Biomedical subjects

R J Post

Publications and source records attributed to R J Post.

At least 19 recordsLinked to original sources

PubMed2006

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.

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PubMed2005

Incrimination of Simulium thyolense (Diptera: Simuliidae) as the anthropophilic blackfly in the Thyolo focus of human onchocerciasis in Malawi.

The sibling species of the Simulium damnosum complex that are potential vectors in the foci of human onchocerciasis found in southern Malawi (the southern-most foci in Africa) were identified, using a combination of cytotaxonomic and molecular techniques. Simulium thyolense was found to be the most abundant species within the foci (representing 91% of the larvae collected), whilst S. kilibanum and cytoforms of the 'Ketaketa' subcomplex were the dominant members of the complex found outside of the foci. As all 188 biting female flies collected from the Thyolo, Mwanza and Mulanje foci were identified as S. thyolense, this species is probably the only significant vector of Onchocerca volvulus in the area, and the other Simulium species collected are assumed to be zoophilic. The results confirm the diagnostic value of the size of the amplicon(s) produced in PCR based on the sequences coding for the internal-transcribed-spacer region of the nuclear ribosomal DNA (ITS-1 rDNA), with a characteristic 340bp amplicon for S. thyolense. Chromosomal polymorphisms within S. thyolense indicate some degree of geographical isolation of the 'Thyolo-Mulanje' focus from the Mwanza focus.

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PubMed2004

A new cytotype of Simulium squamosum from south-west Cameroon.

A new cytotype of Simulium squamosum Enderlein (Diptera: Simuliidae) (here named S. squamosum'D') is described from around Mount Cameroon in south-west Cameroon on the basis of sex-chromosome differences on the short arm of chromosome I. Two polymorphic inversions, IS-To (a new inversion) and IS-2, are partially Y linked. These inversions usually occur together, although IS-To has a higher frequency and is more strongly Y linked than IS-2. This sex-chromosome system has not been previously described and the occurrence and evolution of sex-chromosome systems in S. squamosum is discussed.

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PubMed2004

Two new cytoforms of the Simulium damnosum complex (Diptera: Simuliidae) from Malawi and Tanzania and potential onchocerciasis vectors.

During a distribution survey of Simulium damnosum s.l. around the Tukuyu onchocerciasis focus at the northern tip of Lake Malawi/Nyasa (Tanzania), we discovered two new cytoforms of the S. damnosum complex in onchocerciasis-free areas. The Nyika form is related to Simulium thyolense, a vector of onchocerciasis, and can be identified by the new inversion 3L-L on the long arm of chromosome 3. It was found breeding in five rivers in northern Malawi and neighbouring Tanzania and is assumed to be zoophilic. The Njombe form represents a member of the Sanje group of the complex and is characterized by the new diagnostic inversion 2L-35 on chromosome 2. So far, it is only known from around Njombe town in southern Tanzania, where it breeds at remarkably high altitudes. Anthropophily for the Njombe form is well known. The medical importance and systematic position of the new forms within the S. damnosum complex are discussed.

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PubMed2004

The cytotaxonomy and morphotaxonomy of Simulium mengense (Diptera: Simuliidae).

Human onchocerciasis is hyper-endemic in south-western Cameroon and the island of Bioko. The vector on the island is the endemic 'Bioko' form of Simulium yahense. On the mainland, S. damnosum s.s., S. squamosum and S. mengense are recorded around Mount Cameroon but their contribution to transmission in the area is unclear. As elimination of onchocerciasis, through vector eradication, is a possibility on the island, it is important to be able to identify vector flies which might migrate to Bioko from the mainland. The morphologies of S. damnosum s.s. and S. squamosum are already well known. New cytotaxonomic and morphotaxonomic descriptions of S. mengense, a species which is less well known, are presented. Simulium mengense can be distinguished from the other cytospecies in the area by the presence of tufts of hair-like setae on the larval thorax, the presence of hairs on the subcostal vein of the adult female, and by the scutal pattern of the adult male. Although Vajime and Dunbar described seven fixed inversions in S. mengense, in 1977, nine were observed in the present study. Of the nine, three were the same as Vajime and Dunbar's but the other six were either missed or misinterpreted by them.

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PubMed2004

A molecular marker for the identification of Simulium squamosum (Diptera: Simuliidae).

All except one of the important groups of West African vectors of Onchocerca volvulus that lie within the Simulium damnosum complex can be distinguished from each other using morphological characteristics. The exception is S. squamosum, which overlaps with other species, and this results in significant levels of misidentification. Variation in the untranscribed H3-H4 histone intergenic spacer region of flies of the S. damnosum complex has now been investigated. Although a CAA/CAG microsatellite was found to be hyper-variable and unsuitable for species diagnosis, a 10-bp indel seemed to vary in a species-specific manner. This indel was apparently absent from S. squamosum but present in all of the other species tested (S. damnosum s.s., S. sirbanum, S yahense, S. sanctipauli and S. leonense/konkourense). It should now be possible to identify individual, adult, female S. squamosum from the absence of the indel, using a PCR-based amplification and agarose- or polyacrylamide-gel electrophoresis, thus removing the major barrier to the routine identification of unknown samples.

Africa, Western↗

PubMed2003

The morphological discrimination of microfilariae of Onchocerca volvulus from Mansonella ozzardi.

There is no published account which allows the morphological discrimination of microfilariae of Onchocerca volvulus and M. ozzardi from each other. However, they occur together in parts of Brazil and Venezuela, and presumably there is always the possibility that migration could establish new sympatric populations in the future. The objective of this study was to evaluate simple morphological characters that might be used for species-diagnosis of microfilariae. The conclusions were that the location of microfilariae in the blood or skin, the body size and the nucleation of the nerve ring are expected to be useful first indications of species identity, but cannot be used for confident diagnosis. The structure of the cephalic armature (stained with alcian blue) seems to be species specific, but is of limited application because it is often difficult to see. However, the pattern of nucleation of the tail (as expressed by the ratio of the length of the terminal nucleus compared with the length of the tail space) is distinctive and is expected to be diagnostic.

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PubMed2003

Cytotaxonomy, morphology and molecular systematics of the Bioko form of Simulium yahense (Diptera: Simuliidae).

Cytotaxonomic analysis of the polytene chromosomes from larvae of the Simulium damnosum Theobald complex from the island of Bioko in Equatorial Guinea is reported, and a new endemic cytoform is described. Chromosomally this cytoform is close to both S. squamosum (Enderlein) and S. yahense Vajime & Dunbar, but is not identical to either. However, it is morphologically and enzymatically identical to S. yahense. The Bioko form was also found to differ from other cytoforms of the S. damnosum complex in West Africa in the copy number or RFLP pattern of several different repetitive DNA sequences. It is clear that the Bioko form is genetically distinct from other populations of the S. damnosum complex, and whilst it is closest to S. yahense, it shows features that suggest a high degree of geographical and genetic isolation. Such isolation is an important consideration in the assessment of the potential for onchocerciasis vector eradication on Bioko.

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PubMed2002

Alternative polymerase chain reaction method to identify Plasmodium species in human blood samples: the semi-nested multiplex malaria PCR (SnM-PCR).

A simplified protocol for the identification of Plasmodium species by semi-nested multiplex polymerase chain reaction (SnM-PCR) in human blood samples is compared with microscopical examination of thin and thick blood films in 2 field trials in Côte d'Ivoire and Cameroon. Also, dried blood spots or liquid blood collected from Dutch soldiers returning from Goma, Zaire (n = 141), Angola (n = 40), and from Marechaussee (Dutch border police) returning from various parts of the world (n = 161) were examined, together with miscellaneous other material obtained from laboratories and hospitals. The method is based on features of the small subunit nuclear ribosomal ribonucleic acid (RNA) gene (ssrDNA), a multicopy gene which possesses both highly conserved domains and domains characteristic for each of the 4 human malaria parasites. The first reaction of the SnM-PCR includes a universal reverse primer with 2 forward primers specific for Plasmodium and mammals, respectively. The mammalian-specific primer was included as a positive control to distinguish uninfected cases from simple PCR failures. The second PCR reaction includes a Plasmodium-specific forward primer plus species-specific reverse primers for P. vivax, P. ovale, P. falciparum and P. malariae. The technique worked better with samples collected in the field as dried blood spots on filter paper and heparinized blood rather than with frozen pelleted blood; it was more sensitive and more specific than the standard microscopical examination.

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PubMed2002

Assessment of rDNA IGS as a molecular marker in the Simulium damnosum complex.

For five cytospecies of the Simulium damnosum Theobald complex of blackflies (Diptera: Simuliidae) from West Africa, both ends of the intergenic spacer region (IGS) of the rDNA have been sequenced with the aim of developing specific molecular markers. No specific differences in these two regions were detected between Simulium sanctipauli V. & D., Simulium sirbanum V. & D., Simulium soubrense V. & D., Simulium squamosum Enderlein and Simulium yahense V. & D., except in the number of A subrepeats at the 5' end of the IGS (two in S. squamosum and four or five in the others) and in position 310 of the 3' end (a C in S. squamosum and a G in the others). However, genetic distances within and between species overlapped. These DNA sequences had no strong phylogenetic signal, and the trees obtained were mostly unresolved. Although most sequences from S. squamosum clustered together, a few of them were more similar to those in other cytospecies. These results could be explained either by hybridization with genetic introgression or by ancestral polymorphism and recent speciation.

Africa, Western↗

PubMed2002

Completion of the sequence of the nuclear ribosomal DNA subunit of Simulium sanctipauli, with descriptions of the 18S, 28S genes and the IGS.

We describe the IGS-ETS, 18S and 28S ribosomal gene sequences of Simulium sanctipauli Vajime & Dunbar, a member of the S. damnosum Theobald (Diptera: Simuliidae) complex of blackflies (Diptera: Simuliidae). These regions, together with the ITS-1, ITS-2 and 5.8S rDNA presented elsewhere (accession number U36206), constitute the composite sequence of the entire rDNA unit, making S. sanctipauli the second dipteran species of medical importance for which the entire rDNA has been sequenced. Despite the lack of sequence identity, the IGS of S. sanctipauli showed some structural similarities to other Diptera, i.e. the mosquito Aedes albopictus Skuse (Culicidae), the fruitfly Drosophila melanogaster Meigen (Drosophilidae) and the tsetse Glossina (Glossinidae). Two blocks of tandemly repeated subunits were present in the IGS of S. sanctipauli and, unlike other species of Diptera, they contained no duplications of promoter-like sequences. However, two promoter-like sequences were identified in the unique DNA stretches of the IGS by their sequence similarity to the promoter of Aedes aegypti L. (Diptera: Culicidae). The observed sequence variation can be explained, as in the case of Drosophila spp., by the occurrence of slippage-like and point mutation processes, with unequal crossing-over homogenizing (to a certain extent) the region throughout the gene family and blackfly population. The 18S and 28S rDNA genes show more intraspecific variability within the expansion segments than in the core regions. This is also the case in the interspecific comparison of these genes from S. sanctipauli with those of Simulium vittatum, Ae. albopictus and D. melanogaster. This pattern is typical of many eukaryotes and likely to be the result of a more relaxed functional selection in the expansion segments than on the core regions. The A + T content of the S. sanctipauli genes is high and similar to those of other Diptera. This could be the result of a change in the mutation pressure towards AT in the Diptera lineage.

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PubMed2001

Characterisation of nuclear ribosomal DNA sequences from Onchocerca volvulus and Mansonella ozzardi (Nematoda: Filarioidea) and development of a PCR-based method for their detection in skin biopsies.

The internal transcribed spacer region (ITS1, 5.8S gene and ITS2) of the two filarial nematodes Onchocerca volvulus and Mansonella ozzardi was sequenced, and two species-specific primers designed in the ITS2 to develop a PCR-based method for their specific detection and differentiation. When used with a universal reverse primer, the two species-specific primers gave amplification products of different size, which were readily separated in an agarose gel. The PCR was tested on skin biopsies from 51 people from three localities in Brazil where M. ozzardi is present, and results have been compared with those of parasitological examination of blood. The species-specific PCR gave a higher percentage of detection of infection by M. ozzardi than the parasitological examination of blood. No infection with O. volvulus was detected by PCR. This PCR-based assay may assist in determining the nature of infection in areas where both filarial species exist in sympatry.

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PubMed2001

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.

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PubMed2001

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.

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PubMed2000

Regional genetic variation in the major sperm protein genes of Onchocerca volvulus and Mansonella ozzardi (Nematoda: Filarioidea).

Onchocerca volvulus and Mansonella ozzardi are two human filarial parasites present in South and Central America. In the Brazilian Amazonia they are found in sympatry, and the lack of clear morphological diagnostic characters in the microfilariae hinders their identification. The major sperm protein (MSP) gene of both species has been sequenced and characterised to determine its potential as a molecular diagnostic character. The length of the MSP gene is different in each species, and this could be used to detect and differentiate them by running the polymerase chain reaction (PCR) product in an agarose gel. Two major gene groups were identified in O. volvulus with a genetic distance of 6% between them. In M. ozzardi only one major group of genes was observed. The high similarity between the protein amino acid sequence of both filarial species confirms that the MSP has been highly conserved through nematode evolution.

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PubMed2000

The cytotaxonomy of Simulium damnosum s.l. (Diptera: Simuliidae) from the Thyolo onchocerciasis focus in Malawi and description of a new member of the complex.

Larvae of the Simulium damnosum complex were collected in southern Malawi, in particular from the Thyolo onchocerciasis focus. Cytological identifications revealed the occurrence of S. (Edwardsellum) thyolense sp.nov., S. kilibanum Gouteux, 'Ketaketa', a new cytoform tentatively designated 'Mwamphanzi', and two unassigned specimens. Simulium thyolense sp.nov. is presumably the major vector of Onchocerca volvulus in the area, but it remains possible that the other cytospecies are secondary vectors. Simulium thyolense sp.nov. is most appropriately classified near the 'Sanje' subcomplex, whereas 'Mwamphanzi' and the unassigned specimens are members of the 'Ketaketa' subcomplex.

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PubMed1998

Distribution of the Simulium damnosum complex on Bioko island, Equatorial Guinea, and the potential for onchocerciasis elimination by vector eradication.

Onchocerciasis is endemic on the island of Bioko, Equatorial Guinea, where it is transmitted by the 'Bioko form' of the Simulium damnosum complex, a cytospecies unique to the island. To determine the distribution of vector breeding, three dry season and two wet season expeditions were made in 1989, 1996 and 1997, and 226 of the island's 247 rivers (91.5%) were visited. Of these 226 rivers, 130 (58%) were flowing during the dry season, forty-five (20%) supported aquatic stages of Simuliidae of any species and twenty-five (11%) contained larvae or pupae of the S. damnosum complex. The twenty-one rivers not prospected were in the mountainous south of the island, where an additional seventeen rivers were reached but not satisfactorily prospected. Of these thirty-eight rivers, twenty-nine were considered highly likely to support vector breeding, bringing the total number of rivers which could harbour the vector during the dry season to fifty-four (21.9% of the island's total). Breeding was believed to be limited to river stretches below 1000 m altitude, and during the dry season the total length of those stretches which could support breeding on Bioko was estimated to be 1020 km. A combination of factors, including low river discharges during the dry season, the relatively low water temperature on Bioko, the suitability of limited stretches of most rivers as vector breeding sites and the close proximity of many rivers within a small geographical area, render the vector vulnerable to eradication by aerial treatment of rivers with insecticide. The isolation of the Bioko form of the S. damnosum complex suggests that reinvasion following treatment would be unlikely, and eradication of the vector might be achieved by a dry season larviciding programme in one or two years.

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PubMed1997

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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