Establishment of a novel and highly permissive cell line for the efficient replication of cyprinid herpesvirus 2 (CyHV-2)

Ma, Jie; Jiang, Nan; LaPatra, Scott E.; Jin, Ling; Xu, Jin; Fan, Yuding; Zhou, Yong; Zeng, Lingbing

doi:10.1016/j.vetmic.2015.04.006

Cited by 38 publications

(42 citation statements)

References 33 publications

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“…Cell line derivation was confirmed by PCR and sequencing. The obtained sequence was identical to that of the published black carp mitochondrial 16s rRNA (Figure a‐b) (Ma et al., ). In addition, the MPK cells were cultured in ESM4 medium, which is an established medium for medaka embryonic stem cells (Hong et al., ).…”

Section: Resultssupporting

confidence: 83%

“…Here, 61% of the MPK cells possessed a diploid chromosome number of 48 (2N = 48), which is identical to the number of black carp (Lou et al., ), MAF (Zhu et al., ) and ELGH (Guo et al., ). Mitochondrial DNA, particularly the 16s rRNA gene, has frequently been used in species identification and classification of various aquatic and mammalian species (Li et al., ; Ma et al., ; Yu et al., ). The sequencing data indicated that the cells originated from black carp.…”

Section: Discussionmentioning

confidence: 99%

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Establishment of a cell line from the kidney of black carp and its susceptibility to spring viremia of carp virus

Xue

Wang

Pan

et al. 2017

Journal of Fish Diseases

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In this study, we established and characterized a cell line derived from the kidney of black carp (Mylopharyngodon piceus), which is an important freshwater aquaculture species. The cell line was designated as MPK and subcultured for more than 70 passages in DMEM medium containing 10% fetal bovine serum (FBS) at 28°C. MPK had a modal diploid chromosome number of 48. Moreover, a transient MPK transfection efficiency was up to 18% using a green fluorescent protein plasmid by a modified electroporation. In addition, the MPK cells showed susceptibility to spring viremia of carp virus (SVCV), as demonstrated by the presence of severe cytopathic effects (CPEs) and increased viral RNA. Unexpectedly, the MPK cells expressed pluripotency-associated genes such as nanog, oct4 and vasa, indicating that these are possibly adult stem cells. Taken together, we have established a stable cell line from kidney that may potentially be utilized as an in vitro platform for genetic modifications and host-pathogen analysis in black carp.

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Section: Resultssupporting

confidence: 83%

Section: Discussionmentioning

confidence: 99%

Establishment of a cell line from the kidney of black carp and its susceptibility to spring viremia of carp virus

Xue

Wang

Pan

et al. 2017

Journal of Fish Diseases

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“…At 7 days post‐infection, three fish from each group were sampled for pathogen detection in liver, spleen and kidney tissue. IHNV was re‐isolated by standard methodologies (Batts & Winton, ; Ma et al, ). For F. psychrophilum re‐isolation, tissue samples were suspended in 1 ml of phosphate‐buffered saline (PBS) and the number of CFU of F. psychrophilum per mg of tissue was determined by plating 10‐fold serial dilutions on TYES plates, which were incubated for at least 4 days at 15°C.…”

Section: Methodsmentioning

confidence: 99%

Co‐infection of rainbow trout (Oncorhynchus mykiss) with infectious hematopoietic necrosis virus and Flavobacterium psychrophilum

Bruce

Oliver

et al. 2019

Journal of Fish Diseases

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Co-infection of rainbow trout with infections haematopoietic necrosis virus (IHNV)and Flavobacterium psychrophilum is known to occur, and it has been speculated that a combined infection can result in dramatic losses. Both pathogens can persist in fish in an asymptomatic carrier state, but the impact of co-infection has not been well characterized or documented. In this study, it was hypothesized that fish co-infected with F. psychrophilum and IHNV would exhibit greater mortality than fish infected with either pathogen alone. To test this, juvenile rainbow trout were co-infected with low doses of either IHNV or F. psychrophilum, and at 2 days post-initial challenge, they were given a low dose of the reciprocal pathogen. This combined infection caused high mortality (76.2%-100%), while mortality from a single pathogen infection with the same respective dose was low (5%-20%). The onset of mortality was earlier in the co-infected group (3-4 days) when compared with fish infected with F. psychrophilum alone (6 days) or IHNV (5 days), confirming the synergistic interaction between both pathogens. Co-infection led to a significant increase in the number of F. psychrophilum colony-forming units and IHNV plaque-forming units within tissues. This finding confirms that when present together in co-infected fish, both pathogens are more efficiently recovered from tissues. Furthermore, pathogen genes were significantly increased in co-infected groups, which parallel the findings of increased systemic pathogen load. Extensive tissue necrosis and abundant pathogen present intracellularly and extracellularly in haematopoietic tissue. This was pronounced in co-infected fish and likely contributed to the exacerbated clinical signs and higher mortality. This study provides novel insight into host-pathogen interactions related to co-infection by aquatic bacterial and viral pathogens and supports our hypothesis. Such findings confirm that mortality in fish exposed to both pathogens is greatly elevated compared to a single pathogen infection. K E Y W O R D S co-infection, Flavobacterium psychrophilum, IHNV, rainbow trout How to cite this article: Ma J, Bruce TJ, Oliver L, Cain KD. Co-infection of rainbow trout (Oncorhynchus mykiss) with infectious haematopoietic necrosis virus and Flavobacterium psychrophilum. J Fish Dis. 2019;42:1065-1076. https ://doi.

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“…The major economic aquaculture market of China is dominated by cyprinidae fish, CyHV-2 infection is a problem not only on-farm but also for held in markets, at retail sites and by private owners. Animal experiments Ma et al, 2015). One report found that formalin-inactivated vaccine that was inoculated by intraperitoneal injection and immersion were both effective to against CyHV-2 infection in goldfish, this operable vaccination method provides us with an effective prevention method .…”

Section: Discussionmentioning

confidence: 99%

“…With increasing development of interior and international trade of aquaculture, geographical scope of CyHV‐2 might spread further unless establishing more effective quarantine methods, prevention and control technologies. On account for infeasibility of CyHV‐2 culture through conventional training, several experiments have established susceptible cell lines obtained from goldfish and gibel carp, providing potent tools for researching virology, immunology, genetics and toxicology between CyHV‐2 and hosts (Ito et al, ; Ma et al, ). One report found that formalin‐inactivated vaccine that was inoculated by intraperitoneal injection and immersion were both effective to against CyHV‐2 infection in goldfish, this operable vaccination method provides us with an effective prevention method ( Ito & Ototake, ).…”

Section: Discussionmentioning

confidence: 99%

New detection of Cyprinid herpesvirus 2 associated with mass mortality in colour crucian carp ( Carassius auratus ), in China

Zhao

Geng

et al. 2019

Aquac Res

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During October 2016, a mass mortality of colour crucian carp (Carassius auratus), which the affected fish were lethargic, inappetence and anoxic, was occurred in a fish farm located in Chengdu, Sichuan province, China. To elucidate the aetiology of this outbreak, histological and electron microscope examination, molecular investigation were conducted. Pathologic examination revealed multi foci necrosis on haematopoietic organs, gills, hearts and pancreas. Transmission electron microscopy observations exhibited sphere herpesvirus‐like particles distributed amongst the tissues of gill, spleen and kidney. Molecular analysis is verified that the causative agent of this outbreak was Cyprinid herpesvirus 2 (CyHV‐2). This report first report CyHV‐2 in colour crucian carp, which increases the concern about damage of CyHV‐2 and its potential role in species.

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Establishment of a novel and highly permissive cell line for the efficient replication of cyprinid herpesvirus 2 (CyHV-2)

Cited by 38 publications

References 33 publications

Establishment of a cell line from the kidney of black carp and its susceptibility to spring viremia of carp virus

Establishment of a cell line from the kidney of black carp and its susceptibility to spring viremia of carp virus

Co‐infection of rainbow trout (Oncorhynchus mykiss) with infectious hematopoietic necrosis virus and Flavobacterium psychrophilum

New detection of Cyprinid herpesvirus 2 associated with mass mortality in colour crucian carp ( Carassius auratus ), in China

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