PCR and in situ hybridization for the detection and localization of a new pathogen Francisella-like bacterium (FLB) in ornamental cichlids

Hsieh, Chi-Ming; Wu, Zong-Bing; Tung, Ming-Chen; Tsai, Shinn-Shyong

doi:10.3354/dao075029

Cited by 37 publications

(36 citation statements)

References 22 publications

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“…nov. (Ottem et al 2009) (syn = F. asiatica sp. nov. [Mikalsen & Colquhon in press]) is the cause of disease in farmed tilapia in Asia and South America (Hsieh et al 2006, Mauel et al 2007), three-lined grunt Parapristipoma trilineatum in Japan (Kamaishi et al 2005) and a range of cichlid species (Hsieh et al 2007). …”

Section: Introductionmentioning

confidence: 99%

An outbreak of disease caused by Francisella sp. in Nile tilapia Oreochromis niloticus at a recirculation fish farm in the UK

Jeffery¹,

Stone²,

Feist³

et al. 2010

Dis. Aquat. Org.

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This study details the first diagnosis of Francisella sp. in tilapia in the United Kingdom. Losses of tilapia fry at a recirculation fish farm in England were investigated, giving a presumptive positive diagnosis of infection with Francisella sp. by histopathological examination. Most fish sampled showed moderate to marked pathology of the major organs, with lesions being present in most tissues. The most obvious host response was granuloma formulation. A subsequent follow-up visit provided further evidence for the presence of a Francisella species. PCR amplicons were obtained using Francisella spp.-specific primers that shared 100% sequence identity with the 16S rRNA gene of the type strain of the species F. asiatica previously described as the cause of disease in tilapia in Southeast Asia and Central America. This outbreak and the subsequent investigation emphasise the importance of strict biosecurity at fish farms and the care that needs to be taken when using a new supplier of fish.

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Section: Introductionmentioning

confidence: 99%

An outbreak of disease caused by Francisella sp. in Nile tilapia Oreochromis niloticus at a recirculation fish farm in the UK

Jeffery¹,

Stone²,

Feist³

et al. 2010

Dis. Aquat. Org.

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“…During the past 5 yr, bacteria of the genus Francisella have caused significant mortalities in cultured tilapia Oreochromis spp., Atlantic cod Gadus morhua, Atlantic salmon Salmo salar L., hybrid striped bass Morone chrysops × M. saxatilis, three-line grunt Parapristipoma trilineatum, and ornamental cichlids, both in warm-and cold-water environments (Kamaishi et al 2005, Nylund et al 2006, Olsen et al 2006, Ostland et al 2006, Birkbeck et al 2007, Hsieh et al 2007, Mauel et al 2007, Mikalsen et al 2007, Ottem et al 2007, Soto et al 2009a.…”

Section: Introductionmentioning

confidence: 99%

Development of a real-time PCR assay for identification and quantification of the fish pathogen Francisella noatunensis subsp. orientalis

Soto¹,

Bowles²,

Fernandez³

et al. 2010

Dis. Aquat. Org.

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Members of the genus Francisella are small Gram-negative facultative intracellular bacteria that cause francisellosis in a wide variety of fish species worldwide. F. noatunensis subsp. orientalis has been recently described as a warm-water pathogen of tilapia Oreochromis spp. In this study, a quantitative real-time polymerase chain reaction (qPCR) TaqMan probe assay was developed to rapidly and accurately detect and quantify F. noatunensis subsp. orientalis from fish tissue. The target region of the assay was the F. tularensis iglC gene homologue previously found in F. noatunensis subsp. orientalis. Probe specificity was confirmed by the lack of signal and cross-reactivity with 12 common fish pathogens, 2 subspecies of F. tularensis, F. noatunensis subsp. noatunensis, and tilapia tissue. The range of linearity was determined to be 50 fg to 1.4 mg, and the lower limit of detection was 50 fg of DNA (equivalent to ~25 genome equivalents) per reaction. A similar sensitivity was observed with DNA extracted from a mixture of F. noatunensis subsp. orientalis and fish tissue. The assay was also able to detect and quantify F. noatunensis subsp. orientalis from the spleens of experimentally infected tilapia. No signal was observed in the control groups. In conclusion, we have developed a highly sensitive and specific assay that can be used for the specific identification and quantification of F. noatunensis subsp. orientalis. KEY WORDS: Francisella · Tilapia · qPCRResale or republication not permitted without written consent of the publisher

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“…The subspecies, F. noatunensis subsp noatunensis and F. noatunensis subsp orientalis (FNO; syn. Francisella asiatica), have been shown to be associated with francisellosis in the chicken grunt (Parapristipoma trilineatum), hybrid striped bass (Morone saxatilis x M. chrysops), Atlantic salmon (Salmo salar), Atlantic cod (Gadus morhua), tilapia (Oreochromis spp), ornamental cichlid, and damselfish (Chromis viridis) (Hsieh et al, 2007;Birkbeck et al, 2011;Camus et al, 2013). Francisellosis primarily affects the fry and fingerling stages of fish, which exhibit systemic granulomatosis with multifocal granulomas in the internal organs, including the liver, spleen, and kidney (Soto et al, 2009(Soto et al, , 2012a.…”

Section: Introductionmentioning

confidence: 99%

Outbreaks and genetic diversity of Francisella noatunensis subsp orientalis isolated from farm-raised Nile tilapia (Oreochromis niloticus) in Brazil

2014

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ABSTRACT. Francisella noatunensis subsp orientalis (FNO) is an emerging pathogen of warm water tilapia in a number of different countries. The disease caused by this bacterium in fish is characterized by a systemic granulomatous infection that causes high mortality rates during outbreaks. FNO has been previously described in Asia, Europe, and Central and North America. Its occurrence in South America has never been described. Since 2012, outbreaks of a granulomatous disease have been recorded in cage farms of Nile tilapia (Oreochromis niloticus L.) in Brazil. The current study aimed to identify the etiologic agent of recent francisellosis outbreaks at Brazilian tilapia farms, and to characterize the genetic diversity of the pathogen from farms with distinct geographic origins and without epidemiological connections. Bacteriological analysis Francisellosis in cage-raised Nile tilapia of 44 diseased Nile tilapia collected from five cage farms in Brazil was performed during 2012 and 2013. The farms were in different locations and had no recent history of animal or biological material transport between each other. Sixty-two FNO isolates were identified on the basis of FNO-specific qPCR. The main predisposing factors for the occurrence of outbreaks on Brazilian farms were lower water temperature (<22°C) and life stage of fish, affecting mainly fry, fingerlings and young adults (live weight <100 g). The genetic diversity of the Brazilian FNO isolates was evaluated using repetitive extragenic palindromic-PCR. The isolates from different origins were shown to be clonally related. This is the first report of the occurrence and genetic diversity of FNO in South America.

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PCR and in situ hybridization for the detection and localization of a new pathogen Francisella-like bacterium (FLB) in ornamental cichlids

Cited by 37 publications

References 22 publications

An outbreak of disease caused by Francisella sp. in Nile tilapia Oreochromis niloticus at a recirculation fish farm in the UK

An outbreak of disease caused by Francisella sp. in Nile tilapia Oreochromis niloticus at a recirculation fish farm in the UK

Development of a real-time PCR assay for identification and quantification of the fish pathogen Francisella noatunensis subsp. orientalis

Outbreaks and genetic diversity of Francisella noatunensis subsp orientalis isolated from farm-raised Nile tilapia (Oreochromis niloticus) in Brazil

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