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Biomedical subjects

R J Post

Publications and source records attributed to R J Post.

At least 37 records · Page 2Linked to original sources

Molecular population studies of Simulium damnosum s.l. (Diptera: Simuliidae) using a novel interspersed repetitive DNA marker.

Simulium damnosum s.l., the vector of West African onchocerciasis, has been the target of a major insect control initiative for the past 20 years. However, attempts to study the migration patterns of reinvading infective flies into controlled areas have been restricted by the lack of suitable genetic markers. Here, the results of the first population-level study of S. damnosum s.l. using a repetitive DNA marker, pSO11, are presented. Sequence analysis of pSO11 revealed a complex internal pattern of repetition and an open reading frame with 56.8 per cent similarity to a mouse retrotransposon protein. Combined with an interspersed genomic distribution, the internal structure of pSO11 suggested that it represents the 5' half of a transposable element. The genomic diversity of the sequence was analysed using Southern blot analysis of genomic DNA. Data were collected from 475 individuals from the island of Bioko (in Equatorial Guinea), Cameroon, Côte d'Ivoire and Sierra Leone. The results indicate that pSO11 is useful as an indicator of genomic diversity and that the experimental design used here permits comparison of an approximately random sample of chromosomal loci. Making this assumption, estimates of homozygosity based on pSO11 diversity were used to study inter- and intraspecific variability. The results indicate that pSO11 is potentially useful for studying population-level processes in S. damnosum s.l.

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Cytotaxonomic identifications of the Simulium damnosum complex (Diptera:Simuliidae) from Nigeria.

Cytotaxonomic analysis of Simulium damnosum s.l. larvae collected from 23 sites across 4 bioclimatic zones in Nigeria revealed the presence of 5 cytospecies (S. damnosum s.str., S. sirbanum, S. squamosum, S. yahense and S. soubrense (including the Beffa form). The biogeographical distribution of the sibling species was mostly similar to that observed further west in the OCP area, although geographic cytotaxonomic variation was recorded in S. squamosum, S. damnosum s.str. and S. sirbanum. Some aspects of the chemical and physical environment of the breeding habitats were investigated.

Animals↗

Detection of Plasmodium sporozoites in mosquitoes by polymerase chain reaction and oligonucleotide rDNA probe, without dissection of the salivary glands.

Dried Anopheles gambiae mosquito head+thorax portions, infected with Plasmodium falciparum sporozoites, were processed by the polymerase chain reaction. The PCR product was hybridized to an oligonucleotide probe (known as 114R or AW34) diagnostic for Plasmodium. The detection level by autoradiography was ten sporozoites per mosquito. Head+thorax of mosquitoes that contained mature P.falciparum oocysts, without sporozoites, gave no positive signal, indicating that the test detects only infective mosquitoes. This test can be applied to wild mosquito specimens collected, prepared and processed at different time intervals. The technique is convenient, highly sensitive, and could be used with a non-radioactive detection system and specific probes to differentiate Plasmodium spp.

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Multivariate morphotaxonomy in the identification of adult females of the Simulium damnosum Theobald complex (Diptera: Simuliidae) in the Onchocerciasis Control Programme area of West Africa.

Adult female Simulium damnosum s.l. flies obtained in nine countries of the Onchocerciasis Control Programme were examined for 14 morphological characters with the aim of selecting specific characters to identify the main species that form the complex. Two approaches were used, the first involved discriminant function analysis of the species groups using all the characters which were both qualitative (the colour of the fore coxa, the antennae, the wing tuft, the arculus, the ninth abdominal tergite setae and the scutellar setae) and quantitative (the lengths of thorax, antenna, and femur, tibia and first basitarsus of both the fore and hind legs) in nature. The second sought colour characters unique to the S. damnosum subcomplex and to S. yahense, to separate them from the rest, and progressively applied discriminant analyses in the dichotomous stepwise manner to within the subcomplex level. The combination of pale arculus and pale antennae with pale or dark fore coxae were unique to the S. damnosum subcomplex, whilst those of black arculus together with wholly dark or mixed ninth abdominal tergite setae and scutellar setae were unique to S. yahense. Further discrimination was achieved between subcomplexes and species by entering the qualitative and quantitative characters into discriminant function analysis. The two schemes were both successful at separating the main species subcomplexes of the S. damnosum complex, with 90-100% correct classification. Within the subcomplex level, correct identification of sibling species was 85-100% successful.(ABSTRACT TRUNCATED AT 250 WORDS)

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How long do the effects of ivermectin on adult Onchocerca volvulus persist?

The persistence of the effects of ivermectin on the viability, morphology and reproductivity of adult Onchocerca volvulus was examined eighteen months after treatment with a single or five six-monthly doses of ivermectin and compared with untreated controls. Treated nodules were removed from patients participating in a randomised controlled trial of ivermectin in Sierra Leone. Adult filariae, 545 females and 348 males, were isolated by collagenase digestion. The nodules were significantly smaller, contained fewer young worms and supported lower microfilarial production in those treated with five doses of ivermectin. The productivity index, a measure of the reproductive potential of a worm population, was still reduced by 83% eighteen months after five doses and by 63% after a single dose compared to controls. These results show that worms recover their fertility even after multiple doses of ivermectin, but do so slowly compared to standard dosage intervals. In addition ivermectin may have a partial chemoprophylactic effect which contributes to the maintenance of low microfilarial production in conditions of on-going transmission.

Adult↗

Absence of sperm from the seminal receptacle of female Onchocerca volvulus following multiple doses of ivermectin.

It is already known that multiple doses of ivermectin have a profound effect on embryonic development in Onchocerca volvulus and that this appears to operate mainly at the single cell stage. To investigate this further, we examined adult female O. volvulus originating from patients treated either with placebo or one, four or five doses of ivermectin. The reproductive organs were dissected out of the worm and examined for evidence of oogenesis and for the quantity of sperm and oocytes in the seminal receptacles. A single dose of ivermectin had no measurable effect on oogenesis or on the amount of sperm and oocytes compared to placebo. However after multiple doses of ivermectin a significantly lower proportion of seminal receptacles contained sperm and there was a significantly higher proportion of ovaries with impaired oogenesis compared to placebo. It is concluded that the reduction in the number of multicellular embryonic stages from worms exposed to multiple doses of ivermectin is due, at least in part, to a major reduction in the effective insemination of female worms and to a minor impairment of oogenesis.

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Studies on environmentally-induced colour variation in Simulium sirbanum (Diptera: Simuliidae) using a portable rearing system.

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.

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DNA probes for the identification of members of the Simulium damnosum complex (Diptera: Simuliidae).

Genomic libraries in plasmid have been constructed from various sibling species of blackflies of the Simulium damnosum complex from West Africa. Three cloned repetitive sequences, which show variation in copy number between sibling species, have been isolated. These clones can be used as probes for the dot-blot identification of larvae, pupae or adults into the three main West African subcomplexes, i.e. damnosum, squamosum and sanctipauli subcomplexes. The sequences also show some intraspecific variation in copy number.

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The effect of repeated doses of ivermectin on adult female Onchocerca volvulus in Sierra Leone.

The effects of single and multiple doses of ivermectin on mortality and morphology were assessed in over 700 female Onchocerca volvulus worms and the effects on embryogenesis were assessed in 490. Nodules were surgically removed from Sierra Leoneans recruited from a double-blind placebo controlled study of ivermectin given at six-monthly intervals. Nodules were digested in collagenase to isolate whole adult worms. After four or five doses of ivermectin there were significant increases in the numbers of discoloured and calcified worms and possibly a trend towards increased mortality, but this was not seen consistently. There was no evidence of a prophylactic effect of the drug. Worms were then homogenised and embryograms constructed. A single dose of ivermectin produced large numbers of degenerating intrauterine microfilariae, but embryonic development occurred normally. After multiple doses we observed almost complete cessation of embryogenesis, with a highly significant decrease in the numbers of viable multicellular embryonic stages, while oocytes appeared to be produced normally. Development is probably impeded at the single cell stage, possibly because of reduced fertilization. In planning the future role of ivermectin as a control measure for onchocerciasis it is crucial to determine if these effects on embryogenesis are reversible.

Adolescent↗

Chromosome numbers in Onchocerca dukei and O. tarsicola.

Testicular examination of male Onchocerca dukei and German O. tarsicola revealed five pairs of chromosomes, which is typical of most filariae. In contrast to German O. tarsicola, specimens from Sweden had only three pairs of chromosomes.

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Simulium (Edwardsellum) squamosum kitetense ssp. n., a new chromosomal and morphological subspecies within the Simulium damnosum complex from east Zaire.

A new subspecies within the Simulium damnosum complex is described from the Lualaba river, East Zaire, on the basis of polytene chromosome analysis and larval morphology, under the name Simulium (Edwardsellum) squamosum kitetense ssp. n. Chromosomally it is most similar to S. squamosum, from which it differs by three new fixed inversions. Morphological differences include a reduced number of rays on the premandibular fans, the particular aspect of the combs on those rays, and the general colour of the body which is a very light yellow. A second population from a nearby river was morphologically and chromosomally quite different, although more samples are needed to clarify its taxonomic position.

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Cloning and characterization of a species-specific repetitive DNA sequence from Onchocerca armillata.

Two clones, pOA1 and pOA5, have been isolated from a genomic DNA library prepared from pools of Onchocerca armillata adults in the plasmid vector pUC12. In dot-blot hybridisations, these two clones do not cross-hybridise significantly with total genomic DNA from O. volvulus, O. gutturosa, O. ochengi, O. gibsoni, O. lienalis, bovine, human, Culicoides nubeculosus, Simulium species or Brugia pahangi, but do hybridise with as little as 100 pg of DNA from two separate geographic isolates of O. armillata. The sequence of pOA1 and pOA5 has been determined and found to contain a repetitive DNA sequence 147 bp in length. These clones can be used as specific and sensitive DNA probes for the identification of O. armillata capable of identifying a single L3 larva.

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