Marta Silvia Contigiani

Genotype III Saint Louis Encephalitis Virus Outbreak, Argentina, 2005

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.

Human outbreak of St. Louis encephalitis detected in Argentina, 2005

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.

Age-related seroprevalence study for St. Louis encephalitis in a population from Cordoba, Argentina

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.

Silent Circulation of St. Louis Encephalitis Virus Prior to an Encephalitis Outbreak in Cordoba, Argentina (2005)

St. Louis encephalitis virus is a complex zoonoses. In 2005, 47 laboratory-confirmed and probable clinical cases of SLEV infection were reported in Córdoba, Argentina. Although the causes of 2005 outbreak remain unknown, they might be related not only to virological factors, but also to ecological and environmental conditions. We hypothesized that one of the factors for SLE reemergence in Córdoba, Argentina, was the introduction of a new SLEV genotype (SLEV genotype III), with no previous activity in the area. In order to evaluate this hypothesis we carried out a molecular characterization of SLEV detections from mosquitoes collected between 2001 and 2004 in Córdoba city. A total of 315 mosquito pools (11,002 individuals) including 12 mosquitoes species were analyzed. Overall, 20 pools (8 mosquitoes species) were positive for SLEV. During this study, genotypes II, V and VII were detected. No mosquito pool infected with genotype III was detected before the 2005 outbreak. Genotype V was found every year and in the 8 sampled sites. Genotypes II and VII showed limited temporal and spatial activities. We cannot dismiss the association of genotype II and V as etiological agents during the outbreak. However, the silent circulation of other SLEV strains in Córdoba city before the 2005 outbreak suggests that the introduction of genotype III was an important factor associated to this event. Not mutually exclusive, other factors such as changes in avian hosts and mosquitoes vectors communities, driven by climatic and environmental modifications, should also be taken into consideration in further studies.

Vertical transmission of St. Louis encephalitis virus in Culex quinquefasciatus (Diptera: Culicidae) in Córdoba, Argentina.

Arbovirus vector transmission is interrupted when vector abundance decreases or disappears in temperate regions during the winter season. Although the primary overwintering mechanism for many arboviruses in nature remains unknown, vertical transmission is one potential mechanism. Vertical transmission functions as an overwintering mechanism for St. Louis encephalitis virus (SLEV) in temperate areas of Argentina, where SLEV is endemic. The aim of this project was to detect vertical transmission of SLEV in Culex quinquefasciatus mosquitoes. Under laboratory conditions, SLEV vertical transmission (3.4 minimum infection rates) was detected in larvae (1:256) and adults F₁ (1:406). There were no positive larvae for SLEV among over 2011 analyzed individuals collected in nature. This is the first study to confirm experimental vertical transmission of SLEV in Cx. quinquefasciatus populations from Argentina, though additional overwintering mechanisms (e.g., nontraditional vectors such as ticks and nondiapausing female mosquitoes) should be considered.

Enzootic activity of pixuna and Rio Negro viruses (Venezuelan equine encephalitis complex) in a neotropical region of Argentina.

Venezuelan Equine Encephalitis complex viruses cause epidemics and epizootics periodically in some regions of the Americas. In Argentina, only enzootic Rio Negro virus (AG80-663) (RNV) has been isolated. To survey and identify activity of viruses that belong to Venezuelan Equine Encephalitis complex in a template region of the country, a generic Alphavirus RT-Nested PCR was performed in 99 mosquito pools collected in Chaco province. Five pools were positive, and amplicons were sequenced: four of them clustered with RNV(AG80-663) and one with Pixuna virus. This is the first report of the circulation of Pixuna virus in Argentina, and it confirms enzootic and endemic activity of RNV(AG80-663) in neotropical regions of this country.

Reliable detection of St. Louis encephalitis virus by RT-nested PCR

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 < 10 plaque-forming units. 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.

Comparison of Argentinean Saint Louis Encephalitis Virus Non-Epidemic and Epidemic Strain Infections in an Avian Model

St. Louis encephalitis virus (SLEV, Flavivirus, Flaviviridae) is an emerging mosquito-borne pathogen in South America, with human SLEV encephalitis cases reported in Argentina and Brazil. Genotype III strains of SLEV were isolated from Culex quinquefasciatus mosquitoes in Cordoba, Argentina in 2005, during the largest SLEV outbreak ever reported in South America. The present study tested the hypothesis that the recent, epidemic SLEV strain exhibits greater virulence in birds as compared with a non-epidemic genotype III strain isolated from mosquitoes in Santa Fe Province 27 years earlier. The observed differences in infection parameters between adult House sparrows (Passer domesticus) that were needle-inoculated with either the epidemic or historic SLEV strain were not statistically significant. However, only the House sparrows that were infected with the epidemic strain achieved infectious-level viremia titers sufficient to infect Cx. spp. mosquitoes vectors. Furthermore, the vertebrate reservoir competence index values indicated an approximately 3-fold increase in amplification potential of House sparrows infected with the epidemic strain when pre-existing flavivirus-reactive antibodies were present, suggesting the possibility that antibody-dependent enhancement may increase the risk of avian-amplified transmission of SLEV in South America.

Transmission of endemic St Louis encephalitis virus strains by local Culex quinquefasciatus populations in Córdoba, Argentina

BACKGROUND St Louis encephalitis virus (SLEV) is a re-emerging human pathogen widely distributed in the American continent. Although it is not fully understood, the SLEV transmission network may involve Culex quinquefasciatus mosquitoes as vectors and Columbidae species as hosts. METHODS To calculate infection rates, we inoculated Cx. quinquefasciatus mosquitoes from Córdoba, Argentina by feeding them on viremic chicks. RESULTS We observed differences in infection rate among the viral strains, the highest rate (78/87 mosquitoes, 90.8%) being seen in strain 78V-6507. After re-feeding on susceptible chicks, mosquitoes were able to transmit the virus. CONCLUSION Our findings suggest that Cx. quinquefasciatus populations are susceptible to and able to transmit different SLEV strains.

Serological detection of St. Louis encephalitis virus and West Nile virus in equines from Santa Fe, 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.