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Biology subjects

Buathongkam, P.

Publications and source records attributed to Buathongkam, P..

2 recordsLinked to original sources

Shrimp endogenous viral elements (EVE) correlate with survival in white spot syndrome virus (WSSV) challenges

Shrimp and other arthropods are capable of specific, adaptive immune responses to viruses based on viral copy DNA (vcDNA) fragments in the host genome called endogenous viral elements (EVE). These may produce negative sense RNA transcripts leading to an RNA interference (RNAi) defense response against cognate viruses. We first reported high-frequency-read sequences (HFRS) of white spot syndrome virus EVE (named WSSV-EVE 4,6,8) in a WSSV-free breeding stock of whiteleg shrimp (Penaeus vannamei). Here we describe screening for the same HFRS-EVE in a captured giant tiger shrimp (Penaeus monodon) breeding stock, also free of WSSV. WSSV-EVE 4,6,8 was detected in some of the P. monodon stock individuals with positive or negative RNA expression. Eight broodstock individuals were selected for mating in 4 crosses. The offspring from these crosses were grown sufficiently to allow tagging and pleopod sampling for DNA and RNA analysis prior to challenge with WSSV. This allowed for Mendelian analysis of EVE inheritance and for its expression or not in the offspring, together with analysis of their relationships to survival and WSSV infection level after challenge. The results revealed that EVE inheritance was Mendelian, but that their RNA expression or not was independently controlled. In Crosses 1 and 2, all the offspring died and none of them carried 2 or more of the expressed EVE in their parental shrimp. In contrast, 100% of 10 arbitrarily selected surviving shrimp from Cross 3 and 90% from Cross 4 carried and expressed 2 or more of the 3 expressed EVE transmitted from the parental shrimp. These results reveal a potential protocol for development of viral tolerant shrimp stocks.

zoology↗

Shrimp parvovirus circular DNA fragments arise from both endogenous viral elements (EVE) and the infecting virus

Some insects use endogenous reverse transcriptase (RT) to make variable viral copy DNA (vcDNA) fragments from viral RNA in linear (lvcDNA) and circular (cvcDNA) forms. The latter form is easy to extract selectively. The vcDNA produces small interfering RNA (siRNA) variants that inhibit viral replication via the RNA interference (RNAi) pathway. The vcDNA is also autonomously inserted into the host genome as endogenous viral elements (EVE) that can also result in RNAi. We hypothesized that similar mechanisms occurred in shrimp. We used the insect methods to extract circular viral copy DNA (cvcDNA) from the giant tiger shrimp (Penaeus monodon) infected with a virus originally named infectious hypodermal and hematopoietic necrosis virus (IHHNV). Simultaneous injection of the extracted cvcDNA plus IHHNV into whiteleg shrimp (Penaeus vannamei) resulted in a significant reduction in IHHNV replication when compared to shrimp injected with IHHNV only. Next generation sequencing (NGS) revealed that the extract contained a mixture of two general IHHNV-cvcDNA types. One showed 98 to 99% sequence identity to GenBank record AF218266 from an extant type of infectious IHHNV. The other type showed 98% sequence identity to GenBank record DQ228358, an EVE formerly called non-infectious IHHNV. The startling discovery that EVE could also give rise to cvcDNA revealed that cvcDNA provided an easy means to identify and characterize EVE in shrimp and perhaps other organisms. These studies open the way for identification, characterization and use of protective cvcDNA as a potential shrimp vaccine and as a tool to identify, characterize and select naturally protective EVE to improve shrimp tolerance to homologous viruses in breeding programs.

molecular biology↗