Emerging hantaviruses in Central Argentina: First case of Hantavirus Pulmonary Syndrome caused by Alto Paraguay virus, and a novel orthohantavirus in Scapteromys aquaticus rodent.

Orthohantaviruses are emerging rodent-borne pathogens that cause Hantavirus Pulmonary Syndrome in humans. They have a wide range of rodent reservoir hosts and are transmitted to humans through aerosolized viral particles generated by the excretions of infected individuals. Since the first description of HPS in Argentina, new hantaviruses have been reported throughout the country, most of which are pathogenic to humans. We present here the first HPS case infected with Alto Paraguay virus reported in Argentina. Until now, Alto Paraguay virus was considered a non-pathogenic orthohantavirus since it was identified in a rodent, Holochilus chacarius. In addition to this, with the goal of identifying potential hantavirus host species in the province of Santa Fe, we finally describe a novel orthohantavirus found in the native rodent Scapteromys aquaticus, which differed from other hantaviruses described in the country so far. Our findings implicate an epidemiological warning regarding these new orthohantaviruses circulating in Central Argentina as well as new rodent species that must be considered as hosts from now on.

Home range variability, spatial aggregation, and excursions of Akodon azarae and Oligoryzomys flavescens in Pampean agroecosystems.

Rodents are reservoirs of various types of hantavirus, some of which are agents of hantavirus pulmonary syndrome in humans. Each hantavirus is associated with a single rodent host species but successive spill-over events may eventually lead to host-switching and new species' becoming host of a given pathogen. This study aims to gain an understanding of the spatial ecology of two hantavirus-host species, Akodon azarae, and Oligoryzomys flavescens, by identifying factors modulating their home range sizes and stability, and by evaluating intra- and interspecific spatial aggregation for these species and a third one-Oxymycterus rufus-living in sympatry. For this, eleven capture-mark-recapture surveys were carried out, spanning 22 months. We found that A. azarae males have larger and more mobile home ranges than females, independently of the season. Consequently, males could likely have a more relevant role in the transmission of hantavirus because of their greater exposure both to a higher number of contacts between individuals and viral contamination of the environment. Contrasting, O. flavescens individuals showed negligible displacements of their home range through time, which could limit the range of hantavirus spread in host populations. Since O. flavescens is host to Lechiguanas hantavirus (pathogenic to humans) this result encompasses epidemiological relevance, for it may imply the existence of local foci of infection. Additionally, individuals of both species performed excursions outside their home ranges. These events could enable hantavirus spread over distances beyond the normal range of movements and lead to new hantavirus outbreaks in formerly non-infected rodent populations, favoring the persistence of the virus in nature.

Hantavirus Pulmonary Syndrome Risk in Entre Ríos, Argentina.

Hantavirus pulmonary syndrome (HPS) is a severe emerging endemic disease of the Americas. Because hantavirus reservoirs are sylvatic rodents, HPS risk has been associated with occupational and recreational activities in natural and rural environments. The aim of this study was to analyze the risk of HPS in an endemic province of Argentina. For this, we explored the relationship between HPS cases occurring in Entre Ríos province between 2004 and 2015 and climate, vegetation, landscape, reservoir population, and rodent community characteristics by means of generalized linear models. We modeled HPS occurrence at each site, and both the incidence and number of cases grouped by department. The resulting best model of each analysis was applied in a GIS to build HPS risk maps. Risk of occurrence of HPS increased with tree cover and decreased with distance to rivers. We identified the south of Entre Ríos as the area with higher HPS risk, and therefore, where HPS prevention measures should be more urgently applied. Risk maps based on data available in the public domain are a useful tool that should be used by decision makers to concentrate surveillance and control efforts in those areas with highest HPS risk.

A Novel Hepacivirus in Wild Rodents from South America.

The Hepacivirus genus comprises single-stranded positive-sense RNA viruses within the family Flaviviridae. Several hepaciviruses have been identified in different mammals, including multiple rodent species in Africa, Asia, Europe, and North America. To date, no rodent hepacivirus has been identified in the South American continent. Here, we describe an unknown hepacivirus discovered during a metagenomic screen in Akodon montensis, Calomys tener, Oligoryzomys nigripes, Necromys lasiurus, and Mus musculus from São Paulo State, Brazil. Molecular detection of this novel hepacivirus by RT-PCR showed a frequency of 11.11% (2/18) in Oligoryzomys nigripes. This is the first identification of hepavivirus in sigmondonine rodents and in rodents from South America. In sum, our results expand the host range, viral diversity, and geographical distribution of the Hepacivirus genus.

Effective Population Size Differences in Calomys musculinus, the Host of Junín Virus: Their Relationship with the Epidemiological History of Argentine Hemorrhagic Fever.

Argentine hemorrhagic fever (AHF) is a serious endemic disease in Argentina, produced by Junín virus, whose host is the Sigmodontinae rodent Calomys musculinus. Within the endemic area, human incidence and proportion of infected rodents remains high for 5-10 years after the first appearance of the disease (epidemic [E] zone) and then gradually declines to sporadic cases (historic [H] zone). We tested the hypothesis that host populations within the E zone are large and well connected by gene flow, facilitating the transmission and maintenance of the virus, whereas those in the H and nonendemic (NE) zones are small and isolated, with the opposite effect. We estimated parameters affected by levels of gene flow and population size in 14 populations of C. musculinus: population effective size (Ne), genetic variability, and mean relatedness. Our hypothesis was not supported: the lowest levels of variability and of Ne and the highest genetic relatedness among individuals were found in the H zone. Populations from the NE zone displayed opposite results, whereas those in the E zone showed intermediate values. If we consider that populations are first NE, then E, and finally H, a correlative decrease in Ne was observed. Chronically infected females have a low reproductive success. We propose that this would lower Ne because each cohort would originate from a fraction of females of the previous generation, and affect other factors such as proportion of individuals that develop acute infection, probability of viral transmission, and evolution of virulence, which would explain, at least partly, the changing incidence of AHF.

A novel polyomavirus in sigmodontine rodents from São Paulo State, Brazil.

The nearly complete genome sequence of a novel polyomavirus from blood samples of Akodon montensis and Calomys tener collected in Brazil was determined by high-throughput sequencing. This virus showed a typical polyomaviruses genome organization, and it was classified as a member of the genus Betapolyomavirus. Our results expand the host range and viral diversity of the family Polyomaviridae.

Distributional ecology of Andes hantavirus: a macroecological approach.

BACKGROUND: Hantavirus pulmonary syndrome (HPS) is an infection endemic in Chile and Argentina, caused by Andes hantavirus (ANDV). The rodent Oligoryzomys longicaudatus is suggested as the main reservoir, although several other species of Sigmodontinae are known hosts of ANDV. Here, we explore potential ANDV transmission risk to humans in southern South America, based on eco-epidemiological associations among: six rodent host species, seropositive rodents, and human HPS cases. METHODS: We used ecological niche modeling and macroecological approaches to determine potential geographic distributions and assess environmental similarity among rodents and human HPS cases. RESULTS: Highest numbers of rodent species (five) were in Chile between 35° and 41°S latitude. Background similarity tests showed niche similarity in 14 of the 56 possible comparisons: similarity between human HPS cases and the background of all species and seropositive rodents was supported (except for Abrothrix sanborni). Of interest among the results is the likely role of O. longicaudatus, Loxodontomys micropus, Abrothrix olivaceus, and Abrothrix longipilis in HPS transmission to humans. CONCLUSIONS: Our results support a role of rodent species' distributions as a risk factor for human HPS at coarse scales, and suggest that the role of the main reservoir (O. longicaudatus) may be supported by the broader rodent host community in some areas.

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.

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.

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.

Spatial-Temporal Distribution of Hantavirus Rodent-Borne Infection by Oligoryzomys fulvescens in the Agua Buena Region--Panama.

BACKGROUND: Hotspot detection and characterization has played an increasing role in understanding the maintenance and transmission of zoonotic pathogens. Identifying the specific environmental factors (or their correlates) that influence reservoir host abundance help increase understanding of how pathogens are maintained in natural systems and are crucial to identifying disease risk. However, most recent studies are performed at macro-scale and describe broad temporal patterns of population abundances. Few have been conducted at a microscale over short time periods that better capture the dynamical patterns of key populations. These finer resolution studies may better define the likelihood of local pathogen persistence. This study characterizes the landscape distribution and spatio-temporal dynamics of Oligoryzomys fulvescens (O. fulvescens), an important mammalian reservoir in Central America. METHODS: Information collected in a longitudinal study of rodent populations in the community of Agua Buena in Tonosí, Panama, between April 2006 and December 2009 was analyzed using non-spatial analyses (box plots) and explicit spatial statistical tests (correlograms, SADIE and LISA). A 90 node grid was built (raster format) to design a base map. The area between the nodes was 0.09 km(2) and the total study area was 6.43 km(2) (2.39 x 2.69 km). The temporal assessment dataset was divided into four periods for each year studied: the dry season, rainy season, and two months-long transitions between seasons (the months of April and December). RESULTS: There were heterogeneous patterns in the population densities and degrees of dispersion of O. fulvescens that varied across seasons and among years. The species typically was locally absent during the late transitional months of the season, and re-established locally in subsequent years. These populations re-occurred in the same area during the first three years but subsequently re-established further south in the final year of the study. Spatial autocorrelation analyses indicated local populations encompassed approximately 300-600 m. The borders between suitable and unsuitable habitats were sharply demarcated over short distances. CONCLUSION: Oligoryzomys fulvescens showed a well-defined spatial pattern that evolved over time, and led to a pattern of changing aggregation. Thus, hot spots of abundance showed a general shifting pattern that helps explain the intermittent risk from pathogens transmitted by this species. This variation was associated with seasonality, as well as anthropogenic pressures that occurred with agricultural activities. These factors help define the characteristics of the occurrence, timing, intensity and duration of synanthropic populations affected by human populations and, consequently, possible exposure that local human populations experience.

Pinhal Virus, a New Arenavirus Isolated from Calomys tener in Brazil.

Arenavirus Sabiá was originally isolated from a fatal human infection in Brazil, and after the occurrence of the second fatal human case in São Paulo state, epidemiologic and virologic studies were performed in the area where the patient lived, aiming at the identification of the Sabiá natural rodent reservoir. A broadly cross-reactive enzyme-linked immunosorbent assay (ELISA) was used to screen for antibody-positive samples. Antibodies to arenavirus were detected in two of the 55 samples of Calomys tener, and from these results, samples of rodents were analyzed by a broad RT-PCR assay. RT-PCR amplification detected arenavirus sequences in five of the 55 C. tener samples, and sequencing showed that this virus is a distinct form of Sabiá virus. Thus, we describe here the evidence for the circulation of a new arenavirus in Brazil (proposed name Pinhal virus) and its genetic characterization compared to other arenaviruses. This study also suggests C. tener as a probable rodent reservoir for this virus and associates this new virus with the lineage C of New World arenaviruses. Although we have defined some characteristics of this virus, so far, there is no evidence of its involvement in human disease.

Phylogenetic Relationship of Necoclí Virus to Other South American Hantaviruses (Bunyaviridae: Hantavirus).

The results of a previous study suggested that Cherrie's cane rat (Zygodontomys cherriei) is the principal host of Necoclí virus (family Bunyaviridae, genus Hantavirus) in Colombia. Bayesian analyses of complete nucleocapsid protein gene sequences and complete glycoprotein precursor gene sequences in this study confirmed that Necoclí virus is phylogenetically closely related to Maporal virus, which is principally associated with the delicate pygmy rice rat (Oligoryzomys delicatus) in western Venezuela. In pairwise comparisons, nonidentities between the complete amino acid sequence of the nucleocapsid protein of Necoclí virus and the complete amino acid sequences of the nucleocapsid proteins of other hantaviruses were ≥8.7%. Likewise, nonidentities between the complete amino acid sequence of the glycoprotein precursor of Necoclí virus and the complete amino acid sequences of the glycoprotein precursors of other hantaviruses were ≥11.7%. Collectively, the unique association of Necoclí virus with Z. cherriei in Colombia, results of the Bayesian analyses of complete nucleocapsid protein gene sequences and complete glycoprotein precursor gene sequences, and results of the pairwise comparisons of amino acid sequences strongly support the notion that Necoclí virus represents a novel species in the genus Hantavirus. Further work is needed to determine whether Calabazo virus (a hantavirus associated with Z. brevicauda cherriei in Panama) and Necoclí virus are conspecific.

Daily Movements and Microhabitat Selection of Hantavirus Reservoirs and Other Sigmodontinae Rodent Species that Inhabit a Protected Natural Area of Argentina.

Abundance, distribution, movement patterns, and habitat selection of a reservoir species influence the dispersal of zoonotic pathogens, and hence, the risk for humans. Movements and microhabitat use of rodent species, and their potential role in the transmission of hantavirus were studied in Otamendi Natural Reserve, Buenos Aires, Argentina. Movement estimators and qualitative characteristics of rodent paths were determined by means of a spool and line device method. Sampling was conducted during November and December 2011, and March, April, June, October, and December 2012. Forty-six Oxymycterus rufus, 41 Akodon azarae, 10 Scapteromys aquaticus and 5 Oligoryzomys flavescens were captured. Movement patterns and distances varied according to sex, habitat type, reproductive season, and body size among species. O. flavescens, reservoir of the etiologic agent of hantavirus pulmonary syndrome in the region, moved short distances, had the most linear paths and did not share paths with other species. A. azarae had an intermediate linearity index, its movements were longer in the highland grassland than in the lowland marsh and the salty grassland, and larger individuals traveled longer distances. O. rufus had the most tortuous paths and the males moved more during the non-breeding season. S. aquaticus movements were associated with habitat type with longer distances traveled in the lowland marsh than in the salty grassland. Hantavirus antibodies were detected in 20% of A. azarae and were not detected in any other species. Seropositive individuals were captured during the breeding season and 85% of them were males. A. azarae moved randomly and shared paths with all the other species, which could promote hantavirus spillover events.

Patawa Virus, a New Arenavirus Hosted by Forest Rodents in French Guiana.

Molecular screening of rodents from French Guiana has detected a new arenavirus, named "Patawa," in two Oecomys species (Muridae, Sigmodontinae). Further investigations are needed to better understand the circulation of this virus in rodent and human populations and its public health impact.

Pinhal virus, a new arenavirus isolated from calomys tener in Brazil

Arenavirus Sabiá was originally isolated from a fatal human infection in Brazil, and after the occurrence of the second fatal human case in São Paulo state, epidemiologic and virologic studies were performed in the area where the patient lived, aiming at the identification of the Sabiá natural rodent reservoir. A broadly cross-reactive enzyme-linked immunosorbent assay (ELISA) was used to screen for antibody-positive samples. Antibodies to arenavirus were detected in two of the 55 samples of Calomys tener, and from these results, samples of rodents were analyzed by a broad RT-PCR assay. RT-PCR amplification detected arenavirus sequences in five of the 55 C. tener samples, and sequencing showed that this virus is a distinct form of Sabiá virus. Thus, we describe here the evidence for the circulation of a new arenavirus in Brazil (proposed name Pinhal virus) and its genetic characterization compared to other arenaviruses. This study also suggests C. tener as a probable rodent reservoir for this virus and associates this new virus with the lineage C of New World arenaviruses. Although we have defined some characteristics of this virus, so far, there is no evidence of its involvement in human disease.

[Conditions for the transmission of Hantavirus in Rio Negro, Argentina]. / Condiciones para la transmision del hantavirus en zona andina de Río Negro, Argentina.

Hantavirus Pulmonary Syndrome (HPS) is a disease of viral etiology that affects humans causing severe acute respiratory symptoms. In Patagonia the disease is caused by the Andes Virus (AND) and transmitted by the rodent Oligoryzomys longicaudatus. The aim of this study was to identify those human activities that increase the risk of exposure to rodents, what we call "contagious scenarios". A retrospective study was performed with data obtained from cases in Rio Negro, which included clinic-epidemiological records and ecological/environmental assessment reports. The following variables were considered: age, sex, season, percentage of urbanization, geographic location, human settlements in rodent infested areas, probable source of exposure, type of activity and level of sanitary development. In total 32 cases were studied. Exposure was verified in 18 (56.2 %) cases in rural areas and 10 cases (31.3%) in small rural towns. In relation to anthropogenic environment, 24 (75%) cases were reported in developed settlements and 8 cases (25%) were related to slightly modified areas. Major exposition in El Bolson identified 8 cases of indoor activities of the total 18 reported in the area (44.5%), while in Bariloche 8 (57.1%) cases out of 14 were reported in outdoor surroundings. In general, activities that generated greater risk were work-related, accounting for 23 (71.9%) cases while 7 were related to recreational activities (28.1%). The identification of "contagious scenarios" at local level provided information for an effective application of available resources in terms of prevention and sanitary education.

Condiciones para la transmision del hantavirus en zona andina de Río negro, Argentina

El Síndrome Pulmonar por Hantavirus (SPH) es una enfermedad de etiología viral que causa en el hombre un cuadro respiratorio grave. En Patagonia, la enfermedad es causada por el virus Andes Sur (AND), transmitido por el roedor Oligoryzomys longicaudatus. El objetivo del presente trabajo fue identificar las actividades del hombre que favorecen su exposición a roedores, denominados escenarios de contagio. Se realizó un estudio retrospectivo a partir de información recolectada en investigaciones de casos ocurridos en Río Negro, mediante Fichas Clínico-Epidemiológicas e informes de evaluación ecológico/ambiental. Se definieron como variables a ser consideradas: edad, sexo, época del año, grado de urbanización, localización geográfica, integración del hombre al hábitat de roedores, fuente probable de exposición, actividad humana y nivel de saneamiento. Se estudiaron 32 casos. La exposición rural se verificó en 18 (56.2%) de los casos y 10 (31.3%) en paraje rural (grupo de viviendas en zona rural). En relación al ambiente antropogénico 24 (75%) resultaron en ambientes modificados por el hombre y 8 (25%) en áreas poco modificadas. El sitio de exposición de mayor importancia en El Bolsón fue el interior de edificaciones en 8 de los 18 casos allí registrados (44.5%), mientras que en Bariloche fueron ambientes de exterior con 8/14 (57.1%) casos. La actividad de riesgo fue laboral en 23 (71.9%) de los casos y recreacional en 7 (28.1%). Determinar los escenarios de contagio a nivel local ha aportado información para aplicar todos los recursos disponibles en materia de prevención y educación sanitaria.

Hantavirus Pulmonary Syndrome (HPS) is a disease of viral etiology that affects humans causing severe acute respiratory symptoms. In Patagonia the disease is caused by the Andes Virus (AND) and transmitted by the rodent Oligoryzomys longicaudatus. The aim of this study was to identify those human activities that increase the risk of exposure to rodents, what we call "contagious scenarios". A retrospective study was performed with data obtained from cases in Rio Negro, which included clinic-epidemiological records and ecological/environmental assessment reports. The following variables were considered: age, sex, season, percentage of urbanization, geographic location, human settlements in rodent infested areas, probable source of exposure, type of activity and level of sanitary development. In total 32 cases were studied. Exposure was verified in 18 (56.2 %) cases in rural areas and 10 cases (31.3%) in small rural towns. In relation to anthropogenic environment, 24 (75%) cases were reported in developed settlements and 8 cases (25%) were related to slightly modified areas. Major exposition in El Bolson identified 8 cases of indoor activities of the total 18 reported in the area (44.5%), while in Bariloche 8 (57.1%) cases out of 14 were reported in outdoor surroundings. In general, activities that generated greater risk were work-related, accounting for 23 (71.9%) cases while 7 were related to recreational activities (28.1%). The identification of "contagious scenarios" at local level provided information for an effective application of available resources in terms of prevention and sanitary education.

Rapid enzyme-linked immunosorbent assay for the detection of hantavirus-specific antibodies in divergent small mammals.

We assessed the utility of an enzyme-linked immunosorbent assay (ELISA) for the detection of hantavirus-specific antibodies from sera of Oligoryzomys longicaudatus, the principal reservoir of Andes virus (ANDV), using an antigen previously developed for detection of antibodies to Sin Nombre virus (SNV) in sera from Peromyscus maniculatus. The assay uses a protein A/G horseradish peroxidase conjugate and can be performed in as little as 1.5 hours. Serum samples from Oligoryzomys longicaudatus collected in central-south Chile were used and the assay identified several that were antibody positive. This assay can be used for the rapid detection of antibodies to divergent hantaviruses from geographically and phylogenetically distant rodent species.