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Aedes (Stegomyia) aegypti (L.) (Diptera: Culicidae) is the vector of multiple arboviruses. To evaluate the association between environmental factors and the oviposition activity of Ae. aegypti in Argentina, data on the presence and abundance of eggs were collected using ovitraps, between September of 2018 and May of 2019, in the cities of Villa María, Río Cuarto and Salsipuedes (Córdoba province, Argentina). We analysed the relationships between oviposition and five environmental factors: Temperature, precipitation, vegetation cover, human population density and distance to sites with a potential high density of larval habitats, like cemeteries and trash dumps. Environmental factors' data were collected using satellite image products. The oviposition activity was randomly distributed in three cities. Using generalized linear mixed models, we show that the house where each ovitrap was placed was a source of variability in oviposition, suggesting the relevance of microsite factors and the importance of domestic control actions. Ae. aegypti oviposition was positively correlated with night-time temperature of the previous 3 weeks, and in a context-dependent manner, it was positively correlated with human population density, vegetation cover and precipitation. The consistency and magnitude of these relationships varied between cities, indicating that oviposition is related to a complex system of environmental variables.

We evaluated the seroprevalence of Saint Louis encephalitis virus (SLEV) and West Nile virus (WNV) in dogs and cats in Córdoba, Argentina. Monotypic and heterotypic serological patterns were differentiated by means of a neutralization test. The SLEV seroprevalence in dogs was 14.6% (44/302; 100% monotypic). Two out of 94 (2.1%, 100% monotypic) cats were positive for WNV only. Four dogs (1.3%) exhibited neutralizing antibody titers against SLEV and WNV. During the study, three dogs seroconverted to SLEV. Our study demonstrates that pets were useful for detecting viral activity and could be considered as sentinels in the local surveillance of SLEV and WNV.

Flaviviruses are responsible for several emerging mosquito-borne diseases in Argentina and in the Americas. Some of these diseases are zoonosis and were detected in recent years: West Nile virus (WNV) and Saint Louis encephalitis virus (SLEV) among them. WNV originated in Africa was detected for the first time in America in 1999 and it was first isolated from diseased and dead horses in 2006 in Argentina. Neutralizing antibodies from wild birds, humans and horses have been also detected in later years. SLEV re-emerged as a human pathogen in Argentina with the first epidemic in 2005. During this outbreak, viral strains were isolated from Culex mosquitoes and several studies have detected neutralizing antibodies from birds, human and horses, in the last ten years.The aim of this work is to provide information on the activity of the Flavivirus SLEV and WNV, identifying the infection in horses in three localities of the province of Chaco. From May 2013 to May 2014, serological surveys from 68 asymptomatic horses were conducted. Antibodies against SLEV (25 %) and WNV (8.82%) were detected. This is the first report of WNV circulation in equines from Chaco province and circulation of both viruses during the study period was demonstrated. Fil: Oria, Griselda Inés. Universidad Nacional del Nordeste. Instituto de Medicina Regional; Argentina. Fil: Spinsanti, Lorena Ivana. Universidad Nacional de Córdoba. Facultad de Ciencias Médicas; Argentina. Fil: Pirota, Verónica Laura. Ministerio de la Producción. Dirección de Fauna; Argentina. Fil: Martínez, María Silvana. Universidad Nacional del Nordeste. Facultad de Ciencias Económicas; Argentina. Fil: Stechina, Ornela Sofía. Universidad Nacional del Nordeste. Instituto de Medicina Regional; Argentina. Fil: Stechina, Ornela Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste; Argentina. Fil: Etchepare, Eduardo Gabriel. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas y Naturales y Agrimensura; Argentina. Fil: Contigiani, Marta Silvia. Universidad Nacional de Córdoba. Facultad de Ciencias Médicas; Argentina. Fil: Stein, Marina. Universidad Nacional del Nordeste. Instituto de Medicina Regional; Argentina. Fil: Stein, Marina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste; Argentina.

Saint Louis encephalitis virus (SLEV) and West Nile virus (WNV) are two of the major causes of arboviral encephalitis in the Americas. The co-circulation of related flaviviruses in the Americas and prior vaccination against flaviviruses pose problems to the diagnostic specificity of serological assays due to the development of cross-reactive antibodies. An accurate diagnosis method capable of differentiating these related viruses is needed. NS1 is a glycosylated, nonstructural protein, of about 46 kDa which has a highly conserved structure. Anti-NS1 antibodies can be detected within 4?8 days after the initial exposure and NS1 is the least cross-reactive of the flaviviral antigens. This study was aimed to generate SLEV and WNV NS1 recombinants proteins for the development of a flavivirus diagnostic test. Local Argentinian isolates were used as the source of NS1 gene cloning, expression, and purification. The protein was expressed in Escherichia coli as inclusion bodies and further purified by metal-chelating affinity chromatography (IMAC) under denaturing conditions. Human sera from SLEV and WNV positive cases showed reactivity to the recombinant NS1 proteins by western blot. The unfolded NS1 proteins were also used as immunogens. The polyclonal antibodies elicited in immunized mice recognized the two recombinant proteins with differential reactivity. Fil: Lorch, Matías Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; Argentina Fil: Collado, Maria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; Argentina Fil: Argüelles, Marcelo Horacio. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Rota, Rosana Paola. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Spinsanti, Lorena Ivana. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología Dr. J. M. Vanella; Argentina Fil: Lozano, Mario Enrique. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Goñi, Sandra Elizabeth. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Saint Louis encephalitis virus (SLEV) reemerged in South America, and caused encephalitis outbreaks at the beginning of the 21st century. To enhance our knowledge about SLEV virulence, we performed comparative pathogenesis studies in Swiss albino mice inoculated with two different variants, the epidemic strain CbaAr-4005 and the non-epidemic strain CorAn-9275. Only the infection of mice with SLEV strain CbaAr-4005 resulted in high viremia, invasion of peripheral tissues including the lungs, kidney, and spleen, and viral neuroinvasion. This was associated with inflammatory pathology in the lungs, spleen, and brain as well as morbidity and mortality. In contrast, neither signs of desease nor viral replication were observed in mice infected with strain CorAn-9275. Interestingly, important loss of B cells and development of altered germinal centers (GC) were detected in the spleen of mice infected with strain CbaAr-4005, whereas mice infected with SLEV CorAn-9275 developed prominent GC with conserved follicular architecture, and neutralizing antibodies. Fil: Rivarola, María Elisa. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología ; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina Fil: Albrieu Llinás, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología ; Argentina Fil: Pisano, María Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología ; Argentina Fil: Tauro, Laura Beatriz. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología ; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina Fil: Gorosito Serran, Melisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas; Argentina Fil: Beccaria, Cristian Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas; Argentina Fil: Diaz, Luis Adrian. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología ; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina Fil: Vázquez, Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología ; Argentina Fil: Quaglia, Agustín Ignacio Eugenio. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología ; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina Fil: López, Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Médicas; Argentina Fil: Spinsanti, Lorena Ivana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología ; Argentina Fil: Gruppi, Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas; Argentina Fil: Contigiani de Minio, Marta Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología ; Argentina

St. Louis encephalitis virus (SLEV) and West Nile virus (WNV) present ecological and antigenic similarities and are responsible for serious human diseases. In addition, WNV is a significant pathogen in terms of equine health. The purpose of our study was to analyse the seroprevalence of SLEV and WNV in equine sera collected in Santa Fe Province, Argentina. The seroprevalence determined using the plaque reduction neutralisation test was 12.2% for SLEV, 16.2% for WNV and 48.6% for a combination of both viruses. These results provide evidence of the co-circulation of SLEV and WNV in equines in Santa Fe.

In the central area of Argentina, epidemiological and molecular characteristics of human enterovirus infections are still unknown. RT-nested PCR of the highly conserved 5'NCR was used to detect enteroviruses in 168 samples of cerebrospinal fluid from hospitalized patients with suspected infection of the central nervous system (2007-2008), and 13 (7.7%) were positive. Molecular typing was performed by sequencing of the 3'-half VP1 region. Echovirus 30 was the predominant type detected, followed by coxsackie viruses A9 and B4. All echovirus 30 strains of 2007 clustered in lineage H, whereas the echovirus 30 isolate obtained in 2008 was more distantly related, possibly representing a new lineage.

St. Louis encephalitis virus (SLEV) is endemic in Argentina. The first outbreak was recorded in Córdoba during the late summer of 2005. This study analyzes the association between social and demographic factors and other variables related to exposure to mosquitoes with SLEV infection in 264 individuals who attended two health centers in the city of Córdoba during the period December 2004-January 2005. Multiple logistic regression models were used to estimate the risk coefficients. The seroprevalence was 12.9%, similar to that in previous reports. Risks of infection were associated with the presence of garbage dumps near dwellings, the practice of outdoor activities at night and place of residence. Risk for older people (60-80 years old) was moderate. The identification of risk factors related to SLEV infection would be useful to improve programs for vector control and community health.

A serological screening was performed in 615 individuals aged 0-87 years, living in the city of Cordoba, Argentina to study the relationship between antibody prevalence for the SLE virus and age. A 13.98% prevalence of neutralizing antibodies was obtained and its relation to age was significantly high (p = 0.045). The highest seroprevalence was noted on individuals over 60 years old (>20%), whereas no subject under 10 was seropositive for this virus. Our results confirm that the agent is endemic in this area and neurological pathology studies should be performed on those individuals aged 60 since they represent the most susceptible group to SLE virus.

To the Editor: West Nile virus (WNV), genus Flavivirus, family Flaviviridae has been rapidly dispersing through the Americas since its introduction in 1999 in New York (1). By 2004, serologic studies detected WNV-specific antibodies in birds and horses from Canada to northern South America (2–4). The first report of WNV activity in the Southern Cone of South America surfaced in April 2006, when 3 horses died in Argentina (5). However, established transmission foci in Argentina are unknown. We report evidence for the introduction and establishment of WNV in Argentina as early as January 2005. Serum samples from free-ranging birds were collected from 5 locations in Argentina and screened for generic flavivirus antibodies by using a blocking ELISA with monoclonal antibody 6B6C-1 (6). Positive serum specimens were further characterized by plaque-reduction neutralization test (PRNT). We identified the etiologic agent responsible for the previous flavivirus infection by using the following criteria: 80% neutralization of reference virus (WNV NY99-4132 or an Argentinean strain of St. Louis encephalitis virus [SLEV CbaAr4005]) in serum diluted at least 1:40 and 4-fold greater titer compared with the other virus. Overall, 474 (25.6%) of 1,845 serum specimens from 117 bird species collected from January to June 2006 tested positive when using the blocking ELISA; 30% inhibition was the threshold for a positive test. SLEV infections were confirmed in 105 birds by PRNT; WNV infections were confirmed in 43 birds. Anti-WNV antibody titers ranged from 40 to 2,560 in birds collected as early as January 2005 in Cordoba City and as late as June 2006 in Mar Chiquita (Table). Recent WNV activity was indicated by seroconversion in 3 banded rufous hornero in Cordoba City between January and March 2005. Although 659 (1.5%) of serum samples were positive for SLEV, no WNV infection was detected in free-ranging birds collected in 2004. As early as January 2005, WNV was detected in a seroconversion so we suspect WNV was introduced before 2005 at the end of 2004 in all 5 sampling locations and in a variety of ecosystems: Cordoba, periurban (1.1%, 6/543); Mar Chiquita, thorn forest (5.1%, 16/313); Monte Alto, semidry chaco forest (9.8%, 8/82); Montecristo, cropland (9.5%, 2/21); and San Miguel de Tucuman, periurban yungas foothills (4.9%, 12/227). Table Prevalence of West Nile virus–neutralizing antibodies among birds grouped by taxonomic family, sampled in Chaco, Cordoba, and Tucuman Provinces, Argentina, 2004–2006* In 2006, WNV was isolated from equines in Buenos Aires province (5). WNV transmission to resident birds collected further north in Cordoba, Chaco, and Tucuman provinces was detected in 2005 and 2006. Our data suggest that WNV was introduced into Argentina before 2005 and maintained naturally in enzootic foci where numerous bird species from many families were exposed. Presumably, as in North America, locally abundant passerine birds such as turdids (thrushes) are amplifying hosts. If common species of the Furnariidae (a family absent from temperate North America) prove to be competent hosts, they could play an important role in WNV transmission in Argentina because of their frequent exposure to WNV. Twelve (12.5%) of 96 F. ruffus sampled in 2005 and 2006 tested positive. How WNV reached Argentina may never be known. Dispersal by migrating birds is a popular hypothesis, although relatively few North American breeding birds migrate to Argentina, and austral migrants number fewer than boreal migrants. Komar and Clark (2) suggested that bird species in the order Charadriiformes, such as shorebirds and terns, are candidates for carrying WNV from North America to South America due to long lasting high-level viremias, occasional persistent infectious viral loads in skin, and direct, long-distance flights. WNV spread southward from the United States to northern South America between 1999 and 2004 following a stepping stone pattern, consistent with spread by birds. Moreover, introduction of WNV into Argentina by migratory birds could explain the presence of the virus in many places in a brief period. However, for migratory birds (211 serum samples tested) in this study, serologic test results were negative. The high titers of WNV-reactive antibody are strongly indicative of WNV infections. Overall, 216 serum specimens reacted by PRNT test against SLEV, WNV or both at titers ≥20. Sixty-eight serum samples remain unidentified. The large number of unidentified flavivirus-positive samples detected by PRNT, ELISA, or both (148/474) could be due to 1) false positives; 2) cross-reactions between WNV- and SLEV-reactive antibodies that prevented definitive diagnosis by PRNT; 3) cross-reactive antibody and multiple, heterologous flavivirus infections; 4) previous infections by both WNV and SLEV; and/or 5) presence of other flaviviruses circulating in Argentina. SLEV is endemic throughout Argentina and, like WNV, belongs to the Japanese encephalitis virus serocomplex. Hemagglutination-inhibiting antibodies against several Brazilian flaviviruses (e.g., Bussuquara, Ilheus, Rocio viruses) have been reported in the neotropical region of extreme northern Argentina (7), but these viruses have not been isolated in Argentina. Our serologic data suggest that WNV has established itself in 4 ecologic regions in Argentina in a brief period. Additional studies are needed to define the reservoir hosts and vectors of WNV in Argentina, and most importantly, to define the public health risk this virus represents.

Twenty-six years after it was last detected, Saint Louis encephalitis virus (SLEV) genotype III reemerged in 2005 in Córdoba, Argentina, where it caused an outbreak. Two genotype III SLEV strains were isolated from Culex quinquefasciatus. A 71.43% prevalence for neutralizing antibodies was found in domestic fowl in the homestead of a patient with encephalitis.

Hepatitis A virus (HAV) has shown intermediate endemicity in Argentina, but notification of clinical cases has decreased since the introduction of the vaccine in 2005.In order to get insight into the local circulation of this virus after four years of the official introduction of the vaccine, the aims of this study were to provide information on HAV immune status of the adult population of Córdoba city and to conduct environmental surveillance of HAV in sewage and river samples in the same region.The prevalence of anti-HAV was determined by EIA in 416 samples of people (without prior vaccination) from Córdoba city (2009-2010). Spline regression models were estimated under generalized additive models. Environmental surveillance was conducted in river and sewage samples collected in the same period. Viral detection was performed by RT-Nested PCR of the 5'UTR.In Córdoba, the global prevalence of anti-HAV was 73.5%. It increased with age (p0.0001) and it was associated with the low-income population (OR: 1.14; 95% CI 1.05-1.25). This prevalence decreased in younger age groups, especially in the high-income population. Environmental monitoring revealed the presence of HAV (IA) in 20.8% and 16.1% of wastewater and river samples, respectively.As a consequence of a decrease in HAV circulation due to improvements in immunization, socio-economic and hygienic conditions, young adults are becoming increasingly susceptible to HAV infections. Environmental monitoring demonstrated that HAV circulates in the local population; therefore, health care systems should consider the implementation of preventive measures for susceptible adults in order to reduce the risk of HAV infection.

To the Editor: St. Louis encephalitis is a mosquito-borne viral disease that affects humans. The causative agent, SLEV (formal name: Saint Louis encephalitis virus), is a member of the Flaviviridae family. Severity of the clinical syndromes increases with age, and persons >60 years old have the highest frequency of encephalitis. The primary transmission cycle involves wild passeiform, columbiform birds, and Culex sp. mosquitoes (1). In Argentina, an urban cycle may involve Cx. quinquefasciatus, which is a source of a viral isolate, and abundant birds (house sparrows, doves, or chickens) (2). The distribution of SLEV in Argentina is wide; seroprevalence ranges from 3% to 50% of the country’s population (3). Spinsanti et al. reported results of a serologic screening in persons ages 0–87 years who live in the city of Cordoba; antibodies were most frequently found in persons >60 years of age (4). However, cases of St. Louis encephalitis reported in Argentina are very rare. Two cases with serologic diagnosis were reported in 1964 and 1968, respectively (2). In 1971, two more cases were diagnosed on the basis of viral isolation (5). Finally, the last case reported was a patient with meningoencephalitis diagnosed in the province of Buenos Aires by hemagglutination inhibition assay (6). Herein, we report a case of Saint Louis encephalitis that occurred in the province of Cordoba, Argentina. A 61-year-old man was admitted to the hospital in February 2002, complaining of headache, fever, and diplopia. He had been well until 3 months before admission, when ophthalmic herpes zoster was diagnosed. He underwent therapy with oral acyclovir and had a good clinical outcome. Ten days before admission, he developed unstable gait with misbalance and hand tremors, mainly at his left side. On admission, he had occipital headache, diplopia, and nausea and vomiting associated with high fever and chills. Somnolence appeared a few hours before the consultation. The patient was a right-handed businessman, a native of Cordoba. He was married and had no risk factors for sexually transmitted diseases. He had not traveled inside or outside the country during the last year. He lived near a river with a high-density population of mosquitoes. Vital signs on admission showed axillary temperature of 39°C, pulse of 90 beats per minute, respiratory frequency of 20 per minute, and blood pressure of 110/70 mmHg. Physical examination demonstrated a somnolent patient who was easily aroused and oriented. His speech was slurred. Results of a fundoscopic examination appeared normal. Results of a cranial-nerve examination showed horizontal left diplopia with left sixth nerve paresia. A resting, postural, and intentional hand tremor was evident. Motor strength was 5/5 throughout with normal bulk and tone, tendon reflexes, and coordination. Examination of sensitivity showed no abnormalities. A slight neck rigidity was detected. Routine laboratory analysis was unremarkable, and results of serologic tests for coxsackie virus, echovirus, and HIV were negative. HIV-1 RNA by polymerase chain reaction (PCR) and p24 antigen were also negative. Cerebrospinal fluid study revealed a leukocyte count of 18/mm3 (80% lymphocytes), a glucose level of 48 mg/dL, and a protein level of 87 mg/dL. Cryptococcal antigen, antibodies for syphilis, Human herpesvirus 1 and 2, and PCR for varicella-zoster virus 1 and Human herpesvirus were also negative. Results of an electroencephalogram and a chest radiograph were normal. Therapy with intravenous acyclovir was initiated. A magnetic resonance imaging (MRI) scan of the brain showed a striking signal change on T2 in the substantia nigra of the midbrain, mainly at the right side. The patient continued febrile, diplopia disappeared, and meningeal signs progressed with frank cervical stiffness, positive Kerning sign, and photophobia. Diffuse tremulousness and axial rigidity appeared. Upper extremities showed rigidity with cogwheel phenomenon. Conversely, lower extremities showed spasticity with bilateral Babinski sign. Tendon reflexes became enhanced. His gait showed retropulsion with wide base sustentation. Dysdiadochokinesia appeared. On the third day, a new lumbar puncture showed worse results: a leukocyte count of 210/mm3 (82% lymphocytes), a glucose level of 51 mg/dL, and a protein level of 106 mg/dL. Another electroencephalographic examination showed unspecific centroparietal disorganization with right side predominance. Intravenous acyclovir was stopped. On the 5th day, the patient began to recover; he was discharged on the 10th day. After 3 months of follow-up, only left arm rigidity and a left hand tremor persisted. Acute- and convalescent-phase serum samples (taken 10 and 16 days after onset of illness, respectively) were sent to the Arbovirus and Arenavirus Disease Laboratory, Instituto de Virologia, Cordoba. SLEV immunoglobulin (Ig) M antibodies were positive by indirect immunofluorescence assay (IFA). Seroconversion for IgG antibodies was demonstrated by IFA (7) and hemagglutination inhibition assay, with titers of 640 and 80 in the first sample and 2,560 and 320 in the second sample. These results were confirmed by neutralization test using the reduction of plates technique in Vero cells culture, as described (8). Eastern equine encephalomyelitis virus and Western equine encephalomyelitis viruses with known circulation in Argentina were included in the assay with negative results (3). An increase in antibodies titers between acute- (320) and convalescent-phase (1,280) samples was found only for SLEV. Among other flaviviruses, dengue, yellow fever, and Ilheus circulate only in subtropical areas of Argentina (the province of Cordoba is not included in this area); only dengue virus was investigated (by neutralization test) because of a current epidemiologic surveillance program; results were negative. No evidence that West Nile virus is currently circulating or has entered Argentina was found, so we did not perform tests to detect it (2,9). Isolation of SLEV from the cerebrospinal fluid and blood was attempted in newborn mice and Vero cell cultures with negative results. While the typical clinical manifestations of viral encephalitis (fever, headache, and altered level of consciousness) are indistinguishable from each other, tremor and other extrapyramidal signs are described in St. Louis encephalitis and Japanese encephalitis (10). The typical MRI finding of patients with St. Louis encephalitis is localized in the substantia nigra (11). In summary, the occurrence of St. Louis encephalitis in a 61-year-old patient, after >10 years of no reports in Argentina, along with specific epidemiology, suggest that further studies are needed to assess the risk for human infection by SLEV in Argentina and the role of several mosquitoes species in its transmission.

Saint Louis encephalitis virus (SLEV) is endemic in Argentina. During 2005 an outbreak occurred in Córdoba. From January to April of 2010 a new outbreak occurred in Córdoba city with a lower magnitude than the one reported in 2005. Understanding the association of different landscape elements related to SLEV hosts and vectors in urban environments is important for identifying high risk areas for human infections, which was here evaluated. The current study uses a case-control approach at a household geographical location, considering symptomatic and asymptomatic human infections produced by SLEV during 2010 in Córdoba city. Geographical information systems and logistic regression analysis were used to study the distribution of infected human cases and their proximity to water bodies, vegetation abundance, agricultural fields and housing density classified as high/low density urban constructions. Population density at a neighborhood level was also analyzed as a demographic variable. Logistic regression analysis revealed vegetation abundance was significantly associated with the presence of human infections by SLEV. A map of probability of human infections in Córdoba city was derived from the logistic model. The model highlights areas that are more likely to experience SLEV infections. Landscape variables contributing to the outbreak were the proximity to places with vegetation abundance (parks, squares, riversides) and the presence of low density urban constructions, like residential areas. The population density analysis shows that SLEV infections are more likely to occur when population density by neighborhood is lower. These findings and the predictive map developed could be useful for public health surveillance and to improve prevention of vector-borne diseases. Fil: Vergara Cid, Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología "Dr. J. M. Vanella"; Argentina Fil: Estallo, Elizabet Lilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina Fil: Almiron, Walter Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina Fil: Contigiani de Minio, Marta Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología "Dr. J. M. Vanella"; Argentina Fil: Spinsanti, Lorena Ivana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología "Dr. J. M. Vanella"; Argentina

In response to the first human outbreak (January - May 2005) of Saint Louis encephalitis (SLE) virus in Córdoba province, Argentina, we developed an environmental SLE virus risk map for the capital, i.e. Córdoba city. The aim was to provide a map capable of detecting macro-environmental factors associated with the spatial distribution of SLE cases, based on remotely sensed data and a geographical information system. Vegetation, soil brightness, humidity status, distances to water-bodies and areas covered by vegetation were assessed based on pre-outbreak images provided by the Landsat 5TM satellite. A strong inverse relationship between the number of humans infected by SLEV and distance to high-vigor vegetation was noted. A statistical non-hierarchic decision tree model was constructed, based on environmental variables representing the areas surrounding patient residences. From this point of view, 18% of the city could be classified as being at high risk for SLEV infection, while 34% carried a low risk, or none at all. Taking the whole 2005 epidemic into account, 80% of the cases came from areas classified by the model as medium-high or high risk. Almost 46% of the cases were registered in high-risk areas, while there were no cases (0%) in areas affirmed as risk free. Fil: Rotela, Camilo Hugo. Comision Nacional de Actividades Espaciales; Argentina Fil: Spinsanti, Lorena Ivana. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología "Dr. J. M. Vanella"; Argentina Fil: Lamfri, Mario. Comision Nacional de Actividades Espaciales; Argentina Fil: Contigiani de Minio, Marta Silvia. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología "Dr. J. M. Vanella"; Argentina Fil: Almiron, Walter Ricardo. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Investigaciones Entomológicas de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina Fil: Scavuzzo, Carlos Marcelo. Comision Nacional de Actividades Espaciales; Argentina

Introduction St. Louis encephalitis virus (SLEV) is a re-emerging arbovirus in South America, with reported cases in humans in Argentina and Brazil. This fact indicates that there is an urgent need to increase the current knowledge about this virus in order to control and prevent future cases. Exhaustive epidemiological and laboratory investigation is required to ensure fast, accurate identification of the viral agent and allow prompt surveillance action by health authorities. Herein, we report the development of a species-specific RT-nested PCR to detect SLEV. Material and methods After selecting the SLEV genomic region providing the greatest information on the natural genetic variability of this virus, degenerated oligonucleotide primers were designed to amplify a 234-bp fragment of the envelope gene from nine SLEV strains (Parton, BeH356964, SPAN11916, AN9275, AN9124, 78V6507 and 3 SLEV strains obtained from naturally infected mosquito pools). Results The method was able to identify the genome of all the SLEV strains tested and did not amplify unrelated RNA viruses, such as yellow fever virus, Ilheus virus, dengue-2 virus, Bussuquara virus, West Nile virus, Japanese encephalitis virus and Murray Valley encephalitis virus. The method was specific and sensitive, with a lower detection limit of Conclusion This molecular assay is a reliable procedure with a wide spectrum for detecting the natural diversity of SLEV and may be useful for ecological studies, clinical and laboratory settings and virological surveillance.

Background: An outbreak of flavivirus encephalitis occurred in 2005 in Córdoba province, Argentina. Objectives: To characterize the epidemiologic and clinical features of that outbreak and provide the serologic results that identified St. Louis encephalitis virus (SLEV) as the etiologic agent. Study design: From January to May 2005, patients with symptoms of encephalitis, meningitis, or fever with severe headache were evaluated and an etiologic diagnosis achieved by detection of flavivirus-specific antibody sera and cerebrospinal fluid. Results: The epidemic curve of 47 cases showed an explosive outbreak starting in January 2005 with one peak in mid-February and a second peak in mid-March; the epidemic ended in May. Cases occurred predominantly among persons 60 years and older. Nine deaths were reported. SLEV antibodies, when detected in 47 patients studied, had a pattern characteristic of a primary SLEV infection. Conclusions: Even though isolated cases of St. Louis encephalitis have been reported in Argentina, this is the first description of a large SLEV encephalitis outbreak in Argentina. Fil: Spinsanti, Lorena Ivana. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología "Dr. J. M. Vanella"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Diaz, Luis Adrian. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología "Dr. J. M. Vanella"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Glatstein, Nora Viviana. Provincia de Córdoba. Ministerio de Salud; Argentina Fil: Arselán, Sergio. Clínica Privada Velez Sársfield; Argentina Fil: Morales, María A.. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbran". Instituto Nacional de Enfermedades Virales Humanas; Argentina Fil: Farias, Adrian Alejandro. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología "Dr. J. M. Vanella"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Fabbri, Cintia. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología "Dr. J. M. Vanella"; Argentina Fil: Aguilar, Juán J.. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología "Dr. J. M. Vanella"; Argentina Fil: Ré, Viviana Elizabeth. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología "Dr. J. M. Vanella"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Frías, María. Provincia de Córdoba. Ministerio de Salud; Argentina Fil: Almiron, Walter Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina Fil: Hunsperger, Elizabeth. Centers for Disease Control and Prevention. Dengue Branch; Puerto Rico Fil: Siirin, Marina. University of Texas Medical Branch; Estados Unidos Fil: Da Rosa, Amelia Travassos. University of Texas Medical Branch; Estados Unidos Fil: Tesh, Robert B.. University of Texas Medical Branch; Estados Unidos Fil: Enría, Delia. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbran". Instituto Nacional de Enfermedades Virales Humanas; Argentina Fil: Contigiani de Minio, Marta Silvia. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología "Dr. J. M. Vanella"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

To determine hepatitis C virus (HCV) genotypes circulating in the central region of Argentina, 96 consecutive anti-HCV positive subjects were studied. The presence of HCV RNA was detected in 60 samples by RT-nested PCR of the 5' noncoding region (5' NCR). Genotyping was performed by restriction fragment length polymorphism analysis of 5' NCR region combined with PCR using type-specific primers of the core region. The groups of individuals in this study included hemophilia and hemodialysis patients, injecting drug users, screened blood donors, and patients with acute or chronic liver disease, all from Córdoba, Argentina. Overall, genotype 2 was the most prevalent (55.0%), followed by genotypes 1 (38.3%), and 3 (5.0%). Within genotype 1, subtype 1b was the most prevalent. An unexpected high prevalence of genotype 2 (61.9%) was found among patients with acute or chronic HCV infection (without known risk factors). These figures differ from other cohorts from East-Argentina where genotype 1 has been found as the most prevalent. This indicates that regional differences of genotype distribution might exist between Central and East Argentina.

Una virus (UNAV), Togaviridae family, is widely distributed in South America, where infections have been detected in mosquitoes and vertebrate hosts (humans, birds and horses). We analyzed human sera from Córdoba inhabitants aged 44 to 89 years and using a neutralization test, we found a prevalence of UNAV antibodies of 3.8% (3/79). The low titers detected suggest past infections probably acquired in rural areas of the Province of Córdoba (central Argentina). None sera were found positive for MAYV neutralizing antibodies. This is the first report of human infections by UNAV in Argentina.

This is the first evidence of a possible association with Bun- 54 yamwera serogroup virus (CbaAr426 strain or a closely related 55 strain) with febrile syndrome in Córdoba city. Fil: Tauro, Laura Beatriz. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología ; Argentina. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina Fil: Venezuela, Raul Fernando. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología ; Argentina Fil: Spinsanti, Lorena Ivana. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología ; Argentina Fil: Contigiani de Minio, Marta Silvia. Universidad Nacional de Córdoba. Facultad de Medicina. Instituto de Virología ; Argentina