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Acid phosphatase activity in the larval stages of Onchocerca volvulus developing in the vector Simulium damnosum.

The staining patterns of acid phosphatase activity of the various larval stages of Onchocerca volvulus from experimentally infected Simulium damnosum s.l. have been described. The enzyme activity was localized in the anal vesicle/pore area of the first-stage larva, in the anal vesicle and intestine of the secong-stage larva and all over the body in the third-stage larva. Early third-stage larvae could be differentiated from late third-stage infective larvae by the presence of pronounced enzyme activity in the rectum area of the former. There were no detectable differences in the staining patterns between the forest and Sudan-savanna strains of O. volvulus developing in S. damnosum.

Acid Phosphatase↗

The effect of drug treatment on the pattern of emergence of Onchocerca volvulus microfilariae from skin snips.

A controlled study was carried out to determine the effect of some known anti-filarial drugs on the pattern of emergence of Onchocerca volvulus microfilariae from skin snips into saline. The results show that before treatment, 81.7 to 92.1% of microfilariae emerge in all the groups within 24 hours. The differences between the groups were not significant. After treatment with oral diethylcarbamazine, metrifonate and topical diethylcarbamazine (DEC) the percentage of microfilariae emerging from the snips dropped significantly to 48.4, 56.0 and 60.1 respectively, whereas no significant change occurred following treatment with levamisole (88.1%) and yeast tablets (89.1%). A pair-wise comparison of the post-treatment microfilarial emergence in all the groups showed statistically significant differences when (i) oral DEC is compared to levamisole, topical DEC and yeast, (ii) metrifonate is compared to levamisole and yeast and (iii) topical DEC is compared to yeast. It is suggested that microfilariae which fail to emerge from snips after treatment are those that have been killed or paralysed by the action of the drugs. The study has thus established that drug action in vivo is one of the factors which affects the proportion of microfilariae which emerge from skin snips and provides an additional parameter for measuring the antimicrofilarial potential of drugs in clinical chemotherapeutic trials.

Animals↗

Limited fly load and development of Onchocerca volvulus microfilariae in Guatemalan Simulium ochraceum.

Fly load of Onchocerca volvulus microfilariae and their early development in Simulium ochraceum were examined at intervals of 10, 30 and 60 min after ingestion and then hourly up to 72 hours. Many microfilariae ingested were damaged by the cibarial armature of the vector, limiting the fly load, but some of these damaged microfilariae were able to penetrate the gut before they died. Thus, the mean number of the living parasites per fly in the head and thorax decreased gradually with elapse of time after ingestion. The parasites ingested by S. ochraceum did not develop beyond the microfilarial stages during the first 24 hours; thereafter only thoracic forms (1st-stage larvae) were found to be alive. The high mortality of the infected fly might be caused by the high intakes of microfilariae and also by normal and/or partly wounded microfilariae invading various organs. The number of microfilaria ingested from different biting sites were usually high in comparison with the densities observed in each site; this indicates that the microfilariae might be attracted by the biting females.

Animals↗

Observations on Onchocerca volvulus in experimentally infected chimpanzees.

The paper describes observations made on 32 chimpanzees experimentally infected with Onchocerca volvulus. The mean pre-patent intervals for the Cameroon forest and the Guatemalan strains of O. volvulus were 13-16 months and 12-15 months respectively. That for the Cameroon Sudan-savanna strain was much longer, i.e. 22-23 months. The numbers of microfilariae found in the skins of animals infected with the Cameroon Sudan-savanna strain were also much lower than in animals infected with the other two strains. Long-term observations on infected animals showed that microfilarial infections had virtually died out 6.5-9 years after the last inoculation with infective larvae. Those animals which were inoculated with infective larvae in the head or above the waist tended to show a higher proportion of microfilariae in the upper parts of the body, than did those inoculated with infective larvae below the waist. In animals which showed adult worm-bundles on only one side of the body, the concentration of microfilariae was usually greater on that side of the body. Worm-bundles in the chimpanzee varied in size from 8 x 5 x 2 mm to 4 x 3 x 2 cm. Out of 47 worm-bundles found, only two were subcutaneous. The remainder lay deep in the tissues, most commonly adjacent to the posterior surface of the capsule of the hip joint. No onchocerciasis eye lesions were seen in any of the infected animals.

Animals↗

Experimental infection of laboratory rodents and calves with microfilariae of Onchocerca gibsoni.

Uterine microfilariae of Onchocerca gibsoni migrated to skin of all parts of the body when inoculated into rats and mice. Higher microfilarial densities were obtained in the skin of mice compared with rats. In guinea pigs very few microfilariae migrated to the skin and densities of microfilariae were very low. Densities of skin microfilariae were lower in rats when inoculated intraperitoneally compared with inoculation by 2 subcutaneous routes. In the latter case, higher densities of microfilariae were obtained in areas of skin closer to the injection sites. Persistence of microfilariae in the skin of rats was variable but only low numbers were recovered between 21 and 70 days after infection. Microfilariae also migrated to the skin in calves experimentally infected by subcutaneous injection into the dewlap. Microfilaria were recovered consistently and in high numbers from skin snips along the ventral midline. Small numbers of microfilariae were occasionally found in skin snips from the dorsal midline.

Animals↗

Analysis of enzymatically isolated adults of Onchocerca volvulus after treatment of patients with suramin or metrifonate.

The application of a new technique is described permitting a better recognition of activities of drugs on macrofilariae of Onchocerca volvulus. In two villages of a hyperendemic focus in Liberia onchocercomata were excised from untreated persons and from patients treated with conventional doses of suramin or metrifonate. The adult worms were enzymatically isolated with collagenase and microscopically examined for degenerative alterations including calcifications and for the presence of eggs and embryos in the female parasites. Six weeks after the last dose of suramin the degenerative alterations could not be distinguished qualitatively from those in untreated worms, but a statistically significant macrofilaricidal efficacy could be shown by quantitative analyses, although more than half of the parasites were still alive at this early date. Metrifonate did not show any macrofilaricidal effect.

Animals↗

[Onchocerca sweetae occurring in the buffalo in Thailand (author's transl)].

The intradermal Onchocerca from Bubalus bubalis of Thailand is the same as that which was described as O. sweetae from intradermal nodules in B. bubalis from Australia. This identification suggests there has been no speciation between the parasites of Australian and Asiatic Buffaloes. The taxon O. indica Sweet 1915 consists of two distinct species; one in perimuscular nodules of Bos indicus is most probably O. gibsoni (and thus, the species O. indica, based essentially on the description of this material, should fall in synonymy); the other in intradermal nodules of B. bubalis does not differ from O. sweetae. To confirm the distinction between O. cebei, a parasite in the same host from Hanoï, and O. sweetae, it must be demonstrated that the presence or absence of clearly precloacal papillae is a valid character for separating these species.

Animals↗

Life span and distribution of Onchocerca volvulus microfilariae in mice.

Microfilariae of Onchocerca volvulus obtained from onchocercomas were inoculated subcutaneously into the inguinal region of mice. Microfilariae disseminated rapidly, invading the tail and lungs within 1 hr and the ears within 3 hr postinoculation. During the early period of infection microfilariae appeared in the eyes, ears, viscera, pelt, carcass, and tail; later, the microfilariae tended to accumulate in the tail. Liver microfilariae were recovered up to 12 wk postinoculation. Intraperitoneal inoculation and subcutaneous inoculation into the inguinal region resulted in the concentration of microfilariae in the tail, whereas inoculation into the scalp caused microfilarial accumulation in the ears.

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↗

Seasonal prevalence of Onchocerca-like microfilaria in camels in Kuwait.

Infection of the camel (Camelus dromedarius) with Onchocerca-like microfilaria is recorded for the first time in Kuwait. Throughout a period of 14 months, the blood of 284 camels were examined, of these 32 (11%) were found infected with the microfilaria. Infections were recorded in all months except during summer, July to September, and in February. The highest monthly incidence (27%) was detected in June at the end of spring. It is suggested that harsh environmental conditions in Kuwait during summer either arrest development of microfilariae or influence their diurnal distribution in tissues of the camel.

Animals↗

Evaluation of suramin, ivermectin and CGP 20376 in a new macrofilaricidal drug screen, Onchocerca ochengi in African cattle.

To aid the development of a macrofilaricidal agent for Onchocerca volvulus, the African bovine parasite, O. ochengi, was evaluated as a drug screen by testing three known filaricidal drugs. Groups of five Zebu cattle, naturally infected with more than 15 palpable O. ochengi nodules in the ventral skin, were treated with either suramin (10 mg/kg/day i.v. for 6 days), ivermectin (200 micrograms/kg, s.c.), CGP 20376 (20 mg/kg orally) or left untreated as controls and examined at intervals up to 137 days post-treatment (d.p.t.). After ivermectin treatment, microfilarial densities in the skin decreased within one week to virtually zero and remained at a very low level. A similar rapid and profound reduction was seen after CGP 20376 treatment, but by 137 d.p.t. microfilarial skin densities were approaching pre-treatment levels. With suramin, skin microfilarial densities fell to very low levels after 12 weeks but rose slightly by 137 d.p.t. Effects on the macrofilariae were assessed by sequential nodulectomies at -3 and 28, 84 and 137 d.p.t. By 137 d.p.t. embryogenesis was almost completely interrupted in the CGP 20376 and ivermectin treated animals, although not in the suramin treated group, but in all three groups the majority of remaining intrauterine microfilariae were pathologically altered. Degenerating intrauterine microfilariae accumulated in the ivermectin and in the CGP 20376, but not in the suramin treated worms. The motility of male and female worms was not reduced by any treatment except for female worms at 84 d.p.t. with CGP 20376. Viability of the worms as indicated by the MTT-formazan reduction assay was not reduced in any of the treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Africa↗

Effect of semiannual treatments of ivermectin on the prevalence and intensity of Onchocerca volvulus skin infection, ocular lesions, and infectivity of Simulium ochraceum populations in southern Mexico.

The effect of semiannual ivermectin treatment along with nodulectomy on filarial transmission levels were estimated during the three dry seasons of 1991-1993 in a hyperendemic village in southern Mexico. Parasitologic and ophthalmologic examinations were carried out every six months until five drug treatments were completed. Ivermectin mass treatment with a coverage of approximately 80% had a significant impact (P < 0.05) on the prevalence of skin infection and the mean microfilarial skin density (CMFL), which were reduced 38% and 89%, respectively. A gradual and significant (P < 0.05) decrease in the mean microfilariae number in the anterior chamber of the eye and in corneal opacities was also observed as the CMFL was reduced. After three treatments, these were reduced 84% and 69%, respectively. However, after two years of continuous intervention, no significant differences (P > 0.05) were observed in either the daily mean infective biting density and the daily mean transmission potential. This was probably due to the remaining microfilarial load provided by the untreated resident population and migrant groups. On the whole, our results confirm both the efficacy of ivermectin to alleviate the clinical manifestations of the disease and its minimal impact on Onchocerca volvulus transmission, and indicate the need both to achieve higher levels of drug coverage and to incorporate other measures to stop transmission until a macrofilaricide drug is found.

Adolescent↗

Efficacy of moxidectin oral gel against Onchocerca cervicalis microfilariae.

During a series of dose-titration experiments designed to evaluate the efficacy of moxidectin oral gel against equine gastrointestinal parasites, infection with Onchocerca cervicalis was diagnosed in 25 of 82 ponies prior to treatment. Microfilariae were identified in full-thickness skin biopsies taken from the ventral midline. Treatment with moxidectin in single doses of 300, 400, or 500 micrograms/kg of body weight was 100% effective in eliminating microfilariae from 20 skin biopsies taken 14 days posttreatment, whereas 5 microfilaria-positive ponies in 2 control groups remained positive following placebo treatment. No adverse reaction was seen in any pony following treatment with moxidectin or the vehicle control.

Administration, Oral↗

Genomic fingerprinting of Onchocerca species using random amplified polymorphic DNA.

A method based on amplification of genomic DNA by decamer primers of random nucleotide sequence was used to obtain DNA fingerprints from different species of the genus Onchocerca. Each of the 20 primers tested allowed a clear distinction between the different species of the genus on an agarose gel. The technique offers the potential to construct species or strain specific probes and oligonucleotides PCR primers from the species specific fragments. A combination of these primers or others could be useful as population markers.

Animals↗

[ Onchocerciasis control program in West Africa: socioeconomic development and risk of recrudescence of transmission. 1. Experimental study of the transmission of Onchocerca volvulus strains from Southwestern Sierra Leone by Simulium sirbanum].

As part of the return of savanna migrants installed since a long time in forest regions, in the south of Sierra Leone, we carried out an experimental study about a cross-transmission between Simulium sirbanum from Missira (West-Mali) and the forest strain of Onchocerca volvulus in the south-west of Sierra Leone. This study will allow to know if there is a risk of onchocerciasis transmission recrudescence in relation to the reinstallation of these migrants in their native region. Because of the very high limitation to the forest strain of O. volvulus microfilariae output of the peritrophic membrane reduction with savanna black-flies and according to the very low mature parasite out put of S. sirbanum with this strain observed along this experimentation, the forest strain of O. volvulus from the south Sierra Leone appears maladjusted to S. sirbanum, the main vector of onchocerciasis in savanna regions. This observation implicates a very low intensity of transmission for this forest strain by savanna onchocerciasis vectors. The return of savanna migrants in their native region, installed in the south Sierra Leone since several decades, could not be, in a short time, an origin of onchocerciasis recrudescence in savanna regions of the Onchocerciasis Control Programme area cleaned by an effective vector control carried out since 1975 sustained now by a chemotherapeutic treatment reducing the human parasite reservoir. However, the preservation of this acquired necessitates an epidemiological supervision increased, because the interactions between the vector and the parasite for a long time could carry away a mutual adaptation and a sickness recrudescence.

Animals↗

[Onchocerciasis control program in West Africa: socioeconomic development and risk of recrudescence of transmission. 2. Experimental study of the transmission of Onchocerca volvulus strains from Southwestern Sierra Leone by Simulium yahense and Simulium squamosum].

The movements of human populations towards the mining wealth of the northern parts of Sierra Leone are favorable to a high contact rate between onchocerciasis patients coming from the south-western area of this country and the vector species Simulium yahense and Simulium squamosum which assume the essential of onchocerciasis transmission in the above-mentioned mining area. In fact, the Onchocerca volvulus strains concerned by this contact seem to be more pathogenic than those locally transmitted. In order to assess the danger it could represent for the Onchocerciasis Control Programme in West Africa, we carried out the experimental study of transmission which may result from this contact when more or less infected onchocerciasis patients are involved. The results indicated that this transmission by S. yahense may reach high proportions only when heavily infected onchocerciasis patients are implicated. We took also notice of the low capacity of S. squamosum to transmit the O. volvulus strains from the south-western Sierra Leone, irrespective of the microfilarial load of patients. Thus, in the most favorable conditions of a high parasite-vector contact of the study, involvement of S. yahense and onchocerciasis patients with high skin microfilarial loads is the only occurrence to which a high risk of intensive transmission may be related. The authors consider that the probability of such a risk occurring will be drastically reduced, due to the considerable decrease of skin microfilarial loads in human communities which regularly have the advantage of ivermectin (Mectizan) mass treatments.

Animals↗

Histochemical distribution of esterases in adult Onchocerca fasciata (Filarioidea: Onchocercidae).

The localization and distribution patterns of non-specific esterases (NSE) and acetylcholinesterase (AChE) activities in adult female Onchocerca fasciata (Filarioidea: Onchocercidae) have been determined using enzyme histochemical techniques. NSE activity was highest in the hypodermis and mild to moderate in the somatic musculature and uterine epithelium. The intensity of the reaction was decreased but not eliminated by sodium fluoride. AChE activity was found in the hypodermis and cuticle of the female worm, highest in fully formed microfilariae in the uterus, whereas early oocytes and embryonic forms were negative. Butyrylcholinesterase activity showed similar but less intense staining reactions at the same sites. Eserine-treated sections showed no AChE activity in all areas where it is usually present. The present techniques failed to demonstrate AChE activity in the nervous tissue including the hypodermal nerve trunks and neuromuscular junctions of the worm. The possible functional significance of the demonstrable enzymes was discussed with respect to their location. NSE activity may be involved in lipid metabolism, whereas AChE activity may have a role in host-parasite relationship.

Acetylcholinesterase↗

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↗