方斑东风螺“脱壳病”病原及病理初步研究
|
摘要
为明晰方斑东风螺"脱壳病"的病原或病理,对东风螺"脱壳病"进行了病原分离纯化、病原菌形态观察、组织病理观察、人工感染试验,并对病原菌16S rDNA序列进行PCR扩增和测序,以及系统发育树分析。试验中分离纯化了3种纯菌W、B、Y。致病菌W杆状,端生单鞭毛。致病菌Y杆状,两端略尖,周生菌毛。致病菌B短杆状,无鞭毛。结果显示,患病个体和人工感染个体的腹足及连壳肌肉组织均出现明显病变,肌纤维受损,出现空腔,并存在细菌;肝胰腺组织无明显病变,但血细胞显著减少。腹足肌注射人工感染试验结果表明,致病菌W在3种细菌中毒力最强。混合菌感染的毒力在混合比例接近原始比例时达到最大。致病菌B与报道的鲍希瓦氏菌相似度为99%;致病菌Y与报道的哈维弧菌相似度为98%;致病菌W与报道的变形假单胞菌相似度为99%。推测致病菌W、B、Y可能是此次方斑东风螺脱壳病的重要病原菌。
The pathogens in shell and flesh separating disease in Fujian were isolated and purified from ivory shell Babylonia areolata,and artificially infected,whose sequences were analized by 16 SrDNA,and whose histopathology was observed.Three dominant bacterial strains(B,W and Y)were isolated from the proboscis,muscle and hepatopancreas of moribund ivory shell collected from a farm in Jinjiang,Fujian by aseptic methods,physiological and biochemical methods combined with molecular biology methods to classify and identify the pathogen.The results showed that the organs of ivory shell including foot muscle,connecting shells muscle,digestive gland,and hepatopancreas were severely damaged and a series of serious lesions were caused,with muscle fibers arranged disorderly,hollow myofibers,and blood cell decrease in hepatopancreas and connective tissue.The bacterial pathogen B was postulated to be Shewanella haliotis,the bacterial pathogen Y was postulated to be Vibrio harveyi,and the bacterial pathogen W was postulated to be Pseudomonas plecoglossicida.It was shown that the bacterial pathogen W has strong toxiferous to ivory shell,and the maximum toxicity in mixed bacteria when the proportion of the mixed bacteria was close to the original proportion.In summary,the findings provide the theoretical basis and guide in preventing and controlling of the shell and flesh separating disease in ivory shell.
The pathogens in shell and flesh separating disease in Fujian were isolated and purified from ivory shell Babylonia areolata,and artificially infected,whose sequences were analized by 16 SrDNA,and whose histopathology was observed.Three dominant bacterial strains(B,W and Y)were isolated from the proboscis,muscle and hepatopancreas of moribund ivory shell collected from a farm in Jinjiang,Fujian by aseptic methods,physiological and biochemical methods combined with molecular biology methods to classify and identify the pathogen.The results showed that the organs of ivory shell including foot muscle,connecting shells muscle,digestive gland,and hepatopancreas were severely damaged and a series of serious lesions were caused,with muscle fibers arranged disorderly,hollow myofibers,and blood cell decrease in hepatopancreas and connective tissue.The bacterial pathogen B was postulated to be Shewanella haliotis,the bacterial pathogen Y was postulated to be Vibrio harveyi,and the bacterial pathogen W was postulated to be Pseudomonas plecoglossicida.It was shown that the bacterial pathogen W has strong toxiferous to ivory shell,and the maximum toxicity in mixed bacteria when the proportion of the mixed bacteria was close to the original proportion.In summary,the findings provide the theoretical basis and guide in preventing and controlling of the shell and flesh separating disease in ivory shell.
引文
[1]王江勇,王瑞旋,苏友禄,等.方斑东风螺“急性死亡症”的病原病理研究[J].南方水产科学,2014,9(5):93-99.
[2]杨章武,张扬波.东风螺脱壳病活体接触感染试验[J].水产养殖,2010,31(9):1-3.
[3]黄海立,余德光.方斑东风螺幼虫纤毛虫病的防治方法[J].科学养鱼,2007(7):56.
[4]郑养福.方斑东风螺浮游期聚缩虫病的防治[J].福建水产,2007(2):50-51.
[5]王建钢,乔振国.方斑东风螺肉壳分离病病因的初步研究[J].现代渔业信息,2011,26(10):16-18.
[6]彭景书,戈贤平,李明,等.方斑东风螺单孢子虫病的研究[J].水生生物学报,2011,35(5):803-807.
[7]黄郁葱,简纪常,吴灶和,等.方斑东风螺吻管水肿病病原菌的初步研究[J].渔业现代化,2009,36(4):37-41.
[8]张新中,文万侥,冯永勤,等.方斑东风螺肿吻症病原菌的分离鉴定及药敏分析[J].海洋科学,2010,34(5):7-12.
[9]裴琨.方斑东风螺工厂化养殖的关键技术[J].水产科技情报,2006,33(3):107-111.
[10]贾春红,李淑芳,张继东,南药组方对方斑东风螺病原性弧菌的抗菌作用研究[J].水产科学,2013,32(8):459-462.
[11]王国福,张瑞姿,曾令明,等.方斑东风螺肉壳分离病的防治方法[J].河北渔业,2008(8):37-40.
[12]刘巧红,王世锋,蔡岩,等.海南养殖方斑东风螺暴发性疾病病原分离鉴定及药敏分析[J].渔业科学进展,2014,35(1):74-81.
[13]罗杰,杜涛,梁飞龙,等.方斑东风螺养殖方式的初步研究[J].海洋科学,2004,28(7):39-43.
[14]钟春雨,张锡佳,曲于红,等.泥东风螺的人工育苗及养成技术[J].齐鲁渔业,2005,22(6):5-6.
[15]黄进光,李活,谢成营,等.方斑东风螺工厂化养殖[J].海洋与渔业,2006(2):20-21.
[16]郑雅友,杨章武,李正良,等.方斑东风螺人工育苗技术[J].福建水产,2005(2):58-60.
[17]黄郁葱.东风螺三种病害的病原及组织病理学研究[D].湛江:广东海洋大学,2006.
[18]沈萍,范秀容,李广武.微生物学实验[M].3版.北京:高等教育出版社,1999.
[19]张汉华,吴进锋,陈利雄,等.东风螺的人工养殖实用技术和产业发展展望[J].农业网络信息,2006(1):81-83.
[20]王志成,梁志辉,彭银辉,等.方斑东风螺室内水泥池养殖研究[J].水产科学,2010,29(1):40-43.
[21]冯永勤,周水灿,李芳远,等.方斑东风螺规模化苗种繁育技术研究[J].水产科学,2009,28(4):209-213.
[22]王晓勤.腹足类血淋巴细胞的研究[J].国外医学寄生虫病分册,1994,21(2):52-55.
[23]Jiang Q,Shi L,Ke C,et al.Identification and characterization of Vibrio harveyi associated with diseased abalone Haliotis diversicolor[J].Diseases of Aquatic Organisms,2013,103(2):133-139.
[24]赵旺,吴开畅,王江勇,等.方斑东风螺“翻背症”的病原及初步治疗研究[J].水产科学,2016,35(5):552-556.
[25]Li S F,Zhang J,Qiu D,et al.Biological characteristics and pathogenicities of Shewanella algaand Shewanella abalone fromBabylonia[J].Agricultural Science&Technology,2015,16(9):1845-1850,1859.
[26]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.
[27]胡娇,张飞,徐晓津,等.大黄鱼(Pseudosciaena crocea)内脏白点病的病原分离鉴定及致病性研究[J].海洋与湖沼,2014,45(2):409-417.
[28]Mao Z J,Li M F,Chen J G.Draft genome sequence of Pseudomonas plecoglossicida strain NB2011,the causative agent of white nodules in large yellow croaker(Larimichthys crocea)[J].Genome Announc,2015,1(4):e00586-13.
[2]杨章武,张扬波.东风螺脱壳病活体接触感染试验[J].水产养殖,2010,31(9):1-3.
[3]黄海立,余德光.方斑东风螺幼虫纤毛虫病的防治方法[J].科学养鱼,2007(7):56.
[4]郑养福.方斑东风螺浮游期聚缩虫病的防治[J].福建水产,2007(2):50-51.
[5]王建钢,乔振国.方斑东风螺肉壳分离病病因的初步研究[J].现代渔业信息,2011,26(10):16-18.
[6]彭景书,戈贤平,李明,等.方斑东风螺单孢子虫病的研究[J].水生生物学报,2011,35(5):803-807.
[7]黄郁葱,简纪常,吴灶和,等.方斑东风螺吻管水肿病病原菌的初步研究[J].渔业现代化,2009,36(4):37-41.
[8]张新中,文万侥,冯永勤,等.方斑东风螺肿吻症病原菌的分离鉴定及药敏分析[J].海洋科学,2010,34(5):7-12.
[9]裴琨.方斑东风螺工厂化养殖的关键技术[J].水产科技情报,2006,33(3):107-111.
[10]贾春红,李淑芳,张继东,南药组方对方斑东风螺病原性弧菌的抗菌作用研究[J].水产科学,2013,32(8):459-462.
[11]王国福,张瑞姿,曾令明,等.方斑东风螺肉壳分离病的防治方法[J].河北渔业,2008(8):37-40.
[12]刘巧红,王世锋,蔡岩,等.海南养殖方斑东风螺暴发性疾病病原分离鉴定及药敏分析[J].渔业科学进展,2014,35(1):74-81.
[13]罗杰,杜涛,梁飞龙,等.方斑东风螺养殖方式的初步研究[J].海洋科学,2004,28(7):39-43.
[14]钟春雨,张锡佳,曲于红,等.泥东风螺的人工育苗及养成技术[J].齐鲁渔业,2005,22(6):5-6.
[15]黄进光,李活,谢成营,等.方斑东风螺工厂化养殖[J].海洋与渔业,2006(2):20-21.
[16]郑雅友,杨章武,李正良,等.方斑东风螺人工育苗技术[J].福建水产,2005(2):58-60.
[17]黄郁葱.东风螺三种病害的病原及组织病理学研究[D].湛江:广东海洋大学,2006.
[18]沈萍,范秀容,李广武.微生物学实验[M].3版.北京:高等教育出版社,1999.
[19]张汉华,吴进锋,陈利雄,等.东风螺的人工养殖实用技术和产业发展展望[J].农业网络信息,2006(1):81-83.
[20]王志成,梁志辉,彭银辉,等.方斑东风螺室内水泥池养殖研究[J].水产科学,2010,29(1):40-43.
[21]冯永勤,周水灿,李芳远,等.方斑东风螺规模化苗种繁育技术研究[J].水产科学,2009,28(4):209-213.
[22]王晓勤.腹足类血淋巴细胞的研究[J].国外医学寄生虫病分册,1994,21(2):52-55.
[23]Jiang Q,Shi L,Ke C,et al.Identification and characterization of Vibrio harveyi associated with diseased abalone Haliotis diversicolor[J].Diseases of Aquatic Organisms,2013,103(2):133-139.
[24]赵旺,吴开畅,王江勇,等.方斑东风螺“翻背症”的病原及初步治疗研究[J].水产科学,2016,35(5):552-556.
[25]Li S F,Zhang J,Qiu D,et al.Biological characteristics and pathogenicities of Shewanella algaand Shewanella abalone fromBabylonia[J].Agricultural Science&Technology,2015,16(9):1845-1850,1859.
[26]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.
[27]胡娇,张飞,徐晓津,等.大黄鱼(Pseudosciaena crocea)内脏白点病的病原分离鉴定及致病性研究[J].海洋与湖沼,2014,45(2):409-417.
[28]Mao Z J,Li M F,Chen J G.Draft genome sequence of Pseudomonas plecoglossicida strain NB2011,the causative agent of white nodules in large yellow croaker(Larimichthys crocea)[J].Genome Announc,2015,1(4):e00586-13.