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PubMed2001

Onchocerca volvulus: genetic diversity of parasite isolates from Sudan.

Onchocerciasis in Sudan exists in three distinct foci which exhibit differing clinical presentations. Previous studies have demonstrated that a tandemly repeated Onchocerca sequence family with a unit repeat length of 150 bp (the O-150 family) is a useful marker for deducing relationships among different O. volvulus populations. In the current study, the O-150 repeat families of O. volvulus from Sudan were analyzed and compared to each other and to those of parasites from West Africa. Similar to West African and American O. volvulus, the O-150 families of the Sudanese parasites could be divided into clusters within which little or no intracluster variation was evident, suggesting that the O-150 family in these parasites was subject to the forces of concerted evolution. Statistical analysis of the O-150 families from the different Sudanese parasite isolates, employing a nested algorithm based on an analysis of variance, revealed that O. volvulus endemic to the northern focus at Abu Hamed were significantly different from all other O. volvulus populations examined to date. In contrast, parasites from the southern and eastern foci of Sudan were indistinguishable from those endemic to the West African savanna. The significance of these data are discussed in light of knowledge of the biogeography and biology of transmission of O. volvulus in Africa.

Africa, Western↗
PubMed1990

Onchocerca raillieti: release from skin snips, maintenance in vitro and periodicity of microfilariae.

Several media were tested for the release of Onchocerca raillieti microfilariae from skin snips and for their subsequent in vitro maintenance. Tyrode's solution containing 20% equine serum and antibiotics was the best medium tested, followed by phosphate buffered saline. Tyrode's solution alone or distilled water were poor media. A temperature 7-12 degrees C lower than the host's body temperature favoured release of the microfilariae from skin snips. The microfilariae were best maintained at 4-10 degrees C, when they remained alive for up to 5 days. O. raillieti microfilariae had an evening periodicity which could be related to a possible vector's peak of feeding activity.

Animals↗
PubMed1987

Electron-microscopic observations on the female worms of six Onchocerca species from cattle and red deer.

The midbody regions of female worms of six Onchocerca species (O. flexuosa, O. tarsicola, O. lienalis, O. gutturosa, O. armillata, O. gibsoni) were studied by transmission electron microscopy. The cuticular layering was rather similar in all species with the ridges built up by the cortical layers and the inner cuticular striations by the median or basal layers. Differences in the epicuticular morphology were considerable. O. flexuosa and O. lienalis had a thin epicuticle without protuberances, the epicuticle of O. armillata carried small knobs, and O. tarsicola, O. gutturosa, and O. gibsoni had a thick trilaminar epicuticle with long protuberances. Extreme hypertrophy of hypodermis and reductions of somatic musculature were observed in O. flexuosa and O. gibsoni. Less extended thickenings of the hypodermis were observed in the other species. No degenerative alterations were found in the muscle cells of O. gutturosa and O. lienalis. The intestinal lumen of most of the species was in a central position, but in O. tarsicola and O. gibsoni the lumen was reduced to small clefts between the intestinal cells. In these species, numerous electron-dense, concentric granules were observed in the cytoplasm of the intestinal cells. The proportions of the various organs differed considerably from species to species, e.g., the uteri contained the embryos filed one behind the other in O. tarsicola, whereas 50 or more embryos were found beside one another in cross-sections of the uterus of O. gibsoni. The comparative study showed that O. gibsoni and O. volvulus have many derived morphological characteristics in common and that in the other species more primitive stages of development of these morphological marks can be observed.

Animals↗
PubMed1988

Studies on glucose-6-phosphate dehydrogenase from the human parasite, Onchocerca volvulus.

Glucose-6-phosphate dehydrogenase (E.C. 1.1.1.49) was partially purified from the extracts of adult Onchocerca volvulus by affinity chromatography on 2'5'ADP-Sepharose-4B. Kinetic studies revealed a typical bell-shaped pH profile with an optimum lying between pH 7.3 and 7.8. The apparent Km for glucose-6-phosphate was 5.66 x 10(-5) M, whereas that for NADP was 2.17 x 10(-6) M. Suramin, a filaricidal drug, inhibited the enzyme competitively with respect to NADP as a substrate: the apparent Ki values were 2.23 x 10(-6) M and 4.21 x 10(-7) M, respectively, for the crude and purified enzyme preparations. Glucose-6-phosphate dehydrogenase therefore, could be one of the targets of suramin in vivo.

Animals↗
PubMed1983

Screening of some British simuliids for susceptibility to experimental Onchocerca lienalis infection.

Ten species of Simulium from Britain were tested for susceptibility to Onchocerca lienalis infection by intrathoracic injection of microfilariae recovered from the skin of cattle at slaughter. All of the species examined supported development of a proportion of the microfilariae to third-stage larvae, but this was greatest at 53% in S. ornatum s.l., which is a known natural vector in Britain. Inocula of 20, 30 or 50 microfilariae resulted in similar levels of mortality in S. ornatum s.l. (42-53%), but a rise in the dose above 20 microfilariae per fly increased the proportion which developed to third-stage larvae. In a series of comparative experiments employing S. ornatum s.l. as a reference positive for blackflies of unknown susceptibility to infection, 6 other species (S. reptans, S. erythrocephalum, S. lineatum, S. equinum, S. nitidifrons and S. austeni) allowed a proportion of the microfilariae to reach third-stage larvae equivalent to 37-50% of those which developed in S. ornatum s.l. A small sample of S. angustipes supported development as well as its respective control group of S. ornatum s.l., but S. vernum and S. aureum were relatively refractory and were the only species in which third-stage larvae did not accumulate in the head. Fly survival rates varied between the species, but all were below those in S. ornatum s.l. It is concluded that some of these blackflies offer promise as surrogate vectors of O. lienalis in the laboratory and their possible role in natural transmission should receive further attention.

Animals↗
PubMed1983

Isolation of antigen from Litomosoides carinii macrofilariae detecting serum antibodies due to Onchocerca volvulus.

Crude aqueous Litomosoides carinii adult worm extract was used as antigen for the detection of antibodies in sera from African patients with proven onchocerciasis (n = 45) resident in rural endemic areas of Togo and Sierra Leone. In 71% of cases this extract was found to produce 1 to 5 precipitation arcs in immunoelectrophoresis. Using a crude aqueous extract from adult Onchocerca volvulus, precipitation tests were positive in 75% of cases. The complexity of the L. carinii crude extract was shown by PAG-disc electrophoresis, PAG-electrofocusing, immunoelectrophoresis and crossed immunoelectrophoresis with the appropriate rabbit-antiserum. An antigen detecting onchocercal antibodies was isolated by two step preparative flat bed electrofocusing in granulated gel (PEGG). The antigen (pI 6.55, molecular weight 55 to 60 kd as estimated by SDS-PAG electrophoresis) was very suitable for antibody demonstration in double diffusion test and immunoelectrophoresis. Preliminary controls for specificity were performed by diffusing the antigen against sera from human and animal helminthoses including filarial infections. In contrast to the crude L. carinii extract no reaction was observed with sera from helminthic infections others than filariasis.

Animals↗
PubMed1991

Morphological demonstration of essential functional changes after in vitro and in vivo transition of infective Onchocerca volvulus to the post-infective stage.

Transmission electron microscopic investigation of the morphogenesis of Onchocerca volvulus through the third moult to the post-infective stage revealed essential alterations in various larval organs. Complete rebuilding of surface structures, the reduction of secretory granules in the glandular oesophagus, the unfolding of the intestine, an increase in the number of nerve fibres in the nerve ring and novel sensory papillae were significant findings. Transition from third- to fourth-stage larvae (L4) started as early as 48 h after transfer to vertebrate conditions in vivo in surrogate hosts and in vitro. After a resting period of about 60 h to enable a reduction in gland size and the loosening of the old cuticle and formation of the new one, the larvae started to cast the infective-stage cuticle. Young L4 exhibited a thin, monolayered cuticle and did not rebuild a glandular oesophagus. The body cavity widened, the intestine unfolded and the increased number of microvilli indicated the resumption of metabolic activity.

Animals↗
PubMed1991

Thin-layer and liquid column chromatographic analyses of the lipids of adult Onchocerca gibsoni.

Lipids were extracted from adult Onchocerca gibsoni with chloroform/methanol and the total lipid content was characterized. Glycolipids were isolated from other lipid classes by Florisil column chromatography and were then fractionated by DEAE-Sephadex ion-exchange chromatography. HPTLC revealed the presence of 9 neutral glycolipid bands and of 15 acidic glycolipid bands that stained for sialic acid with resorcinol. Lipids that contained no carbohydrates were analyzed by a combination of TLC and amino column chromatography. Triacylglycerols, cholesterol, cholesterol esters, and free fatty acids were found to be major components of the neutral lipid fraction, and diacylglycerols and monoacylglycerols were minor components. Phosphatidylethanolamine and phosphatidylcholine were the predominant phospholipids. Phosphatidylserine, phosphatidylinositol, sphingomyelin, lysophosphatidylcholine, and lysophosphatidylethanolamine were also present in significant amounts, whereas only traces of cardiolipin and phosphatidic acid were detected. Several minor lipids and phospholipids remained unidentified. These results indicate that adult O. gibsoni have a nonglycosylated lipid composition resembling that of other parasitic nematodes as well as a substantial repertoire of glycolipids, including many with the characteristics of gangliosides.

Animals↗
PubMed1995

Comparison of the patterns of codon usage and bias between Brugia, Echinococcus, Onchocerca and Schistosoma species.

Patterns of codon usage and bias were compared among taxa of the genera Brugia, Echinococcus, Onchocerca and Schistosoma by metric multidimensional scaling and three commonly used indices of bias: Nc, GC3S and B. The overall codon usage for each taxon was compared, as was the codon usage for each individual gene within the taxa. Differences in the patterns of codon usage observed between taxa were dependent on the overall base composition of the genes analysed. The codon usage of Echinococcus was distinct from that of the other taxa. Furthermore, the pattern of codon usage detected by the average codon usage summed across all genes for each taxon was not shown by all genes from that taxon.

Animals↗
PubMed1995

Cell-adhesion molecules expressed by activated eosinophils in Onchocerca volvulus infection.

Cell-adhesion receptors mediate the interaction between host effector cells and cellular or multicellular targets, including opsonized parasites. Our recent finding of a deposition of plasma proteins, including fibronectin, on the surface of infective larvae of the helminthic parasite Onchocerca volvulus led us to investigate the possible expression of cell-adhesion molecules (CAM), particularly fibronectin receptors, on eosinophilic granulocytes from persons infected with O. volvulus. Immunofluorescence analyses showed that freshly isolated eosinophils strongly expressed the beta 2-integrin CR3 and exhibited to a lower degree CR4 and the integrin-associated carbohydrate determinant Le(x), as well as antigen p24 (CD9). Eosinophils exposed to the eosinophil-active cytokines recombinant human interleukin 3 (rhIL-3) and granulocyte/macrophage colony-stimulating factor (rhGM-CSF) in addition to CR3, CR4, and CD9 expressed the beta 1-integrins VLA-4 and to a lesser extent VL-5 (both fibronectin receptors) as well as the intercellular adhesion molecule ICAM-1. Low-level expression of these adhesins was also observed on eosinophils cultured in the presence of these interleukins on confluent fibroblasts. The presence of VLA-4 as well as VLA-5 on activated eosinophils was confirmed by demonstration of the formation of immune rosettes using antibody-coated microspheres and by their attachment to fibronectin-coated surfaces. These results indicate the possibility of an involvement of previously unidentified antibody- and complement-independent mechanisms in cellular interactions with the parasite O. volvulus.

Animals↗
PubMed1995

Ultrastructure study of the excretory system and the genital primordium of the infective stage of Onchocerca volvulus (Nematoda:Filarioidea).

The electron microscopic investigation of the anterior part of the infective third-stage juvenile of Onchocerca volvulus provides first insights into the structure of the excretory system of this developmental stage of the parasite. The most anterior part of this system consists of a cell process of the syncytial excretory cells. At this height the excretory cells enclose the cuticle-lined excretory channel. The channel is in the process of elongation in the anterior-posterior direction, indicated by cell division in this region. More posteriad an ampulla-like structure is forming in the cytoplasm of the excretory cells. The inner surface of this ampulla is lined with a small number of single microvilli. In this part of the system the cytoplasm of the excretory cells is rich in Golgi bodies and endocytic vesicles. The ampulla has direct access to the exterior by the excretory duct. The excretory duct is a cuticle-lined structure surrounded by supporting fibres of an additional cell. This duct cell connects the excretory duct to the body-wall cuticle at the excretory pore. Adjacent to the region of the excretory system a cell is found that resembles a gland cell. This cell is in close contact to the ventral nerve cord. The genital primordia of the third-stage juvenile consist of several dividing cells. The female genital primordium is seen at the junction of the muscular with the glandular oesophagus and the male primordium can be found at the junction of the glandular oesophagus with the gut.

Animals↗
PubMed1993

Ultrastructural observations on the nervous system and the sensory organs of the infective stage (L3) of Onchocerca volvulus (Nematoda: Filarioidea).

A detailed morphological investigation of the sensory organs and the nervous system of the third juvenile stage of Onchocerca volvulus was performed at the ultrastructural level. A complex system of different receptor cells is found at the anterior and posterior end of this developmental stage. The eight papillae are arranged in two concentric circles consisting of two types of morphologically different receptors. Accessory nerve processes end free in the tip of the head. The paired amphids contain nine dendritic processes and accessory axons are seen in the surrounding cells. The basic structure of the amphids and of the circumoesophageal nerve ring is similar to that of other filarial nematodes. Two presumably neurosecretory cells are associated with the nerve ring. The reticular cytoplasm of these cells merges with the rough-surfaced endoplasmic reticulum of the lateral hypodermal chord. The paired phasmids at the posterior end of the developmental stages consist of single modified cilia that are embedded in an electron-dense mass. The receptor cell has access to the outside by a channel ending with a cuticular pore.

Animals↗
PubMed1995

Light and electron microscopy studies on Onchocerca jakutensis and O. flexuosa of red deer show different host-parasite interactions.

Adult filariae of two intranodular Onchocerca species of red deer (Cervus elaphus) were examined at the ultrastructural level. In all, 90 nodules of O. flexuosa and 20 nodules of O. jakutensis were studied by histology. O. jakutensis caused cystic and pus-filled nodules in which the female and male worms were motile. Female worms possessed a thick cuticle and a well-developed somatic musculature. The basal lamina of the muscular syncytia was connected with the transhypodermal fibers through long protrusions. The epicuticle had only few protuberances and no visible surface coat. Female O. flexuosa showed morphological features similar to those of other intranodular filariae. A surface coat could not be detected. In 24% of 33 nodules with young live female worms and 40% of 38 nodules with older live female O. flexuosa the worms were surrounded by giant cells apparently attacking a cuticle. This was found even in nodules obtained from animals no older than 9 months. It is probable that the lack of a surface coat protecting the female worm and the inability to move due to the reduced somatic muscles lead to an early elimination of O. flexuosa by the host's immune system.

Animals↗
PubMed1994

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

Histochemical techniques were employed to study the tissue distribution of hydrolytic enzymes in adult female Onchocerca fasciata (Filarioidea: Onchocercidae). Different tissues differed considerably in the localization and distribution of the six enzymes studied. Acid phosphatase (AcPase) activity was detected in the cuticle, hypodermis and reproductive organs. Alkaline phosphatase (AlkPase) activity was largely absent. Adenosine triphosphatase (ATPase) was found in the somatic musculature and muscles of the uterine ducts, whereas 5'-nucleotidase (5'-Nu) was restricted to young oocytes and dividing embryos in the female worm. Strong glucose-6-phosphatase (G-6-Pase) activity was demonstrated in the uterine epithelial cells and microfilariae, as was weak activity in the hypodermis. Naphthylamidase (NAM) activity was detected in the hypodermis, with lower activity occurring in the somatic musculature. The possible functions of these enzymes are discussed with respect to their location. The hydrolytic enzymes AcPase and NAM in the body wall are probably involved in absorptive-digestive functions, NAM in the somatic musculature may be concerned with tissue protein turnover, and ATPase, 5'-Nu and G-6-Pase may have a role in active transport and energy metabolism.

5'-Nucleotidase↗
PubMed2003

Epidemiology of concomitant infections due to Loa loa, Mansonella perstans, and Onchocerca volvulus in rain forest villages of Cameroon.

An epidemiological survey was conducted in 16 remote villages of the rain forest of southern Cameroon to ascertain the prevalence and intensity of three species of filariae: Loa loa, Onchocerca volvulus, and Mansonella perstans. We examined 1458 individuals for blood-dwelling microfilariae and 1255 of these were also for the presence of palpable nodules. All the villages surveyed were found highly endemic for onchocerciasis and mansonellosis with prevalence ranging from 28.44% to 87.17% for O. volvulus and 52.48% to 100% for M. perstans. The intensities of infection were also found high for M. perstans with arithmetic means of microfilaremia ranging from 280.94 to 4947.57 mf/ml. The loiasis prevalence was relatively low with value from 2.22% to 19.23%. Males were found more infected than females for the three species of filariae, and the prevalence and intensities of microfilaremia vary differently in males and females at different ages. The three species of filariae displayed different degrees of association in the inhabitants with a low prevalence of co-occurrence between L. loa/O. volvulus and between L. loa/M. perstans. In contrast, there was a high prevalence of co-occurrence between M. perstans and O. volvulus. The implications of the co-occurrence of the three species of filariae in the populations of these remote villages on the intervention programs based on mass treatment with mectizan are discussed.

Adolescent↗
PubMed2003

An aspartate aminotransferase of Wolbachia endobacteria from Onchocerca volvulus is recognized by IgG1 antibodies from residents of endemic areas.

Wolbachia are intracellular alpha-proteobacteria, closely related to Rickettsia, that infect various arthropods and filarial parasites. In the present study, the cDNA encoding the aspartate aminotransferase (AspAT) of Wolbachia from the human pathogenic filarial parasite Onchocerca volvulus (Ov-WolAspAT) was identified. At the amino acid level, the identity of the Ov-WolAspAT was 56% to Rickettsia prowazekii AspAT and 54% to the AspAT of the nitrogen-fixing bacterium Sinorhizobium meliloti, but the highest degree of identity was found to the putative AspAT of Wolbachia from Brugia malayi and Drosophila melanogaster (85%). All of these bacterial AspATs are members of the AspAT subclass Ib. A 35 kDa fragment of the Ov-WolAspAT was expressed in Escherichia coli, and immunolocalization using polyclonal antibodies against this antigen revealed that Ov-WolAspAT is present in a considerable proportion of the Wolbachia from O. volvulus, as well as in the endobacteria of several other filarial parasites. Western blot analysis using recombinant Ov-WolAspAT as antigen showed that IgG1 antibodies were present in 70 (51%) individuals living in areas endemic for O. volvulus, B. malayi or Wuchereria bancrofti and no IgG4 or IgE antibodies were found. Among 40 sera of persons from Uganda and Liberia who were putatively not infected with human filarial parasites, 11 (28%) individuals presented IgG1 antibodies, while none of the 33 sera from healthy Europeans and none of the 14 sera from patients with proven Rickettsia or Brucella infections reacted with the antigen. These results also show that an intracellular protein of Wolbachia endobacteria (WolAspAT) acts as antigen in human filariasis.

Amino Acid Sequence↗
PubMed1996

Onchocerca fasciata: enzyme histochemistry and tissue distribution of various dehydrogenases in the adult female worm.

Histochemistry studies of key dehydrogenases in the glycolytic pathway and related enzymes and the tricarboxylic acid (TCA)-cycle enzymes were carried out on adult female Onchocerca fasciata. The distribution pattern and enzymatic activities of 6-phosphogluconate dehydrogenase (6-GPDH), lactate dehydrogenase (LDH), mitochondrial glycerol-3-phosphate dehydrogenase (GPDH), nicotinamide adenine dinucleotide (phosphate) [NAD+(P)]-linked isocitrate dehydrogenase (ICDH), and NAD+(P)-linked malate dehydrogenase (MDH) in various tissues of the worm were determined. Moderate to intense enzyme activities were localized in three main areas, namely, the hypodermis, body-wall muscle, and reproductive tissues. Activity of the formazan reaction product was very low, if at all present, in the intestinal epithelium and was completely absent in the cuticle. On the basis of the present results and earlier observations, it is suggested that glycolysis leading to the end product lactate is the main energy-generating pathway in O. fasciata. The presence of significant activity of 6-GPDH indicates that the pentose-phosphate pathway might be operative in O. fasciata. In light of the activity of some of the TCA-cycle enzymes, ICDH and MDH, demonstrable in O. fasciata, it is possible that an additional pathway (pyruvate-succinate) of glucose metabolism via a reverse sequence of the TCA cycle may also be operative in the worm. The possible functional significance of the enzymes detected is discussed with respect to their location.

Alcohol Oxidoreductases↗