Víctor Alberto Olano

Epidemic Venezuelan Equine Encephalitis in La Guajira, Colombia, 1995

In 1995, the first Venezuelan equine encephalitis (VEE) outbreak in Colombia in 22 years caused an estimated 75,000 human cases, 3000 with neurologic complications and 300 fatal, in La Guajira State. Of the states estimated 50,000 equines, 8% may have died. An epizootic IC virus, probably introduced from Venezuela, was rapidly amplified among unvaccinated equines. Record high rainfall, producing high densities of vector Aedes taeniorhynchus, led to extensive epidemic transmission (30% attack rate) in the four affected municipalities. Native Wayuu Indians, constituting 24% of the states population, were at increased risk of infection (risk ratio, 3.3; 95% confidence interval, 2.2-5.3). Epidemiologic studies found no evidence of human-to-human transmission. A higher-than-expected number of abortions during the outbreak confirmed a previously suspected abortifacient role of VEE infection. Pesticide applications and a mass equine vaccination program contributed to preventing the outbreaks spread south of La Guajira.

BITING ACTIVITY AND BREEDING SITES OF ANOPHELES SPECIES IN THE MUNICIPALITY VILLAVICENCIO, META, COLOMBIA

ABSTRACT Villavicencio, the capital city of the Department of Meta, Colombia, is at high risk for the urbanization of malaria because of the regions ecological conditions, as well as the permanent presence of infected human populations arriving from rural areas. From August to November 2002 and in April 2003, anopheline collections were undertaken in the area. Isofamilies were obtained from 331 wild females, which were then recorded according to their abundance as follows: Anopheles marajoara, Anopheles rangeli, Anopheles braziliensis, Anopheles darlingi, and Anopheles apicimacula. Anopoheles darlingi showed the highest biting activity (3.0) between 1800 and 1900 h. Forty-five breeding places were sampled, 64% of which were fish ponds, 6.7% flooded meadows, and 6.7% drainpipes, with these being the most representative locations. All sampled breeding sites were positive for anophelines. Anopheles marajoara could play an important role as an auxiliary vector in Villavicencios urban area. Control measures should be aimed at weeding the marginal areas around fish ponds and at evaluating the use of impregnated bed-nets.

A new continuous cell line from the mosquito Psorophora confinnis (Diptera: Culicidae) and its susceptibility to infections with some arboviruses

A new cell line, PC-0199-BR, was established from embryonated eggs of the mosquito Psorophora confinnis. To date (September 2000) it has had 62 continuous passages. This is the first report of a cell line of mosquitoes belonging to the genus Psorophora. Cell growth initially was achieved in the MM/VP12 medium, supplemented with 20% fetal bovine serum; however, the subcultures were later adapted to Graces medium with 10% fetal bovine serum. Cell morphology in the primary cultures was heterogeneous; but later in the established cell line, the predominant cell type was epithelioid. Cultured cells were predominantly diploid (2n=6); however, chromosome abnormalities were observed in a small proportion of the cells in later passages. C and G band patterns were also determined in the karyotype. The cell line isozyme profiles coincided with pupae and adult samples of the species taken from the same colony. A preliminary arbovirus susceptibility study for the cell line was undertaken. No evidence was observed of contamination of the cell line with bacteria, fungi or mycoplasma.

Development of sporogonic cycle of Plasmodium vivax in experimentally infected Anopheles albimanus mosquitoes

The sporogonic cycle of Plasmodium vivax was established and maintained under laboratory conditions in two different strains of Anopheles albimanus mosquitoes using as a parasite source blood from human patients or from Aotus monkeys infected with the VCC-2 P.vivax colombian isolate. Both the Tecojate strain isolate from Guatemala and the Cartagena strain from the colombian Pacific coast were susceptible to infections with P.vivax. A higher percentage of Cartagena mosquitoes was infected per trial, however the Tecojate strain developed higher sporozoite loads. Intravenous inoculation of Aotus monkeys with sporozoites obtained from both anopheline strains resulted in successful blood infections. Animals infected with sporozoites from the Tecojate strain presented a patent period of 21-32 days whereas parasitemia appeared between days 19-53 in monkeys infected with sporozites from Cartagena strain.

Diarrhea and dengue control in rural primary schools in Colombia: study protocol for a randomized controlled trial

BackgroundDiarrheal diseases and dengue fever are major global health problems. Where provision of clean water is inadequate, water storage is crucial. Fecal contamination of stored water is a common source of diarrheal illness, but stored water also provides breeding sites for dengue vector mosquitoes. Poor household water management and sanitation are therefore potential determinants of both diseases. Little is known of the role of stored water for the combined risk of diarrhea and dengue, yet a joint role would be important for developing integrated control and management efforts. Even less is known of the effect of integrating control of these diseases in school settings. The objective of this trial was to investigate whether interventions against diarrhea and dengue will significantly reduce diarrheal disease and dengue entomological risk factors in rural primary schools.Methods/designThis is a 2×2 factorial cluster randomized controlled trial. Eligible schools were rural primary schools in La Mesa and Anapoima municipalities, Cundinamarca, Colombia. Eligible pupils were school children in grades 0 to 5. Schools were randomized to one of four study arms: diarrhea interventions (DIA); dengue interventions (DEN); combined diarrhea and dengue interventions (DIADEN); and control (C). Schools were allocated publicly in each municipality (strata) at the start of the trial, obviating the need for allocation concealment. The primary outcome for diarrhea is incidence rate of diarrhea in school children and for dengue it is density of adult female Aedes aegypti per school. Approximately 800 pupils from 34 schools were enrolled in the trial with eight schools in the DIA arm, nine in the DEN, eight in the DIADEN, and nine in the control arms. The trial status as of June 2012 was: completed baseline data collections; enrollment, randomization, and allocation of schools. The trial was funded by the Research Council of Norway and the Lazos de Calandaima Foundation.DiscussionThis is the first trial investigating the effect of a set of integrated interventions to control both dengue and diarrhea. This is also the first trial to study the combination of diarrhea-dengue disease control in school settings.Trial registrationCurrent Controlled Trials ISRCTN40195031

Schools as potential risk sites for Vector-Borne disease transmission: mosquito vectors in rural schools in two municipalities in Colombia

Abstract Dengue and other vector-borne diseases are of great public health importance in Colombia. Vector surveillance and control activities are often focused at the household level. Little is known about the importance of nonhousehold sites, including schools, in maintaining vector-borne disease transmission. The objectives of this paper were to determine the mosquito species composition in rural schools in 2 municipalities in Colombia and to assess the potential risk of vector-borne disease transmission in school settings. Entomological surveys were carried out in rural schools during the dry and rainy seasons of 2011. A total of 12 mosquito species were found: Aedes aegypti, Anopheles pseudopunctipennis, Culex coronator, Cx. quinquefasciatus, and Limatus durhamii in both immature and adult forms; Ae. fluviatilis, Cx. nigripalpus, Cx. corniger, and Psorophora ferox in immature forms only; and Ae. angustivittatus, Haemagogus equinus, and Trichoprosopon lampropus in adult forms only. The most common mosquito species was Cx. quinquefasciatus. Classrooms contained the greatest abundance of adult female Ae. aegypti and Cx. quinquefasciatus. The most common Ae. aegypti breeding sites were containers classified as “others” (e.g., cans), followed by containers used for water storage. A high level of Ae. aegypti infestation was found during the wet season. Our results suggest that rural schools are potentially important foci for the transmission of dengue and other mosquito-borne diseases. We propose that public health programs should be implemented in rural schools to prevent vector-borne diseases.

Detection of all four dengue serotypes in Aedes aegypti female mosquitoes collected in a rural area in Colombia

The Aedes aegypti vector for dengue virus (DENV) has been reported in urban and periurban areas. The information about DENV circulation in mosquitoes in Colombian rural areas is limited, so we aimed to evaluate the presence of DENV in Ae. aegypti females caught in rural locations of two Colombian municipalities, Anapoima and La Mesa. Mosquitoes from 497 rural households in 44 different rural settlements were collected. Pools of about 20 Ae. aegypti females were processed for DENV serotype detection. DENV in mosquitoes was detected in 74% of the analysed settlements with a pool positivity rate of 62%. The estimated individual mosquito infection rate was 4.12% and the minimum infection rate was 33.3/1,000 mosquitoes. All four serotypes were detected; the most frequent being DENV-2 (50%) and DENV-1 (35%). Two-three serotypes were detected simultaneously in separate pools. This is the first report on the co-occurrence of natural DENV infection of mosquitoes in Colombian rural areas. The findings are important for understanding dengue transmission and planning control strategies. A potential latent virus reservoir in rural areas could spill over to urban areas during population movements. Detecting DENV in wild-caught adult mosquitoes should be included in the development of dengue epidemic forecasting models.

ESTABLISHMENT AND CHARACTERIZATION OF A CELL LINE FROM THE MOSQUITO ANOPHELES ALBIMANUS (DIPTERA : CULICIDAE)

A new cell line designated LSB-AA695BB, was established from embryos of the mosquito Anopheles albimanus. The primary culture was initiated in April, 1995, and the first passage was made 48 days later. Serial subcultures of the cells have been carried through 90 passage from Abril 1995 to February 1996. The cells were grown at 28 degrees C in MK/VP12 medium, supplemented with 20% fetal bovine serum: the pH tolerance ranged between 6.8 to 7.0. The cells have also been adapted to MM/VP12 medium under the same pH, temperature and serum concentration. The majority of the cells were a fibroblast-type. Isozyme characterization showed a pattern similar to that of An. albimanus pupae and adults but distinct from Ae. taeniorhynchus and Ae. albopictus (C6/36) mosquito cell lines. The culture was shown to be free of mycoplasma, bacteria and fungi. Microsporidia contamination of transovarial transmission was controlled with 6.0 micrograms/ml of albendazole.

Initiation of primary cell cultures from embryos of the mosquitoes Anopheles albimanus and Aedes taeniorhynchus (Diptera: Culicidae)

Primary cell cultures were obtained from eggs of Anopheles albimanus and Aedes taeniorhynchus mosquitoes, vectors of human malaria and of Venezuelan equine encephalitis virus, respectively. The cellular growth of the An. albimanus cells began four weeks after explanting the embryonic tissues in MK/VP12 medium, supplemented with 15% fetal bovine serum. The culture showed heterogeneous cellular morphology. With regard to the Ae. taeniorhynchus culture, growth occurred three weeks after initiating the culture in MM/VP12 medium. The majority of cells were small and round. Karyotypes were examined in the latter species.

A cross-sectional survey of Aedes aegypti immature abundance in urban and rural household containers in central Colombia.

BackgroundAedes aegypti, the major vector of dengue, breeds in domestic water containers. The development of immature mosquitoes in such containers is influenced by various environmental, ecological and socioeconomic factors. Urban and rural disparities in water storage practices and water source supply may affect mosquito immature abundance and, potentially, dengue risk. We evaluated the effect of water and container characteristics on A. aegypti immature abundance in urban and rural areas. Data were collected in the wet season of 2011 in central Colombia from 36 urban and 35 rural containers, which were either mosquito-positive or negative. Immature mosquitoes were identified to species. Data on water and container characteristics were collected from all containers.ResultsA total of 1452 Aedes pupae and larvae were collected of which 81% were A. aegypti and 19% A. fluviatilis. Aedes aegypti immatures were found in both urban and rural sites. However, the mean number of A. aegypti pupae was five times higher in containers in the urban sites compared to those in the rural sites. One of the important factors associated with A. aegypti infestation was frequency of container washing. Monthly-washed or never-washed containers were both about four times more likely to be infested than those washed every week. There were no significant differences between urban and rural sites in frequency of washing containers. Aedes aegypti immature infestation was positively associated with total dissolved solids, but negatively associated with dissolved oxygen. Water temperature, total dissolved solids, ammonia, nitrate, and organic matter were significantly higher in urban than in rural containers, which might explain urban-rural differences in breeding of A. aegypti. However, many of these factors vary substantially between studies and in their degree of association with vector breeding, therefore they may not be reliable indices for vector control interventions.ConclusionsAlthough containers in urban areas were more likely to be infested with A. aegypti, rural containers still provide suitable habitats for A. aegypti. Containers that are washed more frequent are less likely to produce A. aegypti. These results highlight the importance of container washing as an effective vector control tool in both urban and rural areas. In addition, alternative designs of the highly productive washbasins should continue to be explored. To control diseases such as dengue, Zika and chikungunya, effective vector breeding site control must be implemented in addition to other interventions.