Use of PCR-restriction fragment length polymorphism for the identification of zoonotic mycobacteriosis in zebrafish caused by Mycobacterium abscessus and Mycobacterium chelonae.
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.
Seok
Seung-Hyeok
SH
Department of Laboratory Animal Medicine, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
Koo
Hye Cheong
HC
Kasuga
Asako
A
Kim
Yeun
Y
Lee
Eun Gae
EG
Lee
Hyeyoung
H
Park
Jong-Hwan
JH
Baek
Min-Won
MW
Lee
Hui-Young
HY
Kim
Dong-Jae
DJ
Lee
Byeung-Hee
BH
Lee
Yong-Soon
YS
Cho
Sang-Nae
SN
Park
Jae-Hak
JH
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
D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit