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.     

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.     

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.     

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.     

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.     

BOVINE ONCHOCERCIASIS: ASPECTS OF CARCASE INFECTION

SUMMARY The distribution of the three known species of Onchocerca in Australian cattle is described. O. gutturosa was found in the ligamentum nuchae, on the scapular cartilage and in the hip, stifle and shoulder regions. Nodules of O. gibsoni were found in the brisket***, stifle and hip regions, while O. lienalis occurred along the gastrosplenic ligament and above the xiphisternum. O. gutturosa is potentially the most important species to the meat industry since it is widely distributed in the carcase and is easily missed during inspection. Trimming procedures which could be used to ensure removal of known sites of infection are outlined.     

Distribution of microfilariae of Onchocerca lienalis and Onchocerca gutturosa in the skin of cattle in Germany and their development in Simulium ornatum and Culicoides nubeculosus following artificial infestation

Onchocerca microfilariae were isolated form the umbilicus and neck of 438 cow hides at the abattoir in Tübingen, F.R.G. The overall Onchocerca infection rate was 40.4%. The presence of Onchocerca lienalis and O. gutturosa microfilariae, which are difficult to distinguish by morphological criteria, was retrospectively demonstrated after artifically infesting Simulium ornatum and Culicoides nubeculosus and identifying the infective larvae recovered. Nine of 16 samples of umbilical microfilariae fed to C. nubeculosus through a latex membrane developed to O. gutturosa third stage larvae (L3). Six of seven umbilical samples injected into the thorax of S. ornatum yielded O. lienalis L3. In six infestation trials in which microfilariae were introduced both into S. ornatum and C. nubeculosus, O. lienalis L3 were recovered exclusively from simuliids, while O. gutturosa L3 developed only in midges. Of six umbilical skins tested by cross-infestation, one contained exclusively O. gutturosa microfilariae, four only O. lienalis microfilariae and one was infected with both species. Developmental success of O. lienalis microfilariae to L3 in S. ornatum following intrathoracic injection was 22% of the mean inoculum. O. gutturosa microfilariae, ingested by C. nubeculosus through a latex membrane, developed to L3 at a rate of 2.3% of the mean microfilarial uptake.     

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.     

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.     

Ocular Onchocerca infections in two dogs in western United States

Two dogs, one from California and one from Arizona, were found to have aberrant infections caused by filarial nematodes of the genus Onchocerca. In both cases, the parasites are localized in or near the eye. In one case the worm was located in the cornea and was surgically removed. In the second case, a very marked granulomatous reaction was induced in the retrobulbar space, mimicking an abscess. This eye was enucleated. The worms in both instances were female, and were gravid, i.e. contained microfilariae in utero, indicating that a male worm(s) had been present and mating had occurred. The exact identity of the species of Onchocerca responsible cannot be determined, although the features observed are most like Onchocerca lienalis of cattle. These cases represent the fourth and fifth such cases reported from the US, and are especially interesting because of the unusual location of the worms, the small number of recognized cases, and the similarity to a recent zoonotic human infection.     

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.     

Filarioid nematodes in cattle,sheep and horses in Finland

Background

In autumn 2006, Finnish meat inspection data revealed lesions in tendons, muscles and ligaments of bovine hind legs leading to partial condemnation of carcasses. In gross pathological examination at Finnish Food Safety Authority Evira, Oulu (now Fish and Wildlife Health) Research Unit, Onchocerca sp. (Filarioidea; Onchocercidae) nematodes were detected in lesions. Due to this, a pilot study was made in order to find out what filarioid nematodes do occur in cattle, horses and sheep in Finland.

Methods

Ventral skin biopsies from 209 dairy cattle and 42 horses, as well as blood samples from 209 cattle, 146 horses and 193 sheep, were collected from different parts of Finland and examined for microfilariae. Visceral organs and other tissues from 33 cattle with parasitic lesions were studied histopathologically.

Results

Onchocerca sp. microfilariae (mf), 240 μm long, range 225–260 μm, 5.4 μm thick, were found in 37% of the skin biopsies of cattle. All blood samples from cattle, horses and sheep and skin biopsies from horses were negative for mf. Ventral skin microfilaria prevalence in cattle was higher in southern Finland than in the North (p = 0.001). Animal age and sampling time was not associated with mf prevalence. The infection was evenly distributed among young and older animals. Macroscopic lesions on tissues included greenish-grey discolouration and often oedema. In most of the lesions, small pale nodules were seen on the fasciae. Histopathologic examination of the samples revealed mild to intense infiltration with eosinophilic granulocytes and multifocal nodular lymphoplasmacytic aggregations were seen. In some samples, there were granulomatotic lesions with central necrotic tissue and cell detritus, surrounded by eosinophilic granulocytes, lympho-, plasma- and histiocytes and some multinucleated giant cells. Around living nematodes no or only weak inflammatory changes were observed.

Conclusion

Onchocerca sp. infection in cattle was found to be common in Finland, but the amount of pathological changes leading to condemnation of infected parts is low compared to the mf prevalence. Pronounced pathological changes are distinct but rare and mild changes are difficult to distinguish. No other filarioid nematodes were observed from the animals and it appears that horses and sheep may be free from filarioid nematodes in Finland.     

Evidence of Onchocerca fasciata (Filarioidea: Onchocercidae) in camels (Camelus dromedarius): I-prevalence, nodular lesions appearance and parasite morphology

Examination of 3376 imported and 200 local Egyptian camels was carried out during the period extending between September 1997 and August, 1998. These animals were carefully examined for the presence of Onchocerca fasciata nodules and subsequently for O. fasciata adult parasites. Results of this study revealed that imported camels had the higher infection rate (2.75%), while those of local origin showed no palpable or detected Onchocerca nodules. Distribution of these nodules was mainly on the two sides of abdomen, hind limbs (concentrated in thigh region) and fore limbs particularly on the shoulders and nuchal ligament. This distribution varied according to the degree of infection. Searches for microfilariae were also performed using either blood samples, or in the subcutis and fascial sheath near or around the detected nodules.     

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.     

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.     

EPIDEMIOLOGICAL AND GROSS PATHOLOGICAL STUDIES OF ONCHOCERCA GIBSONI INFECTION IN CATTLE

SUMMARY A total of 5712 cattle from 62 properties or localities in northern, central and eastern Australia were examined at abattoirs, and the presence of Onchocerca gibsoni infection was related to region, season when slaughtered, and breed, sex and age of slaughtered cattle. Additionally, these features were also related to nodule characteristics determined from macroscopic examination of all nodules in single (left or right) briskets from 1287 infected cattle selected at random from those examined in abattoirs. Infection rate was 86% and number of nodules per brisket was 3.7 ± 4.1 (Mean ± Standard Deviation). When adjusted for other variables infection rates varied from 18% in central New South Wales to a maximum of 96% in north western Queensland. Number of nodules per brisket in cattle from north western Queensland was 9 times greater than in cattle from central New South Wales. Most nodules (70%) were 1–2 cm in diameter and weighed less than 2 g. The occurrence of changes such as calcification and caseation indicated that 20–30% of nodules studied were undergoing advanced degeneration or resolution. More nodules were detected in autumn than during winter or spring and the proportions of nodules which were hard on palpation, or had thick capsules, both increased during the autumn-spring interval. Although inconclusive, breed differences observed suggested that Bos indicus and B. indicus crossbreds had slightly greater resistance to O. gibsoni infection than cattle of British breed. Infection rate and number of nodules per brisket were greater in bulls than in cows or steers but weight of nodules in steers was greater than in other groups. Percentage of animals infected, and nodule prevalence both increased with increasing age but annual increase in nodule prevalence was less in cattle over 5 years than in younger ones, suggesting development of some immunity in mature cattle.     

Hematological and biochemical studies of stephanofilarial dermatitis in naturally infected cattle of West Bengal, India

This experimental study was conducted to assess the alterations in hematobiochemical values of cattle infected with stephanofilarial microfilariae. Hematological studies of infected cattle revealed significant (P < 0.05) decrease in Hb%, PCV and TEC values whereas TLC values were significantly (P < 0.05) increased. There was insignificant increase in lymphocyte concentration (%), significant (P < 0.05) increase in eosinophil concentration (%) and insignificant decrease in neutrophil concentration (%) in infected cattle in relation to control animal. Biochemical studies revealed that the values of glucose, alkaline phosphatase, AST and ALT were significantly (P < 0.05) increased in infected cattle than control cattle. However, the value of total serum protein was not affected.     

The comparative study digestion and metabolism of nitrogen and purine derivatives in male, Thai, Swamp buffalo and Thai, Brahman cattle

Studies on in vivo digestion, rates of passages, metabolism of nitrogen, urinary purine derivative excretion and blood metabolites were carried out in Thai Brahman cattle and Thai swamp buffaloes (16 months old). The animals were fed mixed diets based on pineapple (Ananas comusus) waste silage containing urea‐N (NPN) and true protein from a concentrate (TP). The Brahman cattle (310 ± 15 kg) were heavier than the swamp buffaloes (195 ± 9.4 kg) and had higher dry matter (DM), organic matter (OM) and neutral detergent fiber (NDF) intakes when compared on the basis of their metabolic body weight (BW^0.75), but these intakes did not differ significantly when the diets of each animal species were compared. The total tract, apparent digestibilities of dry matter (DM) and organic matter (OM) were not significantly different between the animal species when comparing the two types of diets. The NDF digestibility was significantly (P < 0.01) decreased in both animal species when fed the TP diet, but was significantly (P < 0.01) greater in cattle than in swamp buffaloes. The passage rate digesta k₁ (P < 0.01) and the passage rate digesta k₂ (through the caecum and proximal colon) (P < 0.03) were significantly slower, and the total mean retention time (TMRT) (P < 0.01) was significantly longer in swamp buffaloes when compared to Brahman cattle, but the transit time (TT) showed no difference (P = 0.07) between the animal species or the diets. The N intakes were not different in both animal species and diets, but urine‐N was greater (P < 0.05) in Brahman cattle than that in swamp buffaloes. Urine N and digestibility of N were significantly (P < 0.04) higher in animals fed the NPN diet than those fed the TP diet. Urinary purine derivatives (PD) and the creatinine (Cr) excretion of swamp buffaloes were significantly (P < 0.01) lower than those in Brahman cattle. Plasma urea‐N (BUN) concentration was significantly (P < 0.01) higher in swamp buffaloes than that in Brahman cattle, but plasma glucose and insulin concentrations were significantly (P < 0.01) higher in Brahman cattle than in swamp buffaloes. The concentrations of non‐esterified fatty acids (NEFA) were not significantly (P > 0.05) different in animals fed different diets. The present study demonstrated that Brahman cattle were better in fiber digestibility than swamp buffaloes at utilizing pineapple waste silage with both N sources.     

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%.