title: Use of PCR-restriction fragment length polymorphism for the identification of zoonotic mycobacteriosis in zebrafish caused by Mycobacterium abscessus and Mycobacterium chelonae.
abstract: Skin ulcers, scoliosis, and dropsy-like scale edema were observed in laboratory-maintained zebrafish. Affected fish had multifocal granulomas not only in internal organs such as the liver, intestine, genital organs, kidney, muscle, and spleen but also in the fin, epithelium, gills, and sclera of the eyes. Large numbers of acid-fast-rod-shaped bacteria were observed within the necrotic centers of well-demarcated, multifocal granulomas with Gram's stain and Ziehl-Neelson's stain. The size of the Mycobacterium spp. was 1-2 microm x 2-3 microm with a double-layered cell wall, based upon electron-microscopical features. Definitive diagnosis of these outbreaks was obtained by culture on selective media followed by PCR-restriction fragment length polymorphism analysis (PRA) of the rpoB gene for species identification. The amplified 360-bp products of the rpoB gene of mycobacteria isolated from zebrafish were digested with MspI restriction enzyme, which revealed unique band patterns matching those of Mycobacterium abscessus and Mycobacterium chelonae which are responsible for skin and soft tissue infection caused by rapidly growing mycobacteria in humans. This is the first documentation of the precise identification of zoonotic non-tuberculous mycobacteria isolated from laboratory-maintained zebrafish by the PRA of the rpoB gene; this study thus provides a great deal of useful epidemiological information and reduces the likelihood that epizootics will occur.
authors.affiliation: Department of Laboratory Animal Medicine, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
authors.forename: Seung-Hyeok
authors.initials: SH
authors.lastname: Seok
authors.forename: Hye Cheong
authors.initials: HC
authors.lastname: Koo
authors.forename: Asako
authors.initials: A
authors.lastname: Kasuga
authors.forename: Yeun
authors.initials: Y
authors.lastname: Kim
authors.forename: Eun Gae
authors.initials: EG
authors.lastname: Lee
authors.forename: Hyeyoung
authors.initials: H
authors.lastname: Lee
authors.forename: Jong-Hwan
authors.initials: JH
authors.lastname: Park
authors.forename: Min-Won
authors.initials: MW
authors.lastname: Baek
authors.forename: Hui-Young
authors.initials: HY
authors.lastname: Lee
authors.forename: Dong-Jae
authors.initials: DJ
authors.lastname: Kim
authors.forename: Byeung-Hee
authors.initials: BH
authors.lastname: Lee
authors.forename: Yong-Soon
authors.initials: YS
authors.lastname: Lee
authors.forename: Sang-Nae
authors.initials: SN
authors.lastname: Cho
authors.forename: Jae-Hak
authors.initials: JH
authors.lastname: Park
mesh_terms: D000818:Animals; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D004196:Disease Outbreaks / Q000662:veterinary; D005393:Fish Diseases / Q000175:diagnosis / Q000382:microbiology* / Q000473:pathology; D005784:Gene Amplification; D009161:Mycobacterium / Q000235:genetics / Q000302:isolation & purification*; D009164:Mycobacterium Infections / Q000175:diagnosis / Q000382:microbiology / Q000473:pathology / Q000662:veterinary*; D009165:Mycobacterium Infections, Nontuberculous / Q000175:diagnosis / Q000382:microbiology / Q000473:pathology; D016926:Mycobacterium chelonae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D012150:Polymorphism, Restriction Fragment Length; D015027:Zebrafish; D015047:Zoonoses / Q000382:microbiology
chemical_list: D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit