海豚链球菌（Streptococcus iniae）疫苗对淡、海水养殖鱼类的保护性试验
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摘要
近年来我国水产养殖业发展迅猛，随着高密度、集约化、大规模水产养殖业的出现，鱼类疾病频繁爆发。特别是细菌性疾病种类多、发病率和死亡率高，给水产养殖业造成了巨大的经济损失。由于药物的频繁使用，导致了大量耐药性病原菌的出现，同时也造成大量的药物残留，对人类食品安全及人类健康提出巨大挑战。现代水产养殖业，接种疫苗已被作为评价标准渔业养殖计划的一个重要组成部分，开发环保高效的鱼病疫苗成为当今水产养殖产业的当务之急。本研究主要针对给淡、海水鱼类养殖业带来巨大危害的海豚链球菌(Streptococcus iniae S．iniae)病，以制备的海豚链球菌灭活疫苗为研究对象，主要进行了海豚链球菌灭活疫苗对淡水罗非鱼和海水花鲈鱼的保护性试验。通过本试验筛选了最佳疫苗免疫原用量和最佳免疫佐剂，同时也对药物免疫调节剂在鱼类中应用进行了初步探讨。为海豚链球菌灭活疫苗的推广应用打下了基础。
将S．iniae海水株HD-1、HD-4，S．iniae淡水株TBY-1，TBY-5的培养液灭活后，制备弗氏佐剂灭活疫苗，腹腔注射接种罗非鱼(1×10~(10)CFU×mL~(-1)0.1 mL×尾~(-1))，经过两次免疫，初次免疫后35天，分别用对应的菌株攻毒，统计14天内各组鱼的死亡情况，计算各免疫组的相对保护率(RPS)，并测定各组鱼血清抗体效价的变化。结果表明：HD-1株弗氏佐剂灭活疫苗保护率达到93.6％，最高血清抗体凝集效价达到1：64；TBY-1株弗氏佐剂灭活疫苗保护率达到100％，最高血清抗体凝集效价达到1：64。海水株HD-1和淡水株TBY-1免疫效果均较好，可作为海水鱼和淡水鱼制苗菌株的候选。
利用S．iniae海水株HD-1对大亚湾地区主要的海水养殖经济鱼类石斑鱼、红鳍笛鲷、卵圆鲳鲹、花鲈及美国红鱼进行攻毒试验。结果显示花鲈鱼对S．iniae海水株HD-1最为敏感，可以作为S．iniae海水株HD-1灭活疫苗试验用鱼进行进一步的相关试验。
将S．iniae海水株HD-1制备成10~(10)CFU×mL~(-1)、10~9CFU×mL~(-1)及10~8CFU×mL~(-1)三种菌体抗原浓度的白油佐剂灭活疫苗对花鲈鱼进行了免疫保护性试验，攻毒结果显示：10~(10)CFU×mL~(-1)组相对保护率为100％、10~9 CFU×mL~(-1)组相对保护率为92.5％、10~8CFU×mL~(-1)组相对保护率为85％、对照组的死亡率是100％；最高抗体滴度分别为1：16、1：64、1：32。10~(10)CFU×mL~(-1)组相对保护率与10~9 CFU×mL~(-1)组相对保护率比较，无显著性差异(p＞0.05)；与10~8CFU×mL~(-1)组相对保护率为85％比较，10~(10)CFU×mL~(-1)组保护率较高，具有显著性差异(p＜0.05)。综合考虑生产实际等各项指标，本试验采用10~9CFU×mL~(-1)菌体抗原浓度为制苗用菌体抗原量的浓度。
将含10~9CFU×mL~(-1)菌体抗原浓度的S．iniae海水株HD-1灭活并制备成白油佐剂疫苗和铝胶佐剂疫苗，对花鲈鱼进行免疫保护性试验，攻毒结果显示：白油佐剂疫苗获得92.5％的相对保护率，铝胶佐剂疫苗组获得82.5％的相对保护率；两种疫苗的最高抗体滴度分别为1：64、1：32。两者比较具显著性差异(p＜0.05)。白油佐剂疫苗获得较好的保护效果，可以有效防制花鲈鱼海豚链球菌病。
将左旋咪唑和中药芦荟多糖分别以一定的量加入到10~9CFU×mL~(-1)菌体抗原浓度的S．iniae海水株HD-1白油佐剂灭活疫苗中制备成复合佐剂疫苗(compound adjuvant vaccine)，同时设白油佐剂疫苗基础上单独注射芦荟多糖组、白油佐剂组及PBS组，对花鲈鱼进行免疫保护性试验，攻毒结果显示：各试验组相对保护率差异性不显著(p＞0.05)；但白油佐剂灭活疫苗加单独注射芦荟多糖组鱼血清抗体滴度比其他各组要高(p＜0.05)，而左旋咪唑复合佐剂疫苗组鱼血清抗体效价显著低于白油佐剂灭活疫苗组(p＜0.05)，另外各组血清抗体滴度之间比较没有显著性差异。测定各组血清中溶菌酶变化显示，单独注射免疫增强剂芦荟多糖组血清溶菌酶含量明显比其他各组都要高(p＜0.05)，试验结果表明：中药免疫增强剂芦荟多糖能刺激鱼类产生非特异性免疫应答反应并提高其血清抗体效价。
In decades, with rapid development of aquaculture industry with the high-density and the production on a large scale, the diseases of cultured fish took place frequently, especially the bacteriosis, with high variety, morbidity and mortality, caused huge economic loss to aquaculture. A frequent use of chemical and antibiotic resulted in occurrence of drug- resistant pathogenic strains, simultaneously brought about severe public health and sanitation problems, such as drug residue and food security, to which our human being have to faced and challenged with. In modern aquaculture industry, vaccination will offer a good alternative to strategy of fish disease provention in the highly qualitative fishery cultivation. So developing highly effective vaccine for cultured fish have been an urgent matter in aquaculture industry. In present studies, we mainly focused on Streptococcus iniae, as one of important pathogens in cultured freshwater fish and seawater fish, which caused severe economic losses to the aquaculture industries, developing a vaacine to prevent streptococcosis of fish. The efficacy of S. iniae inactivated vaccine in the freshwater fish tilapia (Oreochromis niloticus) and the sea water Japanese sea perch (Lateolabrax japonicus) were evaluated, the best inoculated doses of antigen were balanced and the best immune adjuvant were choosed. Simultaneously the use of immunostimulants in fish was discussed preliminarily. Our experimental results formed a good scientific basic for the preparation and application of Streptococcus iniae inactivated vaccine in cultured fish.
Formalin killed whole-cell vaccines were prepared with sea water strain HD-1, HD-4 and freshwater strain TBY-1, TBY-5 of S. iniae respectively. Tilapia were vaccinated with vaccine (1×10~(10) colony-forming units of bacterial per fish) mixed with equal doses in volume of Freund's ajuvant by intraperitoneal (i.p.) injection, through twice immunization to fish, the protective efficacies of four kinds of vaccines were compared by challenge of fish with the corresponding strain on the 35th day after the first vaccination. The number of fish died in each group was recorded in 14 days, the relative percent survivl (RPS) of each immune group was statisticsed and the titer of serum antibody in each group fish was detected by bacterial agglutination assays to be determined the efficacy of vaccine on fish. The results showed that the Freund's ajuvant vaccine of S. iniae sea water strain HD-1 could present 93.6% RPS for vaccinated fish and the serum antibody titer of immunized fish reached 1:64; TBY-1 strain vaccine had a 100% RPS with high serum antibody titer by 1:64. As a result, we suggested that HD-1 and TBY-1 strain could be used as the candidate strains to prepare vaccine of cultured fish.
To detect the sensitivity of mariculture fish to S. iniae, Epinephelus coioides, Sciaenops ocellatus, Lutjanus erythopterus, Trachinotus ovatus and Lateolabrax japonicus, as main mariculture species in Guangdong, were infected with HD-1 strain. The result showed that Lateolabrax japonicus were most sensitive to S. iniae seawater HD-1 strain, which would be as animal model for inactivated S. iniae vaccine experiment.
Three types of inactivated S.iniae HD-1 strain vaccine in bacterial concentration of 1×10~(10) CFU×rnL~(-1),1×10~9 CFU×mL~(-1) and 1×10~8 CFU×mL~(-1) mixed with equal volume of white oil adjuvant were prepared. After immunized Japanese sea perch (L. japonicus) had been challenged with corresponding virulent strain, the fish vaccinated with 10~(10) CFU×mL~(-1), 10~9 CFU×mL~(-1), 10~8 CFU×mL~(-1) doses showed high immune protection with RPS of 100%, 92.5%, 85% and high antibody titer with 1:16, 1:64, and 1:32 respectively, while all fish in non-immnized control group died. Antigen dose of 10~9 CFU×mL~(-1) concentrations was most suitable for S.iniae HD-1 strain vacine to immunize: maricultured fish.
To detect immunoprotection of S.iniae HD-1 strain vacine to Japanese seap erch (L. japonicus), two type of vacine with a dose of 1×10~9 CFUxmL~(-1) killed HD-1 strain mixed with equal volume of white oil and alhydrogel adjuvant respectively were prepared and used to vacinate 40 tail fish. After experimenal fish were challenged with corresponding virulent stains, the group immunized with white oil and alhydrogel adjuvant vaccine had high immuoprotection with RPS of 92.5% and 82.5%, and high antibody titers with 1:64 and 1:32 respectively, while all fish in non-immnized control group died. So we thought that white oil adjuvant vaccine could elicit good immune effect and protect Japanese sea perch from streptococcicosis.
Respectively added the Levamisole and the aloe polysaccharide of traditional Chinese medicine by a certain quantity into the S.iniae HD-1 strain vacine with a dose of 1×10~9 CFU×mL~(-1) killed HD-1 strain mixed with equal volume of white oil, simultaneously the aloe polysaccharide alone injection group in white oil adjuvant vaccine foundation, the vaccine with white oil adjuvant group and one non-immunized control group. After experimenal fish were challenged with corresponding virulent stains, it was not significant (p＞0.05) difference with RPS in experimental groups. The antibody titer of the aloe polysaccharide alone injection group were higher than the other groups (p＜0.05); the antibody titer of Levamisole group was lower than the white oil adjuvant group (p＜0.05), the other gorups were not significant differently. Measurese the changing tendency of antalzyme in each group's serum showed that the immunoenhancer of aloe polysaccharides alone injection group obviously higher than the other groups (p＜0.05). our experiments proved that the immunoenhancer of aloe polysaccharides can stimulate fish to produce nonspecific immune response.
将S．iniae海水株HD-1、HD-4，S．iniae淡水株TBY-1，TBY-5的培养液灭活后，制备弗氏佐剂灭活疫苗，腹腔注射接种罗非鱼(1×10~(10)CFU×mL~(-1)0.1 mL×尾~(-1))，经过两次免疫，初次免疫后35天，分别用对应的菌株攻毒，统计14天内各组鱼的死亡情况，计算各免疫组的相对保护率(RPS)，并测定各组鱼血清抗体效价的变化。结果表明：HD-1株弗氏佐剂灭活疫苗保护率达到93.6％，最高血清抗体凝集效价达到1：64；TBY-1株弗氏佐剂灭活疫苗保护率达到100％，最高血清抗体凝集效价达到1：64。海水株HD-1和淡水株TBY-1免疫效果均较好，可作为海水鱼和淡水鱼制苗菌株的候选。
利用S．iniae海水株HD-1对大亚湾地区主要的海水养殖经济鱼类石斑鱼、红鳍笛鲷、卵圆鲳鲹、花鲈及美国红鱼进行攻毒试验。结果显示花鲈鱼对S．iniae海水株HD-1最为敏感，可以作为S．iniae海水株HD-1灭活疫苗试验用鱼进行进一步的相关试验。
将S．iniae海水株HD-1制备成10~(10)CFU×mL~(-1)、10~9CFU×mL~(-1)及10~8CFU×mL~(-1)三种菌体抗原浓度的白油佐剂灭活疫苗对花鲈鱼进行了免疫保护性试验，攻毒结果显示：10~(10)CFU×mL~(-1)组相对保护率为100％、10~9 CFU×mL~(-1)组相对保护率为92.5％、10~8CFU×mL~(-1)组相对保护率为85％、对照组的死亡率是100％；最高抗体滴度分别为1：16、1：64、1：32。10~(10)CFU×mL~(-1)组相对保护率与10~9 CFU×mL~(-1)组相对保护率比较，无显著性差异(p＞0.05)；与10~8CFU×mL~(-1)组相对保护率为85％比较，10~(10)CFU×mL~(-1)组保护率较高，具有显著性差异(p＜0.05)。综合考虑生产实际等各项指标，本试验采用10~9CFU×mL~(-1)菌体抗原浓度为制苗用菌体抗原量的浓度。
将含10~9CFU×mL~(-1)菌体抗原浓度的S．iniae海水株HD-1灭活并制备成白油佐剂疫苗和铝胶佐剂疫苗，对花鲈鱼进行免疫保护性试验，攻毒结果显示：白油佐剂疫苗获得92.5％的相对保护率，铝胶佐剂疫苗组获得82.5％的相对保护率；两种疫苗的最高抗体滴度分别为1：64、1：32。两者比较具显著性差异(p＜0.05)。白油佐剂疫苗获得较好的保护效果，可以有效防制花鲈鱼海豚链球菌病。
将左旋咪唑和中药芦荟多糖分别以一定的量加入到10~9CFU×mL~(-1)菌体抗原浓度的S．iniae海水株HD-1白油佐剂灭活疫苗中制备成复合佐剂疫苗(compound adjuvant vaccine)，同时设白油佐剂疫苗基础上单独注射芦荟多糖组、白油佐剂组及PBS组，对花鲈鱼进行免疫保护性试验，攻毒结果显示：各试验组相对保护率差异性不显著(p＞0.05)；但白油佐剂灭活疫苗加单独注射芦荟多糖组鱼血清抗体滴度比其他各组要高(p＜0.05)，而左旋咪唑复合佐剂疫苗组鱼血清抗体效价显著低于白油佐剂灭活疫苗组(p＜0.05)，另外各组血清抗体滴度之间比较没有显著性差异。测定各组血清中溶菌酶变化显示，单独注射免疫增强剂芦荟多糖组血清溶菌酶含量明显比其他各组都要高(p＜0.05)，试验结果表明：中药免疫增强剂芦荟多糖能刺激鱼类产生非特异性免疫应答反应并提高其血清抗体效价。
In decades, with rapid development of aquaculture industry with the high-density and the production on a large scale, the diseases of cultured fish took place frequently, especially the bacteriosis, with high variety, morbidity and mortality, caused huge economic loss to aquaculture. A frequent use of chemical and antibiotic resulted in occurrence of drug- resistant pathogenic strains, simultaneously brought about severe public health and sanitation problems, such as drug residue and food security, to which our human being have to faced and challenged with. In modern aquaculture industry, vaccination will offer a good alternative to strategy of fish disease provention in the highly qualitative fishery cultivation. So developing highly effective vaccine for cultured fish have been an urgent matter in aquaculture industry. In present studies, we mainly focused on Streptococcus iniae, as one of important pathogens in cultured freshwater fish and seawater fish, which caused severe economic losses to the aquaculture industries, developing a vaacine to prevent streptococcosis of fish. The efficacy of S. iniae inactivated vaccine in the freshwater fish tilapia (Oreochromis niloticus) and the sea water Japanese sea perch (Lateolabrax japonicus) were evaluated, the best inoculated doses of antigen were balanced and the best immune adjuvant were choosed. Simultaneously the use of immunostimulants in fish was discussed preliminarily. Our experimental results formed a good scientific basic for the preparation and application of Streptococcus iniae inactivated vaccine in cultured fish.
Formalin killed whole-cell vaccines were prepared with sea water strain HD-1, HD-4 and freshwater strain TBY-1, TBY-5 of S. iniae respectively. Tilapia were vaccinated with vaccine (1×10~(10) colony-forming units of bacterial per fish) mixed with equal doses in volume of Freund's ajuvant by intraperitoneal (i.p.) injection, through twice immunization to fish, the protective efficacies of four kinds of vaccines were compared by challenge of fish with the corresponding strain on the 35th day after the first vaccination. The number of fish died in each group was recorded in 14 days, the relative percent survivl (RPS) of each immune group was statisticsed and the titer of serum antibody in each group fish was detected by bacterial agglutination assays to be determined the efficacy of vaccine on fish. The results showed that the Freund's ajuvant vaccine of S. iniae sea water strain HD-1 could present 93.6% RPS for vaccinated fish and the serum antibody titer of immunized fish reached 1:64; TBY-1 strain vaccine had a 100% RPS with high serum antibody titer by 1:64. As a result, we suggested that HD-1 and TBY-1 strain could be used as the candidate strains to prepare vaccine of cultured fish.
To detect the sensitivity of mariculture fish to S. iniae, Epinephelus coioides, Sciaenops ocellatus, Lutjanus erythopterus, Trachinotus ovatus and Lateolabrax japonicus, as main mariculture species in Guangdong, were infected with HD-1 strain. The result showed that Lateolabrax japonicus were most sensitive to S. iniae seawater HD-1 strain, which would be as animal model for inactivated S. iniae vaccine experiment.
Three types of inactivated S.iniae HD-1 strain vaccine in bacterial concentration of 1×10~(10) CFU×rnL~(-1),1×10~9 CFU×mL~(-1) and 1×10~8 CFU×mL~(-1) mixed with equal volume of white oil adjuvant were prepared. After immunized Japanese sea perch (L. japonicus) had been challenged with corresponding virulent strain, the fish vaccinated with 10~(10) CFU×mL~(-1), 10~9 CFU×mL~(-1), 10~8 CFU×mL~(-1) doses showed high immune protection with RPS of 100%, 92.5%, 85% and high antibody titer with 1:16, 1:64, and 1:32 respectively, while all fish in non-immnized control group died. Antigen dose of 10~9 CFU×mL~(-1) concentrations was most suitable for S.iniae HD-1 strain vacine to immunize: maricultured fish.
To detect immunoprotection of S.iniae HD-1 strain vacine to Japanese seap erch (L. japonicus), two type of vacine with a dose of 1×10~9 CFUxmL~(-1) killed HD-1 strain mixed with equal volume of white oil and alhydrogel adjuvant respectively were prepared and used to vacinate 40 tail fish. After experimenal fish were challenged with corresponding virulent stains, the group immunized with white oil and alhydrogel adjuvant vaccine had high immuoprotection with RPS of 92.5% and 82.5%, and high antibody titers with 1:64 and 1:32 respectively, while all fish in non-immnized control group died. So we thought that white oil adjuvant vaccine could elicit good immune effect and protect Japanese sea perch from streptococcicosis.
Respectively added the Levamisole and the aloe polysaccharide of traditional Chinese medicine by a certain quantity into the S.iniae HD-1 strain vacine with a dose of 1×10~9 CFU×mL~(-1) killed HD-1 strain mixed with equal volume of white oil, simultaneously the aloe polysaccharide alone injection group in white oil adjuvant vaccine foundation, the vaccine with white oil adjuvant group and one non-immunized control group. After experimenal fish were challenged with corresponding virulent stains, it was not significant (p＞0.05) difference with RPS in experimental groups. The antibody titer of the aloe polysaccharide alone injection group were higher than the other groups (p＜0.05); the antibody titer of Levamisole group was lower than the white oil adjuvant group (p＜0.05), the other gorups were not significant differently. Measurese the changing tendency of antalzyme in each group's serum showed that the immunoenhancer of aloe polysaccharides alone injection group obviously higher than the other groups (p＜0.05). our experiments proved that the immunoenhancer of aloe polysaccharides can stimulate fish to produce nonspecific immune response.
引文
[1]张元兴，马悦，孙修勤．海洋鱼病疫苗开发与生物反应器大规模生产[J]．高技术通讯，2000，10(2)：1-5．
[2]Chou H Y. Application of monoclonal antibody against viral nucleoprotein to an aetiological study of infection haematopoietic necrosis[J]. Journal of Fish Diseases, 1993, 16(2): 149-153.
[3]杨先乐，吴明泉．草鱼出血病细胞培养灭活疫苗的研究：生产性免疫试验[J]．水产科技情报，1992，19(1)：10-14．
[4]Anderson D P. Adjuvants and immunostimulants for enhancing vaccine potency in. fish[J]. Developments in Biological Standardization, 1997, 90(7):257-265.
[5]Benmansour A, de Kinkelin P. Live fish vaccines: history and perspectives[J]. Developments in Biological Standardization, 1997, 90(7): 279-289.
[6]陈超然，毛灵娟，陈昌福．鱼类疫苗研制过程中面临的问题与课题[J]．河南水产，2002，1：9-11．
[7]吴乃虎．基因工程原理(第二版)[M]．北京：科学出版社，2001，68-69．
[8]杨先乐，陈远新．鱼用疫苗的现状及其发展趋势[J]．水产学报，1996，20(2)：159-167．
[9]Vaughan L M, Smith P R, Foster T J. An aromatic-dependent mutant of the fish pathogen Aeromonas salmonicida, is attenuated and is effective as a live vaccine against the salmonid diseases furunculosis[J]. Infection and Immunity, 1993, 61(5): 2172-2181.
[10]Marsden M J, Vaughan L M, Foster T J, et al. A live (delta aroA) Aeromonas salmonicida vaccine forfurunculosis preferentially stimulates T-cell responses relative to B-cell responses in rainbow trout (Oncorhynchus mykiss)[J]. Infection and Immunity, 1996, 64(9): 3863-3869.
[11]陈振海，林天龙，夏春．鱼类DNA疫苗研究的新进展[J]．水生生物学报，2003，27(6)：648-653．
[12]Heppell J, Davis H L. Application of DNA vaccine technology to aquaculture[J]. Advanced Drug Delivery Reviews, 2000, 43(1): 29-43.
[13]Anderson E D, Mourich D V, Fahrenkrug S C, et al. Genetic immunization of rainbow trout (Oncorhynchus mykiss) against infectious hematopoietic necrosis virus[J]. Molecular Marine Biology & Biotechnology, 1996, 5(2): 114-112.
[14]Anderson E D, Mourich D V, Lenong J A. Gene expression in rainbow Trout (Oncorhynchus mykiss) following intramuscular injection of DNA[J]. Molecular Marine Biology & Biotechnology, 1996, 5(2): 105-113.
[15]Traxler G S, Anderson E, LaPatra SE, et al. Naked DNA vaccination of Atlantic salmon Salmo salar against IHNV[J]. Disease of Aquatic Organisms, 1999, 38(3): 183-190.
[16]Lorenzen N, Lorenzen E, Einer J K, et al. Genetic vaccination of rainbow trout against viral haemorrhagic septicaemia virus: small amounts of plasmid DNA protect against a heterologous serotype[J]. Advances in Virus Research, 1999, 63(1-2): 19-25.
[17]Kim C H, Johnson M C, Drennan J D, et al. DNA vaccines encoding viral glycoproteins induce nonspecific immunity and Mx protein synthesis in fish[J]. Journal of Virological Methods, 2000, 74 (15): 7048-7054.
[18]王利，谢长勇．鱼类疫苗研究进展[J]．动物医学进展，2003，24(5)：26-28．
[19]Eldar A, Horovitcz A Bercovier H. Development and efficacy of a vaccine against streptococcus iniae infection in farmed rainbow trout[J]. Veterinary Immunology & Immunopathology, 1997, 56(1-2): 175-183.
[20]陈超然，陈昌福．鱼用疫苗的研究现状[J]．水利渔业，2001，21(5)：44-45．
[21]Li S, Woo P T. Efficacy of a live Cryptobia salmositica vaccine and the mechanism of protection in vaccinated rainbow trout, Oncorhynchus mykiss against cryptobiosis[J]. Veterinary Immunology and Immunopathology, 1995, 48(3-4): 343-353.
[22]夏春．鱼病诊断与防治手册[M]．北京：中国农业大学出版社，1999，21-32．
[23]杨先乐，左文功．草鱼出血病细胞培养灭活疫苗的研究：疫苗的稳定性及佐剂和加强免疫对：草鱼免疫应答的影响[J]．水生生物学报，1993，17(1)：46-51．
[24]Stephen G N, John J M. Immunization of Salmonids against furunculosis[J]. Fish Pathology, 1985, 20(2-3): 403-411.
[25]Newman S G. Bacterial vaccines for fish[J]. Annual Review of Fish Diseases, 1993, 32(3): 145-185.
[26]张吉红，陆承平．佐剂对嗜水气单胞菌灭活疫苗免疫效果的作用[J]．中国兽药杂志，2003，7(4)：26-27．
[27]Midtlyng P J. A field study on intraperitoneal vaccination of Atlantic salmon (Salmo salarL.) against furunculosis[J]. Fish & Shellfish Immunology, 1996, 6(8): 553-565.
[28]Midtlyng P J, Reitan L J. Speilberg L. Experimental studies on the efficacy and side-effects of intraperitoneal vaccination of Atlantic salmon (Salmo salarL.) against furunculosis[J]. Fish & Shellfish Immunology, 1996, 6(5): 335-350.
[29]高健编译．免疫激活佐剂在水产养殖中的应用[J]．河北渔业，1994，34．
[30]傅安静，岳华，刘树玲等．疫苗佐剂研究进展[J]．西南民族大学学报(增刊)，2005，73-77．
[31]Donnelly J J. New development in adjuvants[J]. Mechanisms of Ageing and Development, 1997, 93(1-3): 171-177.
[32]Hagan D T, MacKichan M L, Singh M. Recent developments in adjuvants for vaccines against infectious diseases[J]. Biomolecular Engineering, 2001, 18(3): 69-85.
[33]赵建培，史维舟．在草鱼免疫反应中莨菪佐剂性的初步观察[J]．淡水渔业，1995，25(1)：3-5．
[34]吴志新，陈强，陈昌福．左旋咪唑对异育鲫鱼免疫促进作用的初步研究[J]．华中农业大学学报，1998，17(4)：378-381．
[35]Kajita Y, Sakai M, Atsuta S, et al. The immunomodulatory effects of levamisole on rainbow trout, Oncorhynchus mykiss[J]. Fish Pathology, 1990, 25: 93-98.
[36]张庆茹．中草药免疫促进作用的研究进展[J]．中兽医医药杂志，1997，(5)：15．
[37]薛明，孟宪松．生物活性多糖的免疫研究进展与展望[J]．中兽医医药志，1996，(3)：16．
[38]万金志，乔悦昕．芦荟的化学成分及其研究[J]．中草药，1999，30(2)：151-153．
[39]方仲根．芦荟的活性成分与疗效作用[J]．热带农业科学，1999，(1)：59-62．
[40]Ki Hong Kim. Resistance to Vibrio alginolyticus in juvenile rockfish ( Sebastes schlegeli ) fed diets containing different doses of aloe[J]. Aquaculture, 1999, 180(1-2): 13-21.
[41]孙建宏．免疫佐剂的研究进展[J]．黑龙江畜牧兽医，1998，2：40-43．
[42]Sakai M, Atsuta S, Kobayashi M. Protective immune response in rainbow trout, Oncorhynchus mykiss,vaccinated with β-haemolytic streptocococal bacterin[J]. FishPathology, 1989, 24(3): 169-173.
[43]Shoemaker C, Klesius P. Streptococcal disease problems and control: a review In Fitzsimmons, K. Ed., Tilapia Aquaculture vol. 2 Northeast Regional Agricultural Engineering Service-106, Ithaca, NY, USA, 1997: 671-680.
[44]Pier G B, Madin S H, Al-Nakeeb S. Isolation and characterization of a second isolate of Streptococcus iniae[J]. International Journal of Systematic Bacteriology, 1978, 28:311-314.
[45]Sakai M, Atsuta S, Kobayashi M. Protective immune response in rainbow trout, Oncorhynchus mykiss, vaccinated with β-haemolytic streptocococal bacterin[J]. Fish Pathology, 1989, 24(3): 169-173.
[46]Kitao T. Streptococcal infections. In Inglis V, Roberts R J, Bromage NR Eds. Bacterial Diseases of Fish[M]. Oxford: Blackwell Scientific Publications, 1993,196-210.
[47]Evans J J, Klesius P H, Shoemaker C A. Streptococcus iniae in warm-water fish[J]. Aquaculture Health International, 2006, 7: 10-14.
[48]Litt M D, Low D, Cunha C, et al. Surveillance for human infection with Streptococcus iniae[J]. Journal of Clinical Epidemiology, 1997, 50(1): 17.
[49]Eldar A, Bejerano Y, Livoff A, et al. Experimental streptococcal meningo-encephalitis in cultured fish[J]. Veterinary Microbiology, 1995, 43(1): 33-40.
[50]Bromage E S, Owens L. Infection of barramundi Lates calcarifer with Streptococcus iniae: effects of different routes of exposure[J]. Diseasea of aquatic organisms, 2002, 52(3): 199-205.
[51]Evans J J, Shoemaker C A, Klesius P H. Distribution of Streptococcus iniae in hybrid striped bass (Morone chrysops×Morone saxatilis) following nare inoculation[J]. Aquaculture, 2001, 194(3-4): 233-243.
[52]Ana I M, Blanco M M, Dominguez L, et al. Development of a PCR assay for Streptococcus iniae based on the lactate oxidase (lctO) gene with potential diagnostic value[J]. Veterinary Microbiology, 2004, 101 (2): 109-116.
[53]Fuller J D, Bast D J, Nizet V, et al. Streptococcus iniae virulence is associated with a distinct genetic profile[J]. Infection and immunity, 2001, 69(4): 94-100.
[54]柴家前，丁巧玲，王振龙等．罗非鱼链球菌的分离鉴定[J]．中国预防兽医学报，2002，24(1)：18-20．
[55]Elliott J A, Facklam R R, Richter C B. Whole-cell protein patterns of nonhaemolytic group B, type Ⅰb, streptococci isolated from humans, mice, cattle, frogs and fish[J]. Journal of Clinical Microbiology, 1990, 28: 628-630.
[56]Robinson J A, Meyer F P. Streptococcal fish pathogen[J]. Journal of Bacteriology, 1966,92: 512.
[57]Plumb J A, Schachte J H, Gaines J L, et al. Streptococcus sp. from marine fishes along the Alabama and Northwest Florida coast of the Gulf of Mexico[J]. Transactions of the American Fisheries Society, 1974, 103: 358-361.
[58]Baya A M, Lupiani B, Hetrick F M, et al. Association of Streptococcus sp with fish mortalities in the Chesapeake Bay and its tributaries[J]. Journal of Fish Diseases, 1990, 13: 251-253.
[59]Eldar A, Bejerano Y, Bercovier H. Streptococcus shiloi and Streptococcus difficile: two new streptococcal species causing Meningoencephalitis in fish[J]. Current Microbiology, 1994, 28: 139-143.
[60]Wilkinson H W, Thacker L G, Facklam R R. Nonlaaemolytic group B Streptococci of human, bovine and ichthyic origin[J]. Infection and Immunity, 1973, 7: 496-498.
[61]Vandamme P, Devriese L A, Pot B, et al. Streptococcus difficile is a nonhaemolytic group B, type Ⅰb Streptococcus[J]. International Journal of Systematic Bacteriology, 1997, 47: 81-85.
[62]Evans J J, Klesius P H, Glibert P M, et al. Characterisation ofbeta-haemolytic Group B Streptococcus agalactiae in cultured sea bream, Sparus auratus L and wild mullet, Liza klunzingeri (Day), in Kuwait[J]. Journal of Fish Diseases, 2002a, 25: 505-513.
[63]Kawamura Y, Itoh Y, Mishima N, et al. High genetic homogeneity of Streptococcus agalactiae and Streptococcus difficilis: reclassification of S difficilis, Eldar et al 1995 as a junior synonyn of S agalactiae, Lehmann and Neumann 1896[J]. International Journal of Systematic and Evolutionary Microbiology, 2005, 55: 961-965.
[64] Evans J J, Pasnik D J, Klesius P H, et al. First report of Streptococcus agalactiae and Lactococcus garvieae from a wild bottlenose dolphin, Tursiops truncatus[J]. Journal of Wildlife Diseases 2006a, 42: 561-569.
[65] Kitao T. Streptococcal infections. In Inglis V, Roberts RJ, Bromage NR Eds [C]. Bacterial Diseases of Fish, Blackwell Scientific Publications, Oxford, 1993: 196-210.
[66] Evans J J, Pasnik D J, Klesius P H, et al. Identification and epidemiology of Streptococcus iniae and 5 agalactiae in tilapia [C]. Proceedings of the International Symposium of Tilapia Aquaculture ISTA7. Veracruz, Mexico, 2006, 9: 25-42.
[67] Pasnik D J, Evans J J, Panangala V S, et al. Antigenicity of Streptococcus agalactiae extracellular products and vaccine efficacy [J]. Journal of Fish Diseases, 2005,28: 205-212.
[68] Shoemaker C A, Klesius P H, Evans J J. Prevalence of Streptococcus iniae in tilapia, hybrid striped bass and channel catfish from fish farms in the United States [J]. American Journal of Veterinary Research, 2001,62:174-177.
[69] Evans J J, Shoemaker C A, Klesius P H. Distribution of Streptococcus iniae in hybrid striped bass Morone chrysops x Morone saxatilis following nares inoculation [J]. Aquaculture, 2001, 194: 233-243.
[70] Evans J J, Wiedenmayer A A, Klesius P H. A transport system for the maintenance of viability of Acinetobacter calcoaceticus, Streptococcus iniae and S agalactiae over varying time periods [J]. Bulletin of the European Association of Fish Pathologists, 2002b, 22: 238-246.
[71] Evans J J, Wiedenmayer A A, Klesius P H, et al. Survival of Streptococcus agalactiae from frozen fish following natural and experimental infections [J]. Aquaculture, 2004a, 233: 15-21.
[72] Facklam R. What happened to the Streptococci: overview of taxonomic and nomenclature changes [J]. Clinical Microbiology Reviews, 2002, 15: 613-630.
[73] Kimura H, Kusuda R. Studies on the pathogenesis of streptococcal infection in cultured yellowtails, Seriola spp: effect of crude exotoxin fractions from cell-free culture on experimental streptococcal infection [J]. Journal of Fish Diseases, 1982, 5: 471-478.
[74] Boomker J, Imes G J, Cameron C M, et al. Trout mortalities as a result of Streptococcus infection [J]. Onderstepoort Journal of Veterinary Research, 1979, 46: 71-77.
[75] Klesius P H, Evans J J, Shoemaker C A, et al. Rapid detection and identification of Streptococcus iniae using a monoclonal antibody-based indirect fluorescent antibody technique [J]. Aquaculture, 2006a, 258: 180-186.
[76] Berridge B R, Bercovier H, Frelier P F. Streptococcus agalactiae and Streptococcus difficile 16S-23S intergenic rDNA: genetic homogeneity and species-specific PCR [J]. Veterinary Microbiology, 2001, 78:165-173.
[77]Mata A I, Gibello A, Casamayor A, et al. Multiplex PCR assay for detection of bacterial pathogens associated with warm-water streptococcosis in fish[J]. Applied and Environmental Microbiology, 2004, 70: 3183-3187.
[78]Pier G B, Madin S H. Streptococcus iniae sp nov, a beta-haemolytic Streptococcus isolated from an Amazon freshwater dolphin, Inia geoffrensis[J]. International Journal of Systematic Bacteriology, 1976, 26: 545-553.
[79]Eldar A, Bejerano Y, Bercovier H. Streptococcus shiloi and Streptococcus difficile: two new streptococcal species causing Meningoencephalitis in fish[J]. Current Microbiology, 1994, 28: 139-143.
[80]Perera R P, Collins M D, Johnson S K,et al. Streptococcus iniae associated with mortality of Tilapia nilotica x T aurea hybrids[J]. Journal of Aquatic Animal Health, 1994, 6: 335-340.
[81]Shelby R A, Klesius P H, Shoemaker C A, et al. Passive immunisation of tilapia, Oreochromis niloticus L with anti-Streptococcus iniae whole sera[J]. Journal of Fish Diseases, 2002, 25: 1-6.
[82]Evans J J, Shoemaker C A, Klesius P H. Therapeutic and prophylactic immunisation against Streptococcus iniae infection in hybrid striped bass Morone chrysops x M saxatilis[J]. Aquaculture Research, 2006c, 37: 742-750.
[83]Klesius P H, Evans J J, Shoemaker C A, et al. Vaccines to prevent Streptococcus iniae and S agalactiae disease in tilapia, Oreochromis niloticus[C]. Proceedings of the International Symposium of Tilapia Aquaculture ISTA7, Veracruz, Mexico, 2006, 9: 15-24.
[84]Eldar A, Shapiro O, Bejerano Y,et al. Vaccination with whole-cell vaccine and bacterial protein extract protects tilapia against Streptococcus difficile meningoencephalitis[J]. Vaccine, 1995b, (13): 867-870.
[85]Klesius P H, Shoemaker C A, Evans J J. Efficacy of single and combined Streptococcus iniae isolate vaccine administrated by intraperitoneal and intramuscular routes in tilapia (Oreochromis niloticus)[J]. Aquaculture, 2000, 188 (3-4): 237-246.
[86]Eldar A, Horovitcz A, Bercovier H. Development and efficacy of a vaccine against Streptococcus iniae infection in farmed rainbow trout[J]. Veterinary Immunology and Immunopathology, 1997, 56(1-2): 175-183.
[87]Bachrach G, Zlotkin A, Hurvitz A, et al. Recovery of Streptococcus iniae from diseased fish previously vaccinated with a streptococcus vaccine[J]. Applied and Environmental Microbiology, 2001, 67(8): 3756-3758.
[88]Klesius P H, Shoemaker C A and Evans J J. 2000. Efficacy of single and combined Streptococcus iniae isolate vaccine administered by intraperitoneal and intramuscular routes in tilapia (Oreochromis niloticus)[J]. Aquaculture, 188(3-4): 237-246.
[89]Akhlaghi M., Munday B L, Whittington R J. Comparison of passive and active immunization of fish against streptococcosis (enterococcosis)[J]. Journal of fish diseases, 1996, 19: 251-258.
[90]Nakanishi T, Kiryu I, Ototake M. Development of a new vaccine delivery method for fish: percutaneous administration by immersion with application of a multiple puncture instrument[J]. Vaccine, 2002, 20(31): 3764-3769.
[91]Shelby R A, Klesius P H, Shoemaker C A, et al. Passive Immunization of tilapia, Oreochromis Niloticus (L.) with Anti-Streptcoccus iniae whole sera[J]. Journal of Fish Diseases, 2002, 25(1): 1-6.
[92]Shoemaker C A, Vandenberg G W, Desormeaux Andre, et al. Efficacy of a Streptococcus iniae modified bacterin delivered using Oralject~(TM) technology in Nile tilapia(Oreochromis niloticus)[J]. Aquaculture, 2006, 255(1-4): 151-156.
[93]张延龄，张晖．疫苗学[M]．北京：科学出版社，2004，3：323．
[94]Shoemaker C, Klesius P. Streptococcal disease problems and control: a review[C]. In Tilapia Aquaculture Northeast Regional Agricultural Engineering Service, Ithaca, NY, 1997, (2): 671-680.
[95]Eldar A, Shapiro O, Bejerano Y,et al. Vaccination with whole-cell vaccine and bacterial protein extract protects tilapia against Streptococcus difficile meningoencephalitis[J]. Vaccine, 1995b, (13): 867-870.
[96]Klesius P H, Shoemaker C A, Evans J J. Efficacy of single and combined Streptococcus iniae isolate vaccine administrated by intraperitoneal and intramuscular routes in tilapia (Oreochromis niloticus)[J]. Aquaculture, 2000, 188 (3-4): 237-246.
[97]Kenneth E N, Bruce F S, Patricia D I. Protective immunity induced by DNA vaccination of channel catfish with early and late transcripts of the channel catfish herpesvirus (IHV-1)[J].Veterinary immunology and immunopathology, 2002, 84 (4): 151-168.
[98]Klesius P H, Shoemaker C A,Evans J J. Efficacy of single and combined Streptococcus iniae isolate vaccine administered by intraperitoneal and intramuscular routes in tilapia (Oreochrormis niloticus)[J]. Aquaculture, 2000, 188: 237-246.
[99]Ackerman P A, Iwama G K. Physiological and immunological effects of adjuvanted Aeromonas salmonicida vaccines on juvenile rainbow trout[J]. Journal of Aquatic Animal Health, 2000, 12: 157-164.
[100]Toranzo A E, Devesa S, Romaide J L, et al. Efficacy of intraperitoneal and immersion vaccination against Enterococcus sp.infection in turbot[J]. Aquaculture, 1995, 134: 17-27.
[101]Waterstrat P, Ainsworth J, Capley G. Use of an indirect enzyme-linked immunosorbent assay(ELISA) in the detection of channel catfish, lctalurus punctatus (Rafinesque), antibodies to Edwardsiella ictaluri[J]. Journal of Fish Diseases, 1989, 12 (2): 87-94.
[102]Eldar A, Horovitcz A, Bercovier H. Development and efficacy of a vaccine against Streptococcus iniae infection in farmed rainbow trout[J]. Veterinary immunology & Immunopathology, 1997, 56:175-183.
[103]Klesius P H, Shoemaker, C A, Evans, J J. Efficacy of a killed Streptococcus iniae vaccine in tilapia (Oreochromis niloticus)[J]. Bulletin of the European Association of Fish Pathologists. 1999, 19(1): 1-3.
[104]Klesius P H, Shoemaker, C A, Evans, J J.Efficacy of single and combined Streptococcus iniae isolate vaccine administered by intraperitoneal and intramuscular routes in tilapia (Oreochromis niloticus)[J]. Aquaculture, 2000(3-4): 237-246.
[105]Hurvitz A, Bercovier H, Rijn J V. Effect of ammonia on the survival and the immune response of rainbow trout (Oncorhynchus mykiss, Walbaum) vaccinated against Streptococcus iniae[J]. Fish & shellfish immunology, 1997, 7(1): 45-53.
[106]Barnes A C, Young F M, Horne M T, Ellis A E. Streptococcus iniae: serological differences presence of capsule and resistance to immune serum Killing[J]. Diseases of aquatic organisms, 2003, 53(3): 241-247.
[107]Evans J J, Klesius P H, Shoemaker P K, et al. Experimental Streptococcus iniae infection of hybrid striped bass (Morone chrysops×Morone saxatilis) and tilapia (Oreochromis niloticus) by nares inoculation[J]. Aquaculture, 2000, 189(3-4): 197-210.
[108]Kitao T. Streptococcal infections. In Inglis V, Roberts RJ,Bromage NR Eds. Bacterial Diseases of Fish. Oxford: Blackwell Scientific Publications, 1993, 196-210.
[109]Joyce J E, Craig A S, PHillip K. Experimental Streptococcus iniae infection of hybrid striped bass (Morone chrysops×Morone saxatilis) and tilapia (Oreochromis niloticus) by nares inoculation[J]. Aquaculture, 2000, 189: 197-210.
[110]Amir Z, Hannah H, Avi E. Possible transmission of Streptococcus iniae from wild fish to cultured fish[J]. Applied and environmental microbiology, 1998, 4065-4067.
[111]Shorts E B, Blazer V S, and Waltman W D. Pathogenesis of experimental Edwardsiell ictaluri infections in channel catfish (Ictalurus punctatus)[J]. Fish. Sci, 1986,43: 36-42.
[112]Newton J C, Wolfe L G, Grizzle J M, and Plumb J A. Pathology of experimental enteric septcaemia in channel catfish, Ictalurus punctatus (Rafinesque), following immersion-exposure to Edwardsiella ictaluri[J]. Fish Dis, 1989, 12: 335-347.
[113]Morrison E E and Plumb J A. Olfactory organ of channel catfish as a site of experimental Edwardsiella ictaluri infection[J]. Aquat. Anim. Health, 1994, 6: 101-1099.
[114]Hoshina T, Sano T and Morimoto Y. A Streptococcus pathogenic to fish[J]. Tokyo Univ. Fish, 1958, 44: 57-58.
[115]Robinson J A. and Meyer F P. Streptococcal fish pathogen[J]. Bacteriol, 1966, 92(2): 512.
[116]Cook D W and Lofton S R. Pathogenicity studies with a Streptococcus sp. isolated from fishes in an Alabama-Florida fish kill[J]. Trans. Am. Fish. Soc, 1975, 2: 286-288.
[117]Ohnishi K, and Jo Y. Studies of streptococcal infection in pond-cultured fishes. Characteristics of a β-hemolytic Streptococcus isolated from cultured ayu and amago in 1997-1998[J]. Fish Pathol, 1981, 16: 63-67.
[118]Evan J J, Shoemaker C A and Klesius P H. Experimental Streptococcus iniae infection of hybrid striped bass (Morone chrysops×Morone saxatilis) and tilapia (Oreoehromis niloticus) by nares inoculation[J]. Aquaculture, 2000, 189(3-4): 197-210.
[119]Klesius P H, Shoemaker C A, Evans J J.Efficacy of single and combined Streptococcus iniae isolate vaccine administered by intraperitoneal and intramuscular routes in tilapia(Oreochromis niloticus). Aquaculture, 2000, 188: 237-246.
[120]杨先乐，陈远新．鱼用疫苗的现状及其发展趋势[J]．水产学报，1996，20(2)：159-167．
[121]Pdjkers G T. The haemolftic plaque assay in carp, CyprinuscarpioL.[J].Journal of immune Meth, 1980, 33: 79-86.
[122]Eldar A, Horovitcz A, Bercovier H.Development and efficacy of a vaccine against Streptococcus iniae infection in farmed rainbow trout[J]. Veterinary Immunology and Immunopathology, 1997(56): 175-183.
[123]Klesius P H, Shoemaker C A, Evans J J.Efficacy of single and combined Streptococcus iniae isolate vaccine administered by intraperitoneal and intramuscular routes in tilapia(Oreochromis niloticus). Aquaculture, 2000, 188: 237-246.
[124]Eldar A, Horovitcz A, Bercovier H. Development and efficacy of a vaccine against Streptococcus iniae infection in farmed rainbow trout[J]. Veterinary immunology & Immunopathology, 1997, 56: 175-183.
[125]王建英，杨桂文，田雪文等．免疫佐剂及其作用机制[J]．山东师范大学学报，2006，21 (1)：127-129．
[126]Midtlyng P J, Reitan L J. Speilberg L. Experimental studies on the efficacy and side-effects of intraperitoneal vaccination of Atlantic salmon (Salmo salarL.) against furunculosis[J]. Fish & Shellfish Immunology, 1996, 6 (5): 335-350.
[127]Klesius P H, Shoemaker C A, Evans J J. Efficacy of single and combined Streptococcus iniae isolate vaccine administered by intraperitoneal and intramuscular routes in tilapia(Oreochromis niloticus). Aquaculture, 2000, 188: 237-246.
[128]Evans J J, Klesius P H, Shoemaker C A. Efficacy of Streptococcus agalactiae (group B) vaccines in tilapia (Oreochromis niloticus) by intraperitoneal and bath immersion administration[J]. Vaccine, 2004, 22 (27-28): 3769-3773.
[129]Eldar A, Horovitcz A, Bercovier H. Development and efficacy of a vaccine against Streptococcus iniae infection in farmed rainbow trout[J]. Veterinary Immunology and Immunopathology, 1997, 56 (1-2): 175-183.
[130]高冬梅，李健，王群．鳗弧菌灭活疫苗对牙鲆免疫保护效果的研究[J]．海洋水产研究，2004，25(1)：34-40．
[131]李渊，孙红祥．不同类型免疫佐剂的作用比较[J]．浙江江畜牧兽医，2003，(5)：16-18．
[132]Jansen T, Hofmans M M,Theelen J G, et al. Structure-activity relations of water-in-oil vaccine formulations and induced antigen-specific antibody responses[J]. Vaccine, 2005, 23(8):1053-1060.
[133]万金志，乔悦昕．芦荟的化学成分及其研究[J]．中草药，1999，30(2)：151-153．
[134]方仲根．芦荟的活性成分与疗效作用[J]．热带农业科学，1999，(1)：59-62．
[135]苗明三，常章富．芦荟多糖对正常小鼠免疫作用影响的研究[J]．中医药学刊，2003(8)：12-42．
[136]宫朋，王富辉，杨长树．木立芦荟粗多糖对免疫功能的影响[J]．人参研究．2000，12(2)：28-29．
[137]王莉，包旭，陈淑杰等．巴巴多斯芦荟多糖提取物对小鼠免疫功能的影响[J]．华西药学杂志，1999，14(4)：234-235．
[138]王莉，杜俊蓉，蔡绍晖等．AP对正常和内毒素化大鼠脾细胞体外分泌细胞因子的影响[J]．华西药学杂志，1999，14(6)：346-348．
[139]吴志新，陈强，陈昌福等．左旋咪唑对异育鲫鱼免疫促进作用的初步研究[J]．华中农业大学学报，1998，17(4)：378-381．
[140]Kajita Y, Sakai M, Atsuta S, et al. The immunomodulatory effects of levamisole on rainbow trout, Oncorhynchus mykiss[J]. Fish Patholology, 1990, 25: 93-98.
[141]叶丹，连宾．溶菌酶及其应用[J]．贵州科学，2003，21(3)：45-46．
[142]Shih-Chu Chen, Terutoyo Yoshida, Alexandra Adams, et al.Thompson.Immune response of Rainbow Trout to extracellular products of Mycobacterium spp[J]. Journal of Aquatic Animal Health, 1996, 8: 216-222.
[143]王宏田，徐永立，张培军．假雄牙鲆不同组织中溶菌酶比活性的研究[J]．海洋科学，2000，24(10)：7-8．
[144]Crinde B, Lie O, Poppe T, Salte R. Species and individual variation in lysozyme activity in fish of interest in aquaculture[J].Aquaculture, 1988, 68: 299-304.
[145]Paige A A, George K, lwama. Physiological and immunological effects of adjuvanted Aeromonas salmonicida vaccines on juvenile rainbow trout[J]. Journal of Aquatic Animal Health, 2000, 12: 157-164.
[146]安利国，玛程强，邢维贤等．灭活疫苗对鲤鱼血清溶菌酶和腹腔吞噬细胞活性的作用[J]．山东师范大学学报(自然科学版)，1999，14(2)：175-179．
[147]Siwicki A, Studnicka M. The phagocytic ability of neutrophils and serum lysozyme activity in experimentally infected carp, Cyprinus carpio L.[J]. Journal of Fish Biology, 1987, 31 (Supplement A): 57-60.
[2]Chou H Y. Application of monoclonal antibody against viral nucleoprotein to an aetiological study of infection haematopoietic necrosis[J]. Journal of Fish Diseases, 1993, 16(2): 149-153.
[3]杨先乐，吴明泉．草鱼出血病细胞培养灭活疫苗的研究：生产性免疫试验[J]．水产科技情报，1992，19(1)：10-14．
[4]Anderson D P. Adjuvants and immunostimulants for enhancing vaccine potency in. fish[J]. Developments in Biological Standardization, 1997, 90(7):257-265.
[5]Benmansour A, de Kinkelin P. Live fish vaccines: history and perspectives[J]. Developments in Biological Standardization, 1997, 90(7): 279-289.
[6]陈超然，毛灵娟，陈昌福．鱼类疫苗研制过程中面临的问题与课题[J]．河南水产，2002，1：9-11．
[7]吴乃虎．基因工程原理(第二版)[M]．北京：科学出版社，2001，68-69．
[8]杨先乐，陈远新．鱼用疫苗的现状及其发展趋势[J]．水产学报，1996，20(2)：159-167．
[9]Vaughan L M, Smith P R, Foster T J. An aromatic-dependent mutant of the fish pathogen Aeromonas salmonicida, is attenuated and is effective as a live vaccine against the salmonid diseases furunculosis[J]. Infection and Immunity, 1993, 61(5): 2172-2181.
[10]Marsden M J, Vaughan L M, Foster T J, et al. A live (delta aroA) Aeromonas salmonicida vaccine forfurunculosis preferentially stimulates T-cell responses relative to B-cell responses in rainbow trout (Oncorhynchus mykiss)[J]. Infection and Immunity, 1996, 64(9): 3863-3869.
[11]陈振海，林天龙，夏春．鱼类DNA疫苗研究的新进展[J]．水生生物学报，2003，27(6)：648-653．
[12]Heppell J, Davis H L. Application of DNA vaccine technology to aquaculture[J]. Advanced Drug Delivery Reviews, 2000, 43(1): 29-43.
[13]Anderson E D, Mourich D V, Fahrenkrug S C, et al. Genetic immunization of rainbow trout (Oncorhynchus mykiss) against infectious hematopoietic necrosis virus[J]. Molecular Marine Biology & Biotechnology, 1996, 5(2): 114-112.
[14]Anderson E D, Mourich D V, Lenong J A. Gene expression in rainbow Trout (Oncorhynchus mykiss) following intramuscular injection of DNA[J]. Molecular Marine Biology & Biotechnology, 1996, 5(2): 105-113.
[15]Traxler G S, Anderson E, LaPatra SE, et al. Naked DNA vaccination of Atlantic salmon Salmo salar against IHNV[J]. Disease of Aquatic Organisms, 1999, 38(3): 183-190.
[16]Lorenzen N, Lorenzen E, Einer J K, et al. Genetic vaccination of rainbow trout against viral haemorrhagic septicaemia virus: small amounts of plasmid DNA protect against a heterologous serotype[J]. Advances in Virus Research, 1999, 63(1-2): 19-25.
[17]Kim C H, Johnson M C, Drennan J D, et al. DNA vaccines encoding viral glycoproteins induce nonspecific immunity and Mx protein synthesis in fish[J]. Journal of Virological Methods, 2000, 74 (15): 7048-7054.
[18]王利，谢长勇．鱼类疫苗研究进展[J]．动物医学进展，2003，24(5)：26-28．
[19]Eldar A, Horovitcz A Bercovier H. Development and efficacy of a vaccine against streptococcus iniae infection in farmed rainbow trout[J]. Veterinary Immunology & Immunopathology, 1997, 56(1-2): 175-183.
[20]陈超然，陈昌福．鱼用疫苗的研究现状[J]．水利渔业，2001，21(5)：44-45．
[21]Li S, Woo P T. Efficacy of a live Cryptobia salmositica vaccine and the mechanism of protection in vaccinated rainbow trout, Oncorhynchus mykiss against cryptobiosis[J]. Veterinary Immunology and Immunopathology, 1995, 48(3-4): 343-353.
[22]夏春．鱼病诊断与防治手册[M]．北京：中国农业大学出版社，1999，21-32．
[23]杨先乐，左文功．草鱼出血病细胞培养灭活疫苗的研究：疫苗的稳定性及佐剂和加强免疫对：草鱼免疫应答的影响[J]．水生生物学报，1993，17(1)：46-51．
[24]Stephen G N, John J M. Immunization of Salmonids against furunculosis[J]. Fish Pathology, 1985, 20(2-3): 403-411.
[25]Newman S G. Bacterial vaccines for fish[J]. Annual Review of Fish Diseases, 1993, 32(3): 145-185.
[26]张吉红，陆承平．佐剂对嗜水气单胞菌灭活疫苗免疫效果的作用[J]．中国兽药杂志，2003，7(4)：26-27．
[27]Midtlyng P J. A field study on intraperitoneal vaccination of Atlantic salmon (Salmo salarL.) against furunculosis[J]. Fish & Shellfish Immunology, 1996, 6(8): 553-565.
[28]Midtlyng P J, Reitan L J. Speilberg L. Experimental studies on the efficacy and side-effects of intraperitoneal vaccination of Atlantic salmon (Salmo salarL.) against furunculosis[J]. Fish & Shellfish Immunology, 1996, 6(5): 335-350.
[29]高健编译．免疫激活佐剂在水产养殖中的应用[J]．河北渔业，1994，34．
[30]傅安静，岳华，刘树玲等．疫苗佐剂研究进展[J]．西南民族大学学报(增刊)，2005，73-77．
[31]Donnelly J J. New development in adjuvants[J]. Mechanisms of Ageing and Development, 1997, 93(1-3): 171-177.
[32]Hagan D T, MacKichan M L, Singh M. Recent developments in adjuvants for vaccines against infectious diseases[J]. Biomolecular Engineering, 2001, 18(3): 69-85.
[33]赵建培，史维舟．在草鱼免疫反应中莨菪佐剂性的初步观察[J]．淡水渔业，1995，25(1)：3-5．
[34]吴志新，陈强，陈昌福．左旋咪唑对异育鲫鱼免疫促进作用的初步研究[J]．华中农业大学学报，1998，17(4)：378-381．
[35]Kajita Y, Sakai M, Atsuta S, et al. The immunomodulatory effects of levamisole on rainbow trout, Oncorhynchus mykiss[J]. Fish Pathology, 1990, 25: 93-98.
[36]张庆茹．中草药免疫促进作用的研究进展[J]．中兽医医药杂志，1997，(5)：15．
[37]薛明，孟宪松．生物活性多糖的免疫研究进展与展望[J]．中兽医医药志，1996，(3)：16．
[38]万金志，乔悦昕．芦荟的化学成分及其研究[J]．中草药，1999，30(2)：151-153．
[39]方仲根．芦荟的活性成分与疗效作用[J]．热带农业科学，1999，(1)：59-62．
[40]Ki Hong Kim. Resistance to Vibrio alginolyticus in juvenile rockfish ( Sebastes schlegeli ) fed diets containing different doses of aloe[J]. Aquaculture, 1999, 180(1-2): 13-21.
[41]孙建宏．免疫佐剂的研究进展[J]．黑龙江畜牧兽医，1998，2：40-43．
[42]Sakai M, Atsuta S, Kobayashi M. Protective immune response in rainbow trout, Oncorhynchus mykiss,vaccinated with β-haemolytic streptocococal bacterin[J]. FishPathology, 1989, 24(3): 169-173.
[43]Shoemaker C, Klesius P. Streptococcal disease problems and control: a review In Fitzsimmons, K. Ed., Tilapia Aquaculture vol. 2 Northeast Regional Agricultural Engineering Service-106, Ithaca, NY, USA, 1997: 671-680.
[44]Pier G B, Madin S H, Al-Nakeeb S. Isolation and characterization of a second isolate of Streptococcus iniae[J]. International Journal of Systematic Bacteriology, 1978, 28:311-314.
[45]Sakai M, Atsuta S, Kobayashi M. Protective immune response in rainbow trout, Oncorhynchus mykiss, vaccinated with β-haemolytic streptocococal bacterin[J]. Fish Pathology, 1989, 24(3): 169-173.
[46]Kitao T. Streptococcal infections. In Inglis V, Roberts R J, Bromage NR Eds. Bacterial Diseases of Fish[M]. Oxford: Blackwell Scientific Publications, 1993,196-210.
[47]Evans J J, Klesius P H, Shoemaker C A. Streptococcus iniae in warm-water fish[J]. Aquaculture Health International, 2006, 7: 10-14.
[48]Litt M D, Low D, Cunha C, et al. Surveillance for human infection with Streptococcus iniae[J]. Journal of Clinical Epidemiology, 1997, 50(1): 17.
[49]Eldar A, Bejerano Y, Livoff A, et al. Experimental streptococcal meningo-encephalitis in cultured fish[J]. Veterinary Microbiology, 1995, 43(1): 33-40.
[50]Bromage E S, Owens L. Infection of barramundi Lates calcarifer with Streptococcus iniae: effects of different routes of exposure[J]. Diseasea of aquatic organisms, 2002, 52(3): 199-205.
[51]Evans J J, Shoemaker C A, Klesius P H. Distribution of Streptococcus iniae in hybrid striped bass (Morone chrysops×Morone saxatilis) following nare inoculation[J]. Aquaculture, 2001, 194(3-4): 233-243.
[52]Ana I M, Blanco M M, Dominguez L, et al. Development of a PCR assay for Streptococcus iniae based on the lactate oxidase (lctO) gene with potential diagnostic value[J]. Veterinary Microbiology, 2004, 101 (2): 109-116.
[53]Fuller J D, Bast D J, Nizet V, et al. Streptococcus iniae virulence is associated with a distinct genetic profile[J]. Infection and immunity, 2001, 69(4): 94-100.
[54]柴家前，丁巧玲，王振龙等．罗非鱼链球菌的分离鉴定[J]．中国预防兽医学报，2002，24(1)：18-20．
[55]Elliott J A, Facklam R R, Richter C B. Whole-cell protein patterns of nonhaemolytic group B, type Ⅰb, streptococci isolated from humans, mice, cattle, frogs and fish[J]. Journal of Clinical Microbiology, 1990, 28: 628-630.
[56]Robinson J A, Meyer F P. Streptococcal fish pathogen[J]. Journal of Bacteriology, 1966,92: 512.
[57]Plumb J A, Schachte J H, Gaines J L, et al. Streptococcus sp. from marine fishes along the Alabama and Northwest Florida coast of the Gulf of Mexico[J]. Transactions of the American Fisheries Society, 1974, 103: 358-361.
[58]Baya A M, Lupiani B, Hetrick F M, et al. Association of Streptococcus sp with fish mortalities in the Chesapeake Bay and its tributaries[J]. Journal of Fish Diseases, 1990, 13: 251-253.
[59]Eldar A, Bejerano Y, Bercovier H. Streptococcus shiloi and Streptococcus difficile: two new streptococcal species causing Meningoencephalitis in fish[J]. Current Microbiology, 1994, 28: 139-143.
[60]Wilkinson H W, Thacker L G, Facklam R R. Nonlaaemolytic group B Streptococci of human, bovine and ichthyic origin[J]. Infection and Immunity, 1973, 7: 496-498.
[61]Vandamme P, Devriese L A, Pot B, et al. Streptococcus difficile is a nonhaemolytic group B, type Ⅰb Streptococcus[J]. International Journal of Systematic Bacteriology, 1997, 47: 81-85.
[62]Evans J J, Klesius P H, Glibert P M, et al. Characterisation ofbeta-haemolytic Group B Streptococcus agalactiae in cultured sea bream, Sparus auratus L and wild mullet, Liza klunzingeri (Day), in Kuwait[J]. Journal of Fish Diseases, 2002a, 25: 505-513.
[63]Kawamura Y, Itoh Y, Mishima N, et al. High genetic homogeneity of Streptococcus agalactiae and Streptococcus difficilis: reclassification of S difficilis, Eldar et al 1995 as a junior synonyn of S agalactiae, Lehmann and Neumann 1896[J]. International Journal of Systematic and Evolutionary Microbiology, 2005, 55: 961-965.
[64] Evans J J, Pasnik D J, Klesius P H, et al. First report of Streptococcus agalactiae and Lactococcus garvieae from a wild bottlenose dolphin, Tursiops truncatus[J]. Journal of Wildlife Diseases 2006a, 42: 561-569.
[65] Kitao T. Streptococcal infections. In Inglis V, Roberts RJ, Bromage NR Eds [C]. Bacterial Diseases of Fish, Blackwell Scientific Publications, Oxford, 1993: 196-210.
[66] Evans J J, Pasnik D J, Klesius P H, et al. Identification and epidemiology of Streptococcus iniae and 5 agalactiae in tilapia [C]. Proceedings of the International Symposium of Tilapia Aquaculture ISTA7. Veracruz, Mexico, 2006, 9: 25-42.
[67] Pasnik D J, Evans J J, Panangala V S, et al. Antigenicity of Streptococcus agalactiae extracellular products and vaccine efficacy [J]. Journal of Fish Diseases, 2005,28: 205-212.
[68] Shoemaker C A, Klesius P H, Evans J J. Prevalence of Streptococcus iniae in tilapia, hybrid striped bass and channel catfish from fish farms in the United States [J]. American Journal of Veterinary Research, 2001,62:174-177.
[69] Evans J J, Shoemaker C A, Klesius P H. Distribution of Streptococcus iniae in hybrid striped bass Morone chrysops x Morone saxatilis following nares inoculation [J]. Aquaculture, 2001, 194: 233-243.
[70] Evans J J, Wiedenmayer A A, Klesius P H. A transport system for the maintenance of viability of Acinetobacter calcoaceticus, Streptococcus iniae and S agalactiae over varying time periods [J]. Bulletin of the European Association of Fish Pathologists, 2002b, 22: 238-246.
[71] Evans J J, Wiedenmayer A A, Klesius P H, et al. Survival of Streptococcus agalactiae from frozen fish following natural and experimental infections [J]. Aquaculture, 2004a, 233: 15-21.
[72] Facklam R. What happened to the Streptococci: overview of taxonomic and nomenclature changes [J]. Clinical Microbiology Reviews, 2002, 15: 613-630.
[73] Kimura H, Kusuda R. Studies on the pathogenesis of streptococcal infection in cultured yellowtails, Seriola spp: effect of crude exotoxin fractions from cell-free culture on experimental streptococcal infection [J]. Journal of Fish Diseases, 1982, 5: 471-478.
[74] Boomker J, Imes G J, Cameron C M, et al. Trout mortalities as a result of Streptococcus infection [J]. Onderstepoort Journal of Veterinary Research, 1979, 46: 71-77.
[75] Klesius P H, Evans J J, Shoemaker C A, et al. Rapid detection and identification of Streptococcus iniae using a monoclonal antibody-based indirect fluorescent antibody technique [J]. Aquaculture, 2006a, 258: 180-186.
[76] Berridge B R, Bercovier H, Frelier P F. Streptococcus agalactiae and Streptococcus difficile 16S-23S intergenic rDNA: genetic homogeneity and species-specific PCR [J]. Veterinary Microbiology, 2001, 78:165-173.
[77]Mata A I, Gibello A, Casamayor A, et al. Multiplex PCR assay for detection of bacterial pathogens associated with warm-water streptococcosis in fish[J]. Applied and Environmental Microbiology, 2004, 70: 3183-3187.
[78]Pier G B, Madin S H. Streptococcus iniae sp nov, a beta-haemolytic Streptococcus isolated from an Amazon freshwater dolphin, Inia geoffrensis[J]. International Journal of Systematic Bacteriology, 1976, 26: 545-553.
[79]Eldar A, Bejerano Y, Bercovier H. Streptococcus shiloi and Streptococcus difficile: two new streptococcal species causing Meningoencephalitis in fish[J]. Current Microbiology, 1994, 28: 139-143.
[80]Perera R P, Collins M D, Johnson S K,et al. Streptococcus iniae associated with mortality of Tilapia nilotica x T aurea hybrids[J]. Journal of Aquatic Animal Health, 1994, 6: 335-340.
[81]Shelby R A, Klesius P H, Shoemaker C A, et al. Passive immunisation of tilapia, Oreochromis niloticus L with anti-Streptococcus iniae whole sera[J]. Journal of Fish Diseases, 2002, 25: 1-6.
[82]Evans J J, Shoemaker C A, Klesius P H. Therapeutic and prophylactic immunisation against Streptococcus iniae infection in hybrid striped bass Morone chrysops x M saxatilis[J]. Aquaculture Research, 2006c, 37: 742-750.
[83]Klesius P H, Evans J J, Shoemaker C A, et al. Vaccines to prevent Streptococcus iniae and S agalactiae disease in tilapia, Oreochromis niloticus[C]. Proceedings of the International Symposium of Tilapia Aquaculture ISTA7, Veracruz, Mexico, 2006, 9: 15-24.
[84]Eldar A, Shapiro O, Bejerano Y,et al. Vaccination with whole-cell vaccine and bacterial protein extract protects tilapia against Streptococcus difficile meningoencephalitis[J]. Vaccine, 1995b, (13): 867-870.
[85]Klesius P H, Shoemaker C A, Evans J J. Efficacy of single and combined Streptococcus iniae isolate vaccine administrated by intraperitoneal and intramuscular routes in tilapia (Oreochromis niloticus)[J]. Aquaculture, 2000, 188 (3-4): 237-246.
[86]Eldar A, Horovitcz A, Bercovier H. Development and efficacy of a vaccine against Streptococcus iniae infection in farmed rainbow trout[J]. Veterinary Immunology and Immunopathology, 1997, 56(1-2): 175-183.
[87]Bachrach G, Zlotkin A, Hurvitz A, et al. Recovery of Streptococcus iniae from diseased fish previously vaccinated with a streptococcus vaccine[J]. Applied and Environmental Microbiology, 2001, 67(8): 3756-3758.
[88]Klesius P H, Shoemaker C A and Evans J J. 2000. Efficacy of single and combined Streptococcus iniae isolate vaccine administered by intraperitoneal and intramuscular routes in tilapia (Oreochromis niloticus)[J]. Aquaculture, 188(3-4): 237-246.
[89]Akhlaghi M., Munday B L, Whittington R J. Comparison of passive and active immunization of fish against streptococcosis (enterococcosis)[J]. Journal of fish diseases, 1996, 19: 251-258.
[90]Nakanishi T, Kiryu I, Ototake M. Development of a new vaccine delivery method for fish: percutaneous administration by immersion with application of a multiple puncture instrument[J]. Vaccine, 2002, 20(31): 3764-3769.
[91]Shelby R A, Klesius P H, Shoemaker C A, et al. Passive Immunization of tilapia, Oreochromis Niloticus (L.) with Anti-Streptcoccus iniae whole sera[J]. Journal of Fish Diseases, 2002, 25(1): 1-6.
[92]Shoemaker C A, Vandenberg G W, Desormeaux Andre, et al. Efficacy of a Streptococcus iniae modified bacterin delivered using Oralject~(TM) technology in Nile tilapia(Oreochromis niloticus)[J]. Aquaculture, 2006, 255(1-4): 151-156.
[93]张延龄，张晖．疫苗学[M]．北京：科学出版社，2004，3：323．
[94]Shoemaker C, Klesius P. Streptococcal disease problems and control: a review[C]. In Tilapia Aquaculture Northeast Regional Agricultural Engineering Service, Ithaca, NY, 1997, (2): 671-680.
[95]Eldar A, Shapiro O, Bejerano Y,et al. Vaccination with whole-cell vaccine and bacterial protein extract protects tilapia against Streptococcus difficile meningoencephalitis[J]. Vaccine, 1995b, (13): 867-870.
[96]Klesius P H, Shoemaker C A, Evans J J. Efficacy of single and combined Streptococcus iniae isolate vaccine administrated by intraperitoneal and intramuscular routes in tilapia (Oreochromis niloticus)[J]. Aquaculture, 2000, 188 (3-4): 237-246.
[97]Kenneth E N, Bruce F S, Patricia D I. Protective immunity induced by DNA vaccination of channel catfish with early and late transcripts of the channel catfish herpesvirus (IHV-1)[J].Veterinary immunology and immunopathology, 2002, 84 (4): 151-168.
[98]Klesius P H, Shoemaker C A,Evans J J. Efficacy of single and combined Streptococcus iniae isolate vaccine administered by intraperitoneal and intramuscular routes in tilapia (Oreochrormis niloticus)[J]. Aquaculture, 2000, 188: 237-246.
[99]Ackerman P A, Iwama G K. Physiological and immunological effects of adjuvanted Aeromonas salmonicida vaccines on juvenile rainbow trout[J]. Journal of Aquatic Animal Health, 2000, 12: 157-164.
[100]Toranzo A E, Devesa S, Romaide J L, et al. Efficacy of intraperitoneal and immersion vaccination against Enterococcus sp.infection in turbot[J]. Aquaculture, 1995, 134: 17-27.
[101]Waterstrat P, Ainsworth J, Capley G. Use of an indirect enzyme-linked immunosorbent assay(ELISA) in the detection of channel catfish, lctalurus punctatus (Rafinesque), antibodies to Edwardsiella ictaluri[J]. Journal of Fish Diseases, 1989, 12 (2): 87-94.
[102]Eldar A, Horovitcz A, Bercovier H. Development and efficacy of a vaccine against Streptococcus iniae infection in farmed rainbow trout[J]. Veterinary immunology & Immunopathology, 1997, 56:175-183.
[103]Klesius P H, Shoemaker, C A, Evans, J J. Efficacy of a killed Streptococcus iniae vaccine in tilapia (Oreochromis niloticus)[J]. Bulletin of the European Association of Fish Pathologists. 1999, 19(1): 1-3.
[104]Klesius P H, Shoemaker, C A, Evans, J J.Efficacy of single and combined Streptococcus iniae isolate vaccine administered by intraperitoneal and intramuscular routes in tilapia (Oreochromis niloticus)[J]. Aquaculture, 2000(3-4): 237-246.
[105]Hurvitz A, Bercovier H, Rijn J V. Effect of ammonia on the survival and the immune response of rainbow trout (Oncorhynchus mykiss, Walbaum) vaccinated against Streptococcus iniae[J]. Fish & shellfish immunology, 1997, 7(1): 45-53.
[106]Barnes A C, Young F M, Horne M T, Ellis A E. Streptococcus iniae: serological differences presence of capsule and resistance to immune serum Killing[J]. Diseases of aquatic organisms, 2003, 53(3): 241-247.
[107]Evans J J, Klesius P H, Shoemaker P K, et al. Experimental Streptococcus iniae infection of hybrid striped bass (Morone chrysops×Morone saxatilis) and tilapia (Oreochromis niloticus) by nares inoculation[J]. Aquaculture, 2000, 189(3-4): 197-210.
[108]Kitao T. Streptococcal infections. In Inglis V, Roberts RJ,Bromage NR Eds. Bacterial Diseases of Fish. Oxford: Blackwell Scientific Publications, 1993, 196-210.
[109]Joyce J E, Craig A S, PHillip K. Experimental Streptococcus iniae infection of hybrid striped bass (Morone chrysops×Morone saxatilis) and tilapia (Oreochromis niloticus) by nares inoculation[J]. Aquaculture, 2000, 189: 197-210.
[110]Amir Z, Hannah H, Avi E. Possible transmission of Streptococcus iniae from wild fish to cultured fish[J]. Applied and environmental microbiology, 1998, 4065-4067.
[111]Shorts E B, Blazer V S, and Waltman W D. Pathogenesis of experimental Edwardsiell ictaluri infections in channel catfish (Ictalurus punctatus)[J]. Fish. Sci, 1986,43: 36-42.
[112]Newton J C, Wolfe L G, Grizzle J M, and Plumb J A. Pathology of experimental enteric septcaemia in channel catfish, Ictalurus punctatus (Rafinesque), following immersion-exposure to Edwardsiella ictaluri[J]. Fish Dis, 1989, 12: 335-347.
[113]Morrison E E and Plumb J A. Olfactory organ of channel catfish as a site of experimental Edwardsiella ictaluri infection[J]. Aquat. Anim. Health, 1994, 6: 101-1099.
[114]Hoshina T, Sano T and Morimoto Y. A Streptococcus pathogenic to fish[J]. Tokyo Univ. Fish, 1958, 44: 57-58.
[115]Robinson J A. and Meyer F P. Streptococcal fish pathogen[J]. Bacteriol, 1966, 92(2): 512.
[116]Cook D W and Lofton S R. Pathogenicity studies with a Streptococcus sp. isolated from fishes in an Alabama-Florida fish kill[J]. Trans. Am. Fish. Soc, 1975, 2: 286-288.
[117]Ohnishi K, and Jo Y. Studies of streptococcal infection in pond-cultured fishes. Characteristics of a β-hemolytic Streptococcus isolated from cultured ayu and amago in 1997-1998[J]. Fish Pathol, 1981, 16: 63-67.
[118]Evan J J, Shoemaker C A and Klesius P H. Experimental Streptococcus iniae infection of hybrid striped bass (Morone chrysops×Morone saxatilis) and tilapia (Oreoehromis niloticus) by nares inoculation[J]. Aquaculture, 2000, 189(3-4): 197-210.
[119]Klesius P H, Shoemaker C A, Evans J J.Efficacy of single and combined Streptococcus iniae isolate vaccine administered by intraperitoneal and intramuscular routes in tilapia(Oreochromis niloticus). Aquaculture, 2000, 188: 237-246.
[120]杨先乐，陈远新．鱼用疫苗的现状及其发展趋势[J]．水产学报，1996，20(2)：159-167．
[121]Pdjkers G T. The haemolftic plaque assay in carp, CyprinuscarpioL.[J].Journal of immune Meth, 1980, 33: 79-86.
[122]Eldar A, Horovitcz A, Bercovier H.Development and efficacy of a vaccine against Streptococcus iniae infection in farmed rainbow trout[J]. Veterinary Immunology and Immunopathology, 1997(56): 175-183.
[123]Klesius P H, Shoemaker C A, Evans J J.Efficacy of single and combined Streptococcus iniae isolate vaccine administered by intraperitoneal and intramuscular routes in tilapia(Oreochromis niloticus). Aquaculture, 2000, 188: 237-246.
[124]Eldar A, Horovitcz A, Bercovier H. Development and efficacy of a vaccine against Streptococcus iniae infection in farmed rainbow trout[J]. Veterinary immunology & Immunopathology, 1997, 56: 175-183.
[125]王建英，杨桂文，田雪文等．免疫佐剂及其作用机制[J]．山东师范大学学报，2006，21 (1)：127-129．
[126]Midtlyng P J, Reitan L J. Speilberg L. Experimental studies on the efficacy and side-effects of intraperitoneal vaccination of Atlantic salmon (Salmo salarL.) against furunculosis[J]. Fish & Shellfish Immunology, 1996, 6 (5): 335-350.
[127]Klesius P H, Shoemaker C A, Evans J J. Efficacy of single and combined Streptococcus iniae isolate vaccine administered by intraperitoneal and intramuscular routes in tilapia(Oreochromis niloticus). Aquaculture, 2000, 188: 237-246.
[128]Evans J J, Klesius P H, Shoemaker C A. Efficacy of Streptococcus agalactiae (group B) vaccines in tilapia (Oreochromis niloticus) by intraperitoneal and bath immersion administration[J]. Vaccine, 2004, 22 (27-28): 3769-3773.
[129]Eldar A, Horovitcz A, Bercovier H. Development and efficacy of a vaccine against Streptococcus iniae infection in farmed rainbow trout[J]. Veterinary Immunology and Immunopathology, 1997, 56 (1-2): 175-183.
[130]高冬梅，李健，王群．鳗弧菌灭活疫苗对牙鲆免疫保护效果的研究[J]．海洋水产研究，2004，25(1)：34-40．
[131]李渊，孙红祥．不同类型免疫佐剂的作用比较[J]．浙江江畜牧兽医，2003，(5)：16-18．
[132]Jansen T, Hofmans M M,Theelen J G, et al. Structure-activity relations of water-in-oil vaccine formulations and induced antigen-specific antibody responses[J]. Vaccine, 2005, 23(8):1053-1060.
[133]万金志，乔悦昕．芦荟的化学成分及其研究[J]．中草药，1999，30(2)：151-153．
[134]方仲根．芦荟的活性成分与疗效作用[J]．热带农业科学，1999，(1)：59-62．
[135]苗明三，常章富．芦荟多糖对正常小鼠免疫作用影响的研究[J]．中医药学刊，2003(8)：12-42．
[136]宫朋，王富辉，杨长树．木立芦荟粗多糖对免疫功能的影响[J]．人参研究．2000，12(2)：28-29．
[137]王莉，包旭，陈淑杰等．巴巴多斯芦荟多糖提取物对小鼠免疫功能的影响[J]．华西药学杂志，1999，14(4)：234-235．
[138]王莉，杜俊蓉，蔡绍晖等．AP对正常和内毒素化大鼠脾细胞体外分泌细胞因子的影响[J]．华西药学杂志，1999，14(6)：346-348．
[139]吴志新，陈强，陈昌福等．左旋咪唑对异育鲫鱼免疫促进作用的初步研究[J]．华中农业大学学报，1998，17(4)：378-381．
[140]Kajita Y, Sakai M, Atsuta S, et al. The immunomodulatory effects of levamisole on rainbow trout, Oncorhynchus mykiss[J]. Fish Patholology, 1990, 25: 93-98.
[141]叶丹，连宾．溶菌酶及其应用[J]．贵州科学，2003，21(3)：45-46．
[142]Shih-Chu Chen, Terutoyo Yoshida, Alexandra Adams, et al.Thompson.Immune response of Rainbow Trout to extracellular products of Mycobacterium spp[J]. Journal of Aquatic Animal Health, 1996, 8: 216-222.
[143]王宏田，徐永立，张培军．假雄牙鲆不同组织中溶菌酶比活性的研究[J]．海洋科学，2000，24(10)：7-8．
[144]Crinde B, Lie O, Poppe T, Salte R. Species and individual variation in lysozyme activity in fish of interest in aquaculture[J].Aquaculture, 1988, 68: 299-304.
[145]Paige A A, George K, lwama. Physiological and immunological effects of adjuvanted Aeromonas salmonicida vaccines on juvenile rainbow trout[J]. Journal of Aquatic Animal Health, 2000, 12: 157-164.
[146]安利国，玛程强，邢维贤等．灭活疫苗对鲤鱼血清溶菌酶和腹腔吞噬细胞活性的作用[J]．山东师范大学学报(自然科学版)，1999，14(2)：175-179．
[147]Siwicki A, Studnicka M. The phagocytic ability of neutrophils and serum lysozyme activity in experimentally infected carp, Cyprinus carpio L.[J]. Journal of Fish Biology, 1987, 31 (Supplement A): 57-60.