Inverse density dependence of parity rates in the onchocerciasis vector Simulium damnosum s.l.

A correlation between parity rates and an index of adult numbers of Simulium damnosum s.l. (Diptera: Simuliidae) indicates an association, but does not prove causality or show the direction of any causal relationship. The question of whether adult numbers affect parity rates or vice versa is reminiscent of the age‐old query of which of the chicken and the egg came first. A method for resolving such issues based on analyses of pairs of time series was proposed by Granger in 1969. When Granger's method was applied to monthly numbers of adult female S. damnosum s.l. caught attempting to bite humans at Asubende, Ghana, and their parity rates, a significant relationship (P = 0.005) emerged, clearly showing that parity rates were dependent on adult numbers. Implications of this inverse density dependence and the results of analyses of other similar time series are presented and discussed.     

The species delimitation problem in the Simulium damnosum complex, blackfly vectors of onchocerciasis

The Simulium damnosum Theobald complex (Diptera: Simuliidae) comprises 57 cytoforms grouped into six subcomplexes. Previous phylogenetic studies using gene sequences have not completely resolved the evolutionary relationships of the cytoforms. The present study investigated the systematics of the complex using a phylogeographic approach. The differentiation between eastern and western forms observed in the phylogenetic studies is confirmed in the estimated haplotype networks. However, haplotypes tend to group in geographical clades and not according to cytoforms. Spatial analyses of the molecular variance also resulted in optimal groupings of sequences that did not correspond to cytoform boundaries. Moreover, Mantel tests showed significant correlations, although not strong, between genetic and geographical distances. This suggests an isolation-by-distance model of differentiation. Furthermore, there are instances in which genetic differentiation between cytoforms is low and not significant. These results indicate a lack of clear genetic differentiation between the cytoforms, which may be explained either by a separation of the taxa recent enough to allow the accumulation of few genetic differences or by recombination between the genomes of the cytoforms, which may be the result of hybridization with introgression or of non-independent evolutionary lineages. The results also emphasize the need for further sampling and for the use of more variable markers in order to clarify the evolutionary history of the group.     

An 18S ribosomal DNA barcode for the study of Isomermis lairdi, a parasite of the blackfly Simulium damnosum s.l.

The mermithid parasite, Isomermis lairdi Mondet, Poinar & Bernadou (Nematoda: Mermithidae), is known to have a major impact on populations of Simulium damnosum s.l . Theobald (Diptera: Simuliidae) and on their efficiency as vectors of Onchocerca volvulus (Leuckart) (Nematoda: Filarioidea). However, the value of I. lairdi and other mermithid parasites as potential means of integrated vector control has not been fully realized. This is partly because traditional taxonomic approaches have been insufficient for describing and analysing important aspects of their biology and host range. In total, rDNA barcode sequences have been obtained from over 70 I. lairdi mermithids found parasitizing S. damnosum s.l . larvae in three different rivers. No two sequences were found to vary by more than 0.5%, and cytospecies identification of mermithid hosts revealed that I. lairdi with identical rDNA barcodes can parasitize multiple cytoforms of the S. damnosum complex, including S. squamosum (Enderlein). Phylogenetic analysis using a partial sequence from the 18S ribosomal DNA barcode, grouped I. lairdi in a monophyletic group with Gastromermis viridis Welch (Nematoda: Mermithidae) and Isomermis wisconsinensis Welch (Nematoda: Mermithidae).     

The Beffa form of Simulium soubrense of the S.damnosum complex in Togo and Benin

The Beffa form of Simulium soubrense Vajime & Dunbar, a member of the S. sanctipauli sub-complex of the S. damnosum complex, was found breeding throughout rivers in the Togo-Benin Gap, as far north as 9 degrees 30'N. Its distribution changed with the season. In southern Togo there were seasonal fluctuations in the relative abundancies of the Beffa form and of S. damnosum/S.sirbanum. There was considerable temporal and regional variation in the frequencies of different colour morphs of adult flies. The flies in Benin tended to be darker. Infections with Onchocerca volvulus (Leuckart) appeared to be independent of the host's colour morph category. Larger flies harboured significantly more first stage Onchocerca larvae but no significant relations with fly size were found for second and third stage larvae.     

Oviposition aggregation pheromone in the Simulium damnosum complex

Abstract. Communal oviposition by the Simulium damnosum complex of Afrotropical blackflies (Diptera: Simuliidae) was investigated under controlled laboratory conditions, using wild-caught flies in Sierra Leone. Volatile compounds emitted by Simulium eggs were trapped using a closed collection system, and their attractiveness to gravid flies was tested in a two-choice behavioural bioassay. Significantly more female blackflies oviposited on substrates baited with freshly laid eggs (100% chose the baited substrate), or with the volatiles collected from freshly laid eggs (85% chose the baited substrate), in preference to the relevant control substrates. Substrates baited with volatiles from 12-h-old eggs were not significantly more attractive than controls (only 31% chose the baited substrates; P = 0.33). Gas chromatographic analysis of the egg volatiles consistently showed two peaks emanating from fresh eggs, but significantly lower amounts from 12-h-old eggs ( P <0.05). A novel system for collecting the volatiles from this and other blackfly species, as they laid eggs on a substrate in flowing water, is described. Volatiles collected using this method showed identical gas chromatographic profiles to those of fresh eggs alone, indicating that the flies themselves produced no other volatile chemical signals during oviposition. Evidently communal oviposition by S. damnosum s.l . was mediated by a pheromone emanating from fresh eggs. The role of pheromone-mediated egg aggregation in blackfly ecology is discussed, and its possible manipulation is considered.     

Oviposition pheromone in the Simulium damnosum complex: biological activity of chemical fractions from gravid ovaries

Abstract. Communal oviposition in the Afrotropical blackfly species complex Simulium damnosum Theobald (Diptera: Simuliidae) is mediated by a pheromone emitted by freshly laid eggs. Previously, two compounds (designated peaks A and B) emanating from fresh eggs were shown to be associated with attractiveness to gravid blackflies in bioassay. The present study investigated the role of these compounds by testing the responses of wild-caught Simulium yahense in Ghana to fractionated hexane extracts of gravid ovaries prepared by gas chromatography (GC). Although the fractions were prepared from Sierra Leonean Simulium leonense , GC analysis of the emissions from fresh S.yahense eggs showed that the volatile blends of both species were similar. When tested in a two-choice bioassay, 66% of ovipositing blackflies chose the substrate baited with a mixture of the four fractions recombined. In a series of bioassays testing responses to the four individual fractions presented with a control in a multiple-choice arrangement, only fraction 3 (containing peaks A and B) attracted significantly more ovipositions than the other fractions and control. However, fraction 3 failed to elicit a significant response when presented as the sole attractant with a control in a two-choice bioassay. It was concluded that fraction 3 , though mainly responsible for mediating aggregated oviposition by S.yahense , was acting in tandem with additional cues, probably further chemicals, which remain to be isolated and characterized.     

Management of insecticide resistance in control of the Simulium damnosum complex by the Onchocerciasis Control Programme, West Africa: potential use of negative correlation between organophosphate resistance and pyrethroid susceptibility

1. Resistance of some populations of the Simulium damnosum complex to temephos (100-fold at the LC50 level), with degrees of cross-resistance to chlorphoxim (14-fold) and other organophosphate insecticides, follows intensive larvicidal control of S. damnosum s.l. in West African river systems since 1975 by the WHO Onchocerciasis Control Programme. 2. Larvae of at least three sibling species of the S. damnosum complex have become organophosphate-resistant: these are the forest species S. sanctipauli Vajime & Dunbar and the savanna species S. sirbanum V. & D. and S. damnosum Theobald sensu stricto. 3. Organophosphate-resistant S. damnosum s.l. larvae show increased susceptibility to some organochlorine and pyrethroid insecticides, especially to permethrin (up to 11-fold) and OMS 3002 (up to 17-fold), as compared with organophosphate-susceptible populations. 4. This differential susceptibility is reflected by increased pyrethroid efficacy in operational use for river treatments against organophosphate-resistant field populations of S. damnosum s.l. larvae. Treatment of 100 km of the lower Bandama River in 1985 showed that permethrin at the highly selective dosage of 10 min exposure to 0.01 mg/l caused reversion towards organophosphate-susceptibility of the target population of S. sanctipauli. This effect was less pronounced when the Comoe River was treated at the lower dosage of 0.005 mg/l for 10 min. 5. To overcome temephos-resistance, it is proposed that the most rational usage of currently available larvicides would involve the following annual sequence of treatments: Bacillus thuringiensis serotype H-14 when river discharge is below 75 m3/s; chlorphoxim for about eight weekly treatment cycles after river discharge rises; permethrin (or alternative pyrethroid) for up to six treatment cycles--this should eliminate any incipient selection for chlorphoxim-resistance; resume chlorphoxim (or perhaps carbosulfan) treatments until river discharge falls below 75 m3/s permitting resumed use of B.t. H-14.     

Revision of the Ketaketa subcomplex of blackflies of the Simulium damnosum complex

A revision of the taxonomy of the Ketaketa subcomplex of the Simulium damnosum Theobald complex (Diptera: Simuliidae) is presented including new material from Tanzania, Malawi and South Africa. The cytotaxonomy, morphology and molecular identity of known and new taxa are described. The Ketaketa subcomplex is cytotaxonomically defined by the paracentric inversion 1L-7. We recognize three sibling species, namely Simulium latipollex (Enderlein), Simulium plumbeum Krueger, sp.n. and Simulium kipengere Krueger, sp.n., the latter comprising three cytoforms: 'Typical', 'Linthipe' and 'Mombo'. The cytoforms 'Mwamphanzi', 'Ketaketa' and 'Hammerkopi' are synonymized with S. plumbeum. Identification keys are provided on the basis of chromosomal and morphological characters. In view of their potential role as vectors of human onchocerciasis (river blindness) we also discuss the possible medical importance of the different cytoforms and their geographical distribution.     

Four new cytotypes of the onchocerciasis vector blackfly Simulium guianense in Brazil

Abstract. Simulium (Trichodagmia) guianense is an important Amazonian vector of onchocerciasis. Examination of the polytene chromosome banding patterns of larvae from five sites in Brazil revealed the occurrence of four cytotypes, designated A, B, C and D. The chromosomal standard, Simulium guianense A , occurred at two localities in Goias State (on the Rio Tocantins and Rio Mucambao) where it was the only cytotype. The other three sites examined yielded one different cytotype from each: B from Rio Oyapoque in Amapa State, C from Rio Tocantins in Maranhao State, and D from Rio Xingu in Para State. All cytotypes differed by at least two fixed inversions, but a sex determining system was not evident in any cytotype. As the cytotypes have been found allopatrically it is not certain that they represent sibling species; sampling of sympatric populations would resolve this. During certain times of the year, voracious anthropophagy by S.guianense sensu lato occurs at the localities sampled for cytotypes A, C and D (biting data are not available for the cytotype B locality). In some other areas, however, S.guianense s.l. is entirely zoophilic. Further studies are needed, therefore, to elucidate the biting habits, vectorial capacity, geographic distribution and taxonomic status of these four, and perhaps additional, cytotypes comprising the S.guianense complex.     

Experimental hybridization between members of the Simulium damnosum complex

Abstract. Males of the Beffa form of Simulium soubrense Vajime and Dunbar were successfully crossed with female S.squamosum V. & D. and S.sanctipauli V. & D. Fertile eggs from both crosses were reared to larvae and, in the case of the BeifaXsquamosum cross, to adults.
Larval chromosomes from the Beffa x sanctipauli cross had normally paired homologues but those of the Beffa xsquamosum cross showed extensive asynapsis and large heterozygous loops. The morphology of the larval and adult hybrids from the BeftaXsquamosum cross were more characteristic of squamosum , but the adult males had scutal patterns typical of the Beffa form and retained this form's laboratory-mating propensity.     

Sex chromosome variation and cytotaxonomy of the onchocerciasis vector Simulium squamosum in Cameroon and Nigeria

On the basis of sex chromosome variation, three cytotypes of Simulium squamosum (Enderlein) (Diptera: Simuliidae) are described from Cameroon and Nigeria. Simulium squamosum A is the typical form as originally described by Vajime & Dunbar (1975) with chromosome I as the sex chromosome. It occurs throughout most of Cameroon and south-east Nigeria. A second cytotype, S. squamosum B, is described from the river Sanaga (Cameroon). It also has chromosome I as the sex chromosome, but the nature of the sex differential region is different. Simulium squamosum C has no sex-linked chromosomal rearrangements. It is widespread in Nigeria and occurs near Mount Cameroon, where it seems to hybridize with S. squamosum A.     

Efficacy of DEET and non‐DEET‐based insect repellents against bites of Simulium damnosum vectors of onchocerciasis

Coping strategies including smoke screens are used against nuisance bites of Simulium damnosum Theobald (Diptera:Simuliidae) in onchocerciasis endemic communities. To find more effective alternatives, the efficacy of commercially available N,N‐diethyl‐3‐methylbenzamide (DEET) products with active concentrations of 9.5, 13, 25, 50 and 98.1–100% and ‘NO MAS,’ (active component: para‐menthane‐3,8‐diol and lemon grass oil) were tested at Bui‐Agblekame, Ghana. A Latin square study design was implemented using eight groups of two vector collectors each, who used repellents (treatment), mineral oil or nothing each day until the end of the study. Flies were caught and their numbers each hour recorded using the standard methods for onchocerciasis transmission studies. T‐tests were used to compare the mean duration of protection and a one‐way analysis of variance controlling for catchers and repellents was performed. Tukey's test was used to compare protection by repellents and mineral oil. The highest percentage protection was 80.8% by NO MAS and the least 42.5% by the 13% DEET product. The period of absolute protection was 5 h by NO MAS and 1 h by 50% DEET product. No significant increase in protection was offered beyond 25% active DEET products and no significance was observed in terms of catcher × repellent effect (F = 1.731, d.f. = 48, P = 0.209).     

Elimination of the Djodji form of the blackfly Simulium sanctipauli sensu stricto as a result of larviciding by the WHO Onchocerciasis Control Programme in West Africa

Cytotaxonomic identifications of larvae of members of the Simulium damnosum Theobald (Diptera: Simuliidae) complex collected in forest zones of southeast Ghana and southwest Togo between 1977 and 1996 showed that the Djodji form of Simulium sanctipauli Vajime & Dunbar, a vector of onchocerciasis, was eliminated in 1988 by larvicide operations conducted by the World Health Organization (WHO) Onchocerciasis Control Programme (OCP) in West Africa. No members of the form were identified amongst 997 larvae collected up to 8 years after systematic control operations began in February 1988. The results are discussed in relation to estimates of the numbers of samples required to certify elimination and the possibility that other members of the S. damnosum complex were also eliminated by the OCP.