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Sanguanrut, P.

Publications and source records attributed to Sanguanrut, P..

4 recordsLinked to original sources

Novel midgut smooth muscle necrosis (MSMN) in translucent or glass post-larvae of whiteleg shrimp

From the Asia-Pacific region, the suspected TPD/GPD specimens of postlarvae (PL) preserved in Davidsons fixative and 95% ethanol were sent to our laboratory to prepare for PCR and histological analysis. Interestingly, PCR tests with the specimens were negative with the VPHLVD(vtcc2 and vtcc3 gene) primers, but positive with the VPTPD(vhvp-2 gene) primers. There were no characteristic lesions of acute hepatopancreatic necrosis disease detected in these specimens. However, in the sections of several specimens from two affected tanks, the midgut was cut tangentially in the abdominal region, revealing a novel histopathology characterized by severe necrosis specifically in the thin, smooth muscle tissues underlying the midgut lumen epithelium. This comprised nuclear pyknosis and karyorrhexis limited to the smooth muscle and occasionally affecting the midgut epithelium but not involving the surrounding skeletal muscles. The pathology is called midgut smooth muscle necrosis (MSMN). Such pathology would undoubtedly lead to loss of midgut peristalsis and cessation of feeding, but it can be easily overlooked without careful histological examination using a high magnification microscope objective. No bacterial cells were evident, suggesting that the lesions arose from a virus or from a specific toxin response. This report is an urgent plea for colleagues studying TPD/GPD to review their samples histologically to help in determining whether MSMN is pathognomonic for TPD/GPD.

zoology↗

Production results indicate that parvovirus IHHNV should be delisted as a notifiable shrimp disease

The main purpose of this report is to provide hard evidence that the shrimp parvovirus IHHNV has not resulted "in significant consequences e.g. production losses, morbidity or mortality at a zone or country level" in Thailand since at least 2010. It also reveals that no single PCR test is sufficient to identify IHHNV-infected shrimp. It presents historical evidence and new evidence from 11 commercial ponds cultivating the giant tiger shrimp Penaeus monodon in Thailand. These ponds were selected because they were the ponds that gave positive PCR test results for IHHNV using 2 methods recommended for IHHNV diagnosis by WOAH. However, an additional in-house "IHHNV Long-Amp method" (IHHNV-LA) was also used to amplify 90% of the 4 kb IHHNV genome sequence, and it also gave false-positive test results with 2 of the 11 ponds. Further tests using normal histopathological analysis for the presence of pathognomonic Cowdry A type inclusions (CAI), in situ hybridization (ISH) and immunohistochemistry (IHC) could confirm IHHNV infections in only 2 of the 4 ponds PCR-positive using all 3 PCR methods. In addition, positive detection of CAI alone was equivalent to ISH or IHC in confirming IHHNV infection after a positive test with any of the PCR methods used. In summary, the recommended WOAH PCR methods gave false positive test results for IHHNV infection with 9/11 ponds (82%). All 11 ponds gave profitable harvests despite the confirmation of IHHNV in 2 ponds, where it was accompanied by various additional pathogens. Unfortunately, according to current practice, positive PCR test results with the WOAH methods alone sometimes leads to rejection of traded shrimp products without assurance that the test results are not false positive results that may arise from endogenous viral elements (EVE).

zoology↗

Potential universal PCR method to detect decapod hepanhamaparvovirus (DHPV) in crustaceans

Parvoviruses that infect the hepatopancreas (HP) of the penaeid shrimp Penaeus chinensis, P. monodon, and P. merquiensis were previously called hepatopancreatic parvoviruses (HPV). They are now classified in the family Parvoviridae, sub-family Hamaparvovirinae as members of the same genus called Hepanhamaparvovirus and referred to as decapod hepanhamaparvovirus, designated here as DHPV. However, a virus that causes similar lesions in the HP of the giant river prawn Macrobrachium rosenbergii resembles hepanhamaparvoviruses by microscopy and histochemistry. Unfortunately, no genome information is yet available and PCR detection methods that work for DHPV in P. monodon do not work with M. rosenbergii. For hatchery samples of M. rosenbergii in Thailand with DHPV-like lesions, we hypothesized it might be possible to design primer pairs from 8 full DHPV genome sequences at GenBank for use in PCR detection of DHPV in M. rosenbergii. Using this strategy, we successfully designed a new set of primers and a PCR protocol called the DHPV-U method that gave an amplicon with DNA extracts from larvae of M. rosenberigii samples that showed DHPV-like lesions, while extracts from normal larvae gave none. DNA extracts from P. monodon infected with DHPV also gave amplicons. At the same time, the normal PCR method for DHPV in P. monodon gave no amplicon with the M. rosenbergii DNA extracts. The DHPV-U amplicons from P. monodon and M. rosenbergii shared 99% sequence identity, and in situ hybridization (ISH) assays using the DIG-labeled amplicon gave positive histochemical results in the HP tissue of both P. monodon and M. rosenbergii. The DHPV-U method is now being used in Thailand for detection of DHPV in both P. monodon and M. rosenbergii. Overall, the results support the proposal that the HP virus in M. rosenbergii is also a hepanhamaparvovirus. Based on 100% sequence identity of the target region in the currently published DHPV sequences at GenBank, the DHPV-U method may also work for detection of other DHPV isolates.

molecular biology↗

Full genome characterization of Laem Singh virus (LSNV) in shrimp Penaeus monodon

Laem Singh virus (LSNV) was discovered in 2006 and proposed as a necessary but insufficient cause of retarded growth in the giant tiger shrimp Penaeus monodon. Its closest relatives were plant viruses including an unassigned Sobemovirus and viruses in the family Luteoviridae. During succeeding years, attempts to obtain the full LSNV genome sequence by genome walking failed. However, recent publication of the full sequence of Wenzhou shrimp virus 9 (WZSV 9) at GenBank revealed that LSNV sequences in our database shared 99% sequence identity with it. Thus, we hypothesized that LSNV and WZSV 9 were different isolates of the same virus species. Here we confirm that hypothesis by cloning and sequencing of the full genome of LSNV from P. monodon and by showing that it consists of two fragments each with 99% identity to the matching fragments of WZSV.

microbiology↗