Standardization of the larval migration inhibition test for the detection of resistance to ivermectin in gastro intestinal nematodes of ruminants.

Resistance to anthelmintics is an increasing problem in sheep, goat and cattle industries worldwide. For parasite management on farms reliable methods for the detection of resistance are required and it is important that tests give comparable, reproducible and reliable results in different laboratories. The ability of the larval migration inhibition test (LMIT), to detect ivermectin resistance in cattle and sheep nematodes has been evaluated through ring testing in up to six different laboratories in Europe, supported by an EU sixth Framework Project (PARASOL). Third stage larvae of Ostertagia ostertagi, Cooperia oncophora, and Haemonchus contortus with a known resistance status were obtained from faecal cultures of experimentally infected calves and sheep. Following a series of ring tests using identical protocols, reproducible results were obtained within and between participating laboratories. In all tests dose-response curves with R(2) values >0.90 were obtained by all laboratories. Resistance ratios of 8.3 and 8.4 were found when susceptible and IVM-resistant isolates of C. oncophora and H. contortus were compared and differences in the EC(50) values were highly significant (p<0.0001). Protocols for the LMIT and the preparation of ivermectin solutions are described in a supplementary file.

A 3-year field evaluation of pasture rotation and supplementary feeding to control parasite infection in first-season grazing cattle: dynamics of pasture infectivity.

A 3-year grazing trial (2002-2004) was conducted on a commercial beef cattle farm in south-central Sweden to assess different methods of parasite control. This paper focuses on the dynamics of the free-living larval stages, whereas data on performance and within-host parasitological variables are presented in a complementary paper. Each year in May, 4 groups of 10 first-season grazing (FSG) steers were turned out on to separate 2ha paddocks and subjected to the following strategies: (1) spring turn-out on to pasture which had been grazed the previous year by second-season grazing (SSG) steers (paddock RT), followed by a move to aftermath (paddock AM) after 10 weeks (mid-July), (2) supplementary feeding with concentrate and hay for 4 weeks following turn-out (paddock FD), set stocked, (3) untreated control (paddock UT), set stocked and (4) anthelmintic treated control (paddock DO), set stocked. All paddocks were assigned a new set of FSG cattle each year whereas the treatments remained the same. Pasture infectivity were monitored partly by two tracer calves that grazed each paddock along with the FSG calves for 3 weeks after turn-out and prior to housing, partly by analysis of herbage samples for infective larvae (L3) that were collected from each paddock at monthly intervals between April and October. The predominant genera found were Cooperia and Ostertagia. Tracers grazing paddock RT overall harboured less worms, and in particular less Ostertagia spp., and tracers grazing paddock AM in mid-July harboured insignificant numbers of nematodes compared to tracers on the FD and UT paddocks. Although total worm counts varied between groups, smaller numbers were generally observed early in the grazing-season (May), compared to close to housing (September) when inhibited early L4 larvae were almost exclusively found. Results observed from herbage samples showed high numbers of L3 in spring before the time of turn-out, compared to around housing. In conclusion, the rotation control strategy showed promising results and provided a turn-out pasture that was 'nematode safe' to FSG cattle the following spring, whereas the feeding strategy failed as applied in this experiment.

A 3-year field evaluation of pasture rotation and supplementary feeding to control parasite infection in first-season grazing cattle--effects on animal performance.

To evaluate non-chemical strategies to control pasture-borne parasites in first-season grazing (FSG) cattle, a 3-year grazing trial was conducted during 2002-2004 on naturally infected pastures on a commercial beef cattle farm in Sweden. A uniform pasture was divided in 4 equal 2 ha paddocks onto each of which 10, 5-9 months old dairy breed steer calves were allocated at turn-out in May each year. Two strategies were evaluated: (1) turn-out onto pasture which had been grazed the previous year by second-season grazing (SSG) steers, followed by a move to aftermath in mid-July (RT) and (2) supplementation with concentrate and roughage for 4 weeks from turn-out (FD). Comparisons were made with an untreated (UT), and an anthelmintic treated control group (DO). Animal parasitology and performance were monitored monthly throughout the 20 weeks grazing period. Additional sampling occasions were performed on day 9 (for coccidia) and 10 weeks after turn-out (mid-July). Due to clinical parasitic gastro-enteritis (PGE), salvage treatments were performed on all animals in group FD approximately 7 weeks after turn-out in 2003 and of three animals in group UT 5 weeks after turn-out in 2004. In 2003, the geometric mean oocyst excretion 9 days after turn-out was approximately 150,000 opg of mainly Eimeria alabamensis in group FD, and in 2004 approximately 180,000 opg in group UT. Apart from the DO group, geometric mean faecal egg counts (FEC) were between 80 and 400 epg 4 weeks after turn-out. Mean serum pepsinogen concentrations (SPC) of approximately 3.6 U tyrosine were recorded in the FD and UT groups from late August 2002. In 2003 and 2004, mean concentrations in these groups were between 4.1 and 7.2 U tyrosine 8 weeks after turn-out. By the end of the three grazing seasons the average weight gain difference compared to the DO group was for FD -29, -38 and -5 kg and for RT -4, -21 and +14 kg, and compared to the UT group -18, +2 and +22 for FD and +7, +19 and +41 kg for group RT. In conclusion, the rotation control strategy showed promising results, whereas the strategic feeding was poor from a parasite control standpoint.

Biological control of sheep parasites using Duddingtonia flagrans: trials on commercial farms in Sweden.

Trials were conducted on 3 commercial sheep farms in Sweden to assess the effect of administering spores of the nematode trapping fungus, Duddingtonia flagrans, together with supplementary feed to lactating ewes for the first 6 weeks from turn-out on pastures in spring. Also control groups of ewes, receiving only feed supplement, were established on all 3 farms. Groups were monitored by intensive parasitological investigation. The ewes and their lambs were moved in late June to saved pastures for summer grazing, the lambs receiving an anthelmintic treatment at this time. After approximately 6 weeks on summer pasture the lambs were weaned, treated a second time with anthelmintic, and returned to their original lambing pastures for finishing. Decisions as to when lambs were to be marketed were entirely at the discretion of the farmer co-operators. No difference in lamb performance was found between the two treatments on all three farms. This was attributed to the high levels of nutrition initially of the ewes limiting their post-partum rise in nematode faecal egg counts in spring, which in turn resulted in low levels of nematode infection on pastures throughout the autumn period. Additionally, pastures were of good quality for the lambs during the finishing period, so they grew at optimal rates as far as the farmers were concerned.

Reindeer as hosts for nematode parasites of sheep and cattle.

The reindeer husbandry range of Scandinavia overlaps with sheep, goat, and cattle pastures. The aim of this study was to determine whether reindeer are suitable hosts for ovine or bovine nematode parasites, and thus may spread these parasites into the reindeer husbandry regions. To render worm-free, twelve 4-month-old male reindeer calves, six lambs, and six bovine calves were given ivermectin at 200 microg/kg body weight. Five weeks post-treatment, six reindeer calves were each artificially dosed with 10,000 third-stage larvae (L3) of gastrointestinal nematodes derived from sheep, and an additional six reindeer with L3 derived from cattle. Lambs and bovine calves received the same dose of ovine and bovine larvae as reindeer, from the same larval source, respectively. Faecal samples collected on five occasions after the larval dosing revealed that by the fourth week, all reindeer calves, lambs, and bovine calves were infected. Animals were slaughtered on days 40 (reindeer) or 47 (lambs and bovine calves) after the larval dosing. Reindeer calves were most susceptible to L3 derived from sheep. The overall mean intensity of Haemochus contortus, Trichostrongylus axei, and Teladorsagia circumcincta, did not differ between reindeer and sheep; however, early fourth-stage larvae of H. contortus were more abundant in reindeer (p = 0.002). The establishment of bovine-derived Ostertagia ostertagi was similar in reindeer (62%) and bovine calves (57%), but larval inhibition was much higher in reindeer (91%, p < 0.001) than in cattle (31%). Very poor establishment of bovine derived Cooperia oncophora was recorded in reindeer calves (2%) compared with bovine calves (59%). These results show that young reindeer are susceptible hosts to the important gastrointestinal parasites of sheep (T. circumcincta, H. contortus) and cattle (O. ostertagi), as well as being a suitable host for T. axei.

Towards the eradication of Haemonchus contortus from sheep flocks in Sweden.

Previous studies have shown that between-year transmission of Haemonchus contortus in Swedish sheep flocks is almost entirely as over-wintered populations within housed animals, and not on pasture. This suggests that eradication of this parasite is a realistic possibility. Thus, two sheep farms with a history of H. contortus infection on the Swedish island of Oland were selected for study. During the winter housing period of 2003/2004 all ruminants (sheep and cattle) on both farms were treated with ivermectin. Monitoring by faecal egg counts and infective larval differentials of ewes and lambs for the subsequent two grazing seasons, together with total abomasal worm counts of 10 lambs from each farm at the end of the first grazing year, showed that this objective was achieved.

Evaluation of biological control of sheep parasites using Duddingtonia flagrans under commercial farming conditions on the island of Gotland, Sweden.

Field trials, conducted over 3 consecutive years, were aimed at assessing farmer opinions of the practicality and effectiveness of using Duddingtonia flagrans to control nematode parasites in their flocks on the Swedish island of Gotland. These trials were also monitored by intensive parasitological investigation. On Gotland, lambing occurs in spring, and around mid-summer (late June), ewes and lambs are moved to saved pastures due to pasture deterioration caused by dry conditions. Weaned lambs are then returned to original lambing pastures in early autumn for finishing. One farm (B) was used for 2001-2003 and a second farm (N) was also used in 2002 and 2003. On each farm, two flocks (each of 20 ewes + twin lambs) were managed separately, namely: fungus group which received a daily supplement + fungal spores from lambing until the summer move (6 weeks) and: control group which received supplement only. For Farm B, the numbers of lambs that were marketed prior to the end of the grazing season, were 13, 18, 19 for the fungus treatment whereas corresponding numbers for the control treatment were 8, 16 and 11 for years 2001, 2002 and 2003, respectively. Final weights of the remaining lambs at the end of each year were also consistently heavier, and the numbers of lambs retained for finishing during winter were less, on the fungus treatment compared with the control treatment. On Farm N, similar results were recorded, with more lambs marketed earlier in the fungus group (25 and 19) compared with the control (19 and 15) in 2002 and 2003, respectively. The weights of the remaining lambs at the end of the trial in 2003 showed a 4.5 kg weight gain advantage of the fungus group compared to the controls. Tracer tests during autumn 2001 on Farm B, showed that Teladorsagia circumcincta plus Trichostrongylus spp. levels were significantly less on the fungus treatment (P=0.018). The summer/autumn of 2002 was one of the driest on record for Gotland. This resulted in very low levels of infective larval availability. But on both farms, T. circumcincta numbers were less on the fungus than on the control paddocks (P=0.048 on Farm B). In 2003 very low numbers of infective larvae were recorded in the autumn tracers for both treatments on both farms. Both farmer co-operators were encouraged with these results and consider that biological control of nematode parasites in their flocks, using D. flagrans, is of practical value.

The epidemiology of abomasal nematodes of sheep in Sweden, with particular reference to over-winter survival strategies.

In May 2002, studies on the seasonal patterns of nematode infection of sheep were undertaken on four commercial sheep farms in southern Sweden, which had previously reported problems with nematode parasitism, especially due to Haemonchus contortus. One farm was used for intensive investigation. This entailed the establishment of two replicate groups of sheep, each consisting of 20 ewes and their lambs, on adjacent pasture paddocks. The seasonal patterns of nematode infection were followed by regular (approximately monthly) sampling of both ewes and lambs for nematode faecal egg counts and larval differentiation, and the sequential use of replicate groups of tracer lambs. H. contortus and Teladorsagia circumcincta were the most abundant nematode species, with the former most prevalent in the post-parturient faecal egg counts of ewes. Tracer worm counts showed almost 100% arrested development in the early fourth larval stage for H. contortus as early as mid-summer and the numbers of parasites progressively increased during the season. T. circumcincta also showed high levels of arrested development, but not as early, or as absolute, as for H. contortus. Tracers allocated to the paddocks at the time of turn-out following winter in May 2003, showed virtually a total absence of H. contortus in contrast to exceedingly high infections with T. circumcincta. Results of the three additional monitoring flocks supported these findings. It can be concluded that under Swedish sheep farming conditions, H. contortus has evolved to survive the long, cold winters entirely within the host as the arrested larval stage, relying on the lambing ewe to complete its life cycle. The peri-parturient relaxation of resistance in the ewes triggers the resumption of development to the adult egg-laying parasites to result in pasture contamination and the completion of just one parasite generation/year. In contrast, T. circumcincta can survive well over-winter, both on pasture and within the host.

Evaluation of copper supplementation to control Haemonchus contortus infections of sheep in Sweden.

A pen study was conducted to assess the effect of providing daily copper mineral supplement, or copper wire particle (COWP) capsules, on established or incoming mixed nematode infections in young sheep. For lambs with established (6 week old) infections, COWP resulted in 97% and 56% reduction of the adult and early L4 stages of H. contortus, respectively, compared with controls (p<0.001). Additionally there was a 74% reduction in Teladorsagia circumcincta infections in the COWP lambs compared with controls (p<0.01). However, no effect was observed when COWP were given at the commencement of a larval dosing period of 6 weeks. There was no significant effect of copper mineral supplement (given at the recommended rate to prevent Cu deficiency) on either established, or developing parasite infections. In addition, a field trial was conducted on a commercial farm to assess the effects of COWP in the management of recurrent H. contortus infections, but lack of parasites during the grazing season prevented an adequate assessment from being made. These results indicate that there is little, if any, benefit from a parasite control standpoint in recommending copper therapy, specifically to control parasites in Swedish sheep flocks.