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Determination of a novel parvovirus pathogen associated with massive mortality in adult tilapia.

ABSTRACT: Tilapia is one of the most important economic and fastest-growing species in aquaculture worldwide. In 2015, an epidemic associated with severe mortality occurred in adult tilapia in Hubei, China. The causative pathogen was identified as Tilapia parvovirus (TiPV) by virus isolation, electron microscopy, experimental challenge, In situ hybridization (ISH), indirect immunofluorescence (IFA), and viral gene sequencing. Electron microscopy revealed large numbers of parvovirus particles in the organs of diseased fish, including kidney, spleen, liver, heart, brain, gill, intestine, etc. The virions were spherical in shape, non-enveloped and approximately 30nm in diameter. The TiPV was isolated and propagated in tilapia brain cells (TiB) and induced a typical cytopathic effect (CPE) after 3 days post-infection (dpi). This virus was used to experimentally infect adult tilapia and clinical disease symptoms similar to those observed naturally were replicated. Additionally, the results of ISH and IFA showed positive signals in kidney and spleen tissues from TiPV-infected fish. To identify TiPV-specific sequences, the near complete genome of TiPV was obtained and determined to be 4269 bp in size. Phylogenetic analysis of the NS1 sequence revealed that TiPV is a novel parvovirus, forms a separate branch in proposed genus Chapparvovirus of Parvoviridae. Results presented here confirm that TiPV is a novel parvovirus pathogen that can cause massive mortality in adult tilapia. This provides a basis for the further studies to define the epidemiology, pathology, diagnosis, prevention and treatment of this emerging viral disease.

SUBMITTER: Liu W

PROVIDER: S-EPMC7588064 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature

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Determination of a novel parvovirus pathogen associated with massive mortality in adult tilapia.

Liu Wenzhi W Zhang Yecheng Y Ma Jie J Jiang Nan N Fan Yuding Y Zhou Yong Y Cain Kenneth K Yi Meisheng M Jia Kuntong K Wen Hua H Liu Wei W Guan Wuxiang W Zeng Lingbing L

PLoS pathogens 20200924 9

Tilapia is one of the most important economic and fastest-growing species in aquaculture worldwide. In 2015, an epidemic associated with severe mortality occurred in adult tilapia in Hubei, China. The causative pathogen was identified as Tilapia parvovirus (TiPV) by virus isolation, electron microscopy, experimental challenge, In situ hybridization (ISH), indirect immunofluorescence (IFA), and viral gene sequencing. Electron microscopy revealed large numbers of parvovirus particles in the organs ...[more]

PMID: 32970777

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Project description:Largemouth bass (Micropterus salmoides) is an important and fast-growing aquaculture species in China. In 2017, an epidemic associated with severe mortality occurred in fingerlings of largemouth bass in Guangdong, China. The causative pathogen was identified and named as largemouth bass Birnavirus (LBBV) by virome analysis, viral isolation, electron microscopy, genome sequencing, Western blot, indirect immunofluorescence, experimental challenge, and so on. Virome sequencing results showed that the relative abundance reads related to the family Birnaviridae were the highest, occupied ∼25% of the total viral reads. Electron microscopy revealed large numbers of nonenveloped virus particles in the spleen of diseased fish with a diameter of about 53 nm. LBBV was isolated and propagated in Chinese perch brain cells and induced a typical cytopathic effect. LBBV was stable to chloroform, heat, and 5-bromo-2'-deoxyuridine, but sensitive to acid (pH 3.0). The complete genome of LBBV was comprised of segment A with a size 3525 bp and segment B with a size 2737 bp. Phylogenetic analysis basing on RdRp and VP2 protein sequences revealed that LBBV were clustered into one clade with Lates calcarifer Birnavirus (LCBV), sharing 98.7% or 91.9% sequence identity with LCBV, respectively, but only sharing 59.7% and 52.7% sequence identity with Blosnavirus, suggesting that LBBV and LCBV probably belonged to a new genus. Challenge experiments results indicated that clinical disease symptoms similar to those observed naturally were replicated and the cumulative mortality reached 100% at 3 dpi by i.p. injection. The investigation of prevalence of LBBV infection showed that 41.5% (17/41) sample pools collected from diseased ponds was positive during 2017-2020, indicating that an emerging outbreak of this disease may be spreading within the largemouth bass in China. Above results confirmed that LBBV is a novel Birnavirus associated with massive mortality for fingerlings of largemouth bass. This provides a basis for prevention and control of this emerging viral disease. IMPORTANCE Pathogen isolation and identification are vital for emerging infectious outbreaks. Here we report the isolation, determination and characterization of a novel largemouth bass Birnavirus (LBBV) associated with massive mortality in largemouth bass. And genome of LBBV is determined and analyzed. Based on phylogenetic and alignment analysis of genome, we suggest LBBV belongs to a new genus (designated as Perbirnavirus genus) in Birnaviridae family. Our findings will provide a basis for the further study on prevention and control of this emerging viral disease.

Project description:Viral diseases arising in farmed fish are an ongoing challenge to the aquaculture industry, causing severe mortality and economic losses. Recently, there has been a spike in the incidence of a viral disease caused by Tilapia Parvovirus (TiPV) inflicts irreparable damage, and large-scale fish kills in the farmed tilapia species. We investigated a case of disease outbreak and severe mortality in cage and wetland farms of tilapia in West Bengal and Odisha, India. The symptomatic fish showed clinical signs, including hemorrhage, discoloration, ulcer, and redness in the body surfaces. Further analysis revealed that Tilapia Parvovirus was associated (validated by PCR, phylogenetic analysis, and cell line assay) with the infection and mortality of tilapia. The virus was detected in gill, heart, spleen, liver, and kidney samples collected from apparently healthy (asymptomatic) and symptomatic tilapia samples from cage and wetland farms. At the same time, negative results were found in the brain and skin tissue samples. The histological analysis revealed that TiPV induces severe damage invariably in almost all studied tissue, including the liver, kidney, spleen, gill, heart, and brain of tilapia samples. The viral quantification analysis showed that the viral genome was higher in the liver, spleen, and heart than in the tilapia samples' gill, kidney, or brain tissue. Furthermore, the study indicated that TiPV infection has a significant effect on the health of tilapia. The tilapia exhibited an immune reactivity toward TiPV infection (upregulation of chemokine receptors, CRs and interleukin 1β, IL-1β), the majority of the studied immune genes (interleukin 8, IL-8; Toll-like receptors 7, TLR7; tumour necrosis factor α, TNF-α; major histocompatibility complex II, MHC II and nuclear factor kappa B, NF-kB) were significantly downregulated in the kidney, spleen and liver tissue samples of symptomatic tilapia. Further, the in vivo challenge assay confirms that the isolated TiPV is a novel parvovirus pathogen that causes massive mortality in tilapia. The lessons learned from the first cellular and molecular description associated with TiPV epidemiology from wetland and cage farms of tilapia could be critical to developing the current state of the tilapia farming industry. Additionally, a holistic approach is needed to develop management measures to control the virulence and risk factors of TiPV.

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Determination of a novel parvovirus pathogen associated with massive mortality in adult tilapia.

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Determination of a novel parvovirus pathogen associated with massive mortality in adult tilapia.

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