Onchocerca gutturosa and Onchocerca lienalis in cattle: variation in length of microfilariae by site of recovery

Dermal microfilariae recovered from specimens obtained from umbilical and cervical sites of cattle infected with adult Onchocerca gutturosa alone or with adults of O gutturosa and O lienalis were measured and compared with uterine microfilariae obtained directly from gravid female worms of each species. Uterine microfilariae of O gutturosa were longer than dermal microfilariae obtained from cattle harboring only adults of O gutturosa. Dermal microfilariae were recovered from umbilical and cervical sites in these cattle. Those found at the cervical site had lengths equal to or greater than lengths of microfilariae recovered from the umbilical site. There was a significant (P less than 0.0001) shift in length across populations of microfilariae of O gutturosa from various sites in its bovine host, with a progressive decrease in length between microfilariae recovered from the worm's uterus, microfilariae from the cervical dermis, and microfilariae from the umbilical dermis, respectively. A similar direct comparison was not possible for microfilariae of O lienalis, because none of the cattle was infected with only adult worms of this species. In an indirect comparison, microfilariae of O lienalis were identified at the umbilicus, but their presence in the cervical region could not be determined unequivocally because of confounding of microfilariae length by concurrent infection with O gutturosa. Uterine microfilariae from O lienalis were longer than uterine microfilariae of O gutturosa, although a degree of overlap in the range of measurements existed between species.     

Onchocerca gutturosa and Onchocerca lienalis in cattle: effect of age, sex, and origin on prevalence of onchocerciasis in subtropical and temperate regions of Florida and Georgia

Evaluation of 845 cattle from Florida and Georgia for onchocerciasis indicated a significantly higher prevalence of infection with Onchocerca gutturosa and a significantly lower prevalence of infection with Onchocerca lienalis in the southern portion of Florida (subtropical region) than in the northern portion of Florida or in Georgia (temperature region). In all regions, prevalence of infection with O gutturosa was significantly higher than infection with O lienalis. In the temperate region, bulls were infected more often with O lienalis than were cows. Other sex-related differences in infection with either filariid was not found. Prevalence of infection with O lienalis was similar between age groups in all regions. Rates of infection with O gutturosa differed between age groups within and between subtropical and temperate regions. Examination of cervical and umbilical skin specimens for microfilariae detected an additional 6.6% of infected animals that had not been found by examination for adult worms. However, microfilariae were not found in 21.4% of animals that were positive for adult worms.     

Ingestion of microfilariae from different sources by Culicoides nubeculosus (Diptera: Ceratopogonidae) through an artificial membrane

Laboratory-hatched Culicoides nubeculosus midges were membrane-fed on cattle blood containing various Onchocerca microfilariae which had been isolated from the hides of red deer (Cervus elaphus) and from Australian and native German cattle. All microfilariae were ingested through the membrane except those of O. tarsicola, which were too large. Onchocerca gibsoni and O. lienalis reached the thoracic musculature, but no further development occurred. Microfilariae of O. flexuosa developed to the "sausage" stage. Microfilariae of O. gutturosa developed to the infective larval stage within 9 days at a temperature of 25 degrees C.     

Bovine and equine onchocerciasis in eastern North America with a discussion on cuticular morphology of Onchocerca spp. in cattle.

Skin sections and/or the ligamentum nuchae and ligamentum gastrolienale were examined from twelve bovine carcasses obtained from southern and eastern Ontario and from Quebec. Of these, seven were shown to be infected with Onchocerca gutturosa and/or Onchocerca lienalis. The morphology of the adult female cuticle is discussed. Skin sections from 43 equine carcasses from a slaughter house in Grenville, Quebec were examined and microfilariae of Onchocerca sp. were recovered from 32 (74%). There are probably referable to Onchocerca cervicalis.     

Distribution of Onchocerca spp. microfilariae in the dermis of cattle

A grid was drawn on a lateral half of 10 hides, from cattle infected with both Onchocerca gutturosa and Onchocerca lienalis, which were obtained from an abattoir in central Florida. Paired 7-mm diameter skin cores were taken from 24 grid sites on each hide and microfilariae (mff) extracted to determine their distribution. The only consistent pattern distribution was that mff were recovered from all 10 hides at four sample sites along the ventral midline near the umbilicus. Microfilariae were also frequently found in large numbers along the dorsal cervical midline and in the region of the tarsal and carpal-metacarpal joints.     

Simulium jenningsi Malloch (Diptera: Simuliidae): a vector of Onchocerca lienalis Stiles (Nematoda: Filarioidea) in New York

Biting flies were collected from the umbilical areas of Onchocerca lienalis-infected cattle in New York (state) from June through September of 1980. Of the 766 flies collected, 705 were Simulium jenningsi. Microfilariae were detected in the midguts of 37 (50%) of 73 females dissected immediately after the flies had fed. The mean number of larvae per positive fly (fly with microfilariae) was 15.2. Third-stage larvae were recovered from 25 (21.9%) of 114 S jenningsi dissected 8 to 13 days after they had fed on the infected cattle; the mean number of 3rd-stage larvae per positive fly was 3.5. Dissections of flies performed on days 1 through 7 after feeding yielded various numbers of 1st and 2nd-stage larvae from the thoracic muscles. Ovarian dissections performed on 304 S jenningsi attacking cattle indicated an overall parous rate of 58%. Naturally occurring infections with filarial larvae indistinguishable from O lienalis were found in 7.3% of the parous females. Three of these flies, or 1.7% of the parous collection, harbored 3rd-stage larvae. The onset of naturally occurring filarial infections in the population of S jenningsi coincided with a peak in the parous rate in late June. Thereafter, filarial infections were generally detected when the parous rate was above 50%.     

ONCHOCERCA GUTTUROSA IN Danish cattle. Prevalence, geographic distribution and host-vector relationships.

1. Onchocerca gutturosa is reported in Danish cattle for the first time. Microfilariae were found in 38 (9.4%) of 406 cows that were 2 years or older.
2. Microfilariae were not randomly distributed throughout the skin but were concentrated in the umbilical area.
3. No difference in prevalence was observed between breeds of cattle.
4. Most of the infected cows had grazed on fields close to streams that contained Simulium ornatum, the vector of O. gutturosa.
5. Differences in preferred biting sites of S. ornatum from cow to cow were correlated with the arrangement of hair.

     

Teat onchocercosis in cows with reference to prevalence, species involved and pathology

The present study reports teat onchocercosis in cows in the province of Kars in north eastern Turkey with reference to the prevalence, species involved and pathological findings. In the study, 600 cows of various breeds and ages slaughtered in the local abattoir were examined and 145 (24%) were found to have various lesions on their teats, of which 45 (31%) were infected with three Onchocerca species (Onchocerca gutturosa, O. lienalis and an unidentified Onchocerca spp.), identified according to their morphological characteristics. Gross pathological examination of the teats with lesions revealed scabies and chaps of varying sizes, healed sores and small nodules. Histopathologically, large numbers of microfilariae were commonly observed within the collagenous fibre bundles of the dermis, with only a few microfilariae in the dermis in some cases. The microfilariae were also often found to have accumulated around the perivascular spaces and were frequently associated with infiltration by inflammatory cells, predominantly eosinophil leukocytes and mononuclear cells. In two cases, microfilariae were discovered exclusively in the lumen of the capillaries of the dermal papillaries. In one case, in which microfilarae were not encountered, sections of adult worms surrounded by neutrophil leukocytes were detected free in the teat canal. The presence of extensive teat lesions along with microfilariae of the Onchocerca species as found in this study warrants further assessment of the impact of onchocercosis on dairy farming.     

Molecular genetic comparison of Onchocerca sp. infecting dogs in Europe with other spirurid nematodes including Onchocerca lienalis

In the past 15 years, subconjunctival onchocercosis has been reported from 63 dogs in south-western United States (Arizona, California, Utah) and Southern and Central Europe (Germany, Greece, Hungary, Portugal, Switzerland). To reveal the taxonomic status of the parasite responsible for these infections, fragments of the mitochondrial cytochrome oxidase subunit I (COI) and NADH dehydrogenase subunit 5 (ND5) genes of three European strains of canine Onchocerca sp. and the 16S ribosomal RNA (16S rRNA) gene of their Wolbachia endosymbionts were sequenced and compared to the homologous sequences of other spirurid nematodes. The evolutionary divergence between COI and ND5 gene sequences of Greek, Hungarian and Portuguese strains of canine Onchocerca sp. were similar in magnitude to that seen within Thelazia callipaeda or Onchocerca lienalis. The evolutionary divergence between the sequences of canine Onchocerca sp. and other Onchocerca spp. including O. lienalis were similar or higher in magnitude to that seen between other Onchocerca spp. The results of the current and earlier phylogenetic analyses indicate that canine Onchocerca sp. separated from other Onchocerca spp. early in the evolution. Based on the similar clinical pictures, the identical morphology of nematodes and the sequence analyses of COI and ND5 genes of the worms and 16S rRNA gene of their wolbachiae, the Onchocerca worms isolated from European dogs appear to belong to the same species. The results support the earlier biological and morphological arguments that a distinct species, most likely O. lupi originally described from the subconjunctival tissues of a Caucasian wolf is responsible for canine ocular onchocercosis in Europe.     

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.     

Onchocerca ochengi acquisition in zebu Gudali cattle exposed to natural transmission: parasite population dynamics and IgG antibody subclass responses to Ov10/Ov11 recombinant antigens

Ngaoundere Gudali zebu cattle naturally exposed to Simulium damnosum s.l. and Culicoides spp. bites were examined during 4 years for O. ochengi adult worm acquisition, Onchocerca ochengi and Onchocerca gutturosa skin microfilaria dynamics, and IgG1 and IgG2 antibody subclass responses. Eleven animals acquired a total of 465 O. ochengi nodules (average of 17 per female and 72 per male). The O. ochengi nodule load was highly variable in individual animals and exacerbated in mature male cattle. Three patterns of acquisition of O. ochengi (resistant to new infestation, early susceptibility and late susceptibility), not associated with Simulium biting intensity (P > 0.05), were distinguished. The minimum prepatent periods for O. ochengi nodules, O. ochengi microfilariae and O. gutturosa microfilariae were 10, 20 and 21 months, respectively. The O. ochengi microfilaria density significantly (P < 0.001) increased with age, was higher in young mature bulls than female animals (P < 0.001) and finally reached highest levels (P < 0.005) during the dry season. Antibody responses to Ov10/Ov11 recombinant O. volvulus antigens were predominantly of the IgG1 subclass. High levels of this subclass (not IgG2) observed in new born calves declined to almost zero levels at the age of 5-8 months but IgG1 levels significantly increased (P < 0.05) with age subsequently during patency. Put together the acquisition and accumulation of O. ochengi parasites in zebu cattle, apart from being season, sex (gender) and host age associated, may also suggest a density-dependent regulation of parasite establishment in a proportion of the exposed population.     

Prevalence of microfilariae in cattle at slaughter in Zaria, Nigeria

Skin snips and blood taken from 372 randomly selected slaughtered cattle in Zaria, Nigeria were examined for microfilariae. A total of 293 (78.8%) skin snips and 218 (58.6%) blood samples were positive for microfilariae. Two types of microfilariae, identified as Onchocerca armillata, with a percentage prevalence of 64%, and O. gutturosa with a prevalence of 40% were isolated from the skin. The blood microfilariae were identified as those of Setaria labiatopapillosa.     

Electron microscopic and molecular identification of Wolbachia endosymbionts from Onchocerca lupi: implications for therapy

It was recently demonstrated that Wolbachia intracellular bacteria (alpha 2 proteobacteria, Rickettsiales) living in filarial nematodes are obligatory symbionts of their hosts. Herein, we report the electron microscopic and 16S ribosomal DNA-based (16S rDNA) identification of the endobacteria harboring in Onchocerca lupi. The worm nodules containing the nematodes were removed from three Hungarian dogs naturally infected with O. lupi. Wolbachia-like endobacteria were detected by electron microscopy in the lateral chords of both adult worms and microfilariae. The endosymbionts in O. lupi resemble in location, size, and morphology the wolbachiae found in other filariae. The presence of wolbachiae in O. lupi was also confirmed by PCR amplification of the 16S rDNA of the bacteria. The 16S rDNA-based phylogenetic analysis revealed that the endosymbionts of O. lupi infecting dogs belong to the supergroup C of Wolbachia pipientis and are not identical with those of other Onchocerca spp. sequenced so far. Since intermittent treatment with oxytetracycline has adulticid and microfilaricid activity by depletion of Wolbachia endobacteria, this antibiotic treatment regimen may offer an alternative of ivermectin or diethylcarbamazine in the suppression of postoperative microfilaridermia in Onchocerca-infected dogs and may prevent relapse.     

Onchocerca cervicalis in Horses from Southern Brazil

The objective of this study was to determine the occurrence of Onchocerca cervicalis in 1200 adult horses from rural areas of Rio Grande do Sul, Brazil. Umbilical skin specimens measuring 2 x 2 cm were minced, suspended in 10 ml of distilled water and incubated at room temperature overnight. The liquid volume was centrifuged and the sediment was screened for microfilariae. The ligamentum nuchae were totally removed, preserved in ice and dissected for the detection of adult forms of Onchocerca cervicalis. Microfilariae of Onchocerca cervicalis were detected in midventral skin biopsy samples in 215 (17.9%) of 1200 horses examined and the adult worms were recovered from 200 (16.6%) ligamentum nuchae from the same animals. These findings provide a basis for further studies to determine infection rates in horses from other regions of the state and to identify the intermediate host that transmits the disease.     

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°C lower than the host's body temperature favoured release of the microfilariae from skin snips. The microfilariae were best maintained at 4–10°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.Part of this work was carried out when both authors were at the Department of Parasitology, Faculty of Veterinary Science, University of Khartoum, Khartoum North, Sudan.     

Rearing of Culicoides nubeculosus (Diptera: Ceratopogonidae) by natural or artificial feeding in the laboratory

A technique for the mass rearing of Culicoides nubeculosus in the laboratory is described. Female midges were fed either on fresh or deep-frozen, defibrinated cattle blood (-70 degrees C) through latex membranes, or on anesthetized white mice. Feeding rates of up to 90% were obtained on the latex membrane, whereas only 41% of the midges fed on mice. The best oviposition rates of greater than 50% were achieved after feeding either on the latex membrane with fresh cattle blood or on mice. An average of greater than 100 eggs per female were deposited. The highest larval hatching rate was observed after feeding with fresh blood; about half of the larvae developed to the adult stage. A reproduction index was defined for the colony based on the feeding rate, oviposition rate, larval hatching rate and development to the adult stage. The highest reproduction index was obtained when the midges were fed on fresh cattle blood through the membrane.     

Onchocerca armillata Railliet and Henry, 1909 andOnchocerca gutturosa (Neumann, 1910) in camels (Camelus dromedarius L.) in the Sudan

A study undertaken to determine theOnchocerca species infecting camels that live in the same localities as cattle in the Dinder Region, Blue Nile Province, Sudan revealed concurrent infections with the bovine parasitesOnchocerca armillata in the thoracic aorta, brachiocephalic trunks and brachial arteries andOnchocerca gutturosa in the ventral side of the lamellar parts of theligamentum nuchae. The microfilariae of both species had the same predilection sites in the skin of the ears, head and neck regions. Those ofO. gutturosa outnumber those ofO. armillata but both are smaller than the respective uterine microfilariae. Males and microfilariae of both species are smaller than those of cattle origin.Part of the work was carried out while all the authors were at the Department of Parasitology, Faculty of Veterinary Science, University of Khartoum, Sudan.     

Onchocerca ochengi transmission dynamics and the correlation of O. ochengi microfilaria density in cattle with the transmission potential

The intensity of natural transmission of Onchocerca ochengi and Onchocerco volvulus by anthropo-boophilic Simulium damnosum s.l. was studied longitudinally in two cattle watering sites of a cattle ranch within a predominantly cattle populated area of the Guinea savanna of Cameroon and related to cattle O. ochengi skin microfilaria abundance. During the 12 months study period, a total of 4696 flies was individually dissected to examine the monthly transmission potential (MTP) of O. ochengi and O. volvulus. The estimated Simulium damnosum s.l. annual biting rates (ABR) on human baits were 47529 flies at the bank of the Vina "du sud" river. The ABR at the lake, which was situated at about 2 km upland from the perennial river, was 8579. The monthly parous rate was highly correlated with monthly biting rate. The annual transmission potentials (ATP) of O. ochengi were calculated to be 7732 and 1669 at the riverbank and the lake, respectively. Transmission occurred mainly in the dry season, peaking in the months of January to mid-March when dermal microfilaria density in the animals was also the highest. The O. ochengi microfilaria uptake by the fly vectors was host microfilaria density-dependent. The MTP of O. ochengi was positively correlated with dermal microfilaria density. The mean number of microfilariae per fly taken up during a blood meal was high during the dry season as was the mean number of infective larvae per fly but declined significantly with the onset of the early rains. A similar seasonality of transmission was also observed for O. volvulus that was concurrently transmitted by the same vector flies, but its ATP was comparatively much lower: 1332 infective larvae per man per year at the riverbank and 107 around the lake. The population dynamics of cattle microfilariae therefore plays an important role in the regulation of O. ochengi transmission.     

Equine onchocerciasis in Queensland and the Northern Territory of Australia

Investigations were conducted on the taxonomy, distribution in the carcase, pathology and transmission of Onchocerca spp. in equids from Queensland and the Northern Territory. Examination of small groups of horses and ponies revealed high infection rates with O. cervicalis, while lesser numbers were infected with O. gutturosa. O. reticulata was not found. Neither of the Australian species is likely to be of economic importance to the horsemeat industry. The findings support the belief that O. cervicalis is a pre-disposing factor in the aetiology of equine nuchal disease, most commonly seen clinically as fistulous withers. O. gutturosa is virtually non-pathogenic. Forcipomyia (Lasiohelea) townsvillensis, Austrosimulium pestilens and Culicoides victoriae are suggested as potential vectors, and it is unlikely that C. brevitarsis is involved.