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 Grace's 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.

Unstable, low-level transmission of malaria on the Colombian Pacific Coast.

The development of immune responses to malarial infection in inhabitants of endemic areas differs according to the level of exposure to the parasite. Adults living in a region where the level of malaria transmission is low (Colombia) have been shown to exhibit a similar response to each of the three regions of the circumsporozoite protein (the central repeated NANP region, and the flanking N- and C-termini). Conversely, donors exposed to a frequent sporozoite challenge in areas of high malaria transmission (Mali) exhibit antibodies predominantly to the NANP repeated domain. Malaria in the people of Zacarías, a community on the Pacific Coast of Colombia where malaria transmission is low and unstable, was the subject of the present study. Within a 9-year period, a negative correlation between rainfall and documented malaria cases was recorded for this area. Thick smears of blood samples of 319 individuals revealed that 8.5% had malarial infections. As most (67%) of the smear-positive cases were asymptomatic, it seems that, despite the low prevalence of malaria in this area, the establishment of clinical symptoms is attenuated, probably because of the acquisition of premunition. Within this region, the most commonly found Anopheles species (representing 61.1% of the mosquitoes caught) and that giving the highest monthly biting rate (4.0 bites/man) was An. neivai. Most (90%) of the human sera tested possessed antibodies to blood-stage forms of Plasmodium falciparum, and 18% had antibodies to sporozoites. More than half (58%) of the adults had been in contact with hepatitis B virus, 7.2% carried hepatitis B surface antigen, and syphilis was common but no subject was found to be seropositive for HIV. A better understanding of the dynamics of the different elements influencing malaria in areas of low, unstable transmission, such as the one described here, is essential for the design of new malaria-control strategies.

[Transmission of urban malaria in Buenaventrua, Colombia: entomological features]. / Transmisión de la malaria urbana en Buenaventura, Colombia: aspectos entomológicos.

During the past several years, there has been an increase in the cases of urban malaria in Buenaventura, Colombia, from 576 in 1987 to 3296 in 1991 and 2017 in 1992; therefore, an epidemiological study to identify transmission patterns was carried out in this municipality on Colombia's Pacific coast. This paper describes the entomological findings regarding vectors, potential breeding sites and behavior of Anopheles species during the period from June 1993 to May 1994. Of the 469 potential breeding sites identified in the urban area, 28 were positive for anophelines, while in the neighboring rural zone, 20 out of 80 potential breeding sites were positive for the immature forms of Anopheles. Mining excavations, lakes and breeding ponds for fish or shrimp were the places where A. albimanus was more frequently found. For A. nuñeztovari, the breeding sites were rain puddles and ponds for breeding fish or shrimp. A. neivai was also identified in the urban area and larvae were collected from bromeliaceous plants. Both inside and outside the households mosquito collections were carried out, and 90% of the collected mosquitoes were identified as A. albimanus. Of the female A. albimanus 54.8% were parous. This species was found to have peak peridomiciliary activity between 6 p.m. and 10 p.m. The highest biting rate in the peridomiciliary area was 7.1 and was observed during March. It is expected that this entomological information will permit the launching of a strong community participation process and the implementation of control measures in accordance with the entomological findings related to malaria in the urban area of Buenaventura.

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 state's 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 state's 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 outbreak's spread south of La Guajira.

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.

Establecimiento y mantenimiento de una colonia de Aedes taeniorhynchus, Wiedemann, 1821 (Díptera: Culicidae) cepa Barranquilla, Colombia / Establishment and maintainance of a colony of Aedes taeniorhynchus, Wiedemann, 1821 (Diptera: Culicidae) strain Barranquilla, Colombia

Se estableció una colonia del mosquito Aedes taeniorhynchus, eficiente vector del virus de la encefalitis equina venezolana, tipo epidemo-epizoótico, a partir de formas adultas, recolectados en regiones cercanas a Barranquilla, Atlántico (Colombia). Se estudiaron algunos aspectos de la biología de este insecto (ciclo de vida y logevidad), elaboración de tablas de vida e igualmente se maximizaron algunos parámetros físicos, ambientales y nutricionales que garantizan el mantenimiento y la continuidad de la especie en condiciones de laboratorio. Esta colonia fue establecida con el objeto de proporcionar la materia prima biológica para los explantes de tejidos embrionarios que se han venido utilizando para establecer líneas celulares de crecimiento continuo

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.