大黄鱼人工感染杀香鱼假单胞菌的病理形态学及病原分布的初步研究
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摘要
为了研究杀香鱼假单胞菌在感染大黄鱼体内的分布状况及其所致各器官的病理损伤特点,通过人工回感试验,利用组织切片及免疫组化技术对感染大黄鱼的肝脏、脾脏、肾脏及鳃组织进行了组织病理切片,并进行HE染色和免疫组化分析。结果显示,人工感染发病大黄鱼临床症状与自然感染发病症状一致,病鱼体表无明显病症,但肝、脾、肾等内脏有大量白色结节;组织病理显示,病鱼肝脏、脾脏和肾脏是感染损伤的主要靶器官,内脏组织发生变性、坏死以及出现特征性的病理性结节;免疫组化检测特异性阳性信号出现在病鱼内脏组织器官的血窦或血管以及巨噬细胞内。研究表明,杀香鱼假单胞菌侵入鱼体后,主要分布在机体的血液中并大量增殖,通过血液循环到达鱼体各内脏器官;杀香鱼假单胞菌在鱼组织内的分布与其所致病理损伤之间呈正相关。
The white-spots disease was a serious bacterial disease in cultured Pseudosciaena crocea in recent years. The bacterium Pseudomonas plecoglossicida was responsible for the disease. This test using intraperitoneal injection way artificially infected the P. crocea(40±3 g). The collected diseased fish showed clinical signs of numerous white nodules present in the spleen, liver or kidney internally without any other symp-toms. Sick fishes were examined by means of histopathology and immunohistochemistry. Histopathological anal-ysis revealed severe degeneration and necrotic changes occurred in the liver, spleen and kidneys. The results of immunohistochemical staining showed that the strong positive signal of the pathogen was in the liver, spleen and kidneys, particularly in macrophages of the internal organs of diseased fish. Studies shown that P.plecoglossicida invades the fish body, and is mainly distributed in the blood vessels of the body and proliferates in a large amount. The results of P. plecoglossicida distribution in infected P. crocea tissues and pathological changes showed there were positively correlations between the P. plecoglossicida distribution in P. crocea tissues and pathological damage.
The white-spots disease was a serious bacterial disease in cultured Pseudosciaena crocea in recent years. The bacterium Pseudomonas plecoglossicida was responsible for the disease. This test using intraperitoneal injection way artificially infected the P. crocea(40±3 g). The collected diseased fish showed clinical signs of numerous white nodules present in the spleen, liver or kidney internally without any other symp-toms. Sick fishes were examined by means of histopathology and immunohistochemistry. Histopathological anal-ysis revealed severe degeneration and necrotic changes occurred in the liver, spleen and kidneys. The results of immunohistochemical staining showed that the strong positive signal of the pathogen was in the liver, spleen and kidneys, particularly in macrophages of the internal organs of diseased fish. Studies shown that P.plecoglossicida invades the fish body, and is mainly distributed in the blood vessels of the body and proliferates in a large amount. The results of P. plecoglossicida distribution in infected P. crocea tissues and pathological changes showed there were positively correlations between the P. plecoglossicida distribution in P. crocea tissues and pathological damage.
引文
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[13]连林坤.虾蟹螺原体病免疫组化和病理学比较研究[D].南京:南京师范大学, 2011:11-15.
[14]张艳秋,詹勇,许梓荣.鱼类免疫机制及其影响因子[J].水产养殖, 2005, 26(3):1-5.
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[17]张杰,郭海杰,高丽婷,等.杀香鱼假单胞菌Ⅲ型分泌系统转录调控蛋白ExsA突变株的构建及其对大黄鱼的毒力特征分析[J].水产学报, 2017, 41(4):613-621.
[2]刘家富.大黄鱼养殖与生物学[M].厦门:厦门大学出版社, 2013.
[3]鄢庆枇,王军,苏永全,等.网箱养殖大黄鱼弧菌病研究[J].集美大学学报:自然科学版, 2001, 6(3):191-196.
[4]袁雪梅,葛明峰,安树伟,等.大黄鱼致病香鱼假单胞菌对环境因子的响应及其感染检测的分析[J].应用海洋学学报, 2015,34(3):549-553.
[5]张丹枫,安树伟,周素明,等.大黄鱼(Pseudosciaena crocea)内脏白点病的组织病理和超微病理分析[J].渔业科学进展, 2017, 38(4):11-16.
[6] ZHANG J T, ZHOU S M, AN S W, et al. Visceral granulomas in farmed large yellow croaker, Larimichthys crocea(Richardson),caused by a bacterial pathogen, Pseudomonas plecoglossicida[J]. Journal of Fish Diseases, 2013, 37(2):113-121.
[7]刘年锋,郭睿,江小斌,等.福州地区大黄鱼变形假单胞菌的分离与鉴定[J].渔业研究, 2016, 38(1):14-20.
[8]许斌福,程海华,池洪树,等.大黄鱼内脏白点病的病原分析与鉴定[J].福建农业学报, 2015, 30(7):631-635.
[9]邱杨玉.大黄鱼内脏白点病的研究[D].武汉:华中农业大学, 2012:20-21.
[10] NISHIMORI E, KITA-TSUKAMOTO K, WAKABAYASHI H. Pseudomonas plecoglossicida sp. nov., the causative agent of bacterial haemorrhagic ascites of ayu, Plecoglossus altivelis[J]. International Journal of Systematic and Evolutionary Microbiology, 2000, 50(1):83-89.
[11]陈代雄,盂锦绣,易冰,等.抗日本血吸虫循环抗原单链抗体的抗原定位和初步应用[J].中国人兽共患病杂志, 2003, 19(4):63-65.
[12]张智华.用免疫组化与PCR方法鉴定裂头蚴感染病理标本[D].长沙:中南大学, 2013:21-23.
[13]连林坤.虾蟹螺原体病免疫组化和病理学比较研究[D].南京:南京师范大学, 2011:11-15.
[14]张艳秋,詹勇,许梓荣.鱼类免疫机制及其影响因子[J].水产养殖, 2005, 26(3):1-5.
[15] DORHOI A, KAUFMANN K. Immunity to intracellular bacteria[J]. Veterinary Immunology&Immunopathology, 2013, 3(1-2):5-66.
[16] De HERDT P, HAESEBROUCK F, CHARLIER G, et al. Intracellular survival and multiplication of virulent and less virulent strains of Streptococcus bovis in pigeon macrophages[J]. Veterinary Microbiology, 1995, 45(2/3):157-169.
[17]张杰,郭海杰,高丽婷,等.杀香鱼假单胞菌Ⅲ型分泌系统转录调控蛋白ExsA突变株的构建及其对大黄鱼的毒力特征分析[J].水产学报, 2017, 41(4):613-621.