Mini Review: Virus Interference: History, Types and Occurrence in Crustaceans

Virus interference is a phenomenon in which two viruses interact within a host, affecting the outcome of infection of at least one of such viruses. The effect of this event was first observed in the XVIII century and it was first recorded even before virology was recognized as a distinct science from microbiology. Studies on virus interference were mostly done in the decades between 1930 and 1960 in viruses infecting bacteria and different vertebrates. The systems included in vivo experiments and later, more refined assays were done using tissue and cell cultures. Many viruses involved in interference are pathogenic to humans or to economically important animals. Thus the phenomenon may be relevant to medicine and to animal production due to the possibility to use it as alternative to chemical therapies against virus infections to reduce the severity of disease/mortality caused by a superinfecting virus. Virus interference is defined as the host resistance to a superinfection caused by a pathogenic virus causing obvious signs of disease and/or mortality due to the action of an interfering virus abrogating the replication of the former virus. Different degrees of inhibition of the superinfecting virus can occur. Due to the emergence of novel pathogenic viruses in recent years, virus interference has recently been revisited using different pathogens and hosts, including commercially important farmed aquatic species. Here, some highly pathogenic viruses affecting farmed crustaceans can be affected by interference with other viruses. This review presents data on the history of virus interference in hosts including bacteria and animals, with emphasis on the known cases of virus interference in crustacean hosts.Life Science Identifiers (LSIDs)Escherichia coli [(Migula 1895) Castellani & Chalmers 1919]Aedes albopictus (Skuse 1894)Liocarcinus depurator (Linnaeus 1758): urn:lsid:marinespecies.org:taxname:107387Penaeus duorarum (Burkenroad 1939): urn:lsid:marinespecies.org:taxname:158334Carcinus maenas (Linnaeus 1758): urn:lsid:marinespecies.org:taxname:107381Macrobrachium rosenbergii (De Man 1879): urn:lsid:marinespecies.org:taxname:220137Penaeus vannamei (Boone 1931): urn:lsid:zoobank.org:pub:C30A0A50-E309-4E24-851D-01CF94D97F23Penaeus monodon (Fabricius 1798): urn:lsid:zoobank.org:act:3DD50D8B-01C2-48A7-B80D-9D9DD2E6F7ADPenaeus stylirostris (Stimpson 1874): urn:lsid:marinespecies.org:taxname:58498

Application of RNA Interference (RNAi) against Viral Infections in Shrimp: A Review

Shrimp culture has long been done in Asia and America to provide high quality food to people. Modern aquaculture uses advanced techniques to increase shrimp production but it also has enhanced the occurrence of infectious diseases. Disease is the main pitfall for the development and sustainability of shrimp aquaculture worldwide. In the last decade several methods and strategies have been developed and evaluated under experimental conditions in order to curb the negative impact of viral infections. Among these, RNA interference is the most recent tool against viral diseases in shrimp and it is deemed as a promising biotechnology to boost shrimp production. This paper gives a broad overview of the RNAi methods used to fight viral diseases in shrimp aquaculture compared to the antiviral effect of methods previously evaluated against viruses. It also gives examples of the use of RNAi to learn more on mechanisms of the shrimp defense response. The application of RNAi to fight or treat viral infections in shrimp aquaculture has yet to come and it depends on the efficacy of RNAi against several viral diseases, evaluation of environment and food safety and the development of cheap, massive delivery methods of RNAi molecules to shrimp farming facilities

Prevalencia del protozoario Perkinsus sp. en un cultivo de ostión japonés Crassostrea gigas en Sinaloa, México

Crassostrea gigas es un molusco bivalvo de gran importancia comercial. En el noroeste de México su producción es afectada por mortalidades cuyo origen infeccioso no ha sido determinado claramente. En este trabajo se determinó la prevalencia e intensidad de la infección por Perkinsus sp. en un cultivo de C. gigas en el ciclo 2011-2012. El cultivo se hizo en un sistema de línea suspendida con densidades de 28 y 42 ostiones/canasta y se determinó un tamaño de muestra de 30 ostiones por mes. La detección de Perkinsus sp. se hizo de acuerdo a los protocolos de la Organización Mundial de Sanidad Animal (OIE) para Medio Fluido de Tioglicolato y PCR. Con ambos métodos se determinó la prevalencia de Perkinsus sp., que varió entre 3,3 y 40%. La intensidad de la infección estuvo en niveles 1 y 2, de acuerdo a la escala de Mackin. La mortalidad acumulativa en las densidades de 28 y 42 ostiones por canasta fue del 4 y 6%, respectivamente. Las mayores mortalidades del ostión y las mayores prevalencias de Perkinsus sp. ocurrieron en septiembre (2,7 y 16,6%) y octubre (1,5 y 23,3%), respectivamente, cuando la temperatura fue alta. En conclusión, Perkinsus sp. fue detectado en un cultivo de C. gigas en el estero La Pitahaya con prevalencia moderada, baja intensidad de infección y mayor presencia en los meses más calurosos del ciclo de cultivo

Double-stranded RNA against white spot syndrome virus (WSSV) vp28 or vp26 reduced susceptibility of Litopenaeus vannamei to WSSV, and survivors exhibited decreased susceptibility in subsequent re-infections

Abstract The antiviral effect of vp28 or vp26 double-stranded (ds) RNA upon single or consecutive white spot syndrome virus (WSSV) intramuscular challenges with a high infectious dose was evaluated. The vp28 dsRNA showed the highest protection both in single (LT50 = 145 h at 10 d and 98 h at 20 d post treatment [dpt]) or consecutive (LT50 = 765 h) WSSV challenges compared to vp26 dsRNA (LT50 = 126 h at 10 d and 57 h at 20 dpt vs. consecutive challenge LT50 = 751 h). Single WSSV challenges showed that animals treated with vp28 or vp26 dsRNA gradually lost the antiviral effect as virus challenge occurred at 10 dpt (cumulative mortality 63% vs. 80%, respectively) or 20 dpt (87% vs. 100%, respectively). In contrast, animals treated with vp28 or vp26 dsRNA and consecutively challenged with WSSV showed and extended lower susceptibility to WSSV. All dead animals were WSSV-positive by one-step PCR, whereas all surviving shrimp from single or continuous challenges were WSSV-negative as determined by reverse transcription (RT)-PCR. In conclusion, shrimp treated with a single administration of vp28 or vp26 dsRNA and consecutively challenged with WSSV showed a stronger and longer antiviral response than shrimp exposed once to WSSV at 10 or 20 dpt

Silencing Pacific white shrimp Litopenaeus vannamei LvRab7 reduces mortality in brooders challenged with white spot syndrome virus

White spot syndrome virus (WSSV) is a major threat for farmed shrimp worldwide. RNA interference (RNAi) is the most recent tool against viral diseases. Rab7 silencing effectively inhibited virus infections in juvenile shrimp, but the antiviral effect in brooders remains unknown. This study found a homologue Penaeus monodon Rab7 gene in Litopenaeus vannamei brooders from Mexico. Sequence identity was >99% to a Thai LvRab7 sequence and >94% to Rab7 sequences from P. monodon or Marsupenaeus japonicus. Animals treated with a partial (494 bp) or a complete (618 bp) LvRab7 dsRNA sequences and challenged 48 h post treatment (hpt) with a high WSSV dose showed 80–88% mortality respectively. Shrimp treated with 4 or 20 μg LvRab7 dsRNA and challenged with a WSSV high dose had 80% mortality each, but it was reduced to 33% and 40%, respectively, with a low dose. Efficacy of dsRNA to reduce shrimp mortality was dependent on virus dose used regardless of dsRNA concentration. A significant reduction in LvRab7 mRNA levels was observed at 120 hpt. In conclusion, silencing LvRab7 in brooders showed a mild antiviral effect against a WSSV challenge at 48 h

In vivo titration of white spot syndrome virus (WSSV) in specific pathogen-free Litopenaeus vannamei by intramuscular and oral routes

White spot syndrome virus (WSSV) is a devastating pathogen in shrimp aquaculture. Standardized challenge procedures using a known amount of infectious virus would assist in evaluating strategies to reduce its impact. In this study, the shrimp infectious dose 50% endpoint (SID50 ml–1) of a Thai isolate of WSSV was determined by intramuscular inoculation (i.m.) in 60 and 135 dold specific pathogen-free (SPF) Litopenaeus vannamei using indirect immunofluorescence (IIF) and 1-step polymerase chain reaction (PCR). Also, the lethal dose 50% endpoint (LD50 ml–1) was determined from the proportion of dead shrimp. The median virus infection titers in 60 and 135 d old juveniles were 106.8 and 106.5 SID50 ml–1, respectively. These titers were not significantly different (p ≥ 0.05). The titration of the WSSV stock by oral intubation in 80 d old juveniles resulted in approximately 10-fold reduction in virus titer compared to i.m. inoculation. This lower titer is probably the result of physical and chemical barriers in the digestive tract of shrimp that hinder WSSV infectivity. The titers determined by infection were identical to the titers determined by mortality in all experiments using both i.m. and oral routes at 120 h post inoculation (hpi), indicating that every infected shrimp died. The determination of WSSV titers for dilutions administered by i.m. and oral routes constitutes the first step towards the standardization of challenge procedures to evaluate strategies to reduce WSSV infection

Pathogenesis of a Thai strain of white spot syndrome virus (WSSV) in juvenile, specific pathogen-free Litopenaeus vannamei

White spot syndrome virus (WSSV) causes disease and mortality in cultured and wild shrimp. A standardized WSSV oral inoculation procedure was used in specific pathogen-free (SPF) Litopenaeus vannamei (also called Penaeus vannamei) to determine the primary sites of replication (portal of entry), to analyze the viral spread and to propose the cause of death. Shrimp were inoculated orally with a low (101.5 shrimp infectious dose 50% endpoint [SID50]) or a high (104 SID50) dose. Per dose, 6 shrimp were collected at 0, 6, 12, 18, 24, 36, 48 and 60 h post inoculation (hpi). WSSVinfected cells were located in tissues by immunohistochemistry and in hemolymph by indirect immunofluorescence. Cell-free hemolymph was examined for WSSV DNA using 1-step PCR. Tissues and cell-free hemolymph were first positive at 18 hpi (low dose) or at 12 hpi (high dose). With the 2 doses, primary replication was found in cells of the foregut and gills. The antennal gland was an additional primary replication site at the high dose. WSSV-infected cells were found in the hemolymph starting from 36 hpi. At 60 hpi, the percentage of WSSV-infected cells was 36 for the epithelial cells of the foregut and 27 for the epithelial cells of the integument; the number of WSSV-infected cells per mm2 was 98 for the gills, 26 for the antennal gland, 78 for the hematopoietic tissue and 49 for the lymphoid organ. Areas of necrosis were observed in infected tissues starting from 48 hpi (low dose) or 36 hpi (high dose). Since the foregut, gills, antennal gland and integument are essential for the maintenance of shrimp homeostasis, it is likely that WSSV infection leads to death due to their dysfunctio

Standardized white spot syndrome virus (WSSV) inoculation procedures for intramuscular or oral routes

In the past, strategies to control white spot syndrome virus (WSSV) were mostly tested by infectivity trials in vivo using immersion or per os inoculation of undefined WSSV infectious doses, which complicated comparisons between experiments. In this study, the reproducibility of 3 defined doses (10, 30 and 90 shrimp infectious doses 50% endpoint [SID50]) of WSSV was determined in 3 experiments using intramuscular (i.m.) or oral inoculation in specific pathogen-free (SPF) Litopenaeus vannamei. Reproducibility was determined by the time of onset of disease, cumulative mortality, and median lethal time (LT50). By i.m. route, the 3 doses induced disease between 24 and 36 h post inoculation (hpi). Cumulative mortality was 100% at 84 hpi with doses of 30 and 90 SID50 and 108 hpi with a dose of 10 SID50. The LT50 of the doses 10, 30 and 90 SID50 were 52, 51 and 49 hpi and were not significantly different (p > 0.05). Shrimp orally inoculated with 10, 30 or 90 SID50 developed disease between 24 and 36 hpi. Cumulative mortality was 100% at 108 hpi with doses of 30 and 90 SID50 and 120 hpi with a dose of 10 SID50. The LT50 of 10, 30 and 90 SID50 were 65, 57 and 50 hpi; these were significantly different from each other (p < 0.05). A dose of 30 SID50 was selected as the standard for further WSSV challenges by i.m. or oral routes. These standardized inoculation procedures may be applied to other crustacea and WSSV strains in order to achieve comparable results among experiments