柱状黄杆菌弱毒株和车轮虫混合感染致金鱼烂鳃病的病原分析

doi:10.16431/j.cnki.1671-7236.2023.11.029

Abstract

Abstract: 【Objective】 The experiment was aimed to confirm the pathogen of gill rot disease in a goldfish farm in Beijing.【Method】 The moribund goldfishes with clinical symptoms were collected, and wet mount of gill tissue were prepared for observing parasites and fungal hypha.Liver, kidney, spleen and gill tissues were collected to detect Goldfish hematopoietic necrosis virus (GFHNV).At the same time, the pathogenic bacteria were isolated from the gill, and the morphological characteristics, physical and chemical characteristics, taxonomic status and drug sensitivity of the pathogenic bacteria were analyzed.The experiments of artificial infection of pathogenic bacteria were carried out by the methods of gill tissue scratch soaking and non-scratch soaking.【Result】 There was a small amount of trichomyces parasitica on the gill filaments of naturally occurring fish, no fungal hypha was observed, and GFHNV nucleic acid test was negative.A large number of rhizoid colonies were isolated from gill tissue.The strain was Gram-negative longobacterium, with a body size of (0.5-0.7) μm×(4-8) μm.It was positive for oxidase and catalase, degraded gelatin and chondroitin sulfate, produced hydrogen sulfide, and did not fermentation glucose and other sugar alcohols.The phylogenetic tree constructed based on 16S rDNA sequence was polymerized with Flavobacterium columnare.The isolated bacteria was identified as Flavobacterium columnare, genotype Ⅰ.All groups of goldfish soaked in scratches experienced death, and the symptoms were similar to those of the naturally infected fish.No death occurred in scratch control group and non-scratch soaking group.The calculated median lethal dose (LD₅₀) of the bacteria to the goldfish was 8.8×10⁶ CFU, which was a mild strain.The bacterium was sensitive to enrofloxacin, doxycycline hydrochloride and sulfonamides, but not to the other antimicrobials tested.【Conclusion】 The weakly virulent strains of Flavobacterium columnare isolated in this study could cause gill tissue damage and death in goldfish.Based on practical production, many goldfish had been infected with a small amount of Trichodina spp., causing gill tissue damage but no deaths.It was determined that this chronic and low mortality rate of goldfish gill rot disease was caused by a mixed infection of Trichodina spp.and weakly virulent strains of Flavobacterium columnare, and the secondary infection of weakly virulent strains of Flavobacterium columnare was the main cause of goldfish death.

Key words: goldfish; gill rot disease; Flavobacterium columnare; drug sensitivity

CLC Number:

S943

WANG Xiaoliang, CAO Huan, WANG Shu, ZHANG Wen, WANG Jingbo, LYU Xiaonan, WANG Peng, XU Lipu. Pathogenic Analysis on Gill Rot Disease in Goldfish Caused by Mixed Infection of Low Virulent of Flavobacterium columnare and Trichodina spp.[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4612-4620.

References

[1] 黄锦炉, 汪开毓, 黄艺丹, 等.水生动物致病菌——柱状黄杆菌研究进展[J].中国水产, 2009, 10:59-61. HUANG J L, WANG K Y, HUANG Y D, et al.Research progress of Flavobacterium columnosum, the pathogenic bacteria of aquatic animal[J].China Fisheries, 2009, 10:59-61.(in Chinese)
[2] AUSTIN B, AUSTIN D A.Bacterial Fish Pathogens:Diseases of Farmed and Wild Fish[M].4 ed.New York-Chichester:Springer-Praxis Publishing, 2007.
[3] 黎雪梅, 李小兵, 郑宗林, 等.水生动物柱状黄杆菌的研究进展[J].饲料与畜牧, 2014, 6:31-37. LI X M, LI X B, ZHENG Z L, et al.The research progress of Flavobacterium columnare isolated from aquatic animals[J].Feed and Animal Husbandry, 2014, 6:31-37.(in Chinese)
[4] 陈爱平.2005年中国水产养殖病害监测报告(一)[J].科学养鱼, 2006, 6:48-49. CHEN A P.2005 China aquaculture disease monitoring report (1)[J].Scientific Fish Farming, 2006, 6:48-49.(in Chinese)
[5] WAGNER B A, WISE D J, KHOOL H, et al.The epidemiology of bacterial diseases in food-size channel catfish[J].Journal of Aquatic Animal Health, 2002, 14(4):263-272.
[6] WANG B C, ZHU F Z, SHI Z C, et al.Molecular characteristics, polymorphism and expression analysis of MHC Ⅱ in Yellow catfish (Pelteobagrus fulvidraco) responding to Flavobacterium columnare infection[J].Fish & Shellfish Immunology, 2022, 125:90-100.
[7] JIA Y F, LI Z S, DU Q Y, et al.Transcriptome analysis of immune-related gene expression in Yellow River carp (Cyprinus carpio var.) after challenge with Flavobacterium columnare[J].Microbial Pathogenesis, 2021, 160:105148.
[8] ZHAO Z C, LIU S X, WU C, et al.Bioinformatics characteristics and expression analysis of TLR3 and its adaptor protein TRIF in largemouth bass (Micropterus salmoides) upon Flavobacterium columnare infection[J].Gene, 2023, 872:147450.
[9] 罗璋.鱼类烂鳃病病原柱状黄杆菌基因工程疫苗的研究[D].武汉:华中农业大学, 2015. LUO Z.Study on possible genetically engineered vaccine against the columnaris disease caused by Flavobacterium columnare[D].Wuhan:Huazhong Agricultural University, 2015.(in Chinese)
[10] THOMAS-JINU S, GOODWIN A.Acute columnaris infection in channel catfish, Ictalurus punctatus (Rafinesque):Efficacy of practical treatments for warm water aquaculture ponds[J].Journal of Fish Diseases, 2004, 27(1):23-28.
[11] SHOEMAKER C A, OLIVARES-FUSTER O, ARIAS C R, et al. Flavobacterium columnare genomovar influences mortality in channel catfish (Ictalurus punctatus)[J].Veterinary Microbiology, 2008, 127(3-4):353-359.
[12] 卢全章, 倪达书, 葛蕊芳.草鱼(Ctenopharyngodon idellus)烂鳃病的研究Ⅰ.细菌性病原的研究[J].水生生物学报, 1975, 5(3):315-334. LU Q Z, NI D S, GE R F. Studies on the gill diseases of the grass carp (Ctenopharyngodon idellus) Ⅰ.Isolation of a myxobacterial pathogen[J].Acta Hydrobiologica Sinica, 1975, 5(3):315-334.
[13] 何君慈, 邓国成.草鱼细菌性烂鳃病病原的研究[J].水产学报, 1987, 11(1):1-9. HE J C, DENG G C.A study on pathogenic bacteria of gill rote disease of grass crap[J].Journal of Fisheries of China, 1987, 11(1):1-9.(in Chinese)
[14] VERMA D K, RATHORE G, PRADHAN P K, et al.Isolation and characterization of Flavobacterium columnare from freshwater ornamental goldfish Carassius auratus[J].Journal of Environmental Biology, 2015, 36(2):433-439.
[15] DECOSTERE A, HAESEBROUCK F, DEVRIESE L A.Shieh medium supplemented with tobramycin for selective isolation of Flavobacterium columnare (Flexibacter columnaris) from diseased fish[J].Journal of Clinical Microbiology, 1997, 35(1):322-324.
[16] 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. DONG X Z, CAI M Y.Manual for Identification of Common Bacterial Systems[M].Beijing:Science Press, 2001.(in Chinese)
[17] 王良发, 谢海侠, 张金, 等.我国淡水鱼类柱形病病原菌柱状黄杆菌的遗传多样性[J].水生生物学报, 2010, 34(2):367-377. WANG L F, XIE H X, ZHANG J, et al.Columnaris disease and genetic diversity of ITS bacterial pathogen Flavobacterium columnare in freshwater fish in China[J].Aata Hydrobiologica Sinica, 2010, 34(2):367-377.(in Chinese)
[18] 邹玉霞, 张培军, 莫照兰, 等.大菱鲆出血症病原菌的分离和鉴定[J].高技术通讯, 2004, 14(4):89-93. ZOU Y X, ZHANG P J, MO Z L, et al.Isolation and identification of a pathogenic bacterium from haemorrhagic turbot (Scophthalmus maximus)[J]. Chinese High Technology Letters, 2004, 14(4):89-93.(in Chinese)
[19] 中华人民共和国国家质量监督检验检疫总局中国国家标准化管理委员会.GB/T 27623.1-2011渔用抗菌药物药效试验技术规范第1部分:常量肉汤稀释法药物敏感性试验[S].2012. GENERAL ADMINISTRATION OF QUALITY SUPERVISION.INSPECTION AND QUARANTINE OF THE PEOPLE'S REPUBLIC OF CHINA, STANDARDIZATION ADMINISTRATION OF THE PEOPLE'S REPUBLIC OF CHINA.GB/T 273623.1-2011.Pharmacodynamic test technical specification for aquaculture antimicrobial agent-Part 1:Antibiotics susceptibility test of macro-broth dilution method[S].2012.(in Chinese)
[20] NATIONAL COMMITTEE FOR CLINICAL LABORATORY STANDARDS.Performance standards for antimicrobial susceptibility testing;Twentieth informational supplement[S].National Committee for Clinical Laboratory Standards, 2010, M100-S20.
[21] PACHA R E, ORDAL E J.Myxobacterial diseases of salmonids[A].A Symposium on Diseases of Fishes and Shellfishes[C].Washinton D C:American Fisheries Society, Special Publication, 1970.
[22] FEIST S W.Fish diseases and disorders, volume 1, protozoan and metazoan infections[J].Aquaculture, 1996, 143(3-4):421-423.
[23] XU D H, SHOEMAKER C A, ZHANG D H.Treatment of Trichodina sp.reduced load of Flavobacterium columnare and improved survival of hybrid tilapia[J].Aquaculture Reports, 2015, 2(C):126-131.
[24] XU D H, SHOEMAKER C A, LAFRENTZ B R.Enhanced susceptibility of hybrid tilapia to Flavobacterium columnare after parasitism by Ichthyophthirius multifiliis[J].Aquaculture, 2014, 430(2):44-49.
[25] EVANS J J, KLESIUS P H, PASNIK D J, et al.Influence of natural Trichodina sp. parasitism on experimental Streptococcus iniae or Streptococcus agalactiae infection and survival of young channel catfish Ictalurus punctatus(Rafinesque)[J].Aquacultrue Research, 2010, 38(6):664-667.
[26] ZHANG C, LI D L, CHI C, et al.Dactylogyrus intermedius parasitism enhances Flavobacterium columnare invasion and alters immune-related gene expression in Carassius auratus[J].Diseases of Aquatic Organisms, 2015, 116(1):11-21.
[27] 陈昌福, 史维舟, 赵桂珍, 等.翘嘴鲡烂鳃病病原菌的分离及初步鉴定[J].华中农业大学学报, 1995, 14(3):263-266. CHEN C F, SHI W Z, ZHAO G Z, et al.Isolation and identification of pathogenic bacteria causing roted gill disease in mandarinfish (Siniperca chuatsi Basilewsky)[J].Journal of Huazhong Agricultural University, 1995, 14(3):263-266.(in Chinese)
[28] DECOSTERE A, HAESEBROUCK F, DEVRIESE L A.Characterization of four Flavobacterium columnare (Flexibacter columnaris) strains isolated from tropical fish[J].Veterinary Microbiology, 1998, 62(1):35-45.
[29] CAI W L, ARIAS C R.Deciphering the molecular basis for attenuation of Flavobacterium columnare strain Fc1723 used as modified live vaccine against columnaris disease[J].Vaccines (Basel), 2021, 9(11):1370.
[30] 杨泽敏, 李双, 金正雨, 等.1株鲤鱼源植物乳杆菌的分离鉴定及其生物学特性分析[J].中国畜牧兽医, 2022, 49(7):2805-2811. YANG Z M, LI S, JIN Z Y, et al.Isolation, identification and biological characteristics of a strain of Lactobacillus plantarum from carp[J].China Animal Husbandry & Veterinary Medicine, 2022, 49(7):2805-2811.(in Chinese)
[31] NGUYEN D T, MARANCIK D, SOTO E.B-glucan immunostilulation against columnaris in a white sturgeon (Acipenser transmontanus) model[J].Fish and Shellfish Immunology Reports, 2022, 3:100067.
[32] CONRAD R A, EVENHUIS J P, LIPSCOMB R S, et al.Flavobacterium columnare ferric iron uptake systems are required for virulence[J].Frontiers in Cellular and Infection Microbiology, 2022, 12:1029833.
[33] DECLERCQ A M, HAESEBROUCK F, BROECK W V, et al.Columnaris disease in fish:A review with emphasis on bacterium-host interactions[J].Veterinary Research, 2013, 44(1):27.
[34] 刘丽娟, 王亚军, 石存斌, 等.水产常见消毒剂对鱼源柱状黄杆菌的抑菌和杀菌效果研究[J].科学养鱼, 2021, 11:47-49. LIU L J, WANG Y J, SHI C B, et al.Study on the bacteriostatic and bactericidal effects of common aquatic disinfectants on Flavobacterium columnare isolated from fish[J].Scientific Fish Farming, 2021, 11:47-49.(in Chinese)
[35] GAUNT P S, GAO D, SUN F, et al.Efficacy of florfenicol for control of mortality caused by Flavobacterium columnare infection in channel catfish[J].Journal of Aquatic Animal Health, 2010, 22(2):115-122.

Pathogenic Analysis on Gill Rot Disease in Goldfish Caused by Mixed Infection of Low Virulent of Flavobacterium columnare and Trichodina spp.

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