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PCR and DNA hybridization indicate the absence of animal filariae from vectors of Onchocerca volvulus in Uganda.

In order to identify Onchocerca volvulus larvae from vectors, DNA of filaria larvae from dissected blackflies was isolated, and a 150-bp long tandemly repeated DNA sequence (0-150), which occurs in many Onchocerca species, was amplified using polymerase chain reaction (PCR). Subsequently, the PCR product was blotted onto a nylon membrane and hybridized with DNA probes specific for O. volvulus or Onchocerca ochengi. Filaria larvae from 395 infected Simulium neavei were examined and 259 samples produced detectable PCR products. Among these samples, 239 (92%) reacted with an O. volvulus-specific oligonucleotide. A sample of 69 PCR products was tested using an O. ochengi DNA probe, but all failed to hybridize. Filaria larvae from 64 infected Simulium damnosum, presumably of the cytotypes "Nyamagasani" and "Nkusi" were studied and 0-150 was amplified from 38 samples. From these samples, 35 (92%) hybridized specifically with an O. volvulus probe but none with the O. ochengi-specific DNA sequence. Nonamplified samples were obtained mainly from blackflies that contained only 1 or 2 filaria larvae, and therefore, an insufficient DNA extraction was assumed. It can be concluded that few, if any, filaria species of animal origin were transmitted by S. neavei and S. damnosum s.l. in Kabarole and Kasese districts in Uganda.

Animals↗

Onchocerca volvulus: comparison of field collection methods for the preservation of parasite and vector samples for PCR analysis.

In recent years, methods for the identification of the filarial worm Onchocerca volvulus and its vector, blackflies of the Simulium damnosum complex (S. damnosum sensu lato (s.l.)), based on the amplification of parasite and vector DNA sequences with the polymerase chain reaction (PCR), have been developed. Routine application of these methods requires techniques for sample collection and preservation that are compatible with the limitations of field collection, yet preserve DNA in a form suitable for PCR. Two different methods for sample preservation were evaluated by the field collection teams and the DNA probe laboratory of the Onchocerciasis Control Programme in West Africa. The most successful involved the preservation of material from O. volvulus and its associated vectors in a dried state on microscope slides. Of over 1200 parasite samples preserved in this manner, more than 93% retained DNA yielding positive results in PCR analysis (1208/1291). Vector material (malpighian tubules and ovaries) preserved in the same manner on the same microscope slides also yielded DNA that was suitable for PCR.

Animals↗

[Window traps and displacement of adult Simulium damnosum s.l. along a line of breeding sites in a forest zone of Côte d'Ivoire].

The displacement of adult black flies along a line of breeding sites has been studied in savannah area and in pre-forest zone. This paper presents the results of studies carried out in forest zone of Côte d'Ivoire. Interception traps made with sticky window traps were vertically displayed cross the river. The results showed that the displacement of adult S. damnosum s.l. along a line of breeding sites is correlated with the river flow. In a period of low discharge, adult black flies present a marked tendency to fly up stream. But in period of heavy discharge, they do not show any tendency; they may fly ever up stream or down stream. They fly essentially at the water level during the period of low flow and at a height between 0.5 to 1.5 m during heavy flow of the river.

Animals↗

[Human onchocerciasis in Africa].

Before the 1980s, the only available method for control of onchocerciasis was elimination of blackfly vector populations. This strategy was used with considerable success in the Onchocerciasis Control Programme in West Africa (OCP). The discovery of ivermectin, the first effective drug suitable for mass treatment of onchocerciasis, has revived international interest not only in fundamental research but also in development of new strategies to control onchocerciasis in the countries outside the OCP area. This report gives an overview of current parasitological, clinical, epidemiological and diagnostic data about onchocerciasis. Although little is known about the early development of Onchocerca volvulus in the human host, significant insight has been gained into the population dynamics of the parasite. The pathogenesis of cutaneous and ocular manifestations in onchocerciasis is now better understood. Epidemiological studies are under way to evaluate the extent of systemic manifestations. Recently developed diagnostic methods are more sensitive than conventional parasitological techniques. A new method for rapid assessment of endemic level has provided a detailed picture of the distribution of onchocerciasis. Species- and strain-specific DNA probes have been developed for identification of parasites in West Africa. New methods for quantifying disability allow evaluation of the socio-economic impact of the cutaneous and ocular complications of onchocerciasis.

Africa↗

Detection of Onchocerca volvulus DNA in pools of wild-caught Simulium ochraceum by use of the polymerase chain reaction.

The presence of Onchocerca volvulus DNA in experimentally infected flies can now be detected by use of the PCR, so that, for example, one infected Simulium damnosum can be detected in a pool of 100 uninfected flies or one S. ochraceum can be detected in pools of 20-40. As this PCR technique is specific for O. volvulus, the results are not confounded by the presence of other, unimportant, Onchocerca species, and the technique could replace time-consuming, manual dissection of flies. In 1996 and 1997, pools of 16-21 Simulium ochraceum were tested by the PCR technique. These flies had been collected biting man, between 1992 and 1994, from two hyperendemic coffee estates (fincas) in Guatemala, and stored in commercial (95%) ethanol. Collections at finca Buena Vista (869 flies in 52 pools) were made 1-2 weeks and 46 weeks after 45% of eligible subjects had been treated with ivermectin for the first time. At finca El Brote, collections (360 flies in 18 pools) were made 13 weeks before and 7 weeks after 97% of eligible subjects had received their first treatment. DNA was easily recovered from simuliids that had been stored in ethanol for up to 4 years. Of the nine pools of flies with visible blood collected at Buena Vista, each of 20 flies, eight tested positive for O. volvulus DNA. In flies without blood, 13 of 22 pools collected at Buena Vista just after treatment tested positive, whereas there were 14 positives in 22 pools taken 46 weeks later (P > 0.05). At El Brote, nine of 10 pre-treatment pools were positive, compared with three of eight taken 7 weeks post-treatment (P = 0.04), indicating that the treatments in this finca had reduced infection in the vector, and possibly transmission, by about 60%. A sub-sample of Buena Vista flies was divided into 19 sets of three separate sub-pools containing heads, thoraces and abdomens. Three pools of heads alone were positive, and had corresponding pools of positive abdomens. Three positive pools of thoraces had negative corresponding pools of heads and abdomens. These results show that PCR can be used to determine the prevalence of O. volvulus DNA in wild-caught S. ochraceum. As the infection rates observed were higher than expected from dissections reported by other workers, PCR-determined rates may not be directly comparable with traditional parameters based on the dissection of flies to reveal O. volvulus larvae.

Animals↗

Blackfly control: what choices after onchocerciasis?

Blackflies are reappearing in areas of West Africa where they used to be controlled with insecticides because they were vectors of the parasite Onchocerca volvulus. Even though they no longer transmit onchocerciasis in these areas they can hinder optimal land use through their biting behaviour. The authors discuss the problems associated with resuming the use of insecticides to control the blackfly and recommend that ground treatment be restricted to areas where it is likely to be effective on a continuing basis. In communities lacking technical and financial resources the only alternative consists of individual protection through the use of repellents or protective clothing.

Africa, Western↗

The impact of Mectizan on the transmission of onchocerciasis.

For many years there was no suitable drug available for the control of onchocerciasis. The advent of Mectizan (ivermectin, MSD; an effective microfilaricide), its registration in October 1987 for the treatment of human onchocerciasis, and its suitability for large-scale application were major break-throughs in the control of human onchocerciasis via chemotherapy. Several studies, both fly-feeding experiments and community trials, have established that Mectizan treatment causes a significant reduction in the transmission of infection. Although long-term treatment in some isolated foci (such as occur in the New World and in some hypo- and meso-endemic areas elsewhere) appears to interrupt transmission, more prolonged treatment is required to prove if transmission can be stopped. Advantage could be taken of the significant impact of Mectizan on transmission by giving treatment while or just before transmission by blackflies is most intense.

Africa, Western↗