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1.
Parasitol Res ; 123(4): 186, 2024 Apr 18.
Artículo en Inglés | MEDLINE | ID: mdl-38634933

RESUMEN

Onchocerciasis is a devastating skin and eye disease that afflicts about 21 million people, most of whom live in sub-Saharan Africa. Its control with the microfilaricidal drug ivermectin is limited, thus necessitating the development of preclinical animal models to aid in the discovery of a macrofilaricide. Previously, we found that Onchocerca ochengi (the closest relative of the human O. volvulus) worm masses survive better in hamsters than in gerbils. The aim of this study was to compare the survival of O. ochengi adult male worms and their susceptibility to flubendazole (FBZ, a macrofilaricide) in gerbils and hamsters. The animals were intraperitoneally implanted with O. ochengi male worms, treated with FBZ, and sacrificed 35 days post-implantation. Unlike gerbils which had some worms moving freely in the peritoneum and some in newly formed nodules (neo-nodules), all the worms in the hamsters were found in neo-nodules. FBZ significantly decreased worm burden, motility, and viability in gerbils whereas it had no significant effect in hamsters. These results highlight a major difference in how O. ochengi adult male worms are sustained and affected by FBZ in gerbils compared to hamsters. Understanding the difference between these two models is important in the development of effective macrofilaricides for onchocerciasis.


Asunto(s)
Mebendazol/análogos & derivados , Onchocerca , Oncocercosis , Adulto , Animales , Cricetinae , Humanos , Masculino , Gerbillinae
2.
PLoS Pathog ; 17(6): e1009601, 2021 06.
Artículo en Inglés | MEDLINE | ID: mdl-34077488

RESUMEN

Onchocerciasis (river blindness), caused by the filarial worm Onchocerca volvulus, is a neglected tropical disease mostly affecting sub-Saharan Africa and is responsible for >1.3 million years lived with disability. Current control relies almost entirely on ivermectin, which suppresses symptoms caused by the first-stage larvae (microfilariae) but does not kill the long-lived adults. Here, we evaluated emodepside, a semi-synthetic cyclooctadepsipeptide registered for deworming applications in companion animals, for activity against adult filariae (i.e., as a macrofilaricide). We demonstrate the equivalence of emodepside activity on SLO-1 potassium channels in Onchocerca volvulus and Onchocerca ochengi, its sister species from cattle. Evaluation of emodepside in cattle as single or 7-day treatments at two doses (0.15 and 0.75 mg/kg) revealed rapid activity against microfilariae, prolonged suppression of female worm fecundity, and macrofilaricidal effects by 18 months post treatment. The drug was well tolerated, causing only transiently increased blood glucose. Female adult worms were mostly paralyzed; however, some retained metabolic activity even in the multiple high-dose group. These data support ongoing clinical development of emodepside to treat river blindness.


Asunto(s)
Enfermedades de los Bovinos/tratamiento farmacológico , Depsipéptidos/uso terapéutico , Filaricidas/uso terapéutico , Canales de Potasio de Gran Conductancia Activados por el Calcio/efectos de los fármacos , Oncocercosis/tratamiento farmacológico , Oncocercosis/veterinaria , Animales , Bovinos , Onchocerca/efectos de los fármacos
3.
Parasitol Res ; 122(7): 1581-1591, 2023 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-37154921

RESUMEN

Onchocerciasis, the second leading infectious cause of blindness, afflicts approximately 21 million people globally. Its control is limited to the use of the microfilaricidal drugs, ivermectin and moxidectin. Both drugs are unable to kill the adult worms which can survive for up to 15 years in patients, justifying the urgent need for potent and novel macrofilaricides that kill adult worms. The development of such drugs has been hindered by the lack of an appropriate small laboratory animal model to evaluate potential drug candidates in vivo. This study assessed the survival of O. ochengi female worms and their embryos over time in two laboratory rodents: gerbils and hamsters and tested using "proof-of-concept" studies, whether known macrofilaricidal drugs can kill these worms. Animals were surgically implanted with mechanical or collagenase-liberated O. ochengi worm masses, and necropsied at various time points to test for survival. Recovered worm masses were assessed for viability by biochemical analysis (MTT/formazan assay) or fecundity (embryogram). Flubendazole (FBZ) administered at 20 mg/kg body weight was used to validate both rodent models. By day 26 post-implantation of 15 worm masses, a median of 7.00 (4.00-10.00) was recovered from hamsters, and 2.50 (2.00-4.00) from gerbils. Worm masses recovered from gerbils were mostly disintegrated or fragmented, with significantly higher fragmentation observed with collagenase-liberated worm masses. FBZ had no significant effect on the number of worm masses recovered, but enhanced embryo degradation in gerbils and reduced worm mass viability in hamsters. This exploratory study has revealed the gerbil and hamster as permissible rodents to adult female worms of O. ochengi. The hamsters appeared to maintain the worms longer, compared to gerbils.


Asunto(s)
Filaricidas , Oncocercosis , Cricetinae , Animales , Femenino , Onchocerca , Gerbillinae , Filaricidas/farmacología , Ivermectina/uso terapéutico , Oncocercosis/tratamiento farmacológico
4.
BMC Vet Res ; 18(1): 66, 2022 Feb 10.
Artículo en Inglés | MEDLINE | ID: mdl-35144598

RESUMEN

BACKGROUND: Onchocerca lupi is a filarial nematode affecting dogs, and occasionally cats and humans, in continental Europe, North Africa, the Middle East, and the USA. Adult worms are usually found in periocular nodules and enucleation is sometimes required if the infection fails to respond to other treatment options. CASE PRESENTATION: Here, we report the presence of O. lupi in the UK for the first time. Of two dogs re-homed from continental Europe, one developed an ocular nodule seven years after arrival from Portugal. The conjunctival perilimbal mass in its left eye was surgically removed but despite anthelminthic treatment, a further nodule developed in the same eye six months later. In the second case - a dog imported from Romania 12 months earlier - a perilimbal mass was excised from the left eye and prior anthelminthic treatment was supplemented with oral prednisolone and doxycycline. However, nodules recurred, and the left globe was subsequently enucleated. Conjunctival hyperaemia then appeared in the right eye and neither additional anthelminthic treatment nor removal of worm masses failed to prevent the further development of lesions. Excised adult worms were identified in both cases as O. lupi based on morphological characteristics, as well as PCR and sequencing of cytochrome c oxidase subunit I and 12S rRNA gene fragments. CONCLUSION: O. lupi parasitosis can apparently remain cryptic in dogs for several years before any clinical signs manifest. Moreover, the progression of infection can be highly aggressive and recalcitrant to both surgical intervention and anthelminthic treatment. Increasingly, former stray dogs of unknown infection status are entering the UK, raising both veterinary and public health concerns.


Asunto(s)
Enfermedades de los Perros , Onchocerca , Animales , Enfermedades de los Perros/diagnóstico , Enfermedades de los Perros/tratamiento farmacológico , Enfermedades de los Perros/cirugía , Perros , Ojo , Onchocerca/genética , Salud Pública , Reino Unido/epidemiología
5.
Parasitol Res ; 121(4): 1199-1206, 2022 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-35006317

RESUMEN

The filarial nematode Onchocerca volvulus causes onchocerciasis (river blindness), a neglected tropical disease affecting 21 million people, mostly in Sub-Saharan Africa. Targeting the endosymbiont Wolbachia with antibiotics leads to permanent sterilization and killing of adult worms. The gold standard to assess Wolbachia depletion is the histological examination of adult worms in nodules beginning at 6 months post-treatment. However, nodules can only be used once, limiting the time points to monitor Wolbachia depletion. A diagnostic to longitudinally monitor Wolbachia depletion from microfilariae (MF) at more frequent intervals < 6 months post-treatment would accelerate clinical trials of antiwolbachials. We developed a TaqMan qPCR amplifying the single-copy gene wOvftsZ to quantify Wolbachia from as few as one MF that had migrated from skin biopsies and compared quantification using circular and linearized plasmids or synthetic dsDNA (gBlock®). qPCR for MF from the rodent nematode Litomosoides sigmodontis was used to support the reproducibility and validate the principle. The qPCR using as few as 2 MF from O. volvulus and L. sigmodontis reproducibly quantified Wolbachia. Use of a linearized plasmid standard or synthesized dsDNA resulted in numbers of Wolbachia/MF congruent with biologically plausible estimates in O. volvulus and L. sigmodontis MF. The qPCR assay yielded a median of 48.8 (range 1.5-280.5) Wolbachia/O. volvulus MF. The qPCR is a sensitive tool for quantifying Wolbachia in a few MF from skin biopsies and allows for establishing the qPCR as a surrogate parameter for monitoring Wolbachia depletion in adult worms of new antiwolbachial candidates.


Asunto(s)
Filarioidea , Onchocerca volvulus , Wolbachia , Animales , Humanos , Microfilarias , Onchocerca , Onchocerca volvulus/genética , Reproducibilidad de los Resultados , Wolbachia/efectos de los fármacos , Wolbachia/genética
6.
BMC Microbiol ; 21(1): 5, 2021 01 06.
Artículo en Inglés | MEDLINE | ID: mdl-33407120

RESUMEN

BACKGROUND: During the last two decades research on animal filarial parasites, especially Onchocerca ochengi, infecting cattle in savanna areas of Africa revealed that O. ochengi as an animal model has biological features that are similar to those of O. volvulus, the aetiological agent of human onchocerciasis. There is, however, a paucity of biochemical, immunological and pathological data for O. ochengi. Galectins can be generated by parasites and their hosts. They are multifunctional molecules affecting the interaction between filarial parasites and their mammalian hosts including immune responses. This study characterized O. ochengi galectin, verified its immunologenicity and established its immune reactivity and that of Onchocerca volvulus galectin. RESULTS: The phylogenetic analysis showed the high degree of identity between the identified O. ochengi and the O. volvulus galectin-1 (ß-galactoside-binding protein-1) consisting only in one exchange of alanine for serine. O. ochengi galectin induced IgG antibodies during 28 days after immunization of Wistar rats. IgG from O. ochengi-infected cattle and O. volvulus-infected humans cross-reacted with the corresponding galectins. Under the applied experimental conditions in a cell proliferation test, O. ochengi galectin failed to significantly stimulate peripheral blood mononuclear cells (PBMCs) from O. ochengi-infected cattle, regardless of their parasite load. CONCLUSION: An O. ochengi galectin gene was identified and the recombinantly expressed protein was immunogenic. IgG from Onchocerca-infected humans and cattle showed similar cross-reaction with both respective galectins. The present findings reflect the phylogenetic relationship between the two parasites and endorse the appropriateness of the cattle O. ochengi model for O. volvulus infection research.


Asunto(s)
Galectinas/administración & dosificación , Galectinas/genética , Inmunoglobulina G/sangre , Leucocitos Mononucleares/inmunología , Onchocerca/inmunología , Animales , Bovinos , Clonación Molecular/métodos , Femenino , Galectinas/inmunología , Perfilación de la Expresión Génica , Proteínas del Helminto/administración & dosificación , Proteínas del Helminto/genética , Proteínas del Helminto/inmunología , Humanos , Inmunización , Leucocitos Mononucleares/parasitología , Onchocerca/genética , Filogenia , Ratas , Ratas Wistar , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/inmunología , Análisis de Secuencia de ADN
7.
Med Vet Entomol ; 35(3): 231-238, 2021 09.
Artículo en Inglés | MEDLINE | ID: mdl-33480060

RESUMEN

Onchocerciasis is a neglected tropical disease (NTD) caused by Onchocerca Diesing 1841 (Spirurida: Onchocercidae) nematodes transmitted by blackflies. It is associated with poverty and imposes a significant health, welfare and economic burden on many tropical countries. Current methods to visualize infections within the vectors rely on invasive methods. However, using micro-computed tomography techniques, without interference from physical tissue manipulation, we visualized in three dimensions for the first time an L1 larva of an Onchocerca species within the thoracic musculature of a blackfly, Simulium damnosum s.l. Theobald 1903 (Diptera: Simuliidae), naturally infected in Ghana. The possibility that thicker peritrophic membranes in savannah flies could account for their lower parasite loads was not supported, but there were limits to our analysis. While there were no statistically significant differences between the mean thicknesses of the peritrophic membranes, in the anterior, dorsal and ventral regions, of forest and savannah blackflies killed 34-48 min after a blood-meal, the thickness of the peritrophic membrane in the posterior region could not be measured. Micro-computed tomography has the potential to provide novel information on many other parasite/vector systems and impactful images for public engagement in health education.


Asunto(s)
Oncocercosis , Simuliidae , Animales , Bosques , Insectos Vectores , Onchocerca , Oncocercosis/veterinaria , Microtomografía por Rayos X
8.
Med Vet Entomol ; 35(4): 646-651, 2021 12.
Artículo en Inglés | MEDLINE | ID: mdl-34013620

RESUMEN

Mosquito Magnet® traps, deployed in widespread parts of England as part of nationwide mosquito surveillance projects, also caught blackflies. As many as 1242 blackflies were caught in a trapping session lasting 4 days. Principal among the species caught were Simulium equinum, Simulium lineatum and Simulium ornatum s.l. As S. ornatum s.l. is a vector that transmits Onchocerca linealis to cattle and S. equinum is responsible for dermatitis ('sweet itch') in cattle and horses, it is suggested that Mosquito Magnet® traps could be used to monitor and partially control these pests, as well as nuisance anthropophilic blackflies such as Simulium posticatum that can cause simuliidosis in southern England.


Asunto(s)
Enfermedades de los Bovinos , Culicidae , Enfermedades de los Caballos , Oncocercosis , Simuliidae , Animales , Bovinos , Vectores de Enfermedades , Caballos , Mosquitos Vectores , Onchocerca , Oncocercosis/veterinaria
9.
Exp Parasitol ; 229: 108143, 2021 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-34437906

RESUMEN

BACKGROUND: Current treatment options for onchocerciasis are sub-optimal, prompting research and development of a safe cure (macrofilaricide). Onchocerca ochengi, a parasite of cattle, is used as a close surrogate for the human parasite O. volvulus in a murine model for pre-clinical screening of macrofilaricides. Skin from naturally infected cattle have been used in previous studies as a reliable source of parasite material. However, there is limited knowledge on how source-related factors such as the microfilaridermia status of the cattle, the nodule load and nodular worm viability may affect survival of male O. ochengi worms implanted in the rodent hosts. Such relationships were investigated in this study. METHODS: Dermal tissue and nodules were obtained from Gudali cattle, dissected and cultured to obtain migrating microfilariae (mf) and male worms. Emerged male worms were implanted into SCID mice and Gerbils (Meriones unguiculatus) and recovery rates were determined upon 42 days post implantation. Finally, nodules were processed for histology and embryogram analyses to assess the nodular worm viability and fertility, respectively. RESULTS: Of the 69 cattle sampled, 24 (34.8%) were mf+ and 45 (65.2%) were mf-. The mean nodule loads were 180.5 ± 117.7 (mf+) and 110.6 ± 102.7 (mf-) (p = 0.0186). The mean male worm harvest from nodules were 76.8 ± 120.3 and 47.2 ± 33.4 (p = 0.2488) for mf+ and mf- cattle, respectively. The number of male worms per 100 nodules were 57/100 and 46/100 nodules for mf+ and mf- cows, respectively. Female worms from nodules of mf- cows had higher counts of both normal and abnormal embryos with higher proportions of dead nodular worms evinced by histology compared to those from mf+ cows. A total of 651 worms were implanted into mice and gerbils, out of which 129 (19.81%) were recovered. Logistic regression analysis indicated that the microfilaridermia status of the cattle (presence of mf) (OR = 4.3319; P = 0.001) is the single most important predictor of the success of male worm recovery after implantation into rodents. CONCLUSION: Microfilaridermic cattle provide a promising source of adult O. ochengi. Male worms from this group of cattle have a better success rate of survival in a murine implant model. Nevertheless, in the programmatic point of view, amicrofilaridermic Gudali cattle would still constitute an important source of O. ochengi male worms with relatively good viability after implantation into rodents.


Asunto(s)
Enfermedades de los Bovinos/parasitología , Onchocerca/fisiología , Oncocercosis/veterinaria , Animales , Bovinos , Modelos Animales de Enfermedad , Femenino , Fertilidad , Gerbillinae , Masculino , Ratones , Ratones SCID , Microfilarias/crecimiento & desarrollo , Microfilarias/fisiología , Análisis Multivariante , Onchocerca/crecimiento & desarrollo , Oncocercosis/parasitología , Análisis de Regresión
10.
Vet Pathol ; 58(2): 401-404, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33356968

RESUMEN

Three donkeys were presented with progressive lameness and distal suspensory ligament breakdown in multiple limbs. Treatment with nonsteroidal anti-inflammatory drugs was only partially effective and eventually the donkeys were euthanized due to further progression of the lameness and concerns for their welfare. At necropsy, the distal part of the suspensory ligaments in multiple limbs, including the suspensory ligament branches, was markedly thickened, enlarged, and mottled white and brown on cut section. In one case, adult Onchocerca sp. nematodes were grossly identified embedded within the suspensory ligaments. Histopathologic examination revealed chronic, multifocal to coalescing, moderate to severe, lymphoplasmacytic, eosinophilic, and fibrosing desmitis and tendinitis with intralesional, coiled adult nematodes of Onchocerca sp., accompanied by osseous and cartilaginous metaplasia. To the authors' knowledge, this is the first histopathologic description of suspensory ligament desmitis and tendinitis associated with Onchocerca sp. in donkeys.


Asunto(s)
Enfermedades de los Caballos , Cojera Animal , Animales , Equidae , Caballos , Ligamentos , Onchocerca
11.
Parasitol Res ; 120(11): 3633-3644, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34519871

RESUMEN

Diverse Onchocerca species are present mostly parasitizing ungulates, with the exception of Onchocerca volvulus (O. volvulus) in humans and O. lupi in canids and cats. The human cases due to the O. lupi have been more highlighted during last years. So, the present review was performed to determine the detailed characteristics of confirmed human O. lupi case reports documented worldwide. Hence, a systematic search was done using English international databases (Scopus, PubMed, Web of Science, Embase, ProQuest, and Google Scholar). Totally, 14 confirmed human cases were documented during the last decade, mostly from the USA and Turkey with 7 and 3 cases, respectively. Most cases (7 individuals) were male with the age range of 22-month-old to 54-year-old. The parasite was frequently isolated from the right eye (5 cases), followed by the left eye (4 cases), cervical spinal canal (3 cases), scalp, and right forearm (one case each). Molecular identification of the isolated agent was the preferred way of diagnosis in most cases (9 records). In conclusion, human O. lupi cases have been more highlighted in recent years, whether due to the improved diagnostics and/or host-switching phenomenon, and both veterinarians and healthcare authorities should be alerted.


Asunto(s)
Onchocerca , Oncocercosis , Animales , Ojo/parasitología , Humanos , Masculino , Onchocerca/aislamiento & purificación , Oncocercosis/epidemiología
12.
Parasitol Res ; 120(6): 2087-2094, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33864105

RESUMEN

Control and treatment of onchocerciasis, a devastating tropical filarial disease caused by Onchocerca volvulus, rely solely on the community directed treatment with ivermectin. However, ivermectin is only microfilaricidal with evidence of resistance of the parasite among other limitations, which necessitate the search for new efficacious and safe filaricides. Ten synthetic thienylazoryl dyes were screened in vitro against adult and microfilariae worm stages of Onchocerca ochengi based on worm motility and MTT formazan assay. Cytotoxicity of active compounds was assessed on monkey kidney epithelial cells (LLC-MK2) using the MTT formazan assay. Seven (7) compounds showed both macrofilaricidal activity against adult male worms and microfilaricidal activity among which three 4a, 4c and 4e recorded the highest activity (IC50 = 4.2 to 8.8µM) against adult male worms, comparable to some standard anthelmintics. Five compounds showed rapid activity against microfilariae with 100% inhibition after 24-h incubation. The active compounds were nontoxic on monkey kidney cells (CC50> 4µg/mL), but their selectivity index values were relatively low (≤ 3). The thienylazoaryls with both macrofilaricidal and microfilaricidal activities may yield molecules which could be used for eradication of onchocerciasis following further medicinal chemistry modification of their structures to enhance their selectivity.


Asunto(s)
Compuestos Azo/farmacología , Filaricidas/farmacología , Microfilarias/efectos de los fármacos , Onchocerca/efectos de los fármacos , Tiofenos/farmacología , Animales , Compuestos Azo/química , Línea Celular , Supervivencia Celular/efectos de los fármacos , Concentración 50 Inhibidora , Masculino , Tiofenos/química
13.
Yale J Biol Med ; 94(2): 331-341, 2021 06.
Artículo en Inglés | MEDLINE | ID: mdl-34211352

RESUMEN

The parasitic filarioid Onchocerca lupi causes ocular disease characterized by conjunctivitis and nodular lesions. This nematode was first described in 1967 in a wolf from Georgia, and since then cases of infection from dogs and cats with ocular onchocercosis and sporadically from humans also with subcutaneous and cervical lesions caused by O. lupi have been reported from the Middle East, Europe, and North America. Due to its zoonotic potential, this parasitic infection has gained attention in the past 20 years. Phylogenetic studies have highlighted the recent divergence of O. lupi from other Onchocerca spp. and the importance of domestication in the evolutionary history of this worm. Moreover, the finding of an O. lupi genotype associated with subclinical and mild infection in the Iberian Peninsula, raises important questions about the pathogenicity of this presently enigmatic parasite.


Asunto(s)
Enfermedades de los Gatos , Enfermedades de los Perros , Oncocercosis Ocular , Animales , Gatos , Perros , Onchocerca/genética , Filogenia
14.
Emerg Infect Dis ; 26(12): 2989-2993, 2020 12.
Artículo en Inglés | MEDLINE | ID: mdl-33219658

RESUMEN

The Onchocerca lupi nematode infects dogs, cats, and humans, but whether it can be spread by coyotes has been unknown. We conducted surveillance for O. lupi nematode infection in coyotes in the southwestern United States. We identified multiple coyote populations in Arizona and New Mexico as probable reservoirs for this species.


Asunto(s)
Coyotes , Enfermedades de los Perros , Oncocercosis , Animales , Arizona/epidemiología , Reservorios de Enfermedades , Enfermedades de los Perros/epidemiología , Perros , New Mexico , Onchocerca/genética , Oncocercosis/epidemiología , Oncocercosis/veterinaria , Sudoeste de Estados Unidos , Estados Unidos/epidemiología , Zoonosis
15.
Parasitology ; 147(14): 1723-1727, 2020 12.
Artículo en Inglés | MEDLINE | ID: mdl-32829715

RESUMEN

Onchocerca lupi is a parasitic filarioid and the causative agent of canine ocular onchocercosis, a zoonotic disease of domestic dogs with sporadic reports in humans. A 13-year-old dog with no travel history outside of Israel was presented to an ophthalmology veterinary clinic in Israel with severe right ocular and periocular disease. After surgical exploration, thin helminths were removed from the dorsal sclera of the eye and identified as Onchocerca lupi by polymerase chain reaction according to the cytochrome c oxidase subunit I (cox1), reduced nicotinamide adenine dinucleotide dehydrogenase subunit 5 (nad5) and 12S rRNA genes. Phylogenetic trees and haplotype networks of the cox1 and nad5 genes confirmed the circulation of two genotypes: genotype 1 with worms from dogs, cats and humans from both the Old and New Worlds, and genotype 2 with specimens from Portugal and Spain. The Israeli sequences clustered in genotype 1 and were identical to O. lupi from the USA. Evidence of two genotypes separated geographically sheds light on the phylogeography and evolution of this zoonotic pathogen, and suggests a diverse pathology observed in different regions of the world.


Asunto(s)
Enfermedades de los Perros/diagnóstico , Genotipo , Onchocerca/genética , Oncocercosis Ocular/veterinaria , Animales , Enfermedades de los Gatos/parasitología , Gatos , Enfermedades de los Perros/parasitología , Enfermedades de los Perros/cirugía , Perros , Proteínas del Helminto/análisis , Humanos , Israel , Onchocerca/aislamiento & purificación , Oncocercosis Ocular/diagnóstico , Oncocercosis Ocular/parasitología , Oncocercosis Ocular/cirugía , Filogenia
16.
Med Vet Entomol ; 34(1): 59-68, 2020 03.
Artículo en Inglés | MEDLINE | ID: mdl-31508843

RESUMEN

Onchocerciasis in camels is caused by adult Onchocerca spp. and results in great economic losses to the camel industry. However, only a few studies on Onchocerca have been conducted, especially regarding the intermediate host and vector(s). In the present study, 192 camels were examined from December and January during 2013 and 2016, and the filarial larvae suspected to be Onchocerca spp. were further identified. Furthermore, aquatic dipteran insects in the living environment of camels were collected from May to September between 2013 and 2017 and dissected. Eventually, onchocercal lesions were observed in 95 of 192 (49%) camels and the captured insects were classified into 49 species from 42 genera in 21 families, among which 18 species were newly recorded in Inner Mongolia and 14 were haematophagous species. The filarial larvae were found in Culicoides puncticollis and identified as Onchocerca fasciata, indicating that C. puncticollis is the vector of O. fasciata in Inner Mongolia. These findings provide an estimate of onchocerciasis infection in camels and an alternative method of identifying insects and screening vectors using molecular methods. Important data are also provided for the diagnosis and control of onchocerciasis, thereby further filling the gap in knowledge regarding transmission vectors in China.


Asunto(s)
Camelus , Ceratopogonidae/parasitología , Onchocerca/fisiología , Oncocercosis/veterinaria , Animales , China/epidemiología , Insectos Vectores/parasitología , Larva/crecimiento & desarrollo , Larva/fisiología , Onchocerca/crecimiento & desarrollo , Oncocercosis/epidemiología , Oncocercosis/parasitología , Prevalencia
17.
Parasitol Res ; 119(3): 985-990, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-31953565

RESUMEN

The present paper deals with the post-mortem diagnostics of onchocerciasis and the molecular detection of causative agents of this disease in wild ruminant ungulates (Cervus elaphus, Dama dama and Capreolus capreolus). The animals were shot in hunting seasons 2017 and 2018, in two regions of the Eastern Slovakia. The total number of examined skins was fifty-eight. The presence of subcutaneous nodules was confirmed in 27.59% (95% CI 16-39) of animals. All positive skins belonged to red deer individuals (47.06%; 95% CI 30-64). The nodules were present mainly in the back area and in the lumbar area, and their sizes ranged from 2.9 to 24.1 mm, with the average count of 10 nodules per animal. Thirteen worms, isolated from the nodules collected from 13 animals, were subjected to molecular identification. Applying the standard PCR method, targeting the mitochondrial 12S rRNA, 16S rRNA and NADH-dehydrogenase gene, and subsequent sequencing, all the worms were identified as Onchocerca flexuosa Wedl, 1856. The sequences were submitted to GenBank under specific accession numbers. Two samples were identified as Onchocerca flexuosa haplotype B, in which T176A and A177T were present. Despite the presence of mutations in the 12S rRNA of the Onchocerca flexuosa, the standardized PCR remains to be a very specific and sensitive method that uses this fragment as a selectable marker for the detection of the studied parasite.


Asunto(s)
Ciervos/parasitología , Onchocerca/aislamiento & purificación , Oncocercosis/veterinaria , Animales , NADH Deshidrogenasa/genética , Onchocerca/clasificación , Onchocerca/genética , Oncocercosis/parasitología , ARN Ribosómico/genética , ARN Ribosómico 16S/genética , Piel/parasitología , Eslovaquia
18.
Molecules ; 26(1)2020 Dec 25.
Artículo en Inglés | MEDLINE | ID: mdl-33375687

RESUMEN

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.


Asunto(s)
Alcaloides/química , Onchocerca/efectos de los fármacos , Oncocercosis/tratamiento farmacológico , Voacanga/química , Alcaloides/farmacología , Animales , Humanos , Onchocerca/patogenicidad , Oncocercosis/parasitología
19.
Parasitology ; 146(14): 1773-1784, 2019 12.
Artículo en Inglés | MEDLINE | ID: mdl-31190665

RESUMEN

Filarial nematodes possess glutathione transferases (GSTs), ubiquitous enzymes with the potential to detoxify xenobiotic and endogenous substrates, and modulate the host immune system, which may aid worm infection establishment, maintenance and survival in the host. Here we have identified and characterized a σ class glycosylated GST (OoGST1), from the cattle-infective filarial nematode Onchocerca ochengi, which is homologous (99% amino acid identity) with an immunodominant GST and potential vaccine candidate from the human parasite, O. volvulus, (OvGST1b). Onchocerca ochengi native GSTs were purified using a two-step affinity chromatography approach, resolved by 2D and 1D SDS-PAGE and subjected to enzymic deglycosylation revealing the existence of at least four glycoforms. A combination of lectin-blotting and mass spectrometry (MS) analyses of the released N-glycans indicated that OoGST1 contained mainly oligomannose Man5GlcNAc2 structure, but also hybrid- and larger oligommanose-type glycans in a lower proportion. Furthermore, purified OoGST1 showed prostaglandin synthase activity as confirmed by Liquid Chromatography (LC)/MS following a coupled-enzyme assay. This is only the second reported and characterized glycosylated GST and our study highlights its potential role in host-parasite interactions and use in the study of human onchocerciasis.


Asunto(s)
Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Onchocerca/enzimología , Onchocerca/genética , Oncocercosis/veterinaria , Secuencia de Aminoácidos , Animales , Bovinos/parasitología , Enfermedades de los Bovinos/parasitología , Cromatografía de Afinidad , Cromatografía Liquida , Femenino , Glicosilación , Espectrometría de Masas , Onchocerca volvulus/enzimología , Onchocerca volvulus/genética , Oncocercosis/parasitología , Polisacáridos/química , Prostaglandina-Endoperóxido Sintasas/metabolismo , Estructura Terciaria de Proteína
20.
Exp Parasitol ; 198: 71-78, 2019 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-30771293

RESUMEN

Despite the efforts employed for the control of onchocerciasis, the latter has remained a significant public health problem, due mainly to the lack of safe and effective adult worm drugs and/or microfilaricides that do not kill Loa loa microfilariae (mf). Serious adverse events have been encountered after administering ivermectin to some onchocerciasis patients coinfected with Loa loa. There is therefore, an urgent need for a macro and/or microfilaricidal drug which kills Onchocerca but not L. loa microfilariae. A total of 12 crude extracts from Milletia comosa and Annona senegalensis were prepared and screened in vitro against the bovine species of Onchocerca, O. ochengi, and L. loa mf from humans. Mf and male worm viabilities were determined by motility scoring using microscopy at 120 h of incubation with drug, while adult female worm viability and cytotoxicity were determined biochemically by MTT/formazan colorimetry after 120 h of incubation with drug. Out of the 12 extracts, all 6 from M. comosa and 4 from A. senegalensis were active against male, female and mf of O. ochengi. The hexane extract from M. comosa leaves (MCL hex) was the most active with IC50 values of 1.38, 0.86 and 17.74 µg/mL for O. ochengi adult males, adult female and the mf, respectively. About 58% of the extracts were more active against O. ochengi than L. loa mf. These results demonstrate that these extracts contain active principles that kill Onchocerca parasite and to a lesser extent L. loa, and suggest that they can be fractionated for isolation of lead molecules for the safe treatment of onchocerciasis.


Asunto(s)
Annona/química , Filaricidas/farmacología , Loa/efectos de los fármacos , Millettia/química , Onchocerca/efectos de los fármacos , Extractos Vegetales/farmacología , Alcaloides/análisis , Animales , Bovinos , Células Cultivadas/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Femenino , Flavonoides/análisis , Masculino , Microfilarias/efectos de los fármacos , Corteza de la Planta/química , Extractos Vegetales/química , Hojas de la Planta/química , Polifenoles/análisis , Saponinas/análisis , Esteroides/análisis
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