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Isolation of living adult Onchocerca volvulus from nodules.

A technique for the isolation of adult Onchocerca volvulus from excised onchocercomata is described. The nodules are incubated in medium 199 containing 1-5 mg collagenase and 0.2 mg gentamicin per ml for 6-48 hours in a waterbath at 30-37 degrees C. A proportion of the worms can be isolated alive.

Animals

The effects of some drugs--pentamidine, stibocaptate, Hoechst 33258, F 151, compound 'E' and Nifurtimox--on Onchocerca volvulus in chimpanzees.

Several drugs were investigated for action against Onchocerca volvulus in chimpanzees. Pentamidine at 4 mg/kg daily for 16 days had no filaricidal action; nor did stibocaptate at 10 mg/kg monthly for 8 months. Hoechst 33258, a substituted bis-benzamidazole, showed micro- and macrofilaricidal activity; F151 (an arsenical compound) showed a purely macrofilaricidal action; and Compound E (a reaction product of Hoechst 33258 and F151) was micro- and macrofilaricidal. The toxicity of the three last-mentioned compounds was high, and further trials were not made. Nifurtimox, at doses of up to 40 mg/kg for 10 days, was shown not be microfilaricidal, but there was a suggestion that this drug may have some macrofilaricidal action.

Animals

Histochemical enzyme-staining patterns of onchocerca volvulus microfilariae and their occurrence in different onchocerciasis areas.

Histochemical differentiation of Onchocerca volvulus microfilariae from 164 patients in West African rain-forest (Liberia), Sudan-savanna (Upper Volta), Guatemala and the Yemen has been carried out using a staining method for the demonstration of acid phosphatase. Intrauterine microfilariae showed considerable changes in their enzyme activity during embryonic development which are probably associated with the maturation of the parasite before migration to the tissues. Five distinct types of staining patterns could be distinguished among microfilariae from the skin according to the localization of the enzyme in specific structures of the microfilaria. Two or more types of staining patterns were found in most persons in the different geographic regions. There were significant differences in the overall distribution of the various staining patterns in persons from the different areas. At the present state of our knowledge, little is known about the nature and significance of these differences in the staining patterns of microfilariae. The question of whether they can be ascribed to an ageing process, strain differences or other factors is discussed.

Acid Phosphatase

Genetic structuring and estimation of reproductive adults in Onchocerca volvulus: A genome-wide analysis across hosts and regions.

Genomic analysis of parasites can deepen our understanding of their transmission, population structure, and important biological characteristics. Onchocerciasis (river blindness), caused by the parasitic nematode Onchocerca volvulus, involves adult worms residing in subcutaneous nodules that produce larval-stage microfilariae (mf), which are routinely detected in the skin for diagnosis. Whole-genome studies of mf are limited; most analyses have focused on the mitochondrial genome. We conducted a genome-wide analysis with 94% median nuclear genome coverage, analyzing 171, 37, and 98 mf from 16, 3, and 5 individuals from Ghana, Liberia, and the Democratic Republic of Congo, respectively. These data were used to investigate population differentiation, estimate the number of reproductive adult worms, and analyze genetic variation across chromosomes. Population genetic analyses across hosts and countries showed that nuclear genome diversity can reveal fine-scale genetic structure, even between geographically close countries, providing more resolution than mitochondrial haplotype data. By reconstructing maternal and paternal sibships, we estimated the number of reproductively active adult filariae. Comparisons between adult worm estimates from genetic data and nodule observations showed that genetics-based estimates were higher or equal to observed worm counts in 8 out of 9 hosts for female worms and 7 out of 9 hosts for male worms. Our analysis also revealed lower-than-expected X chromosome diversity, consistent with neo-X chromosome fusions in filarial species. This study represents an important step in using nuclear genome data from mf to support onchocerciasis elimination efforts and in developing genetic tools that could inform mass drug administration programs.

Onchocerca volvulus

Preliminary studies on the histochemical differentiation of strains of Onchocerca volvulus microfilariae in Togo.

Skin snips were taken from 75 people living in four villages of northern Togo. The 7824 microfilariae that emerged were examined by staining for the presence of acid phosphatase. Four distinct patterns of enzyme staining were observed, and descriptions of the stained microfilariae are given. The study confirms the view that a number of biological strains or variants of Onchocerca volvulus coexist in West Africa, and suggestions are made for further research that could result in the practical application of these observations in onchocerciasis control programmes.

Animals

A simple technique to assess the total number of Onchocerca volvulus microfilariae in skin snips.

A technique for the assessment of the total number of microfilariae in skin snips from onchocerciasis patients is described. For the digestion of the skin tissue the biopsies are incubated at room temperature for 24 hours in medium 199, containing 3 mg/ml collagenase and antibiotics. This process leaves the microfilariae unharmed. A comparative study of the emergence of the microfilariae in different media showed that only 20% of the total number were released after 30 minutes in distilled water. After 24 hours incubation in isotonic saline 80% were found.

Humans

[Immunological diagnosis of filariosis in persons returning from tropical countries and in European and indigenous inhabitants of endemic areas (author's transl)].

Serum samples from persons who lived in areas where onchocerciasis occurred or who had filariasis were examined with the complement fixation test and the indirect hemagglutination test for the presence of antibodies against crude extracts from Dirofilaria immitis, Onchocerca volvulus, Dipetalonema viteae, and Ascaris suum. The results could be interpreted as follows: 1. The indirect hemagglutination test was more sensitive than the complement fixation test for the demonstration of antibodies in sera from European and indigenous inhabitants of endemic areas. 2. There were no differences between the responses to the four crude worm extracts among the groups of 21 patients with Loa loa, 12 patients with Onchocerca volvulus, 11 patients with Dipetalonema perstans, and 22 patients with clinical filariasis in whom no microfilaria had been found. It was concluded that the examination with crude extracts cannot give any information about the antigens that had caused the stimulation of antibodies.

Adolescent

Onchocerciasis transmission potentials of four species of Guatemalan simuliidae.

Two complete days of collections and dissections of all black fly species landing on two persons infected with Onchocerca volvulus were made at monthly intervals for 13 months within a single endemic area of Guatemala. A comparison of the infective biting densities and transmission potentials showed that Simulium ochraceum was the only significant vector.

Adult

Some observations on onchocerciasis including sowda in the Yemen Arab Republic.

A pilot study on onchocerciasis and sowda was carried out in the south-western region of the Yemen Arab Republic. Of 61 persons examined in eight villages 35 (= 57%) were found positive for microfilariae by the skin-snip method, 13 had typical manifestations of sowda, 17 had other onchocercal-suggestive skin lesions and five had subcutaneous nodules. A single larva of Simulium damnosum s.I. was collected in Wadi Barakani. Numerous larvae and pupae of S. ruficorne and S. hargreavesi were taken in fast-flowing streams in four localities. The examination of microfilariae and adult worms by scanning electron microscopy as well as the histochemical staining of microfilariae for the demonstration of acid phosphatase activity confirmed that the concerned filaria belongs to the species Onchocerca volvulus. No morphological differences could be detected between the smaller number of the examined worms from the Yemen and the extensive parasite material from Liberia, West Africa.

Adolescent

Electron microscopical studies on onchocerciasis. III. The onchocerca-nodule.

Six onchocerca-nodules from five Liberian patients were examined by electron microscopy. The bulk of cells in the centre of the nodules consists of lymphocytes and macrophages. The lymphocytes can be observed in various stages of differentiation. A major part of the macrophages shows degenerative changes with decrease of lysosomes and increase of fatty vacuoles. Near the worms epithelioid cells and giant cells can be found. Other cells in the nodule include polymorphonuclear neutrophils, eosinophils, plasma cells, and mast cells. Adult Onchocerca volvulus show the basic nematode body plan. The cuticle is subdivided into cortex and two layers with differently arranged fibrillar structures. The characteristics of the hypodermal cells are best seen in the lateral chords, in the interchordal regions the hypodermis is flattened by a muscle layer. The intestine typically contains pigment granules, the cells of the epithelium have folds of their cell membranes forming a basal labyrinth. The uterus consists of two tubes in which the development of the microfilariae from the early embryonic forms to the mature stages is examined.

Adult