Alkaloids with Anti-Onchocercal Activity from Voacanga africana Stapf (Apocynaceae): Identification and Molecular Modeling.

A new iboga-vobasine-type isomeric bisindole alkaloid named voacamine A (1), along with eight known compounds-voacangine (2), voacristine (3), coronaridine (4), tabernanthine (5), iboxygaine (6), voacamine (7), voacorine (8) and conoduramine (9)-were isolated from the stem bark of Voacangaafricana. The structures of the compounds were determined by comprehensive spectroscopic analyses. Compounds 1, 2, 3, 4, 6, 7 and 8 were found to inhibit the motility of both the microfilariae (Mf) and adult male worms of Onchocerca ochengi, in a dose-dependent manner, but were only moderately active on the adult female worms upon biochemical assessment at 30 µM drug concentrations. The IC50 values of the isolates are 2.49-5.49 µM for microfilariae and 3.45-17.87 µM for adult males. Homology modeling was used to generate a 3D model of the O. ochengi thioredoxin reductase target and docking simulation, followed by molecular dynamics and binding free energy calculations attempted to offer an explanation of the anti-onchocercal structure-activity relationship (SAR) of the isolated compounds. These alkaloids are new potential leads for the development of antifilarial drugs. The results of this study validate the traditional use of V. africana in the treatment of human onchocerciasis.

Impact of Ivermectin Mass Drug Administration for Lymphatic Filariasis on Scabies in Eight Villages in Kongwa District, Tanzania.

Scabies was recently added to the World Health Organization list of neglected tropical diseases. The ability to treat scabies with oral ivermectin makes a mass drug administration (MDA) campaign a feasible option for scabies control. Ivermectin MDA in communities endemic for lymphatic filariasis (LF) or onchocerciasis may already be having an impact on scabies. We examined the effect of ivermectin MDA for LF on scabies prevalence over 4 years in eight Tanzanian villages. At baseline, 4.4% (95% confidence interval [CI]: 3.7-5.4) of individuals tested positive for scabies, decreasing to 0.84% (95% CI: 0.51-1.4) after one round of ivermectin MDA but increased in Year 3 (2.5% [95% CI: 1.9-3.3]) and Year 4 (2.9% [95% CI: 2.2-3.8]). Most scabies cases were seen in children younger than 15 years. The data suggest that single-dose ivermectin MDA may not be effective in attaining long-term decreases when scabies prevalence is less than 5%.

Onchocerca - infected cattle produce strong antibody responses to excretory-secretory proteins released from adult male Onchocerca ochengi worms.

BACKGROUND: The front line molecules from filarial worms and other nematodes or helminthes are their Excretory-Secretory (ES) products. Their interaction with the host cells, proteins and immune system accounts for the skin and eye pathology or hyposensitivity observed in human onchocerciasis. ES products and adult worms' crude extracts from Onchocerca ochengi, a filarial nematode that infects the African zebu cattle, were utilized in the present study as a model for studying Onchocerca volvulus that causes river blindness in man. METHODS: The ES products were generated from adult male and female worms in vitro and analyzed with poly acrylamide gel electrophoresis (PAGE) and enzyme-linked immunosorbent assay (ELISA) using sera from Onchocerca-infected cattle and humans. The cattle sera were collected from a herd that had been exposed for six years to natural transmission of Onchocerca spp. The expressed reactivity was evaluated and differences analyzed statistically using Kruskal-Wallis rank and Chi-square tests. RESULTS: The gel electrophoretic analyses of 156 ES products from O. ochengi female and male worms and of two somatic extracts from three females and 25 males revealed differences in the protein pattern showing pronounced bands at 15, 30-50 and 75 kDa for male ES proteins and 15, 25 and 40-75 kDa for somatic extracts, respectively and less than 100 kDa for female worms. Proteins in the ES products and somatic extracts from female and male Onchocerca ochengi worms were recognized by IgG in sera from both Onchocerca-exposed cattle and humans. Bovine serum antibodies reacted more strongly with proteins in the somatic extracts than with those in the ES products. Interestingly, the reaction was higher with male ES products than with ES products from female worms, suggesting that the males which migrate from one nodule to another are more exposed to the host immune system than the females which remain encapsulated in intradermal nodules. CONCLUSIONS: This study demonstrates that O. ochengi ES products and, in particular, extracts from male filariae may represent a good source of immunogenic proteins and potential vaccine candidates.

International dog travelling and risk for zoonotic Onchocerca lupi.

Onchocerca lupi is a recently recognized threat for the health of animals and humans in European, American, African and Middle Eastern countries. We describe a case of imported O. lupi infection in Italy and report the lifespan of this parasite in a non-endemic area, to advocate increased awareness of the veterinary community for this zoonotic parasitosis.

Development and validation of an Onchocerca ochengi microfilarial hamster model for onchocerciasis drug screens.

BACKGROUND: Onchocerciasis, caused by the parasitic nematode, Onchocerca volvulus afflicts some 37 million people worldwide, and is the second leading infectious cause of blindness globally. The only currently recommended drug for treatment of the disease, ivermectin, is only microfilaricidal and has serious adverse effects in individuals co-infected with high loads of Loa loa microfilariae (mf), prompting the search for new and better drugs. Onchocerciasis drug discovery studies have so far been based on in vivo models using Onchocerca species which are not the closest to O. volvulus, and which may therefore, not adequately mimic the natural infection in humans. Therefore, this study was carried out to develop a better drug screening model for onchocerciasis, based on the use of cow-derived O. ochengi, the closest known relative of O. volvulus. METHODS: Mf of O. ochengi were injected subcutaneously at the nape of Syrian hamsters (Mesocricetus auratus) and BALB/c mice. The skin, and especially the earlobes of the animals were examined for mf 15-31 days after infection. For selected model validation, the hamsters were treated with ivermectin at 150 or 600 µg/kg body weight and examined 30 days after infection for mf. For L. loa studies in hamsters, isolated mf were injected intraperitoneally and animal organs were examined on day 26 for mf. RESULTS: The Syrian hamsters were found to be the more permissive to O. ochengi mf as fully viable mf were recovered from them on day 30, compared to BALB/c mice where such mf were recovered on day 15, but not 30. However, both animals were not permissive to L. loa mf even by day 15. Interestingly, more than 50 % of the total O. ochengi mf recovered were from the earlobes. The number of mf injected was directly proportional to the number recovered. Ivermectin at both concentrations tested completely eliminated the O. ochengi mf from the hamsters. CONCLUSION: This study reveals the Syrian hamster as an appropriate small animal model for screening of novel compounds against O. ochengi, the closest known relative of O. volvulus.

The onchocerciasis chronicle: from the beginning to the end?

The year 2012 marks the 25th anniversary of the donation of ivermectin to fight onchocerciasis and the projected date for elimination of transmission of the disease in the Americas. This review looks at the history of onchocerciasis, from its discovery through to 2025, by which time it is projected that the disease will have been eliminated as a public health problem, except in a handful of sub-Saharan countries, where it should be well on the way towards elimination.

Concurrence of dermatological and ophthalmological morbidity in onchocerciasis.

Prevalence of skin and eye disorders in African onchocerciasis (river blindness) is well documented. However, less is known about their joint occurrence. Information on concurrence may improve our understanding of disease pathogenesis and is required to estimate the disease burden of onchocerciasis. We analysed data from 765 individuals from forest villages in the Kumba and Ngambe Health districts, Cameroon. These data were collected in 1998, as baseline data for the evaluation of the African Programme for Onchocerciasis Control. Concurrence of symptoms was assessed using logistic regression. Onchocerciasis was highly endemic in the study population (63% nodule prevalence among males aged ≥20). Considerable overall prevalences of onchocercal visual impairment (low vision or blindness: 4%), troublesome itch (15%), reactive skin disease (19%), and skin depigmentation (25%) were observed. The association between onchocercal visual impairment and skin depigmentation (OR 9.0, 95% CI 3.9-20.8) was partly explained by age and exposure to infection (OR 3.0, 95% CI 1.2-7.7). The association between troublesome itch and reactive skin disease was hardly affected by adjustment (adjusted OR 6.9, 95% CI 4.2-11.1). Concluding, there is significant concurrence of morbidities within onchocerciasis. Our results suggest a possible role of host characteristics in the pathogenesis of depigmentation and visual impairment. Further, we propose a method to deal with concurrence when estimating the burden of disease.

Infective larvae of five Onchocerca species from experimentally infected Simulium species in an area of zoonotic onchocerciasis in Japan.

Microfilariae of five Onchocerca species, O. dewittei japonica (the causative agent of zoonotic onchocerciasis in Oita, Kyushu, Japan) from wild boar (Sus scrofa), O. skrjabini and O. eberhardi from sika deer (Cenus nippon), O. tienalis from cattle, and an as yet unnamed Onchocerca sp. from wild boar, were injected intrathoracically into newly-emerged black flies of several species from Oita to search the potential vector(s) of these parasites and identify their infective larvae. Development of O. dewittei japonica microfilariae to the infective larvae occurred in Simulium aokii, S. arakowae, S. bidentatum, S. japonicum, S. quinquestriatum, and S. rufibasis while development of infective larvae of O. skrjabini, O. eberhardi, and the unnamed Onchocerca sp. was observed in S. aokii, S. arakawae, and S. bidentatum. Development of O. lienalis microfilaria to infective larvae occurred in S. arakawae. Based on the morphology of infective larvae obtained, we proposed a key of identification of Onchocerca infective larvae found in Oita. We also reconsider the identification of three types of infective larvae previously recovered from Simulium species captured at cattle sheds: the large type I larvae that may be an undescribed species; the small type III identified as O. lienalis may include O. skrjabini too; the intermediary type II that may be O. gutturosa, or O. dewittei japonica, or the unnamed Onchocerca sp. of wild boar.

Onchocercosis mansonella co-infection presenting as a subcutaneous nodule in a child.

A 11-year-old girl presented with a subcutaneous lesion on the forehead, thought to be an implantation dermoid cyst. Microscopic examination revealed an onchocercoma. Microfilariae were found in the blood indicating a co-infection with Mansonella perstans. This case demonstrates the diagnostic difficulties of a parasitic co-infection uncommonly encountered in Europe.

Oncocercosis: una forma de vivir / Onchocerciasis: one way of living

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Pathogenesis and host responses in human onchocerciasis: impact of Onchocerca filariae and Wolbachia endobacteria.

Onchocerca volvulus is a tissue-invasive parasitic nematode causing skin and eye pathology in human onchocerciasis. The filariae habour abundant intracellular Wolbachia bacteria, now recognised as obligatory symbionts, and therefore emerging as a novel target for chemotherapy. Recent research demonstrates that both the filariae and endobacteria contribute to the pathogenesis of onchocerciasis, and molecules have been identified that promote inflammatory or counter-inflammatory immune mechanisms, divert the host's immune response or procure evasion of the parasite.

The role of endosymbiotic Wolbachia bacteria in filarial disease.

In this review, we describe the pathogenic role of Wolbachia endosymbiotic bacteria in filarial diseases, focusing on the host innate immune responses to filarial and Wolbachia products. A description of the host pathogen recognition and early inflammatory responses including TLR4-mediated signalling, chemokine and cytokine responses and inflammatory cell recruitment is provided from human studies and from animal models of filarial disease. Finally, the impact of the discovery and characterization of Wolbachia on filarial research and treatment programmes is discussed.

Association of transient dermal mastocytosis and elevated plasma tryptase levels with development of adverse reactions after treatment of onchocerciasis with ivermectin.

To investigate the role of mast cells in treatment-associated adverse reactions in patients with onchocerciasis, changes in plasma tryptase levels and skin mast cell counts were examined in 2 groups of Onchocerca volvulus-infected subjects after ivermectin treatment. After treatment, an increase in tryptase levels was observed concurrent with the onset of blood eosinopenia and preceding the appearance of plasma eosinophil-derived neurotoxin (EDN) and interleukin-5. Tryptase levels were correlated with development of peripheral eosinopenia and markers of eosinophil activation and degranulation. Dermal mast cell numbers increased transiently at 24 h after treatment, preceding the onset of dermal eosinophil infiltration and the development of clinically apparent inflammation. Local reactions were strongly correlated with levels of plasma tryptase and EDN, and the severity of systemic reactions was correlated with levels of tryptase, EDN, and interleukin-5. The data indicate that mast cells play a role in initiation of tissue inflammatory reactions after ivermectin treatment of onchocerciasis.

Ocular onchocercosis in dogs: a review.

In recent decades, sporadic cases of ocular Onchocerca species infection have been reported in dogs in the USA and Europe. In the acute stage of the disease severe inflammation of the ocular and periocular tissues was observed. In chronic cases, the strongly coiled, gravid nematodes were incorporated in pea- to bean-sized granulomatous nodules in various parts of the eye, including the retrobulbar space, orbital fascia, eyelid, third palpebra, conjunctiva and sclera. Apart from the ophthalmological significance of the disease, the large number of microfilariae in the skin may be responsible for acute and chronic dermatological problems. The geographical distribution and prevalence of the infection may be greater than currently thought, because the lesions may have been erroneously regarded as other ocular diseases. Onchocerciasis is the world's second most prevalent infectious cause of blindness in human beings and parasitologists have long searched for an experimental model of human onchocerciasis; ocular onchocercosis infections in dogs may provide a useful experimental system.

Ocular onchocercosis in dogs: aberrant infection in an accidental host or lupi onchocercosis?

Four adult dogs that had spent their entire life in Hungary, were found to be infected with filaroid nematodes of the genus Onchocerca. The morphology and location of the parasites as well as pathological lesions were similar to those described earlier in the one Hungarian and five US dogs. Only moderate morphological differences were noted between the adults of Onchocerca sp. infecting dogs and O. volvulus of man or O. lienalis of cattle. Nevertheless, the morphology of microfilariae of Onchocerca from dogs is unique within the genus. Their length was less than half the length of microfilariae of other Onchocerca spp. known so far. In addition to size differences, several characteristic morphological features were observed. The unsuccessful attempt to infect dogs with O. lienalis, the absence of O. volvulus and O. lienalis in endemic regions of canine onchocercosis, the different size, morphology, and location of the adults in dogs and cattle, the exceptionally small size and unique morphology of microfilariae of Onchocerca of canids indicate that a distinct species might be responsible for canine onchocercosis. Since the larval concentration in the skin was high (50-3600 microfilariae g(-1)) in all affected dogs, the diagnosis prior to surgical removal of worm nodules can be based on the examination of a small skin snip collected from the head or abdominal region. Infections in dogs may provide a model to study human onchocercosis, therefore, further studies are encouraged on the feasibility of experimental infection of dogs with this Onchocerca species.

Integrin-like RGD-dependent cell adhesion mechanism is involved in the rapid killing of Onchocerca microfilariae during early infection of Simulium damnosum s.l.

Injection trials with compatible and non-compatible Onchocerca species into S. damnosum s.l., the vector of human and bovine onchocerciasis, demonstrated that the rapid killing of microfilariae within the blackfly's haemocoel is species specific. In the presence of the peptide RGDS as a blocking agent for integrin-like receptors of haemocytes, the survival of O. ochengi microfilariae in its natural intermediate host was significantly increased. This increased survival 24 h p.i. correlated with a significant decrease of apoptosis levels in the microfilariae following a 2 h exposure to the haemolymph in vivo. These findings suggest that haemocytes are directly involved in the killing of Onchocerca microfilariae in the blackfly.

[Human onchocerciasis and "sowda" in the Republic of Yemen]. / Onchocercoses humaines et "sowda" en République du Yémen.

The geophysics of the north Yemen, associating a north-south directed mountainous fish bone (rising in more of 2,000 meters), to numerous rivers or "wadis" is convenient to the development of simulium shelters, main vectors for cutaneous filariasis to Onchocerca sp. Following several missions of bio-clinical and epidemiological evaluations in neighbouring villages of wadis, it has been possible to study different clinical aspects: one reminding the classical african onchocerciasis with generalized and diffused dermatitis, and, on an other hand, a hyperreactive dermatitis on one side of the body and associated with a collateral lymphatic ganglion. This disease is well known for local populations as "aswad" meaning "black" or "sowda". Clinically whatever the studied focus, coexists the two types of onchodermatitis (uni or bilateral). Yhe sowda patients are proportionally less numerous than those touched by the generalized type. Frequent eye lesions of the West African onchocerciasis are not found in sowda cases. In classical optical microscopy, microfilaria is morphologically indifferenciable between sowda and onchocerciasis clinical aspects. Skin snips were carried out on patients of both groups. Identification of microfilaria by molecular biology through the study of the DNA genome was done out of 5 skin snips. Microfilaria was kept dry between laminas and the DNA extracted from rehydrated microfilaria. DNA was intensified with specific primers of Onchocerca type (O150PCR). This phase was followed by hybridisation of amplification products by PCR to specific stains: OVS-2 for Onchocerca volvulus species, OCH for Onchocerca ochengi, PFS1 and PSS1-BT respectively for the forest strain and the savannah strain of Onchocerca volvulus as described previously. We can distinguish 2 kinds of answers based on the clinical origin of the snip-tests: the first one concern 3 patients with numerous dermal microfilariae but without any clinical sowda and corresponding to microfilaria O. volvulus type but different from the forest or savannah strains found in sub-Saharan Africa. The second one corresponds to 2 patients with less than 5 microfilaria in their snip-test. They show the typical clinical picture of sowda. They are identified as microfilaria type Onchocerca but they do not belong to species volvulus, or to species ochengi. It seems quite probable that the clinical picture of sowda be the result of developing onchocerciasis of animal origin and not identified as to day. The ivermectin, therapeutic of choice for African onchocerciasis in annual unique cure seems less effective in the coverage of sowda. In that case rehearsal of cures every 3 months would be necessary for mass campaigns to limit the transmission of this filariasis.