养殖刺参（Apostichopus japonicus）腐皮综合症的发生与异养菌区系间的关系
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摘要
随着北方沿海刺参(Apostichopus japonicus)养殖的兴起和养殖规模的日益扩大,在刺参越冬保苗和养成阶段普遍发生腐皮综合症疾病,导致极高死亡率和严重经济损失。该病主要症状表现为:肿嘴、摇头、排脏、身体萎缩、体表溃烂,解体死亡;其传染性强、发病快、流行性广,严重制约了刺参的健康养殖和可持续发展。
目前,关于养殖刺参病害的研究主要集中在疾病的流行病学、病原学以及组织病理学等,而疾病的发生机理特别是疾病发生的环境微生态学特征则未曾见到报道。为了掌握刺参腐皮综合症发生早期水体的水质状况、揭示发病早期病原菌与养殖环境中异养菌区系间的关系,阐明刺参腐皮综合症发生的根源,本文在对青岛周边不同地区三处刺参养殖场进行长期监测跟踪的基础上,首次报道了养殖刺参在低温养殖期间以及渡夏期间刺参腐皮综合症发生早期养殖水体水质、池塘中细菌数量与分布特征以及池塘水体、底泥、刺参肠道中优势菌与刺参病灶组织优势菌之间的相关性,并对该病原进行了鉴定。
2006年1月～5月刺参低温养殖期间,在青岛周边不同地区三处刺参养殖场刺参腐皮综合症发生的早期,对正常及发病池塘的水质、池塘中异养菌总数和弧菌总数的数量与分布以及刺参肠道中异养菌总数和弧菌总数进行了研究。同时,比较分析了发病早期发病池塘水体、表层底泥、刺参肠道中优势菌与刺参病灶组织优势菌之间的相关性。此外,对刺参病灶组织分离的优势菌060102A、060330B进行了人工感染试验证实其为刺参腐皮综合症的致病原并进行了分类鉴定。水质监测内容包括:温度(T)、盐度(S)、pH、溶解氧(DO)、化学需氧量(COD)、氨氮(NH4+-N)和亚硝酸氮(NO2--N)。结果表明:发病池塘水体水温和溶氧形成分层,底层水体逐渐恶化;底层水体异养菌总数和弧菌总数普遍高于表层,但整个池水细菌数量相对较低;池塘表层底泥与刺参肠道中异养菌总数较大,弧菌总数较低,其优势菌落与刺参病灶组织分离的优势菌具有一致性,与水体中优势菌相关不大。通过形态学观察、API半自动化鉴定、常规生理生化试验以及16S rRNA基因序列同源性分析等表明,060102A为假交替单胞菌属细(Pseudoalteromonas sp.);060330B具有假单胞菌属的特征,其表型特征与恶臭假单胞菌(Pseudomonas putida)相似,亲缘关系与恶臭假单胞菌最近,相似率达到99.5%。菌株060330B被鉴定为恶臭假单胞菌,并视为养殖刺参腐皮综合症的致病原之一。通过对多起不同来源患病刺参的调查研究表明,使用年限长、淤泥层厚呈黑色并伴有异味的养殖池塘在该时期容易导致刺参腐皮综合症的发生,刺参腐皮综合症的发生与池塘水体分层引起的水质恶化和底泥中病原菌数量密切相关。
2006年7月～10月养殖刺参渡夏期间,在青岛周边不同地区三处刺参养殖场刺参腐皮综合症发生的早期,对发病池塘的水质、池塘中异养菌总数和弧菌总数的数量与分布以及刺参肠道中的异养菌总数和弧菌总数进行了研究。同时,比较分析了发病早期池塘水体、表层底泥、刺参肠道中优势菌与刺参病灶组织优势菌之间的相关性。此外,对刺参病灶组织分离的优势菌060904E进行了人工感染试验证实其为刺参腐皮综合症的致病原并进行了分类鉴定。水质监测内容包括:温度(T)、盐度(S)、pH、溶解氧(DO)、化学需氧量(COD)、氨氮(NH4+-N)和亚硝酸氮(NO2--N)。结果表明:整个养殖池塘水体出现了不同程度恶化,水体中异养菌总数与弧菌总数平均分别达到3.3×104cfu/mL、2.0×104cfu/mL,水层差异不大;池塘表层底泥中异养菌总数和弧菌总数平均分别为5.9×106cfu/g、1.4×106cfu/g,弧菌数量处于较高水平;刺参肠道中异养菌总数和弧菌总数平均分别为5.8×106cfu/g、1.9×106cfu/g,弧菌数量也处于较高水平;池塘水体、底泥以及刺参肠道中分离出的优势菌与刺参病灶组织分离的优势菌具有一致性。细菌鉴定结果表明菌株060904E为哈维氏弧菌(Vibrio harveyi),哈维氏弧菌被视为渡夏期间养殖刺参腐皮综合症的致病原。刺参腐皮综合症的发生与养殖池塘的水质恶化和池塘水体和底泥中致病性弧菌数量密切相关。
本文研究表明,恶臭假单胞菌(Pseudomonas putida)和哈维氏弧菌(Vibrio harveyi)是青岛地区养殖刺参腐皮综合症的重要致病原,这为丰富养殖刺参的病原学研究奠定了理论基础。同时本文揭示了池塘养殖条件下,刺参腐皮综合症的发生与池塘水体的恶化及底泥中病原菌数量密切相关,对刺参养殖中的健康管理、提高养殖工艺和加强疾病防治提供了指导依据和参考。
As Apostichopus japonicus cultivation began more and more popular and culturing scale increased gradually day by day,skin ulcer syndrome broke out cosmically during winter seed rearing and culturing periods,which resulted in high death rate and serious economic loss.Its dominating symptoms were mouth swelling,shaking head,eviscerating bowels,body shrinking,skin ulcering,body disjointing and death.As it was a acute disease with high infectivity and wide epidemic,it restricted health cultivation and continuable development of sea cucumber seriously.
At present,the study on diseases in cultured sea cucumber are focus on epidemiology,aetiology and tissue pathology,but the mechanism of disease happening,especially microecology characteristics have not seen any reports yet.This study firstly reported the water quality status,the number and distribution characteristics of bacteria in cultural ponds and the relation between the preponderant bacteria that were isolated from ponds water,top layer bottom mud,intestinal tract and focus tissue of sea cucumber during low temperature culturing period and aestivation period on the base of long inspection of three sea cucumber farms in different areas nearby Qingdao,with the purpose of finding out the water quality status,opening out the relation between pathogenic bacteria and heterotrophic bacterial flora in cultural environment at the beginning of cultured sea cucumber suffering from skin ulcer syndrome and clarifying the essential causation of skin ulcer syndrome.In addition,this study also identified the pathogeny of skin ulcer syndrome.
From January to May of year 2006,at the beginning of cultured Apostichopus japonicus suffered from skin ulcer syndrome in three sea cucumber farms in different areas nearby Qingdao,detection of water quality and the number and distribution of heterotrophic bacteria and vibrio bacteria of cultural ponds as well as total count of heterotrophic bacteria and vibrio bacteria in intestinal tract of sea cucumber in normal pond and the pond where sea cucumber were suffered from skin ulcer syndrome in early stage were analyzed with the methods of HPC and VPC.At the same time,this study compared and analyzed the relativity between the preponderant bacteria that were isolated from above abnormal ponds water,top layer bottom mud,intestinal tract and focus tissue of sea cucumber.Besides,artificial infection test was carried out with preponderant bacteria 060102A and 060330B that were isolated from focus tissue of sea cucumber and were verified to be the pathogen causing the skin ulcer syndrome,bacterial classification was carried out samely.The water detection included T,S,pH,DO,COD,NH4+-N and NO2--N.The results showed that as water temperature and dissolved oxygen both formed divided layers in cultural ponds water the bottom layer water began to worsen at some degree;total heterotrophic bacteria and vibrio bacteria number in bottom layer water were both higher than that in surface layer water in general,the whole water bacteria number was in low-level;total heterotrophic bacteria number in top layer bottom mud and intestinal tract were in high-level,whereas total vibrio bacteria number was in low-level relatively.The preponderant bacteria that were isolated from top layer bottom mud and intestinal tract were identical to that in focus tissue of sea cucumber,whereas almost had no relation with ponds water.Morphological observation,semi-automatic identification through API system,traditional physiological and biochemical methods as well as 16S rRNA sequence analysis were applied in the bacterial classification.The results showed that 060102A was Pseudoalteromonas sp.;060330B had same character as Pseudomonas genus bacteria,its characteristics were similar to Pseudomonas putida,with 99.5 % identity.Thus bacterium 060330B was identified as Pseudomonas putida,and regarded as one of the pathogens causing skin ulcer syndrome of cultured sea cucumber.In addition,epidemiology investigation and detections of several samples revealed that the ponds in which cultivation history reached two years or more and having thick black bottom mud with fishy smell were likely to result in the happening of skin ulcer syndrome of cultured sea cucumber at that time.The occurrence of skin ulcer syndrome in cultured sea cucumber was closely related with water deterioration caused by divided water layers and pathogenic bacteria number in ponds bottom mud.
From July to October of year 2006,at the beginning of cultured Apostichopus japonicus suffered from skin ulcer syndrome in three sea cucumber farms in different areas nearby Qingdao,detection of water quality and the number and distribution of heterotrophic bacteria and vibrio bacteria of cultural ponds as well as total count of heterotrophic bacteria and vibrio bacteria in intestinal tract of sea cucumber were analyzed with the methods of HPC and VPC.At the same time,this study compared and analyzed the relativity between the preponderant bacteria that were isolated from ponds water,top layer bottom mud,intestinal tract and focus tissue of sea cucumber.Besides,artificial infection test was carried out with preponderant bacterium 060904E that was isolated from focus tissue of sea cucumber and was verified to be the pathogen causing the skin ulcer syndrome,bacterial classification was carried out samely.The water detection included T,S,pH,DO,COD,NH4+-N and NO2--N.The results showed that the whole cultural ponds water began to worsen at some degree,total heterotrophic bacteria and vibrio bacteria number were 3.3×104cfu/mL and 2.0×104cfu/mL respectively on average,which both the differences were not apparent in water layers.Total heterotrophic bacteria and vibrio bacteria number were 5.9×106cfu/g and 1.4×106cfu/g respectively on average of top layer bottom mud in ponds.Besides,total heterotrophic bacteria and vibrio bacteria number in intestinal tract of sea cucumber were 5.8×106cfu/g and 1.9×106cfu/g.The preponderant bacteria that were isolated from ponds water,top layer bottom mud and intestinal tract were identical to that in focus tissue of sea cucumber.Thus bacterium 060904E was identified as Vibrio harveyi,and regarded as the pathogen causing skin ulcer syndrome of cultured sea cucumber during aestivation period.The occurrence of skin ulcer syndrome in cultured sea cucumber was closely related with the water deterioration and number of pathogenic vibrio bacteria in water and bottom mud of cultural ponds.
This study revealed that Pseudomonas putida and Vibrio harveyi were important pathogenies that causing skin ulcer syndrome of cultured sea cucumber in Qingdao cultivation area,which established theory basis on aetiology research of cultured sea cucumber in future.At the same time,this research opened out that the occurrence of skin ulcer syndrome in cultured sea cucumber was closely related with water deterioration and pathogenic bacteria number in bottom mud under the condition of pond cultivation,which provided supervising references on health management,improving cultivation technology and strengthening prevention and treatment of diseases in cultured sea cucumber.
目前,关于养殖刺参病害的研究主要集中在疾病的流行病学、病原学以及组织病理学等,而疾病的发生机理特别是疾病发生的环境微生态学特征则未曾见到报道。为了掌握刺参腐皮综合症发生早期水体的水质状况、揭示发病早期病原菌与养殖环境中异养菌区系间的关系,阐明刺参腐皮综合症发生的根源,本文在对青岛周边不同地区三处刺参养殖场进行长期监测跟踪的基础上,首次报道了养殖刺参在低温养殖期间以及渡夏期间刺参腐皮综合症发生早期养殖水体水质、池塘中细菌数量与分布特征以及池塘水体、底泥、刺参肠道中优势菌与刺参病灶组织优势菌之间的相关性,并对该病原进行了鉴定。
2006年1月～5月刺参低温养殖期间,在青岛周边不同地区三处刺参养殖场刺参腐皮综合症发生的早期,对正常及发病池塘的水质、池塘中异养菌总数和弧菌总数的数量与分布以及刺参肠道中异养菌总数和弧菌总数进行了研究。同时,比较分析了发病早期发病池塘水体、表层底泥、刺参肠道中优势菌与刺参病灶组织优势菌之间的相关性。此外,对刺参病灶组织分离的优势菌060102A、060330B进行了人工感染试验证实其为刺参腐皮综合症的致病原并进行了分类鉴定。水质监测内容包括:温度(T)、盐度(S)、pH、溶解氧(DO)、化学需氧量(COD)、氨氮(NH4+-N)和亚硝酸氮(NO2--N)。结果表明:发病池塘水体水温和溶氧形成分层,底层水体逐渐恶化;底层水体异养菌总数和弧菌总数普遍高于表层,但整个池水细菌数量相对较低;池塘表层底泥与刺参肠道中异养菌总数较大,弧菌总数较低,其优势菌落与刺参病灶组织分离的优势菌具有一致性,与水体中优势菌相关不大。通过形态学观察、API半自动化鉴定、常规生理生化试验以及16S rRNA基因序列同源性分析等表明,060102A为假交替单胞菌属细(Pseudoalteromonas sp.);060330B具有假单胞菌属的特征,其表型特征与恶臭假单胞菌(Pseudomonas putida)相似,亲缘关系与恶臭假单胞菌最近,相似率达到99.5%。菌株060330B被鉴定为恶臭假单胞菌,并视为养殖刺参腐皮综合症的致病原之一。通过对多起不同来源患病刺参的调查研究表明,使用年限长、淤泥层厚呈黑色并伴有异味的养殖池塘在该时期容易导致刺参腐皮综合症的发生,刺参腐皮综合症的发生与池塘水体分层引起的水质恶化和底泥中病原菌数量密切相关。
2006年7月～10月养殖刺参渡夏期间,在青岛周边不同地区三处刺参养殖场刺参腐皮综合症发生的早期,对发病池塘的水质、池塘中异养菌总数和弧菌总数的数量与分布以及刺参肠道中的异养菌总数和弧菌总数进行了研究。同时,比较分析了发病早期池塘水体、表层底泥、刺参肠道中优势菌与刺参病灶组织优势菌之间的相关性。此外,对刺参病灶组织分离的优势菌060904E进行了人工感染试验证实其为刺参腐皮综合症的致病原并进行了分类鉴定。水质监测内容包括:温度(T)、盐度(S)、pH、溶解氧(DO)、化学需氧量(COD)、氨氮(NH4+-N)和亚硝酸氮(NO2--N)。结果表明:整个养殖池塘水体出现了不同程度恶化,水体中异养菌总数与弧菌总数平均分别达到3.3×104cfu/mL、2.0×104cfu/mL,水层差异不大;池塘表层底泥中异养菌总数和弧菌总数平均分别为5.9×106cfu/g、1.4×106cfu/g,弧菌数量处于较高水平;刺参肠道中异养菌总数和弧菌总数平均分别为5.8×106cfu/g、1.9×106cfu/g,弧菌数量也处于较高水平;池塘水体、底泥以及刺参肠道中分离出的优势菌与刺参病灶组织分离的优势菌具有一致性。细菌鉴定结果表明菌株060904E为哈维氏弧菌(Vibrio harveyi),哈维氏弧菌被视为渡夏期间养殖刺参腐皮综合症的致病原。刺参腐皮综合症的发生与养殖池塘的水质恶化和池塘水体和底泥中致病性弧菌数量密切相关。
本文研究表明,恶臭假单胞菌(Pseudomonas putida)和哈维氏弧菌(Vibrio harveyi)是青岛地区养殖刺参腐皮综合症的重要致病原,这为丰富养殖刺参的病原学研究奠定了理论基础。同时本文揭示了池塘养殖条件下,刺参腐皮综合症的发生与池塘水体的恶化及底泥中病原菌数量密切相关,对刺参养殖中的健康管理、提高养殖工艺和加强疾病防治提供了指导依据和参考。
As Apostichopus japonicus cultivation began more and more popular and culturing scale increased gradually day by day,skin ulcer syndrome broke out cosmically during winter seed rearing and culturing periods,which resulted in high death rate and serious economic loss.Its dominating symptoms were mouth swelling,shaking head,eviscerating bowels,body shrinking,skin ulcering,body disjointing and death.As it was a acute disease with high infectivity and wide epidemic,it restricted health cultivation and continuable development of sea cucumber seriously.
At present,the study on diseases in cultured sea cucumber are focus on epidemiology,aetiology and tissue pathology,but the mechanism of disease happening,especially microecology characteristics have not seen any reports yet.This study firstly reported the water quality status,the number and distribution characteristics of bacteria in cultural ponds and the relation between the preponderant bacteria that were isolated from ponds water,top layer bottom mud,intestinal tract and focus tissue of sea cucumber during low temperature culturing period and aestivation period on the base of long inspection of three sea cucumber farms in different areas nearby Qingdao,with the purpose of finding out the water quality status,opening out the relation between pathogenic bacteria and heterotrophic bacterial flora in cultural environment at the beginning of cultured sea cucumber suffering from skin ulcer syndrome and clarifying the essential causation of skin ulcer syndrome.In addition,this study also identified the pathogeny of skin ulcer syndrome.
From January to May of year 2006,at the beginning of cultured Apostichopus japonicus suffered from skin ulcer syndrome in three sea cucumber farms in different areas nearby Qingdao,detection of water quality and the number and distribution of heterotrophic bacteria and vibrio bacteria of cultural ponds as well as total count of heterotrophic bacteria and vibrio bacteria in intestinal tract of sea cucumber in normal pond and the pond where sea cucumber were suffered from skin ulcer syndrome in early stage were analyzed with the methods of HPC and VPC.At the same time,this study compared and analyzed the relativity between the preponderant bacteria that were isolated from above abnormal ponds water,top layer bottom mud,intestinal tract and focus tissue of sea cucumber.Besides,artificial infection test was carried out with preponderant bacteria 060102A and 060330B that were isolated from focus tissue of sea cucumber and were verified to be the pathogen causing the skin ulcer syndrome,bacterial classification was carried out samely.The water detection included T,S,pH,DO,COD,NH4+-N and NO2--N.The results showed that as water temperature and dissolved oxygen both formed divided layers in cultural ponds water the bottom layer water began to worsen at some degree;total heterotrophic bacteria and vibrio bacteria number in bottom layer water were both higher than that in surface layer water in general,the whole water bacteria number was in low-level;total heterotrophic bacteria number in top layer bottom mud and intestinal tract were in high-level,whereas total vibrio bacteria number was in low-level relatively.The preponderant bacteria that were isolated from top layer bottom mud and intestinal tract were identical to that in focus tissue of sea cucumber,whereas almost had no relation with ponds water.Morphological observation,semi-automatic identification through API system,traditional physiological and biochemical methods as well as 16S rRNA sequence analysis were applied in the bacterial classification.The results showed that 060102A was Pseudoalteromonas sp.;060330B had same character as Pseudomonas genus bacteria,its characteristics were similar to Pseudomonas putida,with 99.5 % identity.Thus bacterium 060330B was identified as Pseudomonas putida,and regarded as one of the pathogens causing skin ulcer syndrome of cultured sea cucumber.In addition,epidemiology investigation and detections of several samples revealed that the ponds in which cultivation history reached two years or more and having thick black bottom mud with fishy smell were likely to result in the happening of skin ulcer syndrome of cultured sea cucumber at that time.The occurrence of skin ulcer syndrome in cultured sea cucumber was closely related with water deterioration caused by divided water layers and pathogenic bacteria number in ponds bottom mud.
From July to October of year 2006,at the beginning of cultured Apostichopus japonicus suffered from skin ulcer syndrome in three sea cucumber farms in different areas nearby Qingdao,detection of water quality and the number and distribution of heterotrophic bacteria and vibrio bacteria of cultural ponds as well as total count of heterotrophic bacteria and vibrio bacteria in intestinal tract of sea cucumber were analyzed with the methods of HPC and VPC.At the same time,this study compared and analyzed the relativity between the preponderant bacteria that were isolated from ponds water,top layer bottom mud,intestinal tract and focus tissue of sea cucumber.Besides,artificial infection test was carried out with preponderant bacterium 060904E that was isolated from focus tissue of sea cucumber and was verified to be the pathogen causing the skin ulcer syndrome,bacterial classification was carried out samely.The water detection included T,S,pH,DO,COD,NH4+-N and NO2--N.The results showed that the whole cultural ponds water began to worsen at some degree,total heterotrophic bacteria and vibrio bacteria number were 3.3×104cfu/mL and 2.0×104cfu/mL respectively on average,which both the differences were not apparent in water layers.Total heterotrophic bacteria and vibrio bacteria number were 5.9×106cfu/g and 1.4×106cfu/g respectively on average of top layer bottom mud in ponds.Besides,total heterotrophic bacteria and vibrio bacteria number in intestinal tract of sea cucumber were 5.8×106cfu/g and 1.9×106cfu/g.The preponderant bacteria that were isolated from ponds water,top layer bottom mud and intestinal tract were identical to that in focus tissue of sea cucumber.Thus bacterium 060904E was identified as Vibrio harveyi,and regarded as the pathogen causing skin ulcer syndrome of cultured sea cucumber during aestivation period.The occurrence of skin ulcer syndrome in cultured sea cucumber was closely related with the water deterioration and number of pathogenic vibrio bacteria in water and bottom mud of cultural ponds.
This study revealed that Pseudomonas putida and Vibrio harveyi were important pathogenies that causing skin ulcer syndrome of cultured sea cucumber in Qingdao cultivation area,which established theory basis on aetiology research of cultured sea cucumber in future.At the same time,this research opened out that the occurrence of skin ulcer syndrome in cultured sea cucumber was closely related with water deterioration and pathogenic bacteria number in bottom mud under the condition of pond cultivation,which provided supervising references on health management,improving cultivation technology and strengthening prevention and treatment of diseases in cultured sea cucumber.
引文
[1]宋修武.大力发展海参健康养殖培育山东海洋渔业经济发展新亮点[J].中国渔业经济,2005,1:9-10.
[2]王印庚,荣小军.我国刺参养殖存在的主要问题与疾病综合防治技术要点[J].齐鲁渔业,2004,21(10):29-31.
[3]张春云,王印庚,荣小军,等.国内外海参自然资源、养殖状况及存在问题[J].海洋水产研究,2004,3:89-97.
[4]王义民.海参养殖热的冷思考[J].齐鲁渔业,2004,21(10):45-46.
[5]于会霆,徐振行,刘勤燕,等.池塘海参养殖存在的问题和对策[J].齐鲁渔业,2004,21(2):25.
[6]王玉堂.海参养殖业管理急待加强[J].中国水产,2004,10:24-25.
[7]王义民,张少华,张秀丽,等.可移动式轻便隐蔽物在池塘养参中的应用研究[J].齐鲁渔业,2004,21(7):9-10.
[8]宋祖德.长岛县推广坑道养参[J].海洋信息,1996,12:11.
[9]王印庚,荣小军,张春云,等.养殖海参主要疾病及防治技术[J].海洋科学,2005,29(3):1-7.
[10]王印庚,荣小军,张春云,等.养殖刺参暴发行疾病—“腐皮综合症”的初步研究与防治[J].齐鲁渔业,2004,21(5):44-47.
[11]乔聚海,程波.刺参人工池塘养殖现状及展望[J].海洋科学,2005,29(9):80-82.
[12]肖树旭,顾功超.刺参南移与人工育苗试验[J].水产学报,1981,6(2):147-153.
[13]杜佳垠.刺参增养殖现状与发展[J].北京水产,2005,3:4-5.
[14]常亚青,隋锡林,李俊.刺参增养殖业现状、存在问题与展望[J].水产科学,2006,25(4):198-201.
[15]邹积波,高广斌,姜洪亮,等.分析刺参养殖发病原因、研讨对策,走可持续发展之路[J].水产科学,2006,25(1):53-54.
[16]袁成玉.海参饲料研究的现状与发展方向[J].水产科学,2005,24(12):54-56.
[17]孙建璋,庄定根,陈兰涛,等.刺参南移养殖技术研究[J].浙江海洋学院学报(自然科学版),2006,25(2):148-153.
[18]王秀菊,王丽敏,杨美桂,等.人工控温工厂化养殖刺参技术[J].齐鲁渔业,2004,21(11):4-5.
[1]王印庚,荣小军,张春云,等.养殖刺参暴发性疾病-“腐皮综合症”的初步研究与防治[J].齐鲁渔业,2004,21(5):44-47.
[2]王印庚,方波,张春云,等.养殖刺参保苗期重大疾病“腐皮综合征”病原及其感染源分析[J].中国水产科学,2006,13(4):610-616.
[3]张春云,王印庚,荣小军.养殖刺参腐皮综合征病原菌的分离与鉴定[J].水产学报,2006,30(1):118-123.
[4]王印庚,荣小军,张春云,等.养殖海参主要疾病及防治技术[J].海洋科学,2005,(3):1-7.
[5]邓欢,隋锡林,陈俅,等.大连地区越冬期刺参一种新病症[J].水产科学,2005,24(2):40-41.
[6]董颖,邓欢,隋锡林,等.养殖仿刺参溃烂病病因初探[J].水产科学,2005,24(3):4-6.
[7]马悦欣,徐高蓉,常亚青,等.大连地区刺参幼参溃烂病细菌性病原的初步研究[J].大连水产学院学报,2006,21(1):13-18.
[8]马悦欣,徐高蓉,张恩鹏,等.仿刺参幼参急性口围肿胀症的细菌性病原[J].水产学报,2006,30(3):377-382.
[9]王印庚,孙素凤,荣小军.仿刺参幼体烂胃病及其致病原鉴定[J].中国水产科学,2006,13(6):908-916.
[10] Bergey’s Manual of Determinative Bacteriology Ninth Edition[M].Maryland:Williams & Wilkins,1994.
[11]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[12]于东祥,孙慧玲,陈四清,等.海参健康养殖技术[M].北京:海洋出版社,2005.
[13]于东祥,张岩,陈四清.养殖水体的双分层是海参发病的普遍原因[J].齐鲁渔业,2004,21(8):8-9.
[1]王印庚,荣小军,张春云,等.养殖刺参暴发性疾病-“腐皮综合症”的初步研究与防治[J].齐鲁渔业,2004,21(5):44-47.
[2]张春云,王印庚,荣小军.养殖刺参腐皮综合征病原菌的分离与鉴定[J].水产学报,2006,30(1):118-123.
[3]王印庚,方波,张春云,等.养殖刺参保苗期重大疾病“腐皮综合征”病原及其感染源分析[J].中国水产科学,2006,13(4):610-616.
[4] Bergey’s Manual of Determinative Bacteriology Ninth Edition[M].Maryland:Williams & Wilkins,1994.
[5]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[6]于东祥,孙慧玲,陈四清,等.海参健康养殖技术[M].北京:海洋出版社,2005.
[7]杨润德,王琳,范欢,等.丹顶鹤恶臭假单胞菌的分离及生物学特性鉴定[J].中国兽医杂志,2005,41(4):55-56.
[8]李庆山,邢瑞云,张芳萍,等.首次发现恶臭假单胞菌引起的食物中毒[J].中国公共卫生,2000,16(1):50-51.
[9]陶保华,石和荣,黄俊文,等.假单胞菌引起罗氏沼虾黄鳃、黑鳃病的研究[J].中山大学学报(自然科学版),2000,39:255-259.
[10]樊海平.恶臭假单胞菌引起的欧洲鳗鲡烂鳃病[J].水产学报,2001,25(2):147-150.
[11] Ilhan Altinok,Sevki Kayis,Erol Capkin.Pseudomonas putida infection in rainbow trout[J].Aquaculture,2006,261:850-855.
[12]江定丰,李槿年,余为一.一株中华绒螯蟹病菌鉴定及其致病机理的研究[J].水利渔业,2003,23(6):51-54.
[1]王印庚,荣小军,张春云,等.养殖刺参暴发性疾病-“腐皮综合症”的初步研究与防治[J].齐鲁渔业,2004,21(5):44-47.
[2]王印庚,荣小军,张春云,等.养殖海参主要疾病及防治技术[J].海洋科学,2005,(3):1-7.
[3]张春云,王印庚,荣小军.养殖刺参腐皮综合征病原菌的分离与鉴定[J].水产学报,2006,30(1):118-123.
[4]王印庚,方波,张春云,等.养殖刺参保苗期重大疾病“腐皮综合征”病原及其感染源分析[J].中国水产科学,2006,13(4):610-616.
[5]于东祥,孙慧玲,陈四清,等.海参健康养殖技术[M].北京:海洋出版社,2005.
[6]贺天笙,杨立艾,陈洪连,等.对虾弧菌病发生时临界菌值的研究[J].水产养殖,1993,2:21-23.
[7]林克冰,周宸,蔡清海,等.班节对虾双季养成期间虾池水质和底泥细菌含量的变化[J].福建水产,1998,12(4):7-12.
[8]邓欢,隋锡林,陈俅,等.大连地区越冬期刺参一种新病症[J].水产科学,2005,24(2):40-41.
[9]董颖,邓欢,隋锡林,等.养殖仿刺参溃烂病病因初探[J].水产科学,2005,24(3):4-6.
[10]马悦欣,徐高蓉,常亚青,等.大连地区刺参幼参溃烂病细菌性病原的初步研究[J].大连水产学院学报,2006,21(1):13-18.
[11]马悦欣,徐高蓉,张恩鹏,等.仿刺参幼参急性口围肿胀症的细菌性病原[J].水产学报,2006,30(3):377-382.
[12]王印庚,孙素凤,荣小军.仿刺参幼体烂胃病及其致病原鉴定[J].中国水产科学,2006,13(6):908-916.
[13] Bergey’s Manual of Determinative Bacteriology Ninth Edition[M].Maryland:Williams & Wilkins,1994.
[14]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[1]张春云,王印庚,荣小军.养殖刺参腐皮综合征病原菌的分离与鉴定[J].水产学报,2006,30(1):118-123.
[2]马悦欣,徐高蓉,张恩鹏,等.仿刺参幼参急性口围肿胀症的细菌性病原[J].水产学报,2006,30(3):377-382.
[3]马悦欣,徐高蓉,常亚青,等.大连地区刺参幼参溃烂病细菌性病原的初步研究[J].大连水产学院学报,2006,21(1):13-18.
[4]王印庚,方波,张春云,等.养殖刺参保苗期重大疾病“腐皮综合征”病原及其感染源分析[J].中国水产科学,2006,13(4):610-616.
[5]王印庚,荣小军,张春云,等.养殖刺参暴发性疾病-“腐皮综合症”的初步研究与防治[J].齐鲁渔业,2004,21(5):44-47.
[6] Bergey’s Manual of Determinative Bacteriology Ninth Edition[M].Maryland:Williams & Wilkins,1994.
[7]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[8]于东祥,孙慧玲,陈四清,等.海参健康养殖技术[M].北京:海洋出版社,2005.
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[12]王国良,金珊,薛良义,等.海水网箱养殖鲈鱼皮肤溃疡病及其病原菌的研究[J].黄渤海海洋,2000,18(3):85-89.
[13]王保坤,余俊红,李筠,等.花鲈弧菌病病原菌(哈维氏弧菌)的分离与鉴定[J].中国水产科学,2002,9(1):52-55.
[14]毛芝娟,刘国勇,陈昌福.大黄鱼溃疡病致病菌的初步分离与鉴定[J].安徽农业大学学报,2002,29(2):178-181.
[15]覃映雪,池信才,苏永全,等.网箱养殖青石斑鱼的溃疡病病原[J].水产学报,2004,28(3):297-302.
[16]陈献稿,吴淑勤,石存斌,等.斜带石斑鱼病原菌(哈维氏弧菌)的分离与鉴定[J].中国水产科学,2004,11(4):313-316.
[17]范文辉,黄倢,王秀华,等.养殖大菱鲆溃疡症病原菌的分离鉴定及系统发育分析[J].微生物学报,2005,45(5):665-670.
[18]潘晓艺,沈锦玉,尹文林,等.哈维氏弧菌黑鲷分离株BK-1培养条件优化研究[J].浙江海洋学院学报(自然科学版),2005,24(3):240-243.
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[20]刘问,钱冬,杨国梁,等.南美白对虾虾苗淡化期间发光病病原研究[J].集美大学学报(自然科学版),2004,9(4):300-304.
[21]邓欢,陈俅,刘权恕,等.养殖海湾扇贝弧菌病的研究[J].应用与环境生物学报,2003,9(5):517-521.
[2]王印庚,荣小军.我国刺参养殖存在的主要问题与疾病综合防治技术要点[J].齐鲁渔业,2004,21(10):29-31.
[3]张春云,王印庚,荣小军,等.国内外海参自然资源、养殖状况及存在问题[J].海洋水产研究,2004,3:89-97.
[4]王义民.海参养殖热的冷思考[J].齐鲁渔业,2004,21(10):45-46.
[5]于会霆,徐振行,刘勤燕,等.池塘海参养殖存在的问题和对策[J].齐鲁渔业,2004,21(2):25.
[6]王玉堂.海参养殖业管理急待加强[J].中国水产,2004,10:24-25.
[7]王义民,张少华,张秀丽,等.可移动式轻便隐蔽物在池塘养参中的应用研究[J].齐鲁渔业,2004,21(7):9-10.
[8]宋祖德.长岛县推广坑道养参[J].海洋信息,1996,12:11.
[9]王印庚,荣小军,张春云,等.养殖海参主要疾病及防治技术[J].海洋科学,2005,29(3):1-7.
[10]王印庚,荣小军,张春云,等.养殖刺参暴发行疾病—“腐皮综合症”的初步研究与防治[J].齐鲁渔业,2004,21(5):44-47.
[11]乔聚海,程波.刺参人工池塘养殖现状及展望[J].海洋科学,2005,29(9):80-82.
[12]肖树旭,顾功超.刺参南移与人工育苗试验[J].水产学报,1981,6(2):147-153.
[13]杜佳垠.刺参增养殖现状与发展[J].北京水产,2005,3:4-5.
[14]常亚青,隋锡林,李俊.刺参增养殖业现状、存在问题与展望[J].水产科学,2006,25(4):198-201.
[15]邹积波,高广斌,姜洪亮,等.分析刺参养殖发病原因、研讨对策,走可持续发展之路[J].水产科学,2006,25(1):53-54.
[16]袁成玉.海参饲料研究的现状与发展方向[J].水产科学,2005,24(12):54-56.
[17]孙建璋,庄定根,陈兰涛,等.刺参南移养殖技术研究[J].浙江海洋学院学报(自然科学版),2006,25(2):148-153.
[18]王秀菊,王丽敏,杨美桂,等.人工控温工厂化养殖刺参技术[J].齐鲁渔业,2004,21(11):4-5.
[1]王印庚,荣小军,张春云,等.养殖刺参暴发性疾病-“腐皮综合症”的初步研究与防治[J].齐鲁渔业,2004,21(5):44-47.
[2]王印庚,方波,张春云,等.养殖刺参保苗期重大疾病“腐皮综合征”病原及其感染源分析[J].中国水产科学,2006,13(4):610-616.
[3]张春云,王印庚,荣小军.养殖刺参腐皮综合征病原菌的分离与鉴定[J].水产学报,2006,30(1):118-123.
[4]王印庚,荣小军,张春云,等.养殖海参主要疾病及防治技术[J].海洋科学,2005,(3):1-7.
[5]邓欢,隋锡林,陈俅,等.大连地区越冬期刺参一种新病症[J].水产科学,2005,24(2):40-41.
[6]董颖,邓欢,隋锡林,等.养殖仿刺参溃烂病病因初探[J].水产科学,2005,24(3):4-6.
[7]马悦欣,徐高蓉,常亚青,等.大连地区刺参幼参溃烂病细菌性病原的初步研究[J].大连水产学院学报,2006,21(1):13-18.
[8]马悦欣,徐高蓉,张恩鹏,等.仿刺参幼参急性口围肿胀症的细菌性病原[J].水产学报,2006,30(3):377-382.
[9]王印庚,孙素凤,荣小军.仿刺参幼体烂胃病及其致病原鉴定[J].中国水产科学,2006,13(6):908-916.
[10] Bergey’s Manual of Determinative Bacteriology Ninth Edition[M].Maryland:Williams & Wilkins,1994.
[11]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[12]于东祥,孙慧玲,陈四清,等.海参健康养殖技术[M].北京:海洋出版社,2005.
[13]于东祥,张岩,陈四清.养殖水体的双分层是海参发病的普遍原因[J].齐鲁渔业,2004,21(8):8-9.
[1]王印庚,荣小军,张春云,等.养殖刺参暴发性疾病-“腐皮综合症”的初步研究与防治[J].齐鲁渔业,2004,21(5):44-47.
[2]张春云,王印庚,荣小军.养殖刺参腐皮综合征病原菌的分离与鉴定[J].水产学报,2006,30(1):118-123.
[3]王印庚,方波,张春云,等.养殖刺参保苗期重大疾病“腐皮综合征”病原及其感染源分析[J].中国水产科学,2006,13(4):610-616.
[4] Bergey’s Manual of Determinative Bacteriology Ninth Edition[M].Maryland:Williams & Wilkins,1994.
[5]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
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[7]杨润德,王琳,范欢,等.丹顶鹤恶臭假单胞菌的分离及生物学特性鉴定[J].中国兽医杂志,2005,41(4):55-56.
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