Variable changes in nematode infection prevalence and intensity after Rabbit Haemorrhagic Disease Virus emerged in wild rabbits in Scotland and New Zealand

The myxoma virus (a microparasite) reduced wild rabbit numbers worldwide when introduced in the 1950s, and is known to interact with co-infecting helminths (macroparasites) causing both increases and decreases in macroparasite population size. In the 1990s Rabbit Haemorrhagic Disease Virus (RHDV) infected rabbits and also significantly reduced rabbit numbers in several countries. However, not much is known about RHDV interactions with macroparasites. In this study, we compare prevalence and intensity of infection for three gastrointestinal nematode species (Trichostrongylus retortaeformis, Graphidium strigosum and Passalurus ambiguus) before and after RHDV spread across host populations in Scotland and New Zealand. During one common season, autumn, prevalence of T. retortaeformis was higher after RHDV spread in both locations, whereas it was lower for G. strigosum and P. ambiguus after RHDV arrived in New Zealand, but higher in Scotland. Meanwhile, intensity of infection for all species decreased after RHDV arrived in New Zealand, but increased in Scotland. The impact of RHDV on worm infections was generally similar across seasons in Scotland, and also similarities in seasonality between locations suggested effects on infection patterns in one season are likely similar year-round. The variable response by macroparasites to the arrival of a microparasite into Scottish and New Zealand rabbits may be due to differences in the environment they inhabit, in existing parasite community structure, and to some extent, in the relative magnitude of indirect effects. Specifically, our data suggest that bottom-up processes after the introduction of a more virulent strain of RHDV to New Zealand may affect macroparasite co-infections by reducing the availability of their shared common resource, the rabbits. Clearly, interactions between co-infecting micro- and macroparasites vary in host populations with different ecologies, and significantly impact parasite community structure in wildlife. Keywords: Co-infection, Community ecology, European rabbit, Helminth, Macroparasite, Microparasite, RHDV, Virus, Within-host ecolog

Measuring Alphavirus Fidelity Using Non-Infectious Virus Particles

Mutations are incorporated into the genomes of RNA viruses at an optimal frequency and altering this precise frequency has been proposed as a strategy to create live-attenuated vaccines. However, determining the effect of specific mutations that alter fidelity has been difficult because of the rapid selection of the virus population during replication. By deleting residues of the structural polyprotein PE2 cleavage site, E3∆56-59, in Venezuelan equine encephalitis virus (VEEV) TC-83 vaccine strain, non-infectious virus particles were used to assess the effect of single mutations on mutation frequency without the interference of selection that results from multiple replication cycles. Next-generation sequencing analysis revealed a significantly lower frequency of transversion mutations and overall mutation frequency for the fidelity mutants compared to VEEV TC-83 E3∆56-59. We demonstrate that deletion of the PE2 cleavage site halts virus infection while making the virus particles available for downstream sequencing. The conservation of the site will allow the evaluation of suspected fidelity mutants across alphaviruses of medical importance

Universal primers that amplify RNA from all three flavivirus subgroups

Background: Species within the Flavivirus genus pose public health problems around the world. Increasing cases of Dengue and Japanese encephalitis virus in Asia, frequent outbreaks of Yellow fever virus in Africa and South America, and the ongoing spread of West Nile virus throughout the Americas, show the geographical burden of flavivirus diseases. Flavivirus infections are often indistinct from and confused with other febrile illnesses. Here we review the specificity of published primers, and describe a new universal primer pair that can detect a wide range of flaviviruses, including viruses from each of the recognised subgroups

Mesoniviruses are mosquito-specific viruses with extensive geographic distribution and host range

Background: The family Mesoniviridae (order Nidovirales) comprises of a group of positive-sense, single-stranded RNA ([+]ssRNA) viruses isolated from mosquitoes. Findings: Thirteen novel insect-specific virus isolates were obtained from mosquitoes collected in Indonesia, Thailand and the USA. By electron microscopy, the virions appeared as spherical particles with a diameter of ~50 nm. Their 20,129 nt to 20,777 nt genomes consist of positive-sense, single-stranded RNA with a poly-A tail. Four isolates from Houston, Texas, and one isolate from Java, Indonesia, were identified as variants of the species Alphamesonivirus-1 which also includes Nam Dinh virus (NDiV) from Vietnam and Cavally virus (CavV) from Côte d’Ivoire. The eight other isolates were identified as variants of three new mesoniviruses, based on genome organization and pairwise evolutionary distances: Karang Sari virus (KSaV) from Java, Bontag Baru virus (BBaV) from Java and Kalimantan, and Kamphaeng Phet virus (KPhV) from Thailand. In comparison with NDiV, the three new mesoniviruses each contained a long insertion (180 – 588 nt) of unknown function in the 5’ region of ORF1a, which accounted for much of the difference in genome size. The insertions contained various short imperfect repeats and may have arisen by recombination or sequence duplication. Conclusions: In summary, based on their genome organizations and phylogenetic relationships, thirteen new viruses were identified as members of the family Mesoniviridae, order Nidovirales. Species demarcation criteria employed previously for mesoniviruses would place five of these isolates in the same species as NDiV and CavV (Alphamesonivirus-1) and the other eight isolates would represent three new mesonivirus species (Alphamesonivirus-5, Alphamesonivirus-6 and Alphamesonivirus-7). The observed spatiotemporal distribution over widespread geographic regions and broad species host range in mosquitoes suggests that mesoniviruses may be common in mosquito populations worldwide

Preliminary characterization of (nucleoside-2′-O-)-methyltransferase crystals from Meaban and Yokose flaviviruses. Corrigendum

A corrigendum to the paper by Mastrangelo et al. (2006), Acta Cryst. F62, 768–770

Negeviruses reduce replication of alphaviruses during co-infection

Negeviruses are a group of insect-specific virus (ISV) that have been found in many arthropods. Their presence in important vector species led us to examine their interactions with arboviruses during co-infections. Wild-type negeviruses reduced the replication of several alphaviruses during co-infections in mosquito cells. Negev virus (NEGV) isolates were also used to express GFP and anti-chikungunya virus (CHIKV) antibody fragments during co-infections with CHIKV. NEGV expressing anti-CHIKV antibody fragments was able to further reduce replication of CHIKV during co-infections, while reductions of CHIKV with NEGV expressing GFP were similar to titers with wild-type NEGV alone. These results are the first to show that negeviruses induce superinfection exclusion of arboviruses and to demonstrate a novel approach to deliver anti-viral antibody fragments with paratransgenic ISVs. The ability to inhibit arbovirus replication and express exogenous proteins in mosquito cells make negeviruses a promising platform for control of arthropod-borne pathogens

Characterization of Farmington virus, a novel virus from birds that is distantly related to members of the family Rhabdoviridae

Background: Farmington virus (FARV) is a rhabdovirus that was isolated from a wild bird during an outbreak of epizootic eastern equine encephalitis on a pheasant farm in Connecticut, USA. Findings: Analysis of the nearly complete genome sequence of the prototype CT AN 114 strain indicates that it encodes the five canonical rhabdovirus structural proteins (N, P, M, G and L) with alternative ORFs (> 180 nt) in the N and G genes. Phenotypic and genetic characterization of FARV has confirmed that it is a novel rhabdovirus and probably represents a new species within the family Rhabdoviridae. Conclusions: In sum, our analysis indicates that FARV represents a new species within the family Rhabdoviridae

Experimental Infection of Potential Reservoir Hosts with Venezuelan Equine Encephalitis Virus, Mexico

Multiple wild rodent species can serve as amplifying reservoir hosts for virus subtype IE

Identifying sources, pathways and risk drivers in ecosystems of Japanese Encephalitis in an epidemic-prone north Indian district

Japanese Encephalitis (JE) has caused repeated outbreaks in endemic pockets of India. This study was conducted in Kushinagar, a highly endemic district, to understand the human-animal-ecosystem interactions, and the drivers that influence disease transmission. Utilizing the ecosystems approach, a cross-sectional, descriptive study, employing mixed methods design was employed. Four villages (two with pig-rearing and two without) were randomly selected from a high, a medium and a low burden (based on case counts) block of Kushinagar. Children, pigs and vectors were sampled from these villages. A qualitative arm was incorporated to explain the findings from the quantitative surveys. All human serum samples were screened for JE-specific IgM using MAC ELISA and negative samples for JE RNA by rRT-PCR in peripheral blood mononuclear cells. In pigs, IgG ELISA and rRT-PCR for viral RNA were used. Of the 242 children tested, 24 tested positive by either rRT-PCR or MAC ELISA; in pigs, 38 out of the 51 pigs were positive. Of the known vectors, Culex vishnui was most commonly isolated across all biotopes. Analysis of 15 blood meals revealed human blood in 10 samples. Univariable analysis showed that gender, religion, lack of indoor residual spraying of insecticides in the past year, indoor vector density (all species), and not being vaccinated against JE in children were significantly associated with JE positivity. In multivariate analysis, only male gender remained as a significant risk factor. Based on previous estimates of symptomatic: asymptomatic cases of JE, we estimate that there should have been 618 cases from Kushinagar, although only 139 were reported. Vaccination of children and vector control measures emerged as major control activities; they had very poor coverage in the studied villages. In addition, lack of awareness about the cause of JE, lack of faith in the conventional medical healthcare system and multiple referral levels causing delay in diagnosis and treatment emerged as factors likely to result in adverse clinical outcomes