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In vitro haemagglutination and attenuation of microfilarial motility by haemolymph from individual blackflies (Simulium ornatum) infected with Onchocerca lienalis.

Two in vitro tests were used to investigate the effect of Onchocerca lienalis Stiles infection on the haemolymph of Simulium ornatum Meigen. The first of these examined the effect of infected haemolymph on the motility of fresh O. lienalis or Brugia pahangi Buckley & Edeson microfilariae. Incubation of haemolymph from individual flies with fresh microfilariae was performed in the wells of Terasaki micro-tissue culture plates. Motility of both species of parasite was found to be significantly attenuated when compared to worms incubated in control haemolymph groups. The second assay was that of agglutination of cat erythrocytes in the presence of haemolymph from individual flies, also performed in Terasaki plates. This test demonstrated significant increases in the rates of haemagglutination in the haemolymph of O. lienalis infected blackflies. The titre appeared to increase during the initial 5 days of infection up to a level of 1/32+, but then fell between day 5 and 7 to a maximum level of 1/2. The proportion of flies exhibiting haemagglutination also rose following infection. Despite the apparent absence of melanization and encapsulation, simuliids may have at least two humoral haemolymph components available to them for parasite regulation; a fast-acting factor responsible for rapid parasite death, and more specific agglutinins, possibly lectins. The role of the latter in defence is as yet unclear.

Animals↗

Population relationships of Simulium yahense and Simulium sanctipauli in the Firestone Rubber Plantation at Harbel, Liberia.

Comparative genetic differences for the phosphoglucomutase and trehalase loci were surveyed in larval and adult blackfly populations of the onchocerciasis vectors Simulium yahense Vajime and Dunbar, and S.sanctipauli Vajime and Dunbar. Genotype frequencies for each stage and all populations were at Hardy-Weinberg equilibrium, indicating that S.yahense and S.sanctipauli remain genetically distinct. S.yahense populations from three different locations were found to be genotypically comparable. The larval population of S.yahense found in closest proximity to a S.sanctipauli population was found to express the phosphoglucomutase allele 1.33, characteristic of S.sanctipauli, with significantly greater frequency than other larval populations of S.yahense. This may constitute evidence of limited genetic introgression of S.sanctipauli with the S.yahense population.

Animals↗

Onchocerciasis transmission levels and Simulium damnosum complex biting activity at riverside and rice field sites in Sierra Leone.

Biting densities of the Simulium damnosum Theobald complex of blackflies and their levels of parasitism by Onchocerca volvulus (Leuckart) were compared at two ecologically contrasted sites in the Southern Province of Sierre Leone, West Africa: by the Tabe riverside close to Gbaiima village where onchocerciasis is hyperendemic (63.1% positive skin-snips) and in open rice fields 2 km from the mesoendemic (54.9%) village of Senehun. Using a combination of morphometric and enzyme electrophoresis techniques, the primary vector was found to belong to the S.sanctipauli subcomplex and was most probably S.soubrense 'B' (sensu Post, 1986). In the wet season biting activity took place throughout the day at the village, but had morning and evening peaks in the rice fields. Transmission of O.volvulus was detected at the village in all months except February-April, while in the rice fields it was detected only during September-November and May at one tenth the level of the village. There was a significant difference between the two sites in the proportion of parous flies carrying third stage larvae in the head, but not of other larval stages.

Animals↗

Long-term storage of eggs of Simulium ornatum.

A method for the storage of non-diapausing eggs of Simulium ornatum Meigen has been developed. High survival rates for 5-6 months depend on the age of eggs when placed at reduced temperature. A period of precooling or conditioning is required for 28 days before storage of the eggs at 1 degree C with lowering of oxygen partial pressure. Induced quiescence does not appear to affect development from egg to adult.

Animals↗

Development of Onchocerca volvulus microfilariae injected into Simulium species from Cameroon.

Microfilariae (mff) of the savanna and forest strains of Onchocerca volvulus (Leuckart) were injected intrathoracically into adult females of Simulium damnosum Theobald sensu stricto, S.sirbanum Vajime & Dunbar, S.squamosum Enderlein and S.mengense Vajime & Dunbar. Nine days post infection (pi) 27-29% of the savanna mff and 31-38% of the forest strain had developed to third-stage larvae (L3), irrespective of the fly species, size or injection dose (5, 10 or 15 mff). Savanna flies supported the development of forest O.volvulus better than forest flies, in contrast to the results after per os infections. Therefore, in these four species of the S.damnosum complex from Cameroon, the peritrophic membrane is considered to be the main factor limiting the success rate of microfilarial development following the ingestion of blood infections, while the fly's haemolymph and intracellular environment play minor roles.

Animals↗

Studies on the possible role of cattle nuisance flies, especially Hydrotaea irritans, in the transmission of summer mastitis in Denmark.

The summer mastitis pathogens Actinomyces pyogenes, Peptococcus indolicus, Bacteroides melaninogenicus ss. levii, Fusobacterium necrophorum and Streptococcus dysgalactiae were isolated from the polyphagous symbovine dipterans Hydrotaea irritans (Fallén) and Morellia sp. caught around dairy heifers on pasture, but not from the haematophagous species Haematobia irritans (L.), Haematobosca stimulans (Meigen), Culicoides sp. and Simulium sp. Secretions from clinical cases of summer mastitis proved to be sources of summer mastitis bacteria for more than 3 weeks despite antibiotic treatment and teat amputation. Taking into account the seasonal activity pattern of Hydrotaea irritans and its topographical distribution on grazing cattle, it appears evident that this fly may play a central role in the establishment and maintenance of the bacterial contamination with summer mastitis pathogens on the teats of healthy cattle. In the present study the survival of A.pyogenes and P.indolicus for 7 days in experimentally infected Hydrotaea irritans, as demonstrated by the recovery of these microorganisms from agar plates exposed to live infected flies, is described. However, experimental transmission of summer mastitis from sick to healthy heifers by Hydrotaea irritans proved unsuccessful.

Actinomyces↗

Cytotaxonomic confirmation of two forms of Simulium sirbanum in the eastern part of the Onchocerciasis Control Programme in West Africa.

An earlier report (Cheke et al., 1987) of morphological differences between populations of Simulium sirbanum Vajime & Dunbar in West Africa was confirmed cytotaxonomically. Populations with adult males having Type III scutal patterns were associated with male larvae heterozygous for the inversion IS-3, whereas adult males with Type IV scutal patterns were associated with larvae homozygous for IS-3. The taxonomic implications are briefly discussed but the evidence is considered to be inappropriate for resurrecting S. sudanense Vajime & Dunbar. Aspects of the biology of the two forms and their importance in onchocerciasis transmission are discussed.

Africa, Western↗

Blood-feeding host preferences of the isomorphic species Simulium venustum and S. truncatum.

Animal-baited traps, using fox, mink, rabbits and ducks, were used in host preference experiments in two habitats (Beach and Woods). A generalized linear model of blood-feeding host preference is presented to test for significant differences between the isomorphic species Simulium venustum and Simulium truncatum. The S. truncatum population peaked before that of S. venustum. On any given day the two species divided their blood-feeding efforts among the different hosts in the same proportions. On the Beach, fox was the preferred host at the beginning of the season, but by the end of the season flies fed with equal frequency on the fox and the mink. In the Woods fox remained the preferred host throughout the season (late May to early July). Possible reasons for these feeding trends are discussed.

Animals↗

Simulium metallicum cytospecies E larval habitat characterization in the Altamira focus of onchocerciasis, northern Venezuela.

Simulium metallicum sibling species E was identified cytotaxonomically from an onchocerciasis focus at Altamira in northern Venezuela. S. metallicum E larvae were sampled monthly from two small mountain streams over a 15-month period (July 1990 to September 1991) and eleven habitat variables were measured at two altitudes. One stream consistently harboured much higher densities of S. metallicum E larvae than the other, with three annual peaks of abundance: during the dry season and at the beginning and end of the rainy season. These peak densities were correlated with high rainfall 4 months previously. Larvae were most abundant on submerged rocks and fallen leaves, in small shallow areas characterized by slow water current, high conductivity and sparse terrestrial vegetation cover. Stream variables which best explained the temporal changes in abundance were water discharge and conductivity. The population dynamics of S. metallicum E appeared to be influenced primarily by interactions between stream discharge and substrate stability. Relevance of these results to vector control with larvicides is discussed.

Animals↗

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

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 Mucambão) 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 Maranhão 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.

Animals↗

Assessment of vector microfilarial uptake as a comparatively non-invasive technique for monitoring onchocerciasis treatment campaigns in the Americas.

Since 1992, efforts have been made to combat onchocerciasis in Guatemala through mass distribution of ivermectin. The impact of the campaign is assessed by taking skin-snips from sentinel groups within selected communities. This method gives an estimate of the prevalence and intensity of infection, and thus the efficacy of the treatment. In some communities people are becoming reluctant to volunteer for skin-snipping, and so there is a need for an alternative technique that will give quantitative results. In most hyperendemic communities in Guatemala, biting blackflies are so ubiquitous that few people object to allowing 10 to 20 flies to engorge upon them. We examined data on the quantitative uptake of microfilariae by Simulium ochraceum before and after ivermectin distribution to see whether results similar to skin-snip data could be obtained. Counts of microfilariae ingested by S. ochraceum are compared to the numbers found in skin-snips from the same volunteers. In a group of 31 untreated infected persons, a skin-snip survey detected 64.5% positive, while feeding flies (vector microfilarial uptake, VmfU) detected 96.8%. Post-treatment, in a sample of 58 of whom 52 (89.7%) had a history of infection, both skin-snips and VmfU detected 54.2%. Vector blood meals contained more microfilariae than a mg of skin before treatment, but both recorded about equal numbers after treatment. When the data set was subdivided to compare samples taken at 2-3, 6-8 and 14-17 months post-treatment, the effect of ivermectin was still apparent at 6-8 months, but had virtually disappeared by 14 months post-treatment. A surprising observation was that the flies ingested fewer microfilariae from treated persons than was expected from the skin densities as estimated by skin-snip. This effect lasted for over 8 months, and could indicate that ivermectin has a greater effect on transmission than previously suspected. We conclude that VmfU could be used as an alternative to skin-snipping, and discuss the ethical implications.

Adolescent↗