Multi-drug resistant Haemonchus contortus in a sheep flock in Antioquia, Colombia.

A sheep farm experiencing high lamb mortality and poor body conditions of ewes due to haemonchosis was tested for the presence of anthelmintic-resistant nematodes in August 2014. A fecal egg count reduction test was performed in Colombia on this farm and a DrenchRite® Larval development Assay (LDA) was shipped to the University of Georgia in the United States for resistance testing. The fecal egg count reduction test (FECRT) was conducted on 70 out of 500 animals allocated into five groups of 14 animals each: untreated control, albendazole (ABZ; 3.5mg/kg sc), ivermectin (IVM; 0.2mg/kg sc), moxidectin (MOX; 0.2mg/kg sc) and levamisole (LEV; 5mg/kg sc) plus triclabendazol/ivermectin (TBZ; 10mg/kg and IVM; 0.2mg/kg po). Individual fecal egg counts (FEC) using the modified McMaster technique and pooled larval cultures were performed 10days after anthelmintic treatment. Following 10days the initial results from the ABZ and IVM groups, 24 animals were treated twice with trichlorfon (TCF; 50mg/kg po) on days 0 and 6, and then checked for FEC on day 10. Mean differences in FEC before and after treatment were negligible (3 to 8% change) for the control, ABZ, LEV-TBZ and IVM groups. The MOX treatment resulted a mean FEC reduction of 76.7%, which was not-significant by t-test (p=0.08). The TCF treatment showed a mean FEC reduction of 42.3%, which was also not-significant (p=0.1). The DrenchRite® LDA was used to evaluate the resistance status to benzimidazole, levamisole, ivermectin and moxidectin on this farm. Coprocultures indicated that the predominant worm species was Haemonchus contortus (Colombia 94-100%) (UGA 77% H. contortus and 23% Trichostrongylus colubriformis). Resistance was evaluated for both H. contortus and T. colubriformis. The LDA results indicated high resistance to all 4 anthelmintics for both parasite species. The current study, together with similar surveys in 4 other Antioquian goat and sheep farms, confirms the presence of multi-drug resistant H. contortus to all three classes of anthelmintics in this region of Colombia.

Chaparro JJ ，Villar D ，Zapata JD ，López S ，Howell SB ，López A ，Storey BE ... -  《-》

Field efficacy of four anthelmintics and confirmation of drug-resistant nematodes by controlled efficacy test and pyrosequencing on a sheep and goat farm in Denmark.

We describe a case of anthelmintic resistance on one of the largest organic small ruminant farms in Denmark. The flock was established in 2007 by purchase of animals from other Danish farms and had history of clinical parasitism, high mortality of young stock and anthelmintic treatment failure. In October 2011, 40 lambs and 40 kids were selected for a faecal egg count reduction test (FECRT) with fenbendazole (FBZ), ivermectin (IVM), moxidectin (MOX) and levamisole (LEV). Lambs were treated with the recommended sheep dose of each product while kids received the sheep dose of IVM, 1.5× sheep dose of MOX and 2× sheep dose of FBZ and LEV. Untreated lambs and kids were also included and three methods for calculating faecal egg count (FEC) reduction were compared. In a subsequent investigation, a controlled efficacy test (CET) with FBZ and IVM was performed in lambs infected with Haemonchus contortus and Trichostrongylus colubriformis isolated from adult goats on the farm. Recovered specimens of H. contortus were subjected to pyrosequencing for detection of single nucleotide polymorphisms (SNPs) related to benzimidazole (BZ) resistance. During the FECRT, FECs in untreated lambs dropped significantly by 47%. No FEC reduction was detected in untreated kids. After FBZ treatments, FEC reductions in lambs and kids ranged from 15 to 54% and 49-56%, respectively, according to the different calculation methods. Post IVM treatments, FEC reductions in lambs and kids varied between 71-90% and 81-83%, correspondingly. LEV and MOX reduced FECs by 98-100% in both species. In the CET, FBZ reduced H. contortus worm counts by 52-56% and no reduction in T. colubriformis counts were detected after treatment. IVM eliminated 100% of H. contortus and reduced T. colubriformis counts by 84-92%, according to different calculation methods. Pyrosequencing of isolated H. contortus revealed increased frequencies of the BZ resistance-related SNP in codon 200 of the β-tubulin isotype 1 gene. Frequency of BZ resistance-related SNPs in codons 167 and 198 were very low and did not exceed levels as obtained in the susceptible reference isolate. Anthelmintic resistance was confirmed in this recently established organic farm and low field efficacy of FBZ was verified by CET and pyrosequencing. BZ-resistant populations of H. contortus and T. colubriformis were isolated for the first time in Denmark. Problems with correct dosing of goats, the observed FEC reduction in untreated lambs and the relevance of including a control group in the FECRT are discussed.

Peña-Espinoza M ，Thamsborg SM ，Demeler J ，Enemark HL ... -  《-》

Multiple resistance to anthelmintics by Haemonchus contortus and Trichostrongylus colubriformis in sheep in Brazil.

The objective of this study was to determine the level of resistance of Haemonchus contortus and Trichostrongylus colubriformis in sheep to levamisole, albendazole, ivermectin, moxidectin, closantel and trichlorfon. The parasites were isolated from sheep naturally infected by gastrointestinal nematodes and were then kept in monospecifically-infected lambs for production of infective larvae (L3) of both species. Forty-two lambs, at three months of age, were simultaneously artificially infected with 4000 L3 of H. contortus and 4000 L3 of T. colubriformis. The animals were allocated into seven groups with six animals each that received one of the following treatments: Group 1--control, no treatment; Group 2--moxidectin (0.2mg/kg body weight (BW)); Group 3--closantel (10mg/kg BW); Group 4--trichlorfon (100mg/kg BW); Group 5--levamisole phosphate (4.7 mg/kg BW); Group 6--albendazole (5.0mg/kg BW); and Group 7--ivermectin (0.2mg/kg BW). Nematode fecal egg counts (FEC) were carried out on the day of treatment and again at 3, 7, 10 and 14 days post-treatment. On the same occasions, composite fecal cultures were prepared for each group for production of L3, which were identified into genus. The animals were sacrificed for worm counts at 14 days after treatment. The efficacy of each treatment was calculated from the arithmetic mean of the FEC or worm burden of the treated group, compared with the values of the control group. Only trichlorfon and moxidectin treatments resulted in a significant reduction of H. contortus recorded at necropsy (73% and 45% respectively). Moxidectin reduced T. colubriformis worm burdens by 82% and albendazole by 19%. All other anthelmintics resulted in no significant reduction in the numbers of worms found at necropsy. In conclusion, the isolates of H. contortus and T. colubriformis showed multiple resistance to all groups of anthelmintics tested. This is the first report, based on the controlled efficacy test, to show resistance of T. colubriformis to macrocyclic lactones in Brazil.

Almeida FA ，Garcia KC ，Torgerson PR ，Amarante AF ... -  《-》

Anthelmintic resistance on goat farms in Georgia: efficacy of anthelmintics against gastrointestinal nematodes in two selected goat herds.

Terrill TH ，Kaplan RM ，Larsen M ，Samples OM ，Miller JE ，Gelaye S ... -  《VETERINARY PARASITOLOGY》

Efficacy of closantel against ivermectin- and fenbendazole-resistant Haemonchus sp. in sheep in Ontario, Canada.

In Ontario, Canada, widespread resistance to ivermectin and fenbendazole, the only readily available ovine anthelmintics, has been documented, primarily in Haemonchus sp. In other parts of the world, closantel has been used to control such infections; however, the drug was not currently licensed for use in Canada and the USA. A randomized controlled trial was conducted on six client-owned farms in Ontario in 2013 and 2014 to determine the efficacy of closantel (Flukiver® 5% Oral Suspension, Elanco Animal Health, 10mg/kg bodyweight) against ivermectin- and fenbendazole-resistant Haemonchus sp. infections in periparturient ewes and grazing lambs. Three farms were randomly assigned to treat all ewes, and three farms were randomly assigned to selectively treat individual ewes at lambing, using predetermined criteria. Fecal samples were collected from a minimum of 15 randomly selected ewes and 13 lambs per group on each farm at the time of treatment and approximately 14days later. Trichostrongyle-type fecal egg counts (FEC) were performed using a modified McMaster technique with a lower detection limit of 8.3 eggs per gram of feces (epg). Haemonchus-specific FECs were determined by multiplying FECs by the proportion of Haemonchus sp. identified from coproculture for each farm; Haemonchus-specific FEC reductions were calculated for each farm. Twenty grazing lambs had FECs conducted monthly, and when mean monthly FECs surpassed 200 epg, all lambs were randomly allocated to either closantel, positive control (ivermectin, fenbendazole, or levamisole) or negative control groups. Pre-treatment Haemonchus-specific mean FECs ranged from 27 to 3359 epg in ewes and 0-5698 epg in lambs. Efficacy of closantel against Haemonchus sp. ranged from 99% (95% CI: 97%-99%) to 100% in recently lambed ewes on all farms in both years (total n=274 ewes), and from 99% (95% CI: 98%-99%) to 100% in grazing lambs in both years on all but one farm (total n=171 lambs). On the latter farm, a whole flock treated farm, closantel efficacy in grazing lambs was 84% (95%CI: 81%-88%) in the first year, but 100% in the second year. Levamisole was effective against overall GIN in lambs on only two farms. Ivermectin and fenbendazole resistance continued to be present, particularly in Haemonchus sp. Closantel had excellent efficacy against Haemonchus sp. over the two year study period, regardless of treatment group, and therefore should be considered one viable component of sustainable integrated parasite control programs for farms with documented anthelmintic resistance and problems with haemonchosis.

Westers T ，Jones-Bitton A ，Menzies P ，Van Leeuwen J ，Poljak Z ，Peregrine AS ... -  《-》