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Characterisation of surface antigens of Strongylus vulgaris of potential immunodiagnostic importance.
When antigens prepared by detergent washes of Strongylus vulgaris and Parascaris equorum were probed in an enzyme-linked immunosorbent assay test with horse sera from single species infections of S. vulgaris and P. equorum, a high degree of cross-reaction between the species was demonstrated. Western blot analysis of four common horse nematode species showed a large number of common antigens when probed with horse infection sera. Antisera raised in rabbits against the four species, including S. vulgaris, were also found to cross-react considerably. Rabbit anti-S. vulgaris sera were affinity adsorbed over a series of affinity chromatography columns, bound with cross-reactive surface antigens, to obtain S. vulgaris-specific antisera and thereby identify S. vulgaris-specific antigens by Western blotting. These studies revealed potentially specific antigens of apparent molecular weights of 100,000, 52,000, and 36,000. Of these bands, only the 52 kDa and 36 kDa appeared to be found on the surface as judged by 125I-labelling of intact worms by the Iodogen method, although neither protein was immunoprecipitated by horse infection sera. Finally, immunoprecipitation of in vitro translated proteins derived from larval S. vulgaris RNA suggests that two proteins may be parasite-derived. These findings are discussed both with respect to the surface of S. vulgaris and to the use of these species-specific antigens in immunodiagnosis.
Strongyle infections of small ruminants in Nigeria.
A survey of strongyle infections was conducted in sheep and goats reared in a traditional extensive husbandry system in two ecological zones of Nigeria. One zone had a seasonal pattern of infection. The majority of animals had faecal worm parasite egg counts of below 500 eggs per gram. Kids, and lambs younger than 3 months did not carry strongyle worm burdens, and the highest infection rate was found in the 7-12 month age group. A high proportion of small ruminants shed strongyle eggs during the postparturient period. The helminth species found by the use of larval culture techniques on the faeces were: Haemonchus contortus, Trichostrongylus colubriformis and Oesophagostomum columbianum. Adults of the same species were found in the few animals necropsied. The significance of the findings is discussed.
Antibody responses of ponies to initial and challenge infections of Strongylus vulgaris.
An indirect fluorescent antibody assay (IFA) was developed using Strongylus vulgaris third stage larvae (L3) as antigens. Observations using the IFA indicate that a species-specific antibody response to S. vulgaris L3 develops in S. vulgaris-infected ponies and that some surface L3 antigens are shared by adult worms. Sequential antibody levels against S. vulgaris were measured in strongyle-naive and in immune ponies following initial and challenge infections using the IFA and an indirect hemagglutination assay (IHA). Antibody levels measured by IFA increased faster following initial infections than did levels measured by IHA. Antibody levels appear to increase following challenge infections of immune ponies when measured with the IFA, but not with the IHA. Significant differences in antibody titers were not seen between ponies which developed colic following challenge infections and those that did not develop colic. Antibodies were not detectable in ponies unexposed to larval migrations, but which received surgical implantation of S. vulgaris adults into the cecum.
Critical tests in equids with fenbendazole alone or combined with piperazine: particular reference to activity on benzimidazole-resistant small strongyles.
Seven critical tests in equids were conducted with single doses of fenbendazole (5 mg kg-1) alone (Panacur--American Hoechst, Somerville, NJ); (2 tests with paste and 1 with suspension formulation) or in combination with piperazine (American Hoechst); (40 mg base kg-1); (4 tests with paste formulation). The main purpose of the tests was evaluation of activity against benzimidazole-resistant small strongyles (Cyathostomum catinatum, Cyathostomum coronatum, Cylicocyclus nassatus, Cylicostephanus goldi, and Cylicostephanus longibursatus). Natural infections of 2 populations of benzimidazole-resistant small strongyles were evaluated; 1 was population B in 2 horses and the other was population S in 5 ponies. Removal of the 5 species of population B was 49-91% in the animal treated with fenbendazole paste alone and 100% (4 of these species present) in the animal treated with the combination. For population S, 2 of the 5 resistant species were present in small numbers in 1 animal treated with fenbendazole paste alone and all were removed; the 1 animal receiving fenbendazole suspension alone had removals of 0-70% for the 5 benzimidazole-resistant species. Also for population S, the 5 resistant species were present in 2 animals treated with the paste combination and removal was 98-100% and of 4 of the 5 resistant species in 1 animal, removal was 76-99%. Removal of large strongyles (Strongylus vulgaris and Strongylus edentatus) was 92-100% for fenbendazole paste alone or in combination with piperazine in the 5 infected animals. For Oxyuris equi, present in 1 animal treated with the combination, there was 91% removal of immature and 100% removal of mature specimens. There probHably was no activity by fenbendazole alone or the combination against bots, tapeworms, and parenteral stages of S. vulgaris and S. edentatus. The combination may have had some activity against immature Habronema spp. and mature abronema muscae.
Control of cyathostome infections in mares treated at parturition with ivermectin.
Six mares were treated on the day of parturition with an intramuscular injection of 0.2 mg kg-1 ivermectin and placed in a pasture free of equine parasites as soon as possible after foaling. The mares and their foals were compared with a similar group of untreated mares and foals on an adjoining pasture. The experimental data was derived from mare and foal fecal egg counts, foal necropsies and pasture larval counts. Ivermectin administered to mares on the day of parturition, when combined with movement to parasite-free pastures, significantly lowered the cyathostome (small strongyle) egg production for 4 months. This reduced cyathostome exposure was reflected in lower worm-burdens in their foals for 5 months. The results indicate that ivermectin will effectively control equine strongyles when mares and their foals are moved to parasite-free pastures.
The elimination of equine strongyles and hematological and pathological consequences following larvicidal doses of thiabendazole.
Twelve horses were divided into three groups and given various doses of a mixed species strongyle inoculum, representing light, moderate, and heavy infections. Three weeks after the larval inoculations, three animals from each group were given larvicidal doses of thiabendazole (TBZ) (440 mg kg-1 on two consecutive days); one animal from each group served as a non-medicated control. Treatment was repeated three weeks later. One treated animal from each group was designated for long-term study; others were necropsied to study adult and larval parasite loads. Six of the twelve animals with strongylosis developed moderate eosinophilia. TBZ given at 440 mg kg-1 on two consecutive days caused depression, lethargy, and anorexia which lasted for five days. Eosinopenia, lymphopenia, and neutrophilia occurred in treated animals, and lasted for three days. During the course of TBZ treatment, one horse died from what appeared to be a mis-dosing or an anaphylactic reaction. At necropsy, active thrombi of the anterior mesenteric artery were seen in parasitized animals, but not in those treated with TBZ. Five out of seven medicated horses were completely free of adult and larval strongyle parasites. One had a few Strongylus edentatus larvae and another had small strongyles. No Strongylus vulgaris larvae or adults were recovered from any horse treated with TBZ.
Pathogenesis of helminths in equines.
This review summarizes information on the clinical signs, gross and microscopic lesions associated with nematode and cestode infections and discusses the development of these conditions in the equine host.
Comparison of two control systems for cyathostome infections in the horse and further aspects of the epidemiology of these infections.
The small strongylid infections of two groups of three yearling female Shetland ponies and one yearling Shetland tracer pony were studied. One group was set stocked from April to November and was treated monthly with 5 mg kg-1 albendazole from two days before turnout until July. The other group grazed similar pasture until July, was treated with 5 mg kg-1 albendazole and subsequently removed to pasture grazed by sheep from April to July. The tracer ponies were added to both groups in September. The efficacy of both methods was not completely satisfactory probably because of low efficacy of anthelmintic treatment. There were no significant differences between the cyathostome burdens of the two groups. A high proportion of the cyathostome populations of all ponies consisted of inhibited early third stage larvae (L3). The finding of low numbers of immature fifth-stage worms in the tracer ponies indicated that the considerable adult burdens in the permanent ponies originated from infection picked up before the tracer ponies were added. In the group which was removed to sheep pasture after treatment in July it was likely that the majority of the adult worm burden had been ingested as infective larvae before treatment.
Control of strongylosis in horses by alternate grazing of horses and sheep and some other aspects of the epidemiology of Strongylidae infections.
Alternate grazing of horses and sheep as a control measure for gastrointestinal helminthiasis was studied in three grazing experiments in 1981, 1982 and 1983. Each year a group of three mare yearling Shetland ponies, which were kept on a small pasture from spring to autumn, were compared with a similar group which grazed a similar or the same pasture until July and were subsequently removed to a similar pasture which had been grazed by sheep from April to July. In addition both groups were treated with an anthelmintic when the latter group was removed to the sheep pasture. Pasture larval counts and worm counts and, in 1982 and 1983, faecal egg counts, clinical condition, total protein, albumin and beta-globulin levels demonstrated that the groups removed to sheep pasture acquired considerably lower burdens of nematodes of the subfamilies Cyathostominae and Strongylinae, but considerably higher burdens of Trichostrongylus axei than the groups which were not moved. These T. axei infections resulted in higher serum pepsinogen levels in the former groups compared to the latter in 1981 and 1982. At necropsy an important part of the T. axei burdens and, in 1982 and 1983, the Cyathostominae burdens consisted of inhibited early third stage larvae. A total of 20 species of the subfamily Cyathostominae and 7 species of the Strongylinae were found. Generally the composition of species was in agreement with other observations in western Europe, the most common species being: Cylicostephanus longibursatus, Cylicostephanus minutus, Cylicostephanus calicatus, Cylicostephanus goldi, Cylicostephanus poculatus, Cyathostomum labratum, Cyathostomum coronatum, Cyathostomum catinatum, Cylicocyclus leptostomus, Cylicocyclus nassatus, Cylicocyclus insigne, Strongylus edentatus and Strongylus vulgaris.
Prevalence and control of benzimidazole-resistant small strongyles on German thoroughbred studs.
The prevalence of benzimidazole-resistant small strongyles was determined in a survey, conducted on 14 thoroughbred studs, which compared the faecal egg counts of groups of horses before and after treatment with the recommended doses of cambendazole (20 mg kg-1 b.w.) or febantel (6 mg kg-1 b.w.). Benzimidazole-resistant cyathostomes were found on all farms examined. Pyrantel pamoate (19 mg kg-1 b.w.), oxibendazole (10 mg kg-1 b.w.) and ivermectin (0.2 mg kg-1 b.w.) reduced the strongyle egg counts on these studs by 97-100% at 2 weeks post-treatment. However, 6 weeks after dosing the reduction of the strongyle egg output had decreased to an average of 67.8% (8.7-97.1%) with pyrantel pamoate and 51.2% (0-95.8%) with oxibendazole, whereas ivermectin still suppressed the egg counts by 98.2% (95-100%).
Estimation of uptake of digestive tract strongyle larvae from pasture, using oesophagus fistulated sheep.
Three Merino of Arles ewes fistulated at the oesophagus were used in autumn and spring in order to assess their larval uptake on infected irrigated pasture. The rate of transmission (ingested/available larvae of digestive tract strongyle) was low when the pasture was heavily infected; it was not otherwise modified by environmental factors.
Development and survival of free-living stages of equine strongyles under laboratory conditions.
In a series of laboratory studies the optimum conditions for the development and survival of the free-living stages of strongyle parasites occurring in horses in tropical north Queensland were determined. No differences in behaviour were noted between the strongyle species. Development to the infective stage occurred only between 10 and 35 degrees C. The rate was affected by temperature, taking 15-24 days and 3 days, respectively, at the lowest and highest temperatures for the developing stages to reach the infective third stage. Yields of infective larvae were very low outside the range 20-33 degrees C, and were highest at 28 degrees C. Survival of infective larvae was good between 20 and 33 degrees C, and large numbers were recovered after 3 months in faeces incubated at 20-28 degrees C. At 33 and 37 degrees C larval survival was affected by the moisture content of the faeces, with infective larvae surviving better in dry than in moist faeces; even a residual moisture level of 40% significantly reduced the number of larvae recovered from faeces incubated at 37 degrees C for 1 month. Moisture also affected larval development, especially at the higher temperatures of 25-39 degrees C. When faecal moisture content fell to less than or equal to 20% by 3 days, larvae which had not yet reached the infective stage were still pre-infective at 7 days, while all larvae in faeces with adequate moisture had reached the infective third stage. It was not possible to determine the critical faecal moisture level below which larval development ceased, however, 28 degrees C (range 25-33 degrees C) was found to be the optimum temperature. Larval development was very rapid and yields of infective larvae highest at this temperature.
Development of free-living stages of equine strongyles in faeces on pasture in a tropical environment.
The development of the free-living stages and yields of infective third stage strongyle larvae in faeces from a horse with a mixed natural infection deposited on pasture plots were studied over a 2-year period in a coastal area in tropical north Queensland. Two sets of faecal masses (one exposed to, and the other protected from the action of a natural population of dung beetles) were deposited monthly and after 7 days faecal samples were taken for larval recovery and counts. Hatching and development of the free-living stages occurred in faeces on pasture throughout the year. Development was rapid as infective stages were reached within a week of faecal deposition in all months. Yields of infective larvae were affected by the season and the action of dung beetles on the faecal masses. Highest yields were obtained from both beetle-exposed and protected faeces during winter (June to August) and lowest yields were in spring (September to November). High temperatures in spring and summer resulted in low yields of larvae, however, the dry conditions in spring made this season the most unfavourable period. In autumn and winter the temperatures were never low enough to stop or markedly slow down the rate of development, and allowed the development of large numbers of infective larvae. Dung beetle activity was observed throughout the year, and exposed faeces were usually completely dispersed within 24 h of deposition. This resulted in lower yields of infective larvae from these than from protected faeces. Though larval yields were lower, the actual numbers were still substantial so as to cast doubt on the usefulness of these beetles as biological control agents for equine strongylosis in the dry tropics.
Small strongyle infections in donkeys from the highveld in Zimbabwe.
In total, 14 male donkeys aged between 2 and 10 years were obtained in July and November 1986, and January and April 1987 from a communal area in the Zimbabwean highveld, with the aim of studying their parasite populations. Thirteen species of small strongyles were found, numbers ranging between 3900 and 222 767 worms. The small strongyle populations consisted predominantly of adult worms in all donkeys. These results suggest that rather stable, small strongyle populations, mainly consisting of long-living adult worms, occur in Zimbabwean donkeys. The species found were, in order of abundance, Cyathostomum montgomeryi, Cylicostephanus minutus, Cylicocyclus nassatus, Cylicocyclus auriculatus, Cyathostomum tetracanthum, Cylicostephanus bidentatus, Triodontophorus nipponicus, Cyathostomum coronatum, Cylicocyclus adersi, Cyathostomum alveatum, Triodontophorus serratus, Cylicocyclus elongatus and Cylicodontophorus bicoronatus.
Seasonal translation of equine strongyle infective larvae to herbage in tropical Australia.
Longevity in faeces, migration to and survival on herbage of mixed strongyle infective larvae (approximately 70% cyathostomes: 30% large strongyles) from experimentally deposited horse faeces was studied in the dry tropical region of North Queensland for up to 2 years. Larvae were recovered from faeces deposited during hot dry weather for a maximum of 12 weeks, up to 32 weeks in cool conditions, but less than 8 weeks in hot wet summer. Translation to herbage was mainly limited to the hot wet season (December-March), except when unseasonal winter rainfall of 40-50 mm per month in July and August allowed some additional migration. Survival on pasture was estimated at 2-4 weeks in the summer wet season and 8-12 weeks in the autumn-winter dry season (April-August). Hot dry spring weather (pre-wet season) was the most unfavourable for larval development, migration and survival. Peak counts of up to 60,000 larvae kg-1 dry herbage were recorded. The seasonal nature of pasture contamination allowed the development of rational anthelmintic control programs based on larval ecology.
Emergence from inhibited development of cyathostome larvae in ponies following failure to remove them by repeated treatments with benzimidazole compounds.
The effect of three albendazole treatments at 5-week intervals, beginning at turnout in April, on cyathostome infections in Shetland ponies was compared with the effect of sequential treatments with albendazole, oxfendazole and oxibendazole. The results showed a substantial reduction in faecal egg output after the first albendazole treatment. Since faecal egg counts remained very low, no estimation of the effect of the second treatment was possible. The third treatment with albendazole and oxibendazole was followed by an increase in faecal egg counts to values of greater than 100 eggs g-1 within 4 weeks. A final albendazole treatment in December, 1 week before necropsy, failed to reduce faecal egg counts. These results suggest resistance to albendazole and oxibendazole in the cyathostome populations of the ponies. The increase in faecal egg counts after the third anthelmintic treatment in July occurred, although overwintered pasture infectivity was very low. The most likely explanation for this increase is resumption of the development of worms which overwintered as inhibited larvae in the host.
Internal parasites of horses on mixed grassveld and bushveld in Transvaal, Republic of South Africa.
Between 1980 and 1982, the gastrointestinal tracts of 17 horses which had been grazing on mixed grassveld at Potchefstroom and bushveld at Onderstepoort in the province of Transvaal, Republic of South Africa, were examined at necropsy and processed for parasite recovery. The large strongyles and their prevalences were as follows: Strongylus vulgaris and associated lesions (88-94%), Strongylus edentatus (24%), Strongylus equinus (30%), Triodontophorus nipponicus (35%) and Craterostomum acuticaudatum (18%). The seven most prevalent and abundant cyathostomes collected were Cylicostephanus longibursatus, Cylicostephanus goldi, Cylicostephanus calicatus, Cylicocyclus nassatus, Cyathostomum catinatum, Cylicostephanus minutus and Cyathostomum coronatum. Gasterophilus intestinalis was the most prevalent and abundant botfly larva recovered. Most of the cyathostome larvae and adults were present during all seasons except winter. The distribution of cyathostome species in the large intestine of the host is discussed.