Gladys E. Calderón

Genetic Diversity and Epidemiology of Hantaviruses in Argentina

Phylogenetic analysis of a 292-nucleotide (nt) fragment of the hantavirus M genome segment from 36 rodent and 13 human samples from three known foci of hantavirus infection in Argentina was conducted. A 1654-nt fragment of the M genome segment was analyzed for 1 representative of 7 genetically distinct hantavirus lineages identified. Additionally, the nt sequence of the complete M genome segments of Lechiguanas, Oran, and Hu39694 hantavirus genotypes was determined. nt sequence comparisons reveal that 7 hantavirus lineages from Argentina differ from each other by 11.5%-21.8% and from Sin Nombre, Bayou, and Black Creek Canal viruses by 23.8%-26.5%. Phylogenetic analyses demonstrate that they form a unique, separate branch within the clade containing other New World sigmodontine-borne hantaviruses. Most Oligoryzomys-borne hantavirus genotypes clearly map together. The Oligoryzomys-borne genotypes Lechiguanas, Oran, and Andes appear to be associated with human disease. Oligoryzomys longicaudatus was identified as the likely rodent reservoir for Andes virus.

Yellow Pygmy Rice Rat (Oligoryzomys flavescens) and Hantavirus Pulmonary Syndrome in Uruguay

During 5,230 trapping nights, 672 small mammals were trapped in the areas where most hantavirus pulmonary syndrome (HPS) cases occur in Uruguay. Yellow pygmy rice rats (Oligoryzomys flavescens) were the only rodents that showed evidence of antibodies to hantavirus, with a seroprevalence of 2.6%. The rodents were trapped in all the explored environments, and most of the seropositive rodents were found in habitats frequented by humans. Nucleotide sequences were obtained from four HPS case-patients and four yellow pygmy rice rats of the M genome segment. Sequence comparison and phylogenetic analysis showed that rodent-borne viruses and viruses from three HPS case-patients form a well-supported clade and share a 96.4% identity with the previously characterized Central Plata hantavirus. These results suggest that yellow pygmy rice rat (O. flavescens) may be the host for Central Plata, a hantavirus associated with HPS in the southern area of Uruguay.[

Hantavirus infection in humans and rodents, northwestern Argentina.

We initiated a study to elucidate the ecology and epidemiology of hantavirus infections in northern Argentina. The northwestern hantavirus pulmonary syndrome (HPS)–endemic area of Argentina comprises Salta and Jujuy Provinces. Between 1997 and 2000, 30 HPS cases were diagnosed in Jujuy Province (population 512,329). Most patients had a mild clinical course, and the death rate (13.3%) was low. We performed a serologic and epidemiologic survey in residents of the area, in conjunction with a serologic study in rodents. The prevalence of hantavirus antibodies in the general human population was 6.5%, one of the highest reported in the literature. No evidence of interhuman transmission was found, and the high prevalence of hantavirus antibody seemed to be associated with the high infestation of rodents detected in domestic and peridomestic habitats.

Temporal and Spatial Host Abundance and Prevalence of Andes Hantavirus in Southern Argentina

Andes virus (AND) is a hantavirus hosted by the sigmodontine rodent Oligoryzomys longicaudatus in southern Argentina, where it is responsible for most cases of hantavirus pulmonary syndrome (HPS). Our study provides data about the spatial variation in abundance of the rodent host of AND hantavirus. We report results of a longitudinal study performed in a locality of the Andean region of Chubut Province. From November 2003 (spring) to July 2006 (winter), O. longicaudatus was the most common species captured (63%) and it showed significant differences in abundance among habitats and seasons. Most antibody-positive rodents were O. longicaudatus (9.2%), followed by A. longipilis (3.6%) and A. olivaceus (1.5%). The highest number of antibody-positive animals was observed for males that belonged to the heaviest mass classes. Antibody-positive O. longicaudatus were more abundant in brush habitats. We found low richness of rodents and abundance of O. longicaudatus in areas affected by anthropogenic activity. The infection seems to be regionally persistent, but the risk to humans in a landscape would be localized. To develop accurate models for predicting HPS outbreaks, further research is needed to characterize rodent movement patterns across the landscape.

Evaluation of an enzyme-linked immunosorbent assay for detection of antibodies to Junin virus in rodents

Junin virus is the etiological agent of Argentine hemorrhagic fever, a serious rodent-borne disease. An enzyme-linked immunosorbent assay (ELISA) to detect Junin virus IgG antibodies in rodents was evaluated using sera from 27 Calomys musculinus and five Calomys laucha, inoculated experimentally with a live attenuated strain of this arenavirus. The test performance was compared against an indirect immunofluorescence assay (IFA). The ELISA had a sensitivity and specificity of 100% and a reproducibility of 87.9% for samples with titers above the selected cut-off value. IFA had lower sensitivity (53%) with the same specificity. The ELISA results were similar, whether carried out on whole blood or serum samples, thus eliminating the need for serum separation. A high correlation (K=0.86) between ELISA and IFA results was obtained from 1011 wild sigmodontine and murine rodents collected within and outside of the Argentine hemorrhagic fever endemic area. These results indicate that Junin virus IgG ELISA is the most suitable assay for detection of Junin virus antibodies in rodent samples.

New strains of Culex flavivirus isolated in Argentina.

ABSTRACT Strains of Culex flavivirus (CxFV), an insect virus isolated initially from Japan, were isolated from different species of Culex sp. mosquitoes collected in Corrientes province, Argentina, during 2009. CxFV was detected by reverse transcription polymerase chain reaction and by isolation in C6/36 cell culture. Phylogenetic analysis of nucleotide sequences showed that these strains are related closely to a CxFV strain isolated from Trinidad. Our study represents the first report of CxFV isolation and characterization in Argentina from the same geographic area where West Nile Virus has been detected. Further evaluation and viral competition studies will be necessary to determine the impact of this insect flavivirus on an infection caused by other pathogenic flaviviruses.

Isolation of Yellow Fever Virus from Mosquitoes in Misiones Province, Argentina

Yellow fever (YF) is a viral hemorrhagic fever endemic to tropical regions of South America and Africa. From 2007 to 2009 an important epidemic/epizootic of YF was detected in different populations of howler monkeys (Alouatta species) in Misiones, a northeastern Argentinian province. Yellow fever virus (YFV) infection was researched and documented by laboratory tests in humans and in dead Alouatta carayá. The objective of that research was to investigate the circulation of YFV in mosquitoes, which could be implicated in the sylvatic transmission of YF in Argentina. The above-mentioned mosquitoes were captured in the same geographical region where the epizootic took place. A YFV strain was isolated in cell culture from pools of Sabethes albiprivus. This study is not only the first isolation of YFV from mosquitoes in Argentina, but it is also the first YFV isolation reported in the species Sabethes albiprivus, suggesting that this species might be playing a key role in sylvatic YF in Argentina.

Detection of the mosquito-borne flaviviruses, West Nile, Dengue, Saint Louis Encephalitis, Ilheus, Bussuquara, and Yellow Fever in free-ranging black howlers (Alouatta caraya) of Northeastern Argentina

Several medically important mosquito-borne flaviviruses have been detected in Argentina in recent years: Dengue (DENV), St. Louis encephalitis (SLEV), West Nile (WNV) and Yellow Fever (YFV) viruses. Evidence of Bussuquara virus (BSQV) and Ilheus virus (ILHV) activity were found, but they have not been associated with human disease. Non-human primates can act as important hosts in the natural cycle of flaviviruses and serological studies can lead to improved understanding of virus circulation dynamics and host susceptibility. From July–August 2010, we conducted serological and molecular surveys in free–ranging black howlers (Alouatta caraya) captured in northeastern Argentina. We used 90% plaque-reduction neutralization tests (PRNT90) to analyze 108 serum samples for antibodies to WNV, SLEV, YFV, DENV (serotypes 1and 3), ILHV, and BSQV. Virus genome detection was performed using generic reverse transcription (RT)-nested PCR to identify flaviviruses in 51 antibody-negative animals. Seventy animals had antibodies for one or more flaviviruses for a total antibody prevalence of 64.8% (70/108). Monotypic (13/70, 19%) and heterotypic (27/70, 39%) patterns were differentiated. Specific neutralizing antibodies against WNV, SLEV, DENV-1, DENV-3, ILHV, and BSQV were found. Unexpectedly, the highest flavivirus antibody prevalence detected was to WNV with 9 (8.33%) monotypic responses. All samples tested by (RT)-nested PCR were negative for viral genome. This is the first detection of WNV-specific antibodies in black howlers from Argentina and the first report in free-ranging non-human primates from Latin-American countries. Given that no animals had specific neutralizing antibodies to YFV, our results suggest that the study population remains susceptible to YFV. Monitoring of these agents should be strengthened to detect the establishment of sylvatic cycles of flaviviruses in America and evaluate risks to wildlife and human health.

On the relationship between the environmental history and the epidemiological situation of Argentine hemorrhagic fever

The aim of this work was to establish the relationship between different Argentine hemorrhagic fever (AHF) epidemiological situations found at different sites and the related large-scale environmental conditions. Large-scale environmental records (vegetation index, temperature, precipitation and elevation) were obtained from a series of monthly NOAA satellite images and global databases considered suitable for modeling climatic and other environmental determinants of large-scale biogeographical regions. The temporal variation in vegetation for cycles of winter-summer showed a greater variation in the nonendemic region than in the other two regions. On the other hand, the average of the temporal variation in precipitation in cycles of spring–autumn was more different in the historic region than in the other two regions, and land surface temperatures in cycles of spring–autumn showed differences between the epidemic region and the other two regions. We found good separation among the epidemic, historic and nonendemic sites, with the greatest difference found between epidemic and nonendemic sites. The classification of sites showed a tendency for grouping according to the epidemiological situation, but there was some variation. It seems possible to establish a close relationship between the state of AHF incidence and the environmental history of sites suggesting the possibility of predicting epidemiological behavior using environmental conditions derived from satellite data.

Dynamics of Oliveros Virus Infection in Rodents in Central Argentina

Oliveros virus (OLV) is an arenavirus hosted by the sigmodontine rodent, Necromys benefactus, in central Argentina. We report a 3-year longitudinal field study of the dynamics of OLV infection in host populations from 15 localities in two provinces on the central Argentine pampa. There was an overall 3-year period immunofluorescent antibody prevalence of 25% in the host population, and infected hosts were found throughout the study area. Spill-over infection into common sympatric species was rare. Infection dynamics exhibited many of the patterns seen for other rodent-borne arenaviruses and hantaviruses, but had some unique characteristics. Host population density was highest in autumn and lowest in spring, while antibody prevalence was highest in spring and lowest in autumn. Virus transmission was horizontal: infection was strongly associated with age, reaching 45% prevalence in the oldest individuals, and prevalence of infection was equal among male and female hosts. Infection may have been associated with scars, which were also approximately equally distributed among male and female Necromys.