Protocolo del Sistema Nacional de Vigilancia Epidemiológica: enfermedades zoonóticas. No. 08 / Protocol of the National Epidemiological Surveillance System: zoonotic diseases. No. 08

Estos protocolos están dirigido a personal médico, paramédico y otros profesionales que realizan acciones gerenciales y operativas de vigilancia epidemiológica en los servicios de salud del país, y están divididos en varios tomos para dar a conocer y actualizar la identificación y medidas de control para diversos padecimientos a fin de continuar con el mejoramiento de las capacidades técnicas de los trabajadores de salud, que permita planificar la prestación de servicios con decisiones partiendo de un enfoque epidemiológico comprobado, para responder a los cambios de tendencias epidemiológicas y con ello contribuir al fortalecimiento de prácticas asertivas de la salud pública de nuestro país.

Acute larvicidal toxicity of five essential oils (Pinus nigra, Hyssopus officinalis, Satureja montana, Aloysia citrodora and Pelargonium graveolens) against the filariasis vector Culex quinquefasciatus: Synergistic and antagonistic effects.

Mosquito vector control is facing a number of important and timely challenges, mainly due to the rapid development of pesticide resistance and environmental concerns. In this scenario, screening of botanical resources for their mosquitocidal activity may offer effective and eco-friendly tools against Culicidae vectors. Culex quinquefasciatus Say (Diptera: Culicidae) is a vector of lymphatic filariasis and of dangerous arboviral diseases, such as West Nile and St. Louis encephalitis. In this study, the chemical composition of five essential oils obtained from different plants, namely Pinus nigra J.F. Arnold var. italica (Pinaceae), Hyssopus officinalis L. subsp. aristatus (Lamiaceae), Satureja montana L. subsp. montana (Lamiaceae), Aloysia citriodora Palau (Verbenaceae) and Pelargonium graveolens L'Hér (Geraniaceae), was investigated by GC-MS analysis. Furthermore, it was evaluated their acute toxicity on larvae of C. quinquefasciatus. Then, the most effective oils were selected, in order to focus on the potential synergistic and antagonistic effects, testing them in binary mixtures on C. quinquefasciatus larvae. Results showed that the higher effectiveness was obtained by S. montana subsp. montana essential oil (LC50=25.6µL·L-1), followed by P. nigra var. italica (LC50=49.8µL·L-1) and A. citriodora (LC50=65.6µL·L-1), while the other essential oils showed LC50 values higher than 90µL·L-1. The larvicidal effectiveness can be enhanced by preparing simple binary mixtures of essential oils, such as S. montana+A. citriodora (ratio 1:1), which showed higher larvicidal toxicity (LC50=18.3µL·L-1). On the other hand, testing S. montana+P. nigra (1:1) an antagonistic effect was detected, leading to a LC50 (72.5µL·L-1) higher than the LC50 values calculated for the two oils tested separately. Overall, our results add useful knowledge to allow the employ of synergistic essential oil blends as effective, cheap and eco-friendly mosquito larvicides.

A comparison of gravid and under-house CO2-baited CDC light traps for mosquito species of public health importance in Houston, Texas.

The relative efficacy of gravid and under-house CO2 traps for monitoring mosquito species of public health importance within the Houston metroplex area was assessed. Gravid and under-house traps were colocated at 10 sites and monitored weekly between 1 March to 31 May 2007. The most numerous species caught was Culex pipiens quinquefasciatus Say. Other species of public health importance caught in gravid and under-house traps included Culex restuans Theobald, Aedes aegypti (L.), and Aedes albopictus Skuse. Adjusting for the week of collection, gravid traps caught significantly more mosquitoes (mean 23.1 per trap) in the study area than under-house traps (mean 3.6 per trap). However, under-house traps caught a greater variety of mosquito species (13) than gravid traps (11). Gravid and under-house traps only caught nine of 15 of the same mosquito species during the study period. In this study area, gravid traps should be used as the primary method of surveillance for mosquito-borne diseases of public health importance during the early part of the season, because of greater catch numbers of mosquitoes that pose a public health risk.

Brote de encefalitis de St. Louis en la provincia de Córdoba

La encefalitis de Saint Louis (ESL) es una enfermedad viral transmitida al humano por mosquitos del género culex spp. El agente causal es un arbovirus miembro de la familia flaviviridae. En Argentina es una infección de distribución amplia con una prevalencia serológica del 10-68%. Está demostrada su circulación en aves. Entre los meses de enero y abril de 2005 se presentaron en la provincia de Córdoba 48 casos de ESL. En nuestra experiencia asistimos 9 casos de esta encefalitis.

Brote de encefalitis de St. Louis en la provincia de Córdoba

La encefalitis de Saint Louis (ESL) es una enfermedad viral transmitida al humano por mosquitos del género culex spp. El agente causal es un arbovirus miembro de la familia flaviviridae. En Argentina es una infección de distribución amplia con una prevalencia serológica del 10-68%. Está demostrada su circulación en aves. Entre los meses de enero y abril de 2005 se presentaron en la provincia de Córdoba 48 casos de ESL. En nuestra experiencia asistimos 9 casos de esta encefalitis.(AU)

Construction of yellow fever/St. Louis encephalitis chimeric virus and the use of chimeras as a diagnostic tool.

St. Louis encephalitis (SLE) and West Nile (WN) flaviviruses are genetically closely related and cocirculate in the United States. Virus neutralization tests provide the most specific means for serodiagnosis of infections with these viruses. However, use of wild-type SLE and WN viral strains for laboratory testing is constrained by the biocontainment requirements. We constructed two highly attenuated yellow fever (YF) virus chimeras that contain the premembrane-envelope (prM-E) protein genes from the virulent MSI-7 (isolated in the United States) or the naturally attenuated CorAn9124 (Argentina) SLE strains. The YF/SLE (CorAn version) virus and the previously constructed YF/WN chimera were shown to specifically distinguish between confirmed human SLE and WN cases in a virus neutralization test using patient sera. These chimeras have the potential for use as diagnostic reagents and vaccines against SLE and WN.

Encephalitis virus persistence in California birds: experimental infections in mourning doves (Zenaidura macroura).

After-hatching and hatching year, mourning doves were infected by inoculation with either western equine encephalomyelitis (WEE) or St. Louis encephalitis (SLE) viruses; some birds in each group also were treated with the immunosuppressant cyclophosphamide before and during infection. Cyclophosphamide treatment significantly increased the WEE viremia but did not alterthe antibody response. In contrast, cyclophosphamide-treated and -untreated doves did not develop a detectable SLE viremia but became antibody positive. Antibody peaked at 10 wk after inoculation for both viruses and remained detectable in most birds throughout the 26-wk study. When treated with cyclophosphamide the following spring, birds did not relapse and develop a detectable viremia. Previously infected birds were protected when challenged with conspecific virus (i.e., none produced a detectable viremia), but there was no anamnestic antibody response to reinfection. In agreement with our failure to detect relapses, all birds were negative for viral RNA when sera, spleen, lung, and kidney tissues were tested by reverse transcriptase-polymerase chain reaction after necropsy. Our results indicated that adult mourning doves were an incompetent host for SLE virus and probably do not serve as a suitable overwintering or dispersal host for either WEE and SLE viruses.

Previous infection protects house finches from re-infection with St. Louis encephalitis virus.

Antibody titers against St. Louis encephalitis virus (SLE) measured by a plaque reduction neutralization test (PRNT) decreased rapidly in house finches (Capodacus mexicanus) after initial infection, whereas antibodies measured by enzyme immunoassay (EIA) remained detectable in all birds for the length of the experiment, indicating long-term persistence and greater assay sensitivity of the EIA. After 52 wk, birds were challenged by subcutaneous inoculation with the same strain of SLE virus. Virus was not detected for 1-4 d postchallenge in blood samples tested by plaque assay and RT-PCR or by xenodiagnosis in Culex tarsalis fed concurrently and then held for 11 d at 26 degrees C. Virus was detected by all three methods in control birds infected concurrently for the first time. Challenge with SLE produced a rapid and marked ananmestic rise in both neutralizing and EIA antibody titers that exceeded the primary response in the same birds or in concurrently inoculated control birds. At necropsy 4 wk postchallenge, 3 of 7 challenged and 1 of 2 positive control birds were chronically infected, with viral RNA detected by RT-PCR in brain, spleen, lung, and/or kidney tissues. Our results indicated that persistence of protective antibody prevents reinfection during the following season and may prevent the recrudescence of infectious virus in chronically infected birds.

California state Mosquito-Borne Virus Surveillance and Response Plan: a retrospective evaluation using conditional simulations.

The California Mosquito-Borne Virus Surveillance and Response Plan recently was developed to provide a semi-quantitative means for assessing risk for western equine encephalomyelitis (WEE) or St. Louis encephalitis (SLE) viruses and to provide intervention guidelines for mosquito control and public health agencies during periods of heightened risk for human infection. West Nile virus recently has arrived in California, and the response plan also will provide a baseline for assessing the risk for human and equine infection with this virus. In the response plan, overall risk is calculated by averaging risk due to 1) environmental conditions, 2) adult mosquito vector abundance, 3) vector infection rates, 4) sentinel chicken seroconversion rates, 5) equine cases (for WEE), 6) human cases, and 7) the proximity of virus activity to populated areas. Overall risk is categorized into three levels: normal season, emergency planning, or epidemic conditions. We evaluated this response plan using historical data from years with no, enzootic, and epidemic activity of WEE and SLE in several areas of California to determine whether calculated risk levels approximated actual conditions. Multiple methods of risk calculation were considered for both viruses. Assessed risk based on cumulative temperature, rainfall, and runoff levels over the entire season provided more or equally accurate assessments than biweekly assessments based solely on the previous half-month. For WEE, during years with enzootic activity or early-season periods of years with WEE epidemic activity, combining horse and human cases as a single risk factor improved the model's ability to forecast pending WEE activity, but separating the two factors allowed a better indication of WEE activity during epidemics and periods with no activity. For SLE, assignment of higher risk to drier conditions as measured by rainfall and runoff yielded the most accurate representation of actual virus activity during all recent study periods.

Wicking assays for the rapid detection of West Nile and St. Louis encephalitis viral antigens in mosquitoes (Diptera: Culicidae).

The recent outbreaks of West Nile (WN) encephalitis and St. Louis encephalitis (SLE) in the United States have highlighted the need for rapid and specific methods of detecting arboviral antigens in mosquitoes. We evaluated rapid, field-usable assays for detecting and differentiating WN and SLE viruses in mosquito pools, based on a patent-pending, immunochromatographic technology (VecTest) formatted on a dipstick. The device provides results in less than 20 min and can be used in laboratories with adequate containment facilities. In laboratory assessments, both the SLE and WN virus tests demonstrated sensitivity comparable with that of an antigen capture ELISA, but less than can be achieved with Vero cell plaque or reverse-transcriptase polymerase chain reaction assays. There was no evidence of cross-reaction when tested with high concentrations of heterologous flavivirus antigens or with Eastern equine encephalitis or Western equine encephalitis viruses. Both the WN and SLE dipstick tests delivered a clear positive result with a single positive specimen in a pool of 50 mosquitoes. This virus assay technology reduces the time required to obtain test results and will allow rapid medical threat assessment and effective targeting of vector control measures.

Multiple genotypes of St. Louis encephalitis virus (Flaviviridae: Flavivirus) circulate in Harris County, Texas.

St. Louis encephalitis (SLE) is endemic in Harris County, Texas. The disease is a public health concern in Houston, the largest city in Harris County, and in the state. Consequently, intensive surveillance for SLE virus in local mosquito populations is carried out by the Harris County Mosquito Control Division each year. In this study, we examined genetic variation among SLE isolates obtained during routine virus surveillance over a 13-year time period (1986-1999). St. Louis encephalitis virus isolates were tested for genetic variation using reverse transcription-polymerase chain reaction followed by single-strand conformation polymorphism (SSCP). The results indicated that multiple genotypes of the virus circulate in Harris County. During several years, the genotypes were restricted in their location, i.e., each general area within the county had a specific genotype of the virus. In other years, the various genotypes were widely distributed throughout the county. The presence of multiple distinct genotypes suggests that viruses with different biological characteristics may be circulating in Harris County, and that discrete foci of SLE virus activity occur simultaneously.

Epidemiological features of and public health response to a St. Louis encephalitis epidemic in Florida, 1990-1.

A St. Louis encephalitis (SLE) epidemic in Florida during 25 weeks in 1990-1, resulted in 222 laboratory-diagnosed cases, an attack rate in the 28 affected counties of 2.25/100,000. Disease risk rose with advanced age, to 17.14/100,000 in persons over 80 years, and all 14 fatal cases were in persons over 55 years (median, 70 years). Community serosurveys in Indian River County, the epicenter of the outbreak (attack rate 21/100,000), showed acute asymptomatic infections in 3.6% of the persons surveyed, with higher rates in persons with outdoor occupational exposure (7.4%) and in clients of a shelter for the indigent (13.3%). A matched case-control study found that evening outdoor exposure for more than 2 h was associated with an increased risk for acquiring illness (odds ratio [OR] 4.33, 95% CI 1.23-15.21) while a number of recommended personal protective measures were protective. Four SLE patients were dually infected with Highlands J virus, the first reported cases of acute infection with this alphavirus. The case-control study provided the first evidence that a public education campaign to reduce exposure had a protective effect against acquiring the disease.

Interferon-alpha protects mice against lethal infection with St Louis encephalitis virus delivered by the aerosol and subcutaneous routes.

In common with other flaviviruses, there is no specific therapy for St Louis encephalitis (SLE) virus infections. A number of cases have occurred where infection may have been acquired by the aerosol route in laboratory accidents. The recombinant human interferon hybrids IFN-alpha A/D (Roche Laboratories) and IFN-alpha B/D (Ciba-Geigy) have activity in murine models. Given for several days around the time of exposure to the virus or shortly after, these compounds reduce the mortality from SLE virus administered to mice subcutaneously by up to 70%. In an aerosol model of SLE disease, the mortality was reduced to 30-50% compared to 100% in controls, depending on the challenge level of virus. These results suggest that interferon-alpha could be used to reduce the mortality from SLE infection after known exposure to the virus.

Immunisation with DNA polynucleotides protects mice against lethal challenge with St. Louis encephalitis virus.

In vivo transfection by intramuscular injection with plasmids expressing the immunogenic proteins of microbial pathogens has considerable potential as a vaccination strategy against many pathogens of both man and animals. Here we report that weanling mice given a single intramuscular injection of 50 micrograms of a plasmid, pSLE1 expressing the St. Louis encephalitis virus (SLE) prM/E protein under the control of the cytomegalovirus immediate early protein promoter produced SLE-specific antibody and were protected against lethal challenge with the virulent virus. Polynucleotide vaccine technology provides a unique opportunity to produce vaccines against flavivirus diseases of low incidence cheaply and rapidly, and to produce multivalent vaccines such as would be required for immunisation against dengue virus disease.

Immunity to St. Louis encephalitis virus by sequential immunization with recombinant vaccinia and baculovirus derived PrM/E proteins.

St. Louis encephalitis (SLE) is an important mosquito-borne disease of great public health concern in parts of the United States. South America and Canada. Protective immunogens of flaviviruses produced in different expression systems have been shown to be effective against virulent virus infection in laboratory animal models. Here we show that the pre-membrane and envelope (PrM/E) of SLE virus expressed in insect and mammalian cell systems using baculovirus and vaccinia virus, respectively, are processed correctly and showed similar antigenic characteristics as the authentic proteins. Immunization with the recombinant proteins individually or in combination resulted in neutralizing and protective immune responses. A schedule consisting of initial immunization with recombinant vaccinia virus followed by a secondary boost with recombinant baculovirus protein resulted in higher levels of neutralizing and protective immune responses. The advantages of the use of such a combined approach as a general immunization strategy are discussed.

Arboviral infections in the United States.

Arboviruses are important considerations in the differential diagnosis of encephalitis and other acute infections. Alterations in the environment and in human behaviors contribute to changing patterns of arboviral transmission. These trends, the periodic epidemic resurgence of arboviral diseases such as St. Louis encephalitis, and the discovery of new arboviruses present a continued challenge to infectious disease clinicians.