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PubMed2002

Investigation of diversity and isotypes of the beta-tubulin cDNA in several small strongyle (Cyathostominae) species.

The diversity of the beta-tubulin cDNAs of the cyathostominae and the occurrence of further isotypes were examined in adult worms isolated from an anthelmintic-naïve horse. cDNAs encoding beta-tubulin from Cyathostomum catinatum, Cylicocyclus nassatus, Cylicocyclus insigne, Cylicocyclus radiatus, Cylicocyclus elongatus, Cyathostomum coronatum, and Cyathostomum pateratum were characterized using specific primers developed from the cDNA sequence of Cc. nassatus. The cDNA sequences span 1,429 bp and show identities ranging from 95.6 to 100%. The deduced protein sequences span 448 amino acids and were 98-100% identical. The amino acid sequences of the 7 species varied within and between species at 10 positions. A 3' Rapid Amplification of cDNA ends using a degenerate forward primer was carried out with cDNA from Cy. pateratum, Cy. coronatum, Cy. catinatum, and Cc. nassatus to investigate the occurrence of further beta-tubulin isotypes. The expected polymerase chain reaction (PCR) product of 400 bp, including 306 bp of coding sequence, was amplified, as was an additional fragment of 600 nucleotides in the case of Cy. pateratum, Cy. coronatum, and Cy. catinatum. Sequencing of the PCR products revealed no evidence for the existence of a second beta-tubulin isotype in cyathostomes. The variation in size was caused by a length polymorphism within the 3' untranslated region, and 2 functional mRNAs seem to be transcribed from the same gene.

3' Untranslated Regions↗
PubMed2002

One season of pasture exposure fails to induce a protective resistance to cyathostomes but increases numbers of hypobiotic third-stage larvae.

The development of acquired resistance to cyathostome challenge after 1 season's exposure to a cyathostome-contaminated pasture was investigated using 17 parasite-naive ponies, which were 2-3 yr of age. These were divided into 3 groups: 1 to graze a cyathostome-contaminated pasture for 4 mo (exposed ponies), 1 to graze a "clean" pasture not previously grazed by parasitized animals (nonexposed ponies), and 1 group to remain in the barn under helminth-free conditions (parasite-free ponies). After pasture exposure all ponies were housed in stalls in the barn dewormed with ivermectin (200 micrograms/kg) and oxibendazole (100 mg/kg), a treatment that eliminated most cyathostomes encysted in the mucosa as well as all luminal parasites, on the basis of necropsies of 5 animals, after 17 days. Remaining ponies were challenged with 100,000 cyathostome-infective third-stage larvae (L3) per os 3 wk after anthelmintic treatment. Necropsies were performed 7 wk after the challenge. Total cyathostome burdens (luminal plus encysted stages) were not significantly different among any of the groups. However, a significantly higher percentage of hypobiotic early L3 (EL3) and a lower percentage of adults were found in exposed ponies. This observation supports the hypothesis that resistance acquired through exposure promotes cyathostome hypobiosis. This increase in EL3 in exposed ponies was associated with a significant increase in weight of cecum and ventral colon biopsies.

Animals↗
PubMed2004

Evidence of p-glycoprotein sequence diversity in cyathostomins.

P-glycoproteins (Pgps) are adenosine triphosphate-binding transporter proteins thought to be associated with multi-drug resistance in mammals and protozoans and have been suggested to be involved in the mechanism of ivermectin (IVM) resistance in Haemonchus contortus. Until now, resistance to IVM has not been reported in cyathostomins in horses in spite of its widespread and frequent use. Reasons for this might be differences in the molecular mechanism of the development of resistance. Based on this hypothesis, the present study was carried out to find homologues of Pgp in cyathostomins. A 416-bp polymerase chain reaction (PCR) product was generated using complementary DNA (cDNA) of Cylicocyclus elongatus and Cylicocyclus insigne and degenerate primers, located in the conserved Pgp nucleotide-binding domains. Resulting PCR products showed interspecific nucleotide and amino acid sequence identities of 73.3 and 76.8%, respectively. Specific primers were designed based on the Cc. elongatus sequence, and a PCR product of 268-bp was amplified from cDNA of single adults of Cylicocyclus radiatus, Cc. insigne, Cylicocyclus nassatus, Cc. elongatus, Cylicostephanus hybridus (2 individuals), Cylicostephanus goldi, Cyathostomum pateratum, Cyathostomum coronatum, and Cyathostomum catinatum. Two clusters of sequences were found representing 2 different internucleotide-binding domains (IBDs). A further distinct IBD is represented by the 416-bp PCR product of Cc. insigne. Therefore, a total of 3 clearly different sequences of the IBD were cloned and sequenced, suggesting that at least 2 Pgp genes exist in cyathostomins.

ATP Binding Cassette Transporter, Subfamily B↗
PubMed2004

Beta-tubulin complementary DNA sequence variations observed between cyathostomins from benzimidazole-susceptible and -resistant populations.

The molecular mechanism of benzimidazole (BZ) resistance in cyathostomins of horses is still unclear. Previous studies revealed that the TTC or TAC polymorphism in codon 200 of the beta-tubulin isotype 1 gene is not as strictly correlated with BZ resistance as in trichostrongyles in sheep. To identify further sites of polymorphism within the beta-tubulin gene related to BZ resistance, complete complementary DNAs (cDNAs) encoding beta-tubulin of adult worms of Cylicocyclus nassatus, Cyathostomum pateratum, Cyathostomum coronatum, Cyathostomum catinatum, Cylicostephanus longibursatus, and Cylicostephanus goldi of a BZ-resistant cyathostomin population were characterized using specific primers. The cDNA sequence of each species spans 1,429 bp, encoding a protein of 448 amino acids. The interspecific identities are 95.2-99.6% at the nucleotide and 98.7-100.0% at the peptide level. The comparison of the amino acid sequences of individuals isolated from the BZ-resistant cyathostomin population with those from individuals of Cc. nassatus, Cy. coronatum, Cy. pateratum, and Cy. catinatum of a BZ-susceptible one showed differing amino acids in 11 positions. The commonness of a phenylalanine to tyrosine mutation at position 167 in all the 6 cyathostomin species isolated from a BZ-resistant population suggests its involvement in the molecular mechanism in BZ resistance.

Amino Acid Sequence↗
PubMed2001

Prevalence and clinical implications of anthelmintic resistance in cyathostomes of horses.

OBJECTIVE: To determine the prevalence and clinical implications of anthelmintic resistance in cyathostomes of horses. DESIGN: Prospective study. ANIMALS: 80 horses on 10 farms in a 5-county region of northeast Georgia. PROCEDURE: On each farm, horses were stratified in descending order according to pretreatment fecal egg count (FEC), blocked into groups of 4, and then randomly assigned to 1 of 4 treatment groups: no treatment (controls), and treatment with pyrantel pamoate, fenbendazole, or ivermectin. Fecal samples were collected 24 hours prior to treatment and 2, 4, and 6 weeks after treatment for determination of FEC. Mean percentage of reduction in FEC was then calculated for each treatment group. For horses from each farm, the efficacy of each anthelmintic was categorized on the basis of mean percentage of reduction in FEC at 2 weeks after treatment (< 80% reduction = ineffective; 80 to 90% reduction = equivocal; and > 90% reduction = effective). RESULTS: Pyrantel pamoate was effective at reducing FEC in horses from 7 farms, ineffective in horses from 2 farms, and equivocal in horses from 1 farm. Fenbendazole was ineffective at reducing FEC in horses from 9 farms and equivocal in horses from 1 farm. Ivermectin was effective at reducing FEC in horses from all 10 farms. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that cyathostome resistance to fenbendazole is highly prevalent, and resistance to pyrantel pamoate is high enough to warrant concern. Resistance to ivermectin was not detected. On the basis of these data, it appears that ivermectin continues to be fully effective in horses. However, too few farms were used in this study to determine the prevalence of cyathostome resistance to ivermectin. Therefore, the efficacy of ivermectin should continue to be monitored closely.

Animals↗
PubMed2003

Management of drug-resistant cyathostominosis on a breeding farm in central North Carolina.

REASONS FOR PERFORMING STUDY: Possible anthelmintic resistance on a breeding farm where a rapid rotation anthelmintic programme had been implemented for 9 years was investigated. Cyathostomins resistant to fenbendazole and pyrantel were documented by faecal worm egg count reduction test (FWECRT). OBJECTIVES: To 1) manage small strongyle transmission in a herd of horses in which resistance to both pyrantel pamoate and fenbendazole was identified and thereby reduce the risk of clinical disease in the individual animal, 2) monitor the change in resistance patterns over time and 3) monitor the efficacy of ivermectin over the study period. METHODS: Targeted ivermectin treatment of horses on the farm was instituted for mature horses with faecal worm egg counts (FWEC) > 200 eggs/g (epg) and for horses < age 2 years with FWEC > 100 epg. RESULTS: Over a 30 month period, targeted ivermectin treatment achieved acceptable control in mares, as judged by FWEC, and improved control of patent cyathostome infection in consecutive foal crops. Egg reappearance time (ERT) after treatment with ivermectin was < 8 weeks in mares and foals more frequently in the second year of the study than in the first year. Numbers of anthelmintic treatments were reduced by 77.6 and 533% in the mare and foal group, respectively. CONCLUSIONS: Targeted ivermectin treatment may be an economically viable method of managing multiple drug resistant cyathostominosis. POTENTIAL RELEVANCE: Use of ivermectin should be monitored closely for development of resistance.

Animals↗
PubMed2000

A study to evaluate the field efficacy of ivermectin, fenbendazole and pyrantel pamoate, with preliminary observations on the efficacy of doramectin, as anthelmintics in horses.

The efficacy of ivermectin, fenbendazole, pyrantel pamoate and doramectin was evaluated under field conditions at 2 sites in the Free State Province of South Africa. The study involved 25 horses at each site, divided into 5 groups of equal size. Ivermectin, fenbendazole and pyrantel pamoate were administered orally at doses of 0.2, 10 and 19 mg/kg respectively. Doramectin was administered by intramuscular injection at a dose of 0.2 mg/kg. Treatment efficacy was based on the mean faecal egg count reduction 14 days post treatment. At site A a faecal egg count reduction of 100% was found after treatment with ivermectin, fenbendazole and doramectin. A 96.1% reduction was found after treatment with pyrantel pamoate. At site B ivermectin and doramectin produced a 100% reduction in faecal egg counts, fenbendazole produced an 80.8% reduction and pyrantel pamoate a 94.1% reduction. Doramectin produced a 100% reduction in faecal egg counts at both sites, despite not being registered for use in horses. In addition, the results indicated reduced efficacy of fenbendazole at site B, which suggested benzimidazole resistance. Larval cultures showed that cyathostomes accounted for between 86 and 96% of pre-treatment parasite burdens at both sites. Other helminths identified in the faecal samples were Strongylus spp. and Trichostrongylus axei.

Animals↗
PubMed1983

The Baermann technique for estimating Protostrongylus infection in bighorn sheep: effect of laboratory procedures.

The modified Baermann funnel technique was evaluated to determine the effects of time of baermannization, fecal preparation, type and size of funnel, and type of filter on the number of first stage larvae of Protostrongylus spp. recovered from feces of Rocky Mountain bighorn sheep (Ovis canadensis). More larvae were recovered when fecal pellets were baermannized for 24 hr compared to 8 hr, and when feces were crushed than when left intact. Use of small funnels resulted in the recovery of more larvae per gram of feces than larger funnels, and glass funnels more than plastic ones. There was no difference in recovery of larvae between cheesecloth filters and cellulose filters.

Animals↗
PubMed1983

Endoparasites of hares (Lepus timidus L. and L. europaeus Pallas) in Finland.

Intestinal and lung parasites of 85 mountain hares (Lepus timidus) and 24 European hares (L. europaeus) were examined. The species of parasites found in both hare species were Trichostrongylus retortaeformis Zeder, 1800; Mosgovoyia pectinata Goeze, 1782 and Protostrongylus pulmonalis Froelich, 1802. Eimeria semisculpta Madsen, 1938 and E. townsendi Carvalho, 1943 were found only in the European hares and Eimeria leporis Nieschulz, 1923 and Dicrocoelium dendriticum Rudolphi, 1819 only in the mountain hares. Ninety-five percent of the mountain hares and 88% of the European hares were infected with parasites. Mountain hares were more commonly infected with P. pulmonalis and D. dendriticum while European hares were more commonly infected with Eimeria spp. and T. retortaeformis. Young mountain hares were more often infected with M. pectinata and adult mountain hares with P. pulmonalis. The management technique of winter-feeding did not increase significantly the percentage of infected animals. Dicrocoelium dendriticum was found only in hares from islands where sheep had grazed during the summers. The absence of cysticerci of Taenia pisiformis Bloch, 1780 in this survey may be a reflection of improved hygiene practices instituted by hunters.

Age Factors↗
PubMed1984

Muellerius capillaris (Mueller, 1889) (Nematoda: Protostrongylidae): an unusual finding in Rocky Mountain bighorn sheep (Ovis canadensis canadensis Shaw) in South Dakota.

Lungs and fecal samples from nine hunter-killed Rocky Mountain bighorn sheep were examined for lungworms. All samples contained adults and/or larvae of Muellerius capillaris (Mueller, 1889). Protostrongylus spp., the lungworms commonly reported from bighorn sheep, were not present in any samples. Larvae of M. capillaris bear a spine on the dorsal side of the posterior end and are shorter than dorsal-spined larvae of other lungworms recorded from North American ungulates. Larvae similar in shape but longer than those of Muellerius were found in free-ranging bighorn sheep in Alberta and British Columbia. In addition, dorsal-spined larvae have been found in bighorn sheep in Montana, North Dakota, and Washington. The identity of the dorsal-spined larvae is known only from sheep in South Dakota. Thus, caution must be taken when diagnosing lungworm infections in Rocky Mountain bighorn sheep.

Animals↗
PubMed1984

Pathologic changes and microorganisms found in bighorn sheep during a stress-related die-off.

An all-age die-off of Rocky Mountain bighorn sheep (Ovis c. canadensis Shaw) occurred from late October 1980 through March 1981 in Waterton Canyon, Colorado, with a loss of 75 to 85% of the sheep. The cause of death was a subacute to chronic bronchopneumonia and the primary etiologic agents isolated from the respiratory system were a Pasteurella sp., P. multocida, Corynebacterium pyogenes, and Protostrongylus stilesi Dikmans, 1931. The underlying predisposing factors that initiated this die-off were believed to be related to multiple chronic environmental stressors associated with the building of a dam which included human contact, vehicular traffic, atmospheric dust, noise and harassment. The die-off was succeeded by a 100% lamb mortality the following summer and a 67% lamb mortality the next two summers. The pneumonia found in these lambs was similar to that found in adult sheep during the previous die-off, except that mature lungworms were absent.

Animals↗
PubMed1985

Prevalence of helminths in a cyclic snowshoe hare population.

Five species of helminths were monitored in a population of snowshoe hares (Lepus americanus) near Rochester, Alberta, during 1961-1977. Prevalence of both Obeliscoides cuniculi and Protostrongylus boughtoni among young hares averaged about 50% by age 2 mo, then tended to level off. Prevalence of Taenia pisiformis (cysticerci) and Dirofilaria scapiceps rose more slowly, but continued to increase steadily beyond their mean levels of 8% and 1% at age 2 mo. There were well defined seasonal (within-year) cycles in prevalence of O. cuniculi and P. boughtoni that were generated evidently to a major degree by arrested development of larvae in fall and renewed development in late winter. It was hypothesized that renewed larval development was triggered (in February) in O. cuniculi by the seasonal rise of circulating pituitary gonadotrophins, and (in April) in P. boughtoni by the seasonal rise of gonadal androgens and estrogens. Indices to gonadal hormone levels in hares indicated that these increased most rapidly among males, and may have accounted for the higher prevalences of P. boughtoni in males during April-May. Neither T. pisiformis nor D. scapiceps exhibited conspicuous seasonal changes in prevalence. Maximum prevalence of T. pisiformis was attained at about 1 yr of age, whereas D. scapiceps increased among adult snowshoes through age 2 yr before stabilizing. Long-term (between-year) changes in prevalence of O. cuniculi, T. pisiformis, and D. scapiceps were correlated significantly with the cyclic hare population which declined from a peak in fall 1961 to a low in 1965-1966, rose to another peak by fall 1970, and declined again to a low in 1975. There was no detectable time lage between this "10-yr" cycle in hare density and the cycles of parasite prevalence among juveniles (less than 1 yr of age). Among adult hares, the cycle of O. cuniculi prevalence was likewise synchronous with that of the hare population, but the cycles of D. scapiceps and T. pisiformis lagged by approximately 1 and 2 yr, respectively. This lag in T. pisiformis prevalence was largely inexplicable to us. Our data on P. boughtoni were not suitable for analyses of between-year trends; nor were those for the fifth helminth, Taenia serialis (coenuri), because mean prevalence was less than 1% among both juveniles and adults. An apparent decline in T. serialis after the early 1950's, and its continued scarcity thereafter, paralleled a major change in numbers of one important definitive host--the red fox (Vulpes vulpes).(ABSTRACT TRUNCATED AT 400 WORDS)

Age Factors↗
PubMed1987

Parelaphostrongylus tenuis in New Brunswick: the parasite in white-tailed deer (Odocoileus virginianus) and moose (Alces alces).

Research was initiated in 1983 to investigate the ecology of Parelaphostrongylus tenuis in New Brunswick. The objectives were to determine the prevalence and intensity of infection in white-tailed deer, and to determine whether or not moose feces contained first stage larvae, signifying the completion of the life cycle of P. tenuis in this host. Forty-nine percent of deer pellet samples were positive and 60% of deer heads contained adults of P. tenuis. None of the moose pellet samples contained first stage larvae.

Animals↗
PubMed1999

Parasites in grizzly bears from the central Canadian Arctic.

Standardized flotation techniques were used to survey 56 grizzly bear (Ursus arctos) fecal samples for parasites. The samples were collected during the spring and autumn of 1995 and 1996 in the central Arctic of the Northwest Territories (Canada). Parasites of the genera Nematodirus, gastrointestinal coccidia, and an unidentified first stage protostrongylid larva are reported for the first time from grizzly bear feces in North America. Parasites of the genera Diphyllobothrium and Baylisascaris also were collected. Prevalence of gastrointestinal parasites were significantly different between the spring and autumn seasons (31% and 58% respectively). Thus, we provide evidence supporting the theory that bears void gastrointestinal parasites before hibernation.

Animals↗