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In the detection of parasitic infection, the traditional methods based on microscopy often have low sensitivity and/or specificity compared with the newer, molecular tests. An assay based on real-time PCR and a reagent strip test for detecting circulating cathodic antigen (CCA) have both now been compared with urine filtration and microscopy, in the detection of Schistosoma haematobium infections. Urine samples, obtained from 74 'cases' in areas of Ghana with endemic S. haematobium and 79 'controls' from non-endemic areas, were each checked using the three methods. With the results of the filtration and microscopy taken as the 'gold standard', real-time PCR was found to be 100% specific and 89% sensitive whereas the CCA strips were 91% specific and 41% sensitive. With the samples found to contain > or =50 eggs/10 ml (indicating relatively intense infections), the sensitivities of the PCR and CCA were higher, at 100% and 62%, respectively. As expected, egg counts were negatively correlated with the number of amplification cycles needed, in the PCR, to give a signal that exceeded the background (r=-0.38; P<0.01). Although the real-time PCR and CCA strip tests are very different, both show promise in the detection of S. haematobium infections. The PCR has optimal specificity and high sensitivity but the specificity of the CCA strips and the sensitivity of both tools could still be improved. A more thorough re-evaluation of the sensitivity and specificity of microscopy and these newer diagnostic methods, with an estimation of the cost-effectiveness of each technique, is recommended.

Although Ethiopia is one of the countries worst affected by human onchocerciasis, the exact taxonomic identity of the blackflies acting as the main vectors in the endemic areas has never been determined. A cytotaxonomic analysis of Simulium damnosum s.l. collected from three endemic sites in south-western Ethiopia has now revealed the existence of the 'Kisiwani' form (a non-anthropophilic cytoform that is common in East Africa) and a newly recognized species, Simulium kaffaense. Simulium kaffaense sp. nov. is differentiated from other members of the S. damnosum complex by six fixed inversions and dozens of 'new' floating inversions. The rearing of egg batches from some of the biting adult females, to larvae or adults, indicated that the human-biting blackflies were all S. kaffaense. As S. kaffaense is not only highly anthropophilic but also, apparently, the only anthropophilic member of the S. damnosum complex present, it is likely to be the main (if not the only) vector of Onchocerca volvulus in the study area. The presence of inversion 1S-1 and a complex inversion possibly involving 1L-3 indicates that S. kaffaense either belongs or is close to the 'Nile' phylogenetic group of S. damnosum s. l. The karyotype frequencies of the inversions in the collections from the three study sites indicate that at least two forms of S. kaffaense, here designated 'Bebeka' and 'Jimma', were caught. The taxonomy and medical importance of S. kaffaense are discussed.

The ability of vector mosquitoes to transmit the microfilariae (mff) of Wuchereria bancrofti, especially when the levels of microfilaraemia in the humans on which the mosquitoes are feeding are very low, is very important for understanding the transmission dynamics of lymphatic filariasis. Data on the correlation between vector competence and the microfilarial load in the human host are also relevant to those trying to improve transmission models for this disease. The majority of the relevant studies have involved culicine rather than anopheline vectors. The competence of populations of Anopheles mosquitoes to transmit W. bancrofti in a district in the Upper East region of Ghana has now been investigated. The wild mosquitoes that fed on 20 volunteers under natural conditions were collected hourly during the night, from 21.00 hours on one day to 06.00 hours on the next. Overall, 1348 fed female mosquitoes--665 Anopheles, 662 Culex and 21 Aedes--were collected. Approximately 50% of the mosquitoes caught were killed immediately post-collection and dissected so that the number of W. bancrofti mff each had ingested could be counted. The remaining mosquitoes where dissected when they died (if this was within 12 days of collection) or when they were killed on day 12 post-collection. With the exception of one Culex mosquito that harboured one microfilaria, none of the Culex and Aedes mosquitoes were found infected with W. bancrofti. All of the other mosquitoes found infected were An. gambiae s.l. or An. funestus. When fingerprick samples of blood, collected hourly from the volunteers during the mosquito infection, were used to estimate the microfilaraemias in the blood on which these mosquitoes had fed, microfilarial uptake and the number of developing larvae were found to vary considerably even when the microfilaraemias in the bloodmeal source were similar. The results of a regression analysis on the pooled data for the Anopheles mosquitoes indicated the process of limitation, although larger samples need to be investigated to determine whether this process occurs only in An. gambiae s.l. or An. funestus or in both of these taxa., (Copyright 2004 The Liverpool School of Tropical Medicine)

The pathogenesis of the sub-optimal response of Onchocerca volvulus to ivermectin was investigated in a 30-month follow-up of 28 individuals who, in a previous study, had been found to show a sub-optimal (N = 15) or adequate response (N = 13) to multiple treatments with the drug. Verbal informed consent was obtained before each subject was given a general clinical and ocular examination. Skin snips were taken from both iliac crests and both calves. Seventeen nodule carriers were hospitalized for nodulectomy. Adult worms were harvested, embryogrammes were constructed and all developmental stages were counted; degenerate, stretched microfilariae were noted separately. All the subjects were in good general health and all except one had received at least one additional treatment with ivermectin since the earlier study. A large proportion of the adult female worms in 10 out of the 11 sub-optimal responders who were nodule carriers were in full embryonic production but most of the stretched microfilariae they carried were degenerate. This picture is similar to that found in adult worms exposed to the first dose of ivermectin. In one subject who had no viable worms in his nodules, the existence of occult but actively reproductive worms was inferred from the high level of microfilaridermia observed less than 12 months after treatment. These observations confirm the existence of populations of adult female O. volvulus that respond poorly to repeated doses of ivermectin. The use of suramin in the treatment of the sub-optimal responders is discussed.

If ivermectin-based programmes for the control of human onchocerciasis are to be successful, the drug must remain effective for as long as necessary. In an open, case-control study, an attempt was made to determine if the persistent, significant, Onchocerca volvulus microfilaridermias seen in some individuals who had received at least nine treatments with ivermectin were the result of the development of drug resistance in the parasite. Twenty-one of these 'sub-optimal' responders (cases) were matched, by age, weight, number of treatments, locality and skin microfilarial counts, with seven amicrofilaridermic responders and 14 ivermectin-naive subjects. The number of treatments taken, any potential drug interactions and significant underlying disease were determined from detailed clinical and laboratory studies. Each subject was treated with ivermectin during the study, so that plasma concentrations of the drug could be determined for 72 h from the time of dosage. The microfilarial and adult-worm responses to this treatment were assessed from skin microfilarial counts (obtained before the treatment and at days 8, 90 and 365 post-treatment), day-90 embryogrammes, and the results of fly-feeding experiments. Parasite-sensitivity criteria for various time-points were derived from earlier data on skin microfilaridermias and the effects of ivermectin on the adult worms. The results indicate that the significant microfilaridermias that persist despite multiple treatments with ivermectin are mainly attributable to the non-response of the adult female worms and not to inadequate drug exposure or other factors. The possibility that some adult female worms have developed resistance to ivermectin cannot be excluded. These results justify the routine monitoring of treatment efficacy in any ivermectin-based programme of disease control.

PCR has recently been studied as a promising tool for monitoring the progress of efforts to eliminate lymphatic filariasis. PCR can be used to test concurrently at least 30 pools, with as many as 40 mosquitoes in each pool, for the presence of filarial larvae. The SspI PCR assay for the detection of Wuchereria bancrofti DNA in pools of mosquitoes has been used since 1994 in a variety of laboratories worldwide. During that time, the original assay has been modified in these different laboratories and no standardized assay currently exists. In an effort to standardize and improve the assay, a meeting was held on 15-16 November 2001, at Emory University in Atlanta, with representatives from most of the laboratories currently using the assay. The first round of testing was designed to test the four most promising methods for DNA extraction from pools of mosquitoes. Two of the four methods stood out as clearly the best and these will be now optimised and evaluated in two further rounds of testing.

Previous studies on the morphological identification of adult female Simulium damnosum s.l. have involved use of colour characters. To determine the degree to which these qualitative characters are influenced by the physical environment, a portable rearing system was constructed. Wild-caught female flies and flies emerging from pupae maintained at ambient temperatures or above showed entirely pale antennae, fore-coxae, wing-tufts and hairs on the scutellum and ninth abdominal tergite, as is normal for S. sirbanum. Adults reared from larvae at below ambient temperatures were also pale, except for the antennae which were dark in all specimens. Adult females emerging from pupae maintained at below ambient temperatures were entirely pale up to the fourth day of emergence and mostly with dark antennae. The biological significance and the implications to the morphological identification of the savannah species of the S. damnosum complex in West Africa are discussed.