Multiple infections caused by white spot syndrome virus and <i>Enterocytozoon hepatopenaei</i> in pond‐reared <i>Penaeus vannamei</i> in India and multiplex PCR for their simultaneous detection

Thamizhvanan, S; Sivakumar, S.; Kumar, S. Santhosh; Kumar, Dilip; Suryakodi, S.; Balaji, Kithiganahalli Narayanaswamy; Rajkumar, T.; Vimal, S.; Majeed, Shahid; Taju, G.; Hameed, A.S. Sahul

doi:10.1111/jfd.12956

Cited by 37 publications

(21 citation statements)

References 32 publications

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“…The incidences of viral diseases in aquaculture species, particularly crustaceans such as shrimps and crabs have been well studied [ 5 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 ]. On the other hand, studies on viral infections in wild marine crustaceans are restricted to few reports [ 23 , 24 , 25 , 26 , 27 , 28 ].…”

Section: Introductionmentioning

confidence: 99%

Co-Infection of Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) and White Spot Syndrome Virus (WSSV) in the Wild Crustaceans of Andaman and Nicobar Archipelago, India

Saravanan

Praveenraj

Kiruba-Sankar

et al. 2021

Viruses

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The present study was intended to screen the wild crustaceans for co-infection with Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) and White Spot Syndrome Virus (WSSV) in Andaman and Nicobar Archipelago, India. We screened a total of 607 shrimp and 110 crab samples using a specific polymerase chain reaction, and out of them, 82 shrimps (13.5%) and 5 (4.5%) crabs were found positive for co-infection of IHHNV and WSSV. A higher rate of co-infection was observed in Penaeus monodon and Scylla serrata than other shrimp and crab species. The nucleotide sequences of IHHNV and WSSV obtained from crab in this present study exhibited very high sequence identity with their counterparts retrieved from various countries. Histopathological analysis of the infected shrimp gill sections further confirmed the eosinophilic intra-nuclear cowdry type A inclusion bodies and basophilic intra-nuclear inclusion bodies characteristics of IHHNV and WSSV infections, respectively. The present study serves as the first report on co-infection of WSSV and IHHNV in Andaman and Nicobar Archipelago, India and accentuates the critical need for continuous monitoring of wild crustaceans and appropriate biosecurity measures for brackishwater aquaculture.

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Section: Introductionmentioning

confidence: 99%

Co-Infection of Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) and White Spot Syndrome Virus (WSSV) in the Wild Crustaceans of Andaman and Nicobar Archipelago, India

Saravanan

Praveenraj

Kiruba-Sankar

et al. 2021

Viruses

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“…The presence of IMNV in the inoculum was confirmed by RT‐PCR using the IMNV‐specific primer set designed by Poulos and Lightner (2006). The homogenate was also tested for other shrimp pathogens such as white spot syndrome virus (WSSV), infectious hypodermal and haematopoietic necrosis virus (IHHNV) and Enterocytozoon hepatopenaei (EHP) by PCR, as multiple viral infections and EHP infection are common in India (Manivannan et al., 2002; Thamizhvanan et al., 2019). The filtrate was stored at −80°C for further analysis.…”

Section: Methodsmentioning

confidence: 99%

In vitro propagation of infectious myonecrosis virus in C6/36 mosquito cell line

Kumar¹,

Sivakumar²,

Majeed³

et al. 2021

Journal of Fish Diseases

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Infectious myonecrosis (IMN) is an important shrimp viral disease caused by infectious myonecrosis virus (IMNV). Based on previous reports, an attempt was made to propagate IMNV in apparently healthy C6/36 subclone of Aedes albopictus cell line. The confirmatory assays such as RT‐PCR, real‐time PCR and bioassay revealed that C6/36 cells were found to be susceptible to IMNV and these cells could be used easily for isolation and propagation of IMNV. The results of real‐time PCR assay showed that a lower CT value of 22.25 in IMNV‐infected cells was obtained on 10 day post‐infection (d p.i.), whereas the higher CT value of 35.21 was obtained in IMNV‐infected cells on 2 d p.i. There is no significant difference between CT values of IMNV production in vitro using C6/36 cell line and in vivo using shrimp. The IMNV propagated in C6/36 cells is capable of infecting shrimp and caused 100% mortality in shrimp. Clinical signs observed in shrimp injected with IMNV propagated in C6/36 cell line were found to be similar to naturally infected shrimp.

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“…When infected, it causes stunted growth, soft shells, lethargy and white feces symptoms (Behera et al, 2019). In Asian countries including India, Thailand and China, EHP caused 10%-20% reduced production of shrimp annually leading to significant economic losses (Thamizhvanan et al, 2019). In the year 2015 in Jiangsu, China, EHP infections had caused 300 million CNY of loss for the shrimp farming industry.…”

Section: Introductionmentioning

confidence: 99%

Rapid Detection of Enterocytozoon hepatopenaei Infection in Shrimp With a Real-Time Isothermal Recombinase Polymerase Amplification Assay

Ma¹,

Fan²,

Wang³

et al. 2021

Front. Cell. Infect. Microbiol.

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Enterocytozoon hepatopenaei (EHP) infection has become a significant threat in shrimp farming industry in recent years, causing major economic losses in Asian countries. As there are a lack of effective therapeutics, prevention of the infection with rapid and reliable pathogen detection methods is fundamental. Molecular detection methods based on polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) have been developed, but improvements on detection speed and convenience are still in demand. The isothermal recombinase polymerase amplification (RPA) assay derived from the recombination-dependent DNA replication (RDR) mechanism of bacteriophage T4 is promising, but the previously developed RPA assay for EHP detection read the signal by gel electrophoresis, which restricted this application to laboratory conditions and hampered the sensitivity. The present study combined fluorescence analysis with the RPA system and developed a real-time RPA assay for the detection of EHP. The detection procedure was completed in 3–7 min at 39°C and showed good specificity. The sensitivity of 13 gene copies per reaction was comparable to the current PCR- and LAMP-based methods, and was much improved than the RPA assay analyzed by gel electrophoresis. For real clinical samples, detection results of the real-time RPA assay were 100% consistent with the industrial standard nested PCR assay. Because of the rapid detection speed and the simple procedure, the real-time RPA assay developed in this study can be easily assembled as an efficient and reliable on-site detection tool to help control EHP infection in shrimp farms.

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Multiple infections caused by white spot syndrome virus and Enterocytozoon hepatopenaei in pond‐reared Penaeus vannamei in India and multiplex PCR for their simultaneous detection

Cited by 37 publications

References 32 publications

Co-Infection of Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) and White Spot Syndrome Virus (WSSV) in the Wild Crustaceans of Andaman and Nicobar Archipelago, India

Co-Infection of Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) and White Spot Syndrome Virus (WSSV) in the Wild Crustaceans of Andaman and Nicobar Archipelago, India

In vitro propagation of infectious myonecrosis virus in C6/36 mosquito cell line

Rapid Detection of Enterocytozoon hepatopenaei Infection in Shrimp With a Real-Time Isothermal Recombinase Polymerase Amplification Assay

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