Evaluating climate-dependent distribution of orthohantaviruses with monitoring wild rodents: One Health Perspective†.

Orthohantaviruses, cause hemorrhagic fever with renal syndrome, nephropathia epidemica, and hantavirus pulmonary syndrome, are major public health problems all over the world. Wild rodent surveillance for orthohantaviruses is of great importance for the preparedness against these human infections and the prediction of possible outbreak regions. Thus, we aimed to screen orthohantaviruses in wild rodents in Southern Anatolia, where the area has some of the glacial period refugia in the Mediterranean Basin, and interpret their current epidemiology with climatic biovariables in comparison with previously positive regions.We trapped muroid rodents between 2015 and 2017, and screened for orthohantaviruses. Then, we evaluated the relationship between orthohantavirus infections and bioclimatic variables. In spite of the long-term and seasonal sampling, we found no evidence for Orthohantavirus infections. The probable absence of orthohantaviruses in the sampling area was further evaluated from the climatic perspective, and results led us suggest that Orthohantavirus epidemiology might be relatively dependent on precipitation levels in driest and warmest quarters, and temperature fluctuations.These initial data might provide necessary perspective on wild rodent surveillance for orthohantaviruses in other regions, and help to collect lacking data for a such habitat suitability study in a bigger scale in the future.

Orthohantaviruses in Misiones Province, Northeastern Argentina.

Few cases of hantavirus pulmonary syndrome have been reported in northeastern Argentina. However, neighboring areas show a higher incidence, suggesting underreporting. We evaluated the presence of antibodies against orthohantavirus in small rodents throughout Misiones province. Infected Akodon affinis montensis and Oligoryzomys nigripes native rodents were found in protected areas of Misiones.

Exposure of free-ranging capybaras (Hydrochoerus hydrochaeris) to the vaccinia virus.

The aim of this study was to evaluate the possibility of free-ranging animals/hunting dogs as sources of infection in the vaccinia virus (VACV) transmission chain. Serological, cell culture and molecular assays were conducted in 56 free-ranging animals and 22 hunting dogs. ELISA/neutralizing assays showed that two (2.5%) capybaras (Hydrochoerus hydrochaeris) had anti-OPV positive antibodies, while all samples tested negative through PCR/cell culture. After being hit by cars on roads, capybaras that exhibited neither clinical signs nor any association with bovine outbreaks had neutralizing antibodies against the Orthopoxvirus, as detected through plaque-reduction neutralizing tests and ELISA. Evidence exists regarding peridomestic capybaras acting as a source of the virus and serving as a link between wild and urban environments, thus contributing to viral maintenance.

A 19 Year Analysis of Small Mammals Associated with Human Hantavirus Cases in Chile.

Small mammals present in areas where hantavirus cardiopulmonary syndrome (HCPS) cases had occurred in central and southern Chile were captured and analyzed to evaluate the abundance of rodents and seroprevalence rates of antibodies to Andes orthohantavirus (ANDV). Sampling areas ranged from the Coquimbo to Aysén regions (30-45° S approx.) regions. Ninety-two sites in peridomestic and countryside areas were evaluated in 19 years of sampling. An antibody against ANDV was detected by strip immunoassay in 58 of 1847 specimens captured using Sherman traps. Of the eleven species of rodents sampled, Abrothrix olivacea, Oligoryzomys longicaudatus and Abrothrix hirta were the most frequently trapped. O. longicaudatus had the highest seropositivity rate, and by logistic regression analysis, O. longicaudatus of at least 60 g had 80% or higher probability to be seropositive. Sex, age and wounds were significantly related to seropositivity only for O. longicaudatus. Across administrative regions, the highest seropositivity was found in the El Maule region (34.8-36.2° S), and the highest number of HCPS cases was registered in the Aysén region. Our results highlight the importance of long term and geographically extended studies, particularly for highly fluctuating pathogens and their reservoirs, to understand the implications of the dynamics and transmission of zoonotic diseases in human populations.

Detection of New World Hantavirus Antibodies in Rodents of Eastern New Mexico, USA.

Hantaviruses, causal agents of the potentially lethal hantavirus pulmonary syndrome, have widely distributed rodent hosts. Using an enzyme-linked immunosorbent assay, we tested blood from 398 wild rodents captured in eastern New Mexico, US in 2015-17 and found 42 antibody-positive samples representing six genera.

Diverse Beta- and Gammaherpesviruses in Neotropical Rodents from Costa Rica.

Neotropical wild rodents from Costa Rica were analyzed for the presence of herpesviruses (order Herpesvirales, family Herpesviridae). Using a broadly generic PCR, herpesvirus sequences were detected in 5% (8/160) of liver and heart samples: seven putative gammaherpesviruses in samples from Talamancan oryzomys (Nephelomys devius), sprightly colilargo (Oligoryzomys vegetus), Mexican deer mouse (Peromyscus nudipes), and Chiriqui harvest mouse (Reithrodontomys creper) and one putative betaherpesvirus in long-tailed singing mouse (Scotinomys xerampelinus). Results from this study could guide ecological investigations targeting the prevalence and host associations of herpesviruses in wild rodents from Costa Rica.

Habitat, species richness and hantaviruses of sigmodontine rodents within the Interior Atlantic Forest, Paraguay.

Four of the nine sigmodontine tribes have species that serve as reservoirs of rodent-borne hantaviruses (RBO-HV), few have been studied in any depth. Several viruses have been associated with human cases of hantavirus pulmonary syndrome often through peridomestic exposure. Jabora (JABV) and Juquitiba (JUQV), harbored by Akodon montensis and Oligoryzomys nigripes, respectively, are endemic and sympatric in the Reserva Natural de Bosque Mbaracayú (RNBM), Paraguay, a protected area of the Interior Atlantic Forest. Rodent communities were surveyed along a 30 km stretch of the RNBM in eight vegetation classifications (Low, High, Bamboo, Riparian and Liana Forests, Bamboo Understory, Cerrado, and Meadow/Grasslands). We collected 417 rodents from which 11 species were identified; Akodon montensis was the predominant species (72%; 95%CI: 64.7%-76.3%), followed by Hylaeamys megacephalus (15% (11.2%-18.2%)) and Oligoryzomys nigripes (9% (6.6%-12.4%)). We examined the statistical associations among habitat (vegetation class) type, rodent species diversity, population structure (age, sex, and weight), and prevalence of RBO-HV antibody and/or viral RNA (Ab/RNA) or characteristic Leishmania tail lesions. Ab/RNA positive rodents were not observed in Cerrado and Low Forest. A. montensis had an overall Ab/RNA prevalence of 7.7% (4.9%-11.3%) and O. nigripes had an overall prevalence of 8.6% (1.8%-23.1%). For A. montensis, the odds of being Ab/RNA positive in High Forest was 3.73 times of the other habitats combined. There was no significant difference among age classes in the proportion of Ab/RNA positive rodents overall (p = 0.66), however, all 11 RNA-positive individuals were adult. Sex and habitat had independent prognostic value for hantaviral Ab/RNA in the study population; age, presence of tail scar/lesion (19% of the rodents) and weight did not. Adjusting for habitat, female rodents had less risk of becoming infected. Importantly, these data suggest habitat preferences of two sympatric rodent reservoirs for two endemic hantaviruses and the importance of including habitat in models of species diversity and habitat fragmentation.

New polyomavirus species identified in nutria, Myocastor coypus polyomavirus 1.

A novel polyomavirus (PyVs) comprising 5,422 bp was identified by high-throughput sequencing (HTS) in pooled organs of nutria (Myocastor coypus). The new genome displays the archetypal organization of PyVs, which includes open reading frames for the regulatory proteins small T antigen (sTAg) and large T antigen (LTAg), as well as for the capsid proteins VP1, VP2 and VP3. Based on the International Committee on Taxonomy of Viruses (ICTV) Polyomaviridae Study Group criteria, this genome comprises a new PyVs species for the Alphapolyomavirus genus and is putatively named "Myocastor coypus Polyomavirus 1" . The complete genome sequence of this Myocastor coypus Polyomavirus 1 (McPyV1) isolate is publically available under the GenBank accession no. MH182627.

Rodent Abundance and Hantavirus Infection in Protected Area, East-Central Argentina.

We captured 3 hantavirus rodent hosts in Otamendi Natural Reserve, Argentina, during 2007-2012. Hantavirus antibodies were found only in Akodon azarae grass mice, mainly in males and old animals. Higher abundance of this species was associated with warm and rainy weather and high water levels, which peaked after a strong El Niño event.

Space Use and Social Mating System of the Hantavirus Host, Oligoryzomys longicaudatus.

The long-tailed mouse, Oligoryzomys longicaudatus (Cricetidae: Sigmodontinae), is the major host of Andes hantavirus, the etiological agent of hantavirus pulmonary syndrome in the south of Argentina and Chile. Studying the ecology of this species is necessary to understand how Andes hantavirus is maintained in nature. In this study, we examine the home range size and intra- and intersexual overlap degree of male and female O. longicaudatus in order to elucidate the mating system of this species. To our knowledge, this research provides the first documentation, obtained from a specific design, of spacing and mating systems in this species in Argentina. The study was conducted seasonally from April (autumn) 2012 to October (spring) 2013 in a shrubland habitat of Cholila, Andean region, Argentina. We studied spacing patterns using 59 and 51 home ranges established by adult males and females, respectively, in two 3.24 ha capture-marked and recapture grids. Significant differences between sexes in home range size and overlap degree were found. Male home ranges were always larger than those of females. We observed exclusive space use both among males and females (13.15 ± 18.67, and 3.60 ± 3.43%, respectively). Considering only those males that get access to receptive females (40%), average intersexual overlap value was about 30.82 ± 19.73%. Sexual differences in home range sizes and the spatial avoidance between breeding males, that would reflect intrasexual competition for receptive females, allows us to propose a polygynous mating system for O. longicaudatus.

Co-circulation of Araraquara and Juquitiba Hantavirus in Brazilian Cerrado.

Hantavirus cardiopulmonary syndrome is an emerging serious disease in the Americas, transmitted from wild rodents to humans through inhalation of aerosol containing virus. Herein, we characterized two distinct hantaviruses circulating in rodent species form Central Plateau, Midwestern region of Brazil in the Cerrado (savanna-like) biome, an area characterized by small trees and grasses adapted to climates with long dry periods. In this study, we identified the co-circulation of the Araraquara virus and a possible new lineage of the Juquitiba virus (JUQV) in Oligoryzomys nigripes. The implications of co-circulation are still unknown, but it can be the key for increasing viral diversity or emergence of new species through spillover or host switching events leading to co-infection and consequently recombination or reassortment between different virus species. Phylogenetic analyses based on the complete S segment indicated that, alongside with Oligoryzomys mattogrossae rodents, O. nigripes species could also have a whole as JUQV reservoir in the Cerrado biome. Although these rodents' species are common in the Cerrado biome, they are not abundant demonstrating how complex and different hantavirus enzootic cycles can be in this particular biome.

Genetic characterization of Cacipacoré virus from ticks collected in São Paulo State, Brazil.

Cacipacoré virus (CPCV) is a potential emerging virus classified in the genus Flavivirus, family Flaviviridae. In the present study, we present the genetic characterization of a CPCV isolated from ticks (Amblyomma cajennense) collected from a sick capybara (Hydrochoerus hydrochaeris) in São Paulo State, Brazil. The CPCV isolate shares the typical genomic organization of flaviviruses with 10,857 nucleotides in length and a single open reading frame of 10,284 nucleotides encoding a polyprotein of 3,427 amino acids. Phylogenetic analysis revealed that CPCV is unique, as a potentially tick-borne virus, in the Japanese encephalitis virus serogroup.

Murine norovirus infection in Brazilian animal facilities.

Murine norovirus (MNV) is a single-stranded positive-sense RNA virus of the Caliciviridae family. MNV has been reported to infect laboratory mice with the ability to cause lethal infections in strains lacking components of the innate immune response. Currently, MNV is considered the most prevalent infectious agent detected in laboratory mouse facilities. In this study, mice in 22 laboratory animal facilities within Brazil were analyzed for MNV infection. Using primers targeting a conserved region of the viral capsid, MNV was detected by RT-PCR in 137 of 359 mice from all 22 facilities. Nucleotide sequencing and phylogenetic analysis of the capsid region from the viral genome showed identity ranging from 87% to 99% when compared to reported MNV sequences. In addition, RAW264.7 cells inoculated with a mouse fecal suspension displayed cytopathic effect after the fifth passage. This study represents the first report of MNV in mouse colonies in Brazilian laboratory animal facilities, emphasizing the relevance of a health surveillance program in such environments.

Evaluation of habitat requirements of small rodents and effectiveness of an ecologically-based management in a hantavirus-endemic natural protected area in Argentina.

Hantavirus pulmonary syndrome is a severe cardio pulmonary disease transmitted to humans by sylvan rodents found in natural and rural environments. Disease transmission is closely linked to the ecology of animal reservoirs and abiotic factors such as habitat characteristics, season or climatic conditions. The main goals of this research were: to determine the biotic and abiotic factors affecting richness and abundance of rodent species at different spatial scales, to evaluate different methodologies for studying population of small rodents, and to describe and analyze an ecologically-based rodent management experience in a highly touristic area. A 4-year study of small rodent ecology was conducted between April 2007 and August 2011 in the most relevant habitats of El Palmar National Park, Argentina. Management involved a wide range of control and prevention measures, including poisoning, culling and habitat modification. A total of 172 individuals of 5 species were captured with a trapping effort of 13 860 traps-nights (1.24 individuals/100 traps-nights). Five rodent species were captured, including 2 hantavirus-host species, Oligoryzomys nigripes and Akodon azarae. Oligoryzomys nigripes, host of a hantavirus that is pathogenic in humans, was the most abundant species and the only one found in all the studied habitats. Our results are inconsistent with the dilution effect hypothesis. The present study demonstrates that sylvan rodent species, including the hantavirus-host species, have distinct local habitat selection and temporal variation patterns in abundance, which may influence the risk of human exposure to hantavirus and may have practical implications for disease transmission as well as for reservoir management.

Molecular evidence of Orthopoxvirus DNA in capybara (Hydrochoerus hydrochaeris) stool samples.

Vaccinia virus (VACV) is responsible for outbreaks in Brazil and has immense potential as an emerging virus. VACV can be found naturally circulating in India, Pakistan and South America, where it causes infections characterised by exanthematic lesions in buffaloes, cattle and humans. The transmission cycle of Brazilian VACV has still not been fully characterised; one of the most important gaps in knowledge being the role of wild animals. Capybaras, which are restricted to the Americas, are the world's largest rodents and have peculiar characteristics that make them possible candidates for being part of a natural VACV reservoir. Here, we developed a method for detecting orthopoxvirus DNA in capybara stool samples, and have described for the first time the detection of orthopoxvirus DNA in capybaras samples from three different regions in Brazil. These findings strongly suggest that capybaras might be involved in the natural transmission cycle of VACV and furthermore represent a public health problem, when associated with Brazilian bovine vaccinia outbreaks. This makes infected animals an important factor to be considered when predicting and managing Brazilian VACV outbreaks.

Experimental infection of Rio Mamore hantavirus in Sigmodontinae rodents

This study shows an experimental spillover infection of Sigmodontinae rodents with Rio Mamore hantavirus (RIOMV). Necromys lasiurus and Akodon sp were infected with 103 RNA copies of RIOMV by intraperitoneal administration. The viral genome was detected in heart, lung, and kidney tissues 18 days after infection (ai), and viral excretion in urine and faeces began at four and six ai, respectively. These results reveal that urine and faeces of infected rodents contain the virus for at least 18 days. It is possible that inhaled aerosols of these excreta could transmit hantavirus to humans and other animals.

Experimental infection of Rio Mamore hantavirus in Sigmodontinae rodents.

This study shows an experimental spillover infection of Sigmodontinae rodents with Rio Mamore hantavirus (RIOMV). Necromys lasiurus and Akodon sp were infected with 103 RNA copies of RIOMV by intraperitoneal administration. The viral genome was detected in heart, lung, and kidney tissues 18 days after infection (ai), and viral excretion in urine and faeces began at four and six ai, respectively. These results reveal that urine and faeces of infected rodents contain the virus for at least 18 days. It is possible that inhaled aerosols of these excreta could transmit hantavirus to humans and other animals.

Hantavirus pulmonary syndrome in a highly endemic area of Brazil.

Hantavirus pulmonary syndrome (HPS) is the most frequently reported fatal rodent-borne disease in Brazil, with the majority of cases occurring in Santa Catarina. We analysed the clinical, laboratory and epidemiological data of the 251 confirmed cases of HPS in Santa Catarina in 1999-2011. The number of cases ranged from 10 to 47 per year, with the highest incidences in 2004-2006. Gastrointestinal tract manifestations were found in >60% of the cases, potentially confounding diagnosis and leading to inappropriate therapy. Dyspnoea, acute respiratory failure, renal failure, increased serum creatinine and urea levels, increased haematocrits and the presence of pulmonary interstitial infiltrate were significantly more common in HPS patients who died. In addition, we demonstrated that the six cases from the midwest region of the state were associated with Juquitiba virus genotype. The case-fatality rate in this region, 19·2%, was lower than that recorded for other mesoregions. In the multivariate analysis increase of serum creatinine and urea was associated with death by HPS. Our findings help elucidate the epidemiology of HPS in Brazil, where mast seeding of bamboo can trigger rodent population eruptions and subsequent human HPS outbreaks. We also emphasize the need for molecular confirmation of the hantavirus genotype of human cases for a better understanding of the mortality-related factors associated with HPS cases in Brazil.

Identification of lymphocytic choriomeningitis mammarenavirus in house mouse (Mus musculus, Rodentia) in French Guiana.

Thirty-seven house mice (Mus musculus, Rodentia) caught in different localities in French Guiana were screened to investigate the presence of lymphocytic choriomeningitis mammarenavirus (LCMV). Two animals trapped in an urban area were found positive, hosting a new strain of LCMV, that we tentatively named LCMV "Comou". The complete sequence was determined using a metagenomic approach. Phylogenetic analyses revealed that this strain is related to genetic lineage I composed of strains inducing severe disease in humans. These results emphasize the need for active surveillance in humans as well as in house mouse populations, which is a rather common rodent in French Guianese cities and settlements.

Serologic Evidence of Flavivirus Infections in Peridomestic Rodents in Merida, Mexico.

We conducted surveillance for flavivirus infection in peridomestic rodents in Merida, Mexico in 2011-12. We captured 161 rodents inside private residences, using Sherman traps, including 86 house mice (Mus musculus) and 75 black rats (Rattus rattus). Serum from each animal was assayed by plaque reduction neutralization test (PRNT) using two vertebrate-specific flaviviruses (Apoi and Modoc viruses) and five mosquito-borne flaviviruses (dengue 2, dengue 4, St. Louis encephalitis virus, West Nile, and yellow fever viruses). Sixty-one (37.9%) rodents had antibodies that neutralized at least one virus. Prevalences for flaviviruses were 64.0% and 15.1% for black rats and house mice, respectively. None of the PRNT90 titers exceeded 80, and often they were highest for Modoc virus. These data suggest that a subset of rodents had been infected with Modoc virus or a closely related flavivirus that was not included in the PRNT analysis.