Molecular phytogeny and typing of blackflies (Diptera: Simuliidae) that serve as vectors of human or bovine onchocerciasis

Abstract. A subregion of the mitochondrial large subunit (16s) rRNA gene was amplified by polymerase chain reaction (PCR) from nine species of blackflies (Diptera: Simuliidae) which serve as natural or experimental vectors of human or bovine Onchocerca parasites. PCR products from each species of blackfly were tested by directed heteroduplex analysis (DHDA), and their genotypes established according to diagnostic banding patterns of the heteroduplex products. Three alleles of mitochondrial 16s rRNA were found to exist in members of the Simulium (Ewardsellum) damnosum sensu lato complex from West Africa, and two alleles were found in the Neotropical Simulium (Psilopelmia) ochraceum Walker complex and the Simulium (Simulium) metallicum Bellardi complex. Different single alleles were detected in Austrosimulium bancrofti , in English S.(S.)noelleri and in two North American laboratory vectors: Simulium (Psilozia) vittatum Zetterstedt and S.(S.)decorum Walker. Phylogenetic analysis of 16s sequences indicated that blackflies from West Africa and the Americas formed distinct clades. Neotropical onchocerciasis vectors were found to be more closely related to Nearctic and Palaearctic non-vector Simulium species than to the African vectors of onchocerciasis.     

Anthropophilic blackflies (Diptera: Simuliidae) and onchocerciasis transmission in southwest Ethiopia

Abstract. From May 1984 to May 1985, the transmission of onchocerciasis by blackflies (Dipteria: Simuliidae) was studied around Gilgel Ghibe and Gojeb rivers in southwest Ethiopia. The predominant anthropophilic species of Simulium at both localities were S.(Edwardsellum) damnosum sensu law and S.(Lewisellum) ethiopiense. Some S.(Anasolen) dentulosum also landed on human bait at Gilgel Ghibe river.
Dissections of S. damnosum from Gilgel Ghibe and Gojeb revealed mean parous rates of 92% (n = 18,291) and 84% (n = 9530), respectively. S. ethiopiense from Gilgel Ghibe and Gojeb showed mean parous rates of 53% (n = 322) and 93% (n = 14), respectively. Of the parous S. damnosum , 1.3% at Gilgel Ghibe and 0.5% at Gojeb harboured infective third-stage larvae (L3) of Onchocerca volvulus (or morphologically indistinguishable from it). Unknown filariae of animal origin, indicative of zoophily, were found in 0.3% and 0.7% of Simulium damnosum complex females from Gilgel Ghibe and Gojeb, respectively. S. ethiopiense harboured developing (L,) larvae only, with 7.7% infection rate in both localities. In contrast, S. dentulosum did not harbour any filaria larvae. The annual infective biting rate (AIBR) and transmission potential (ATP) of the S. damnosum complex at Gilgel Ghibe river were 858 and 5478, respectively. The AIBR and ATP of S. damnosum s.I. at Gojeb river were 519.5 and 1963, respectively. These results emphasize the predominant role of the S. damnosum complex in the transmission of O. volvulus in southwest Ethiopia     

Estimation of preferred water flow parameters for four species of Simulium (Diptera: Simuliidae) in small clear streams in South Africa

Blackfly larvae typically occur in fast-flowing riffle sections of rivers, with different blackfly species showing preferences for different hydraulic conditions. Very little quantitative data exist on hydraulic conditions linked to the blackfly species occurring in South African streams. Stones-in-current biotopes (i.e. fast riffle flows over cobbles) were sampled from four sites in three small clear streams in the Eastern and Western Cape provinces of South Africa. Mean water column velocities at each sampled stone were measured using a mini current meter, while flow velocities closer to the boundary layer where blackfly larvae occurred were estimated using indirect techniques (standard hemispheres and aerating tablets). Standard hemispheres were also used to calculate more complex hydraulic parameters such as Froude and Reynolds numbers. Four species of Simuliid were sampled in sufficient numbers to show trends in flow velocity preferences. Simulium impukane and S. rutherfoordi both occurred at their highest densities at velocities of 0.3m s^?1, while S. merops preferred velocities of 0.7m s^?1. Simulium nigritarse SL attained the highest densities of all the blackfly species sampled, and its relative abundances were greatest at velocities of 0.8–0.9m s^?1. Within the streams surveyed, all blackfly species occurred in subcritical-turbulent flows — based on a classification using Froude and Reynolds numbers — although two of the species were also found in high densities in supercritical flows where these existed at the sites. Local hydraulics within the stones-in-current biotope are complex, but in the absence of fine-scale equipment for measuring micro-velocities, standard hemispheres are a useful, cost-effective technique for the initial quantification of hydraulic parameters in small, clear streams. Such an approach facilitates further understanding of links between hydraulics and aquatic invertebrates in South African streams.     

New species records for the blackfly (Diptera-simuliidae) fauna of Argentina with description of adults, pupa and larva of Simulium oyapockense s. l. And S. seriatum

Two blackfly species Simulium (Cerqueirellum) oyapockense Floch & Abonnenc and S. (Hemicnetha) seriatum Knab are recorded from Argentina, representing the most southern register for both species. S. oyapockense is a species epidemiologically very important, as a vector of onchocerciasis in the Amazonian focus. Both species are described and illustrated and their distribution are reported, in similarity to others like S. roraimense Nunes de Mello and S. ganalesense Vargas et al. in reference to S. oyapockense and S. mexicanum Bellardi similar to S. seriatum are discussed.     

Patterns of covert infection by invertebrate pathogens: iridescent viruses of blackflies

Recently, it has been recognized that blackfly populations may host two forms of infection by iridescent viruses (IVs); a covert (inapparent, nonlethal) form which was common in springtime populations in the River Ystwyth, Wales, and a patent (obvious, lethal) form which was rare. This study aimed to investigate the changes in frequency of the two types of infection in blackfly populations over the reproductive period of the flies, April-September 1992. Blackfly larvae sampled from three different sites along the river were bioassayed for the presence of covert IV infection. Of 870 larvae assayed, 17 were found to be infected. All the infected larvae appeared to be Simnulium variegatum, the dominant species during the sampling period. IV infections were common in the spring (17–37% depending on site) but appeared absent in the S. variegatum population for most of the summer months, reappearing again in the autumn (0–20% infected). These fluctuations were concurrent with biotic and abiotic factors: elevated levels of covert infection occurred at low population densities, high water flow rates, low temperatures (and presumably slower growth rates), although it is not clear if any cause-and-effect relationship exists. Patent infections occurred immediately after the peak of covert infection in the spring, and again in the autumn. Virus characterization of isolates from covertly infected larvae showed that three distinct groups of isolates were present in the blackfly population. Isolates from the springtime populations were mostly variants of an isolate found in patently infected blackfly larvae in the 1970s (Aberystwyth IV). Isolates from the autumn populations were mostly variants of an isolate from a patently infected larva found in September the previous year. A third group comprised a single novel isolate which was detected in a covertly infected larva. The mechanisms by which IVs persist in blackfly populations remain unknown, although the role of alternative hosts is a possibility which needs to be studied.     

Balancing food availability and hydrodynamic constraint: phenotypic plasticity and growth in Simulium noelleri blackfly larvae

Organisms through phenotypic plasticity can cope with multiple changed environmental conditions. Theory predicts that animals in streams and rivers should be able to balance demands of the needs to obtain food efficiently and to adjust response to hydrodynamic variability. This study examined effects of variations in food availability and current velocity on the feeding structure and growth rate of Simulium noelleri blackfly larvae. The larvae developed larger labral fans and more rays under slow current and low food regimes than in fast current and high food conditions. In both fast and slow current regimes, growth rates were higher and development periods to the final-instar stage were shorter in high food treatments. The estimated flux rates of food particles through labral fans under high food treatments for both fast and slow current regimes were higher than those under low food treatments. Although both food and current velocity appeared to have selected for flexibility of feeding structure and growth rate, food availability was a more important factor for phenotypic and developmental plasticity than current velocity. The results indicate a strong link between environmental changes in food availability and current velocity, phenotypic plasticity, and growth rate of S. noelleri. This study suggests that plasticity of ecomorphs with macroevolutionary significance may play a role in the early evolutionary stages of blackfly larvae.     

Spatial and temporal abundance,substrate partitioning and species co-occurrence in a guild of Neotropical blackflies (Diptera: Simuliidae)

Blackfly larvae were sampled monthly from two smallNeotropical mountain streams 500 m apart from each other in northernVenezuela during a 15-month period, and ten habitat variables weremeasured at two altitudes (800 and 900 m) to determine local spatialand temporal variations in abundance, stream variables, speciesco-occurrence, and substrate preference within the blackflyguild. The blackfly species composition of each stream was the same,with five species (in order of abundance): Simulium ochraceumWalker s.l., S. paynei Vargas, S. metallicum Bellardicytospecies E, Simulium sp., and S. horacioi Okazawa &Onishi. There were clear differences in community structurebetween streams (not among altitudes), although S. ochraceums.l. was the most abundant species in both habitats. Principal component andcorrelation analyses revealed that structural (great depth, width,flow, and discharge) and chemical (low conductivity) factors werethe stream variables best associated to the spatial distribution ofmost blackfly species. Blackfly abundance was greater in the largerstream. Larvae were most abundant on submerged rocks and fallenleaves, although the patterns of substrate use within the guildrevealed substrate partitioning among most blackfly species.There was one general annual peak of abundance at the end of therainy season. We suggest that blackfly community structure in theseNeotropical headwaters streams, over the small spatial scaleexamined here, is largely influenced by interactions among streamsize and velocity-related factors (width, streamflow, discharge andsubstrate availability-stability) and the rainfall regime.     

Spatial and temporal distribution of blackflies (Diptera: Simuliidae) in the Itatiaia National Park, Brazil

The distribution of the blackfly larval taxocenoses of different altitudes in the Itatiaia National Park (located in the southeast of Rio de Janeiro state, Brazil 44 degrees 34' - 44 degrees 42' W, 22 degrees 16' - 22 degrees 28' S) was investigated on four campaigns, one in each season of 2003. On each campaign 60 samples were collected, using 30 x 30 cm wooden quadrats, distributed at four sites: Taquaral, Véu da Noiva, Três Picos and Brejo da Lapa. The winter and spring campaigns included two additional sampling sites: Alsene and Agulhas Negras. A total of 3578 larvae and 292 pupae were sampled, representative of six species: Simulium clavibranchium Lutz, S. subnigrum Lutz, S. rappae Py-Daniel, S. incrustatum Lutz, S. stellatum Gil-Azevedo, Figueiró & Maia-Herzog and Lutzsimulium pernigrum Lutz 1910. The highest larvae abundance and diversity were reported at the intermediate altitude site, Brejo da Lapa. Principal Components Analysis (PCA) revealed that L. pernigrum, S. stellatum and S. incrustatum were associated with the smaller breeding sites, the first two species being associated with sunny sites, while S. incrustatum occurred in sites with little sunlight.     

梵净山绳蚋二新种 (双翅目 :蚋科)(英文)

本文首次报道贵州省梵净山自然保护区绳蚋二新种 ,即梵净山绳蚋Simulium (Gomphostilbia) fan jingshanesesp .nov .和孟氏绳蚋S .(G .)mengisp .nov .对其两性成虫、蛹和幼虫进行全面描述并分别与其近缘种进行分类讨论。     

Influence of temperature on microsporidia infections in a natural population of Simulium pertinax Kollar, 1832 (Diptera; Simuliidae).

During the studies involving the correlation between the water temperature of the breeding site of Simulium pertinax larvae and the infection prevalence by microsporidia, developed in the Andorinhas river, Magé, RJ, weekly samples of blackfly larvae were taken within a two-year period (2001-2002 and 2003-2004), and it was noticed that the infections by Amblyospora sp. were more prevalent when compared to infections by Polydispyrenia sp. in larvae. It was also observed that the infections do not follow the same pattern, since the genus Amblyospora was recorded almost every month during the study with the exception of December, 2001. In the results of correlation between the environmental water temperature and the microsporidia infection rates, it was observed that for the first period studied, there was a high negative correlation, while during the second period there was absence correlation. On the other hand, the Amblyospora sp. infection rates prove that the correlation was high and significant in the first period, but was not significant in the second sampling period and Polydispyrenia sp. showed absence correlation in both periods.     

Temephos-resistant larvae of Simulium sanctipauli associated with a distinctive new chromosome inversion in untreated rivers of south-western Ghana

Larvae of the Simulium damnosum Theobald complex (Diptera: Simuliidae) were sampled in June 1996 from two sites in south-west Ghana where larviciding has not been applied: Sutri Rapids on the Tano river (05 degrees 23 minutes N 02 degrees 38 minures W) and Sekyere-Heman on the Pra river (05 degrees 11 minutes N 01 degrees 35 minutes W). All specimens were identified as Simulium sanctipauli Vajime & Dunbar sensu stricto (Diptera: Simuliidae). Bioassays with temephos (organophosphorus larvicide employed by the Onchocerciasis Programme for systematic treatment of most rivers across West Africa since the 1970s) showed about five-fold resistance in the Tano population (LC95 2.37-3.14 mg/L) and slight tolerance to temephos in the Pra population (LC95 0.67-0.76 mg/L), vs. the diagnostic concentration of 0.625 mg/L. Larval salivary polytene chromosomes of S. sanctipauli showed fixed inversions 1S-24/24, standard IIL-6 and a new inversion IL/36 polymorphism at Sutri on the Tano. These karyotype characteristics differ from those of temephos-resistant S. sanctipauli in rivers of C te d'Ivoire and other sites on the Tano in Ghana. Thus, temephos resistance in S. sanctipauli at Sutri is associated with distinct chromosomal configurations, showing that immigration was unlikely. This resistance could have been locally selected by exposure of S. sanctipauli larval populations to agrochemicals run-off from cocoa, coffee and oil plantations flanking the rivers.     

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.     

Effects of Polydispyrenia simulii (Microspora; Duboscqiidae) on development of the gonads of Simulium pertinax (Diptera; Simuliidae).

The effects of the infection caused by the microsporidium Polydispyrenia simulii in gonads of the simuliid Simulium pertinax were evaluated. This blackfly species is the main target of control programmes in southeastern Brazil. Infected and noninfected blackflies were reared from larval stage to adults using a laboratory rearing system. There was no significant difference between the oogenic development and dimensions of ovarian follicles of healthy females compared to those reared from infected larvae. Otherwise, there was a significant difference between the fecundity of healthy females and those reared from infected larvae. In males, there was an evident reduction of motility of spermatozoids of males reared from infected larvae. The results obtained suggest that the microsporidium P. simulii have an important effect on the natural population dynamics of S. pertinax.     

Larval description of Simulium (Notolepria) cuasiexiguum and Simulium (Chirostilbia) obesum and new records of black fly species (Diptera: Simuliidae) in the states of São Paulo and Minas Gerais, Brazil

We describe the larval stage of Simulium cuasiexiguum Shelley, Luna Dias, Maia-Herzog & Lowry and Simulium obesum Vulcano and provide new distribution records of four black fly species in the State of S?o Paulo (Simulium cuasiexiguum, Simulium metallicum s.l. Bellardi, Simulium stellatum Gil Azevedo, Figuer6 & Maia-Herzog and Simulium oyapockense s.l. Floch & Abonnenc) and three species in the State of Minas Gerais (Simulium duodenicornium Pepinelli, Hamada & Trivinho-Strixino, Simulium obesum and Lutzsimulium pernigrum Lutz).     

Complete genome sequence of invertebrate iridovirus IIV22A,a variant of IIV22, isolated originally from a blackfly larva

Members of the family Iridoviridae are animal viruses that infect only invertebrates and poikilothermic vertebrates. The invertebrate iridoviruses 22 (IIV22) and 25 (IIV25) were originally isolated from a single sample of blackfly larva (Simulium spp., order Diptera) collected from the Ystwyth river near Aberystwyth, Wales. Recently, the genomes of IIV22 (197.7 kbp) and IIV25 (204.8 kbp) were sequenced and reported. Here, we describe the complete genome sequence of IIV22A, a variant that was isolated from the same pool of virions collected from the blackfly larva from which the IIV22 virion genome originated. The IIV22A genome, 196.5 kbp, is smaller than IIV22. Nevertheless, it contains 7 supplementary putative ORFs. Its analysis enables evaluation of the degree of genomic polymorphisms within an IIV isolate. Despite the occurrence of this IIV variant with IIV22 and IIV25 in a single blackfly larva and the features of their DNA polymerase, we found no evidence of lateral genetic transfers between the genomes of these two IIV species.     

Laboratory infection of blackflies (Simuliidae) and midges (Chironomidae) by the mosquito mermithid, Romanomermis culicivorax

Penetrating infective juveniles of Romanomermis culicivorax usually killed first-stage larvae of Chironomus maturus Johann., Chironomus sp., Simulium damnosum Theo., and S. venustum Say. Nematodes were melanized and died after they entered fourth stage larvae of 2 chironomid species, but no host reaction was evident after entry into fourth-stage blackfly larvae. In contrast, the nematodes initiated development in the latter hosts, which died before the nematodes completed their development.     

Changing risk of predation for a filter-feeding insect along a current velocity gradient

Flume experiments were carried out to examine whether larval blackflies (Simulium ornatum complex, Diptera: Simuliidae) use microhabitats with a highvelocity current to reduce the risk of predation by some of their main predators, viz. larvae of the stoneflies Isoperla grammatica and Diura nanseni (Plecoptera: Perlodidae), and the caddis-fly Rhyacophila nubila (Trichoptera: Rhyacophilidae). We exposed blackfly larvae to four different current velocities and measured their feeding rate using dye particles. The maximum feeding rate was recorded at intermediate velocities (18.8 and 36.2 cm/s), whereas at low (7.3 cm/s) and high (53.3 cm/s) velocities, the feeding rate was reduced. In separate experiments, we investigated the behaviour and attack success of the different predator species. The two perlodids showed a similar hunting behaviour, which was significantly less successful at higher velocities. Drift of the perlodids from the experimental arena resulted in reduced encounter and attack rates, especially in I. grammatica, which had completely lost efficiency at 36.2 cm/s. R. nubila had a slower mode of hunting and was unaffected by current speed within the velocity gradient studied. Drift in Rhyacophila was rare. Observations on the behaviour of blackfly larvae were performed in the same experiments. The larvae showed no apparent ability to sense the presence of the predators except when these disrupted the flow pattern or were in physical contact, which often resulted in aggressive defence, though without effect on the predators. Escape of blackfly larvae by drift did occur, but this was no more common than being captured. In a current velocity gradient, blackfly larvae showed a weak preference for increasing velocities. Thus, at velocities between 7 and 54 cm/s, blackfly larvae appear to select microhabitats with high current velocities, despite a reduction in feeding optimality, thereby easing the predation impact from perlodids, though not from Rhyacophila. The study demonstrates the importance of microhabitat selection by blackfly larvae both for efficient feeding and predator avoidance.     

Feeding behavior of larval black flies (Diptera: Simuliidae) with and without exposure to Bacillus thuringiensis var. israelensis.

Behavioral time budgets were constructed for larval black flies of the Simulium jenningsi and S. tuberosum groups in a recirculating flume. Larvae spent the majority (54-80%) of time filter feeding. Flick rates of the labral fans, which averaged 0.7-1.9 flicks/sec, did not differ significantly between regimens of 0 mg/l and 10 mg/l of food. Larvae consistently flicked the fan nearest the water surface significantly more often than the lower fan. They also tended to flick the right fan more than the left, although this trend was significant in only one of four treatments. Although handedness cannot be dismissed, the differential flick rate appeared to be related to the position of the fans relative to the water surface, possibly because of different particulate concentrations in the microcurrents received by the upper and lower fans. Cessation of feeding by larvae of the S. jenningsi group from rivers regularly treated with Bacillus thuringiensis var. israelensis was independent of exposure to this biological control agent. Of larvae that ceased to feed, the mean time to cessation of feeding after exposure to Bti was 12-18 s, sufficient time to ingest a lethal dose.     

Factors influencing nuisance blackfly (Diptera: Simuliidae) activity in the Scottish Highlands

Abstract. The nuisance activity of blackflies (Diptera: Simuliidae) was investigated in several habitats on Speyside, near Kincraig (57^o08'N, 3^o56'W), Invernesshire, in central Scotland during May-October 1987-89.
The main blackfly species caught landing/biting on humans were Simulium reptans, S. argyreatum, S. variegetum and the S. tuberosum complex, in order of prevalence. Blackfly biting activity occurred from mid-May to mid-September. Numbers of female blackflies attracted to volunteers were correlated with their body posture, habitat and the season. Overall, Simulium activity was greatest in mixed birch/juniper forest, least in spruce plantations and at intermediate levels on pasture, moorland and in Scots Pine forest.
Compared with an adult, a child experienced twice as many Simulium bites per hour (12.2 v 6.3) in the birch forest. On the child's body, 69% of blackfiies landed on the head, neck and back, whereas the majority landed on the legs (48%) and arms (28%) of adults. Bending over, especially during the exertion of gardening, was more attractive to anthropophilic blackfiies than standing or kneeling.     

Progress in controlling the reinvasion of windborne vectors into the western area of the Onchocerciasis Control Programme in West Africa

Since vector control began in 1975, waves of Simulium sirbanum and S. damnosum s.str., the principal vectors of severe blinding onchocerciasis in the West African savannas, have reinvaded treated rivers inside the original boundaries of the Onchocerciasis Control Programme in West Africa. Larviciding of potential source breeding sites has shown that these 'savanna' species are capable of travelling and carrying Onchocerca infection for at least 500 km northeastwards with the monsoon winds in the early rainy season. Vector control has, therefore, been extended progressively westwards. In 1984 the Programme embarked on a major western extension into Guinea, Sierra Leone, western Mali, Senegal and Guinea-Bissau. The transmission resulting from the reinvasion of northern C?te d'Ivoire and Burkina Faso has been reduced by over 95%, but eastern Mali has proved more difficult to protect because of sources in both Guinea and Sierra Leone. Rivers in Sierra Leone were treated for the first time in 1989 and biting and transmission rates in Sierra Leone and Guinea fell by over 90%. Because of treatment problems in some complex rapids and mountainous areas, flies still reinvaded Mali, though biting rates were approximately 70% lower than those recorded before anti-reinvasion treatments started. It was concluded that transmission in eastern Mali has now been reduced to the levels required to control onchocerciasis.