habitat. The effect of crowding, migration and spawning upon the transmission of this disease is also discussed. Simultaneously conducted microbiological investigations indicated that the bacterium, Vibrio anguillarum was the causative agent of red spot epizootics.