Seasonal variation in anthelmintic response by cattle to dermally applied levamisole

Four experiments in 1978-79, and 2 in 1982 designed to define the optimum dose rate of dermally applied formulations of levamisole are described. These experiments showed that the absorption of levamisole with resultant blood levels and anthelmintic activity is strongly influenced by temperature. In warm to hot conditions percutaneous absorption is rapid and high blood levels with high anthelmintic activity against Haemonchus placei, Ostertagi sp, Trichostrongylus sp, Cooperia sp, Oesophagostomum radiatum, O. venulosum and Dictyocaulus viviparus result from dose rates of 10 mg/kg or more. In cold weather the high efficacy against H. placei, Cooperia sp, and Oesophagostomum sp is unchanged, but efficacy against Ostertagia sp, T. axei and D. viviparus decreases to the extent that a mean dose rate of 20 mg/kg (range 15 to 25 mg/kg) is necessary if the anthelmintic activity of dermally applied levamisole is to match that of either parenterally or orally administered material. These anthelmintic data are supported by the levamisole blood profile which in winter months, peaks at a figure of one quarter or less of that obtained from a similar dose rate in warm conditions. The implications of this variability in action are discussed.     

Naphthalophos combinations with benzimidazoles or levamisole as effective anthelmintics for sheep

Objective: To investigate the relative efficacy and safety of the anthelmintic naphthalophos in sheep, either given alone or in combination with benzimidazole (fenbendazole and albendazole) or levamisole anthelmintics.
Design: A parasitological study using faecal egg count reduction tests, a validating slaughter trial and field safety trials.
Procedure: Faecal egg count reduction tests were carried out on 13 farms. Naphthalophos and combinations of naphthalophos with levamisole and fenbendazole were included in the drench tests. On one property a controlled efficacy study was carried out to validate faecal egg count reduction test findings. In this trial, sheep were slaughtered 10 days after treatment and the remaining parasites recovered from the gastro-intestinal tract. Safety trials were carried out on eight farms where approximately 50 000 sheep were treated with naphthalophos and albendazole that were tank mixed in the backpack.
Results: The efficacy of naphthalophos alone in faecal egg count reduction tests ranged from 59 to 98% with one test showing 95% reduction. The efficacy of naphthalophos and levamisole ranged from 74 to 100%, with 5 farms showing 95% reduction. The efficacy of naphthalophos and fenbendazole ranged between 88 and 100% with 95% reduction achieved on 10 farms. The controlled efficacy study showed a good correlation between the faecal egg count reduction tests and numbers of parasites recovered, except for Nematodirus where the faecal egg count reduction tests overestimated efficacy. The mortality rate in the safety trials was 0.05%, with most fatalities occurring on one farm.
Conclusion: The combination of naphthalophos and fenbendazole was more effective than a combination of naphthalophos and levamisole, and will provide a sufficiently safe drench rotation option.     

Suspected resistance of Ostertagia ostertagi in cattle to levamisole

Observations on a beef cattle farm in Flanders led to the suspicion of resistance to levamisole in a strain of Ostertagia ostertagi. After treating a group of six animals with levamisole (5 mg kg-1 L.W., i.m.) the reduction in the number of trichostrongylid eggs per gram of faeces varied between 0 and 66.6%, whereas a similar group treated with fenbendazole (7.5 mg kg-1 L.W., p.o.) showed a reduction in worm burdens of 100%. Coproculture showed that the remaining eggs in the first treatment group were all Ostertagia sp. The suspected field strain was compared with a reference strain of O. ostertagi by means of the in vitro larval paralysis test. This test showed LC95 values of 9.12 micrograms ml-1 and of 99.04 micrograms ml-1 for the reference and the field strain respectively, which indicates a resistance factor for the latter of 10.9. These results were not unequivocally confirmed by the post mortem findings on a tracer calf necropsied 4 days after treatment with levamisole.     

Field evaluation of a mixture of albendazole sulphoxide and levamisole against Ostertagia and Trichostrongylus spp in sheep

The efficacy of a mixture of albendazole sulphoxide and levamisole, 3.6 and 8.25 mg/kg respectively, at single and double dose rates, was compared with the recommended dose rate of each anthelmintic alone. The comparison was conducted on groups of 6 to 14-week-old lambs on 22 farms, 16 of which had evidence of multiple resistance to benzimidazole and levamisole. A single dose of the mixture reduced mean egg counts by 95% on half the farms with multiple resistance and on all the remaining farms. Consequently, the mixture should be included in an assessment of effective anthelmintics on farms to determine its value for nematode control. A double dose rate of mixture was effective on all but 4 farms. Reductions caused by the mixture were due to the additive actions of the drugs on 18 of 22 farms; synergistic action was noted on only 3 farms. It was concluded that the mixture of albendazole sulphoxide and levamisole offered many farmers an effective anthelmintic for use in preventive control programs. Recommendations for such programs include annual rotation of effective anthelmintics as a means of delaying selection for drug resistance.     

The efficacy of levamisole, and a mixture of oxfendazole and levamisole, against the arrested stages of benzimidazole-resistant Haemonchus contortus and Ostertagia circumcincta in sheep

Sheep were allowed to graze pasture that had been seeded with benzimidazole-resistant Haemonchus contortus and Ostertagia circumcincta in order to acquire a burden of arrested larvae. Following housing, sheep were dosed orally with either oxfendazole at a dose rate of 4.7 mg/kg (to confirm the benzimidazole-resistant status of the species of nematode), levamisole at a dose rate of 7.5 mg/kg, or an oxfendazole/levamisole mixture at a dose rate of 4.6 mg/kg oxfendazole and 8.1 mg/kg levamisole. The efficacies of the treatments were assessed by estimation of the arrested larval burden in the abomasum of each sheep, either at 10 or 11 days (oxfendazole and oxfendazole/levamisole mixture), or 12 or 13 days (levamisole), after treatment. Compared to the untreated controls, the protection afforded by a single dose of either levamisole or the oxfendazole/levamisole mixture was >99% against the arrested stages of both Haemonchus contortus and Ostertagia circumcincta. Treatment with oxfendazole confirmed the benzimidazole-resistance status of the two species.     

Field studies investigating anthelmintic resistance in young cattle on five farms in New Zealand

AIM: To investigate the efficacy of pour-on anthelmintics against field strains of parasitic nematodes in young cattle on five farms in New Zealand. METHODS: Faecal nematode egg count (FEC) reduction (FECR) tests were carried out on five calf-rearing farms using pour-on formulations of levamisole, ivermectin, eprinomectin, and the simultaneous administration of levamisole and ivermectin. Faecal samples were collected per rectum before treatment and about 7, 14, 21 and 28 days after treatment, for FEC and faecal nematode larval culture. RESULTS: Resistance (i.e. <95% reduction in FEC) of Cooperia oncophora to ivermectin and eprinomectin was identified on all five farms. There was limited evidence of possible emerging resistance in Ostertagia spp to ivermectin but not eprinomectin, in short-tailed larvae of Cooperia spp to ivermectin and eprinomectin, and in Trichostrongylus spp to ivermectin, eprinomectin and levamisole used separately. Levamisole was effective against C. oncophora, but had variable efficacy against Ostertagia spp in the calves in this study. Simultaneous treatment with levamisole and ivermectin pour-on formulations were effective against all genera on all farms. CONCLUSIONS: To effectively manage roundworm parasites in their calves farmers need to be aware of the resistance status of the parasites on their farms. Levamisole is likely to be an effective anthelmintic on most farms at times of the year when the impact of Ostertagia spp is not high. Simultaneous administration of levamisole and ivermectin pour-on anthelmintics to cattle is likely to control both ML-resistant C. oncophora and stages of Ostertagia spp that are not controlled by levamisole alone.     

Evidence for multiple anthelmintic resistance in sheep and goats reared under the same management in coastal Kenya

Four experiments, two with sheep and two with goats, were carried out to determine the efficacy of ivermectin, fenbendazole, levamisole, closantel and some of their combinations by faecal egg count reduction tests. In the first experiment, injectable ivermectin, oral ivermectin, fenbendazole and levamisole were tested in 6-month-old lambs, and their reduction percentages were 77%, 13%, 42% and 92%, respectively. In the second experiment, with yearling sheep, the reduction percentages were 35% for injectable ivermectin, 32% for fenbendazole, 99% for levamisole, 48% for closantel, 92% for injectable ivermectin combined with fenbendazole, 99% for injectable ivermectin combined with levamisole, and 100% for fenbendazole combined with levamisole. In the study with 18-month-old goats given the same dose rates as those recommended for sheep, the reduction percentages were 73% for injectable ivermectin, 25% for fenbendazole, and 78% for levamisole. Another group of 14-month-old goats was treated with dose rates 1.5 times those recommended for sheep and the reduction percentages were 93% for levamisole, 92% for injectable ivermectin, and 97% for a combination of levamisole and ivermectin. In all experiments with sheep and goats the gastrointestinal nematode parasites identified by larval cultures were Haemonchus contortus, Trichostrongylus spp. and Oesophagostomum spp. The gastrointestinal nematodes of both sheep and goats on this farm are resistant to ivermectin and fenbendazole, whereas levamisole is still effective in sheep, but not in goats. The results are discussed in relation to the farm as a source of breeding stock to smallholder farmers and its potential to spread anthelmintic resistance.     

Efficacy of Albendazole and Levamisole against Gastrointestinal Nematodes of Sheep and Goats in Morogoro,Tanzania

A study was conducted to determine the efficacy of albendazole after it had been withdrawn from use due to the development of resistant strains of nematodes about ten years ago. The study also aimed to determine the present efficacy of levamisole, which had been recommended to replace albendazole. On one farm, the sheep and goats were divided into two groups, one group of each serving as the untreated control, while the other was treated with levamisole. The sheep on the other farm were divided into three groups, one serving as the untreated control group, the second being treated with levamisole and the third being treated with albendazole. Faecal samples were collected one day before treatment, and again 10 days after treatment. Anthelmintic efficacy was determined by the faecal egg count reduction test. Ten days after treatment, the sheep treated with levamisole on the first farm had a 98% reduction in faecal egg count, with a 95% confidence limit of 76%. The goats on the same farm had a 97% reduction in faecal egg count, with a 95% lower confidence limit of 81%. At the second farm, 10 days after treatment, sheep treated with levamisole had a 99.4% reduction in faecal egg count, with a 95% lower confidence limit of 88.9%, whereas the sheep treated with albendazole only had a 59.4% reduction in faecal egg count, with a 95% lower confidence limit of –19.6%. The study indicated that the gastrointestinal nematodes of sheep at the Department of Animal Science and Production farm were still resistant to albendazole about ten years after this anthelmintic had been withdrawn from use. A reduced efficacy of levamisole was suspected.     

The prevalence of anthelmintic resistance in sheep nematodes on the central tablelands of New South Wales

A survey was undertaken to assess the prevalence of anthelmintic resistance in sheep nematode populations on 40 commercial farms distributed throughout the central tablelands of New South Wales. Representatives of the 2 major groups of broad spectrum anthelmintics with different modes of action (thiabendazole and levamisole) were used at the manufacturer's recommended dose rates. Efficacy was assessed on the basis of the reduction in faecal strongyle egg counts 7 days after treatment. An efficacy of less than 90% using both anthelmintics was obtained on 4 farms. Thiabendazole had an efficacy of less than 90% on a further 21 farms and levamisole had an efficacy of less than 90% on an additional 4 farms. There was no evidence of anthelmintic resistance on 8 farms, while the remaining 3 had insignificant parasite burdens. Based on larval cultures from faeces, Ostertagia and Trichostrongylus were the most significant species in resistant populations. Haemonchus burdens were sporadic and levels of resistance relatively low. Nematodirus burdens were widespread but no evidence of resistance was detected.     

The effectiveness of benzimidazole-levamisole combination drenches in the presence of resistance to both benzimidazole and levamisole anthelmintics in New Zealand sheep

Examination of 25 cases of multiple benzimidazole and levamisole resistance, identified in sheep by faecal egg count reduction testing at the Batchelar, Lincoln and Invermay Animal Health Laboratories, showed that benzimidazole-levamisole combinations provided effective control in eight (47%) of 17 cases in which they were tested. Overall, the use of combination drenches resulted in average improvements in faecal egg count reductions of 25.6% (p<0.01) and 23.2% (p<0.05), respectively, over those achieved by the use of benzimidazole or levamisole drenches alone. The results suggest that instances of multiple resistance in which combination drenches might be effective are unlikely to be predictable either by identification of the parasites involved, or from the levels of benzimidazole or levamisole resistance present.     

Prevalence of anthelmintic resistance on sheep farms in New Zealand

AIM: To establish the prevalence of anthelmintic resistance in parasitic nematodes on sheep farms in New Zealand. METHODS: A cross-sectional prevalence study was conducted, using a standardised faecal nematode egg count (FEC) reduction (FECR) test (FECRT) for ivermectin, at a full (0.2 mg/kg) and half (0.1 mg/kg) dose rate, and albendazole, levamisole and albendazole-levamisole in combination, on 60 lambs (n=10 per group) on farms selected from throughout New Zealand. Farms that conformed with selection criteria were chosen at random (n=80) or with a history of suspected resistance to macrocyclic lactone (ML) anthelmintics (n=32). Resistance to an anthelmintic was inferred when there was <95% reduction in FEC 7-10 days after treatment. Larval cultures were performed for all control groups and for treated groups for which resistance was evident. RESULTS: Of the farms randomly selected, 36% showed > or =95% FECR for all anthelmintics tested; resistance to ivermectin at 0.1 and 0.2 mg/kg liveweight was evident on 36% and 25% of these farms, respectively. Resistance to both ivermectin (0.2 mg/kg) and levamisole was evident on 8/80 (10%) farms, to ivermectin and albendazole on 10/80 (13%) farms, and to ivermectin, levamisole and albendazole on 6/80 (8%) farms. The prevalence of resistance to a half dose of ivermectin tended to be more prevalent on farms with a history of suspected ML resistance (p=0.06). Resistance to albendazole was seen across all the main parasite genera, and to levamisole in Nematodirus, Ostertagia (= Teladorsagia) and Trichostrongylus species. Resistance to ivermectin was dominated by Ostertagia spp, although Cooperia, Nematodirus and Trichostrongylus species were also implicated. CONCLUSION: Anthelmintic resistance in parasitic nematodes of sheep is common in New Zealand. Not only was resistance to albendazole and levamisole common, but resistance to the ML, ivermectin, was at a higher prevalence than expected. Sheep farmers and advisors in New Zealand need to re-evaluate the way they manage parasites, and more research is urgently needed if the steady decline in anthelmintic susceptibility is to be halted.     

Controlled test of anthelmintic activity of levamisole administered to calves via drinking water, subcutaneous injection, or alfalfa pellet premix.

A controlled test of the activity of 3 formulations of levamisole, at the dose level of approximately 8 mg/kg, against naturally occurring infections of gastrointestinal parasites and lungworms was made in 24 calves allotted to 4 groups of 6 calves each. Levamisole was administered to group I calves in the drinking water, to group II calves by subcutaneous injection, and to Group III calves by feeding alfalfa pellets mixed in corn silage; group IV calves were nontreated controls. Group I calves consumed the medicated water between 4 hours and 20 minutes and 9 hours and 40 minutes; group III calves consumed the medicated feed within 2 hours and 15 minutes. For calves of group I, II, and III, removals of 4th-stage Ostertagia sp were 64, 23, and 0%; of mature Ostertagia ostertagi, 90, 93, and 83%; and of mature Trichostrongylus axei, 92, 99, and 92%, respectively. For all 3 treated groups of calves, removal was 100% for 4th-stage Cooperia sp and for mature Trichostrongylus colubriformis, Cooperia oncophora, Cooperia punctata, and Oesophagostomum radiatum. Removals of Dictyocaulus vivipara were 90, 90, and 94% for calves of groups I, II, and III, respectively. There was no evidence of toxicosis. At necropsy, 2 calves in group II had small areas of edema at the sites of injection of levamisole.     

Anthelmintic resistance in nematode parasites of sheep in Denmark with special emphasis on levamisole resistance in Ostertagia circumcincta.

This study was undertaken to elucidate the presence of anthelmintic resistance in nematode parasites of sheep in Denmark. Twenty two flocks of sheep were selected for Faecal Egg Count Reduction (FECR) tests, based on a prior history of either the same anthelmintic, or anthelmintic class having been used 3 times or more over the previous 5 years. Evidence of anthelmintic resistance was detected in 7 flocks. Two flocks showed FECR of 88% and 94% after treatment with thiabendazole, FECR of another 2 were 90% and 94% following treatment with fenbendazole. Three flocks showed FECR of 73%, 89% and 94%, respectively following the use of levamisole. Ostertagia circumcincta was isolated from 1 of the latter flocks and subjected to an in vivo controlled slaughter assay. Following treatment with levamisole at the recommended dose rate of 7.5 mg/kg, FECR was 44.5% and worm counts were reduced by 67.7%. These results were further substantiated by an in vitro egg hatch paralysis assay and by measuring pepsinogen levels in treated and non-treated lambs. This is the first instance of anthelmintic resistance in sheep nematodes in Scandinavia.     

Efficacy of levamisole against immature and mature nematodes in goats with induced infections

Anthelmintic efficacy of levamisole against induced infections with 7- and 21-day-old Haemonchus contortus, Ostertagia circumcincta, Trichostrongylus axei, and T colubriformis was evaluated as an oral drench in goats. Group 1 (n = 8) was not treated, group 2 (n = 8) was given 3.96 mg of levamisole/kg of body weight, group 3 (n = 8) was given 7.92 mg of levamisole/kg, and group 3 (n = 7) was given 11.88 mg of levamisole/kg. Efficacy against all worms was low in goats given 3.96 mg of levamisole/kg, but was high against adult H contortus (99%) and adult T colubriformis (99.7%) in goats given 7.92 mg of levamisole/kg. Although efficacy against adults of all species was high in goats given 11.88 mg of levamisole/kg, some immature worms of all species remained in the abomasa of goats.     

Multiple resistance in goat-derived Ostertagia and the efficacy of moxidectin and combinations of other anthelmintics

Faecal egg count reduction tests were used to identify a strain of Ostertagia circumcincta/trifurcata complex in goats which was resistant to: ivermectin (at 0.2 mg/kg and 0.4 mg/kg orally), oxfendazole (at 5 mg/kg orally), levamisole (at 12 mg/kg orally) and fenbendazole (at 5 mg/kg orally) combined with levamisole (at 9.4 mg/kg orally). The percentage reductions achieved in these faecal egg count reduction tests were respectively 27%, 83%, 82%, 79% and 82%. Moxidectin (at 0.2 mg/kg by subcutaneous injection), fenbendazole (at 10 mg/kg orally) combined with levamisole (at 18.8 mg/kg orally), ivermectin (at 0.4 mg/kg orally) combined with oxfendazole (at 10 mg/kg orally) and ivermectin (at 0.4 mg/kg orally) combined with levamisole (at 12 mg/kg orally) were effective in removing these nematodes in goats as determined by faecal egg count reduction tests. These drenches achieved reductions of 100%, 100%, 98% and 100% respectively.     

The efficiency of levamisole, thiabendazole and fenbendazole against naturally acquired infections of Ostertagia ostertagi in cattle

In two experiments, conducted in cattle with naturally acquired infections of Ostertagia ostertagi, comparative assessments were made of the anthelmintic efficiency of levamisole, thiabendazole and fenbendazole, each at 1.0, 1.5 and 2.0 times the recommended dose rate. Variable efficiencies of 81 and 49 per cent for levamisole, 86 and 56 per cent for thiabendazole were obtained against adult O ostertagi. Neither drug showed substantial activity against early fourth stage larvae. Efficiency of fenbendazole against adult O ostertagi was consistently high; 85 and 89 per cent in the two experiments respectively. In the first experiment in which cattle were slaughtered two to three days after treatment, only 22 per cent of inhibited early fourth stage larvae were removed whereas in the second experiment when slaughter took place 10--11 days after treatment, this efficiency was 89 per cent. There was no increased effect of increased dose rates on treatments with thiabendazole or fenbendazole. The activity of levamisole against adult worms and inhibited larvae was increased at twice the recommended dose rate.     

The comparative efficacy of four broad spectrum anthelmintics against some experimentally induced trichostrongylid infections in sheep and goats

A controlled slaughter trial was undertaken to compare the efficacies of oxfendazole (5 mg/kg), morantel citrate (10 mg/kg), levamisole (8 mg/kg) and ivermectin (0.2 mg/kg) against experimentally induced infections of adult Haemonchus contortus, Ostertagia spp., Trichostrongylus colubriformis and Cooperiacurticei in sheep and goats. Ivermectin and oxfendazole achieved similar levels of efficacy in both hosts against all four worm genera as did levamisole and mortantel against H. contortus and C. curticei. Against Ostertagia spp. and T. colubriformis, however, the latter two drugs were less effective in goats than sheep. Neither the numbers of Ostertagia spp. removed from goats by levamisole (81% reduction) nor the numbers of T. colubriformis removed from goats by morantel (56% reduction) were statistically significant (P>0.05).     

Controlled trials of the anthelmintic oxfendazole in ewes and lambs naturally infected with gastrointestinal nematodes.

Oxfendazole doses at a rate of 5 mg per kg, before or after lambing, reduced nematode egg output to insignificant levels in ewes, most of which were not exposed to re-infection. Ewes treated with oxfendazole had a significantly lower egg output than those treated with levamisole, although the latter anthelmintic was also highly effective. In lambs, oxfendazole at a dose rate of 5 mg per kg, showed 100 per cent efficacy for Ostertagia circumcincta, O trifurcata, Teledorsagia davtiani, Trichostrongylus axei, T vitrinus, T colubriformis, Nematodirus battus, N filicollis, immature Nematodirus, Chabertia ovina, and 93 per cent efficacy for Trichuris spp. Levamisole showed similar efficacies but did not remove Trichuris.     

Seasonal comparisons of anthelmintic activity of levamisole pour-on in cattle in the USA

Anthelmintic activity of a pour-on formulation of levamisole, applied during warm weather (16 degrees to 36 degrees C) at 10 mg/kg bodyweight, was evaluated in groups of naturally parasitised calves. This activity was compared to that obtained in similar groups of calves treated in the winter (-4 degrees to +7 degrees C). Controlled efficacy of the pour-on formulation was determined for each season by comparing mean worm burdens in treated calves sacrificed seven to nine days after treatment to non-treated controls. In these trials, burdens of Bunostomum phlebotomum, Cooperia species, Haemonchus placei, Nematodirus species, Oesophagostomum radiatum, Ostertagia ostertagi and Trichostrongylus axei in treated calves were reduced by 83.3 to 100 per cent in the summer and 89.2 to 100 per cent in the winter. Field investigations at nine locations across the USA compared changes in faecal egg counts for cattle treated and evaluated during warm summer months (27 degrees to 36 degrees C) to those treated during cold winter months (-18 degrees to +10 degrees C). Overall, faecal egg counts were reduced by 90.2 per cent in the summer trials and 94.0 per cent in the winter trials. The results of these trials indicate that there is no seasonal variation in the anthelmintic activity of this pour-on formulation of levamisole.     

Simultaneous resistance to two or more broad-spectrum anthelmintics by gastrointestinal nematode parasites of sheep and goats

The detection, by means of faecal egg count reduction tests and larval cultures, of two cases of multiple anthelmintic resistance in goats and two in sheep, is reported. The former cases appeared to involve resistance to all three broad-spectrum drench groups (benzimidazole, levamisole, ivermectin) by mainly Ostertagia spp., the latter resistance to two of them (benzimidazole, levamisole) by Trichostrongylus spp.