Prolonged anthelmintic effect of closantel and disophenol against a thiabendazole selected resistant strain of Haemonchus contortus in sheep.

Two long-acting anthelmintics, closantel and disophenol, were tested by treatment 90, 60 and 30 days before challenge with a thiabendazole selected laboratory strain of Haemonchus contortus. The sheep were slaughtered 28 days after infection for total worm counts. A significant reduction in the number of adults remaining at autopsy was recorded after treatment with both anthelmintics 30 and 60 days before challenge but there was zero efficacy when the animals were treated 90 days before challenge. No significant difference was exhibited between the two compounds. Variability in response was recorded between animals on the 60 day period of treatment, which for disophenol may be dose related because animals receiving the lower volumes showed higher worm counts. For both anthelmintics a variability in the rate of metabolism may be responsible. The possibility offered by both anthelmintics for treating benzimidazole resistant strains of H contortus is discussed.

Resistance to benzimidazole anthelmintics in equine strongyles. 1. Frequency, geographical distribution and relationship between occurrence, animal husbandry procedures and anthelmintic usage.

A survey was conducted to determine whether benzimidazole resistant populations of equine strongyles are present in New South Wales and north central Victoria; what is their frequency and geographical distribution; which species are involved; and whether different methods of parasite control could be related to the occurrence and frequency of anthelmintic resistant populations. Resistant populations of strongyles were found over wide areas of New South Wales and in north central Victoria. There was no relationship between geographical location and the occurrence of benzimidazole resistance. The species involved were small strongyles of the sub-family Cyathostominae. There was a direct correlation between the occurrence of resistance (including the level at which it is present) and the frequency of use of benzimidazole anthelmintics. Examination of management practices showed that resistance is not an important problem on farms where different chemical classes of anthelmintics were used in a slow rotation programme; combination anthelmintic therapy (for example, benzimidazole/piperazine/organophosphates) was used and anthelmintic treatment was given at intervals of not less than 16 weeks. Tentative suggestions are made for the control of small strongyles in the light of an emerging resistance problem.

Changes in response of a benzimidazole resistant strain of Haemonchus contortus from sheep after passing through calves.

A benzimidazole resistant strain of Haemonchus contortus was passaged through lambs only or from lambs to calves and back into lambs. Changes in response to thiabendazole were monitored by using an egg hatch test at each animal passage and by a controlled experiment on adult worms at the final passage in lambs. An increased level of resistance was shown for the isolate during its passage through calves by the egg hatch test, although this was not supported on the adult worms in sheep using a single dose rate of 66 mg/kg of thiabendazole.

Five generations of selection with benzimidazole and non-benzimidazole anthelmintics against benzimidazole resistant strains of Haemonchus and Ostertagia spp in sheep.

Benzimidazole resistant strains of Haemonchus contortus and Ostertagia spp were subjected to selection pressure over five laboratory generations with the recommended dose rates of either cambendazole, oxfendazole or morantel. A change in response, with larger residual worm burdens remaining after treatment at the fifth generation, was shown for both cambendazole and oxfendazole against both species of nematode. No change in response against either species are shown for morantel. The results are discussed in terms of the problem associated with inefficient removal of the adult parasites after treatment.

Anthelmintic efficacy of low-dose phenothiazine against strains of sheep nematodes susceptible or resistant to thiabendazole, levamisole and morantel tartrate: effect on patent infections.

The anthelmintic efficacy of low-dose phenothiazine therapy, administered as a 1:40 phenothiazine: molasses mixture, was tested against patent infections of strains of Haemonchus contortus, Trichostrongylus colubriformis and Ostertagia spp susceptible or resistant to thiabendazole (an other benzimidazoles), levamisole and morantel tartrate. Phenothiazine reduced faecal egg output for both susceptible and resistant strains of all three nematodes. In daily doses of 0.25 g per sheep per day and above it completely inhibited larval production in susceptible strains. Against resistant strains, there was a reduced efficiency with 82.3 per cent inhibition of egg hatch at the 0.25 g per sheep per day level. Phenothiazine treatment had no significant effect on worm numbers for either susceptible or resistant worms. It is suggested that benzimidazole-resistant strains may be cross-resistant to phenothiazine.

Anthelmintic efficacy of low-dose phenothiazine against strains of sheep nematodes susceptible or resistant to thiabendazole, levamisole and morantel tartrate: efficiency against sequentially administered infections.

Low-dose daily phenothiazine therapy, administered as a 1:40 phenothiazine:molasses mixture, was tested against sequential infections of strains of Haemonchus contortus. Trichostrongylus colubriformis and Ostertagia spp susceptible or resistant to thiabendazole, levamisole and morantel tartrate. Phenothiazine at a dose rate of 0.5 g per sheep per day significantly reduced the numbers of worm eggs per gram of faeces in both susceptible and resistant strains by an average of 48 per cent and 60 per cent respectively. Phenothiazine completely inhibited larval production in susceptible strains. With resistant worms, larval production was depressed by up to 85.7 per cent. Treatment with phenothiazine : molasses mixture reduced the numbers of worms which became established in resistant Ostertagia spp and susceptible T colubriformis but had no effect on the establishment of susceptible H contortus and Ostertagia spp or resistant H contortus and T colubriformis.

Trichostrongylus colubriformis and Ostertagia sp resistant to levamisole, morantel tartrate and thiabendazole: infectivity, pathogenicity and drug efficacy in two breeds of sheep.

Merino and Border Leicester cross Merino sheep, nine months old, were infected with 10,000 third stage larvae of both Trichostrongylus colubriformis (PF4) and Ostertagia circumcincta/O trifurcata (PF5), known to have varying degrees of resistance to levamisole, morantel tartrate and thiabendazole. Crossbred sheep carried heavier Ostertagia sp worm burdens but there was no difference in susceptibility between the two breeds of sheep to infection with T colubriformis. The anthelmintic efficacy of thiabendazole, at 50 mg/kg, against T colubriformis was 81.8 per cent and 92.4 per cent for Merinos and crossbreds respectively while levamisole, at 6.75 mg/kg, was 12.3 per cent and 18 per cent effective. Thiabendazole removed 92.3 per cent and 83.8 per cent of Ostertagia sp in Merinos and crossbreds respectively. However, worm burdens in levamisole treated sheep were not significantly different from controls. No significant breed differences were found in 24 h faecal egg outputs. It is suggested that breed differences previously described in four-month-old sheep may have been due to differences in the rate of development of immune responsiveness. This disparity was no longer detectable in the immunologically mature sheep used in this study.

Trichostrongylus colubriformis and Ostertagia sp resistant to levamisole, morantel tartrate and thiabendazole: isolation into pure strain and anthelmintic titration.

Strains of Trichostrongylus colubriformis (designated PF4) and Ostertagia sp (O circumcincta/O trifurcata, designated PF5), with varying degrees of resistance to levamisole, morantel tartrate and thiabendazole were isolated into pure culture. Detailed dose response studies showed that both T colubriformis and Ostertagia sp were highly resistant to levamisole and morantel with low level resistance to thiabendazole. The effective dose required to remove 80 per cent worm burdens (ED80) was calculated for each anthelmintic. For T colubriformis the ED80 for levamisole and thiabendazole were 12.6 and 40.1 mg/kg respectively. For Ostertagia sp, the ED80 for levamisole, thiabendazole and morantel were 20.4, 45.2 and 35.8 mg/kg respectively. The implications of these results are discussed with reference to alternative means of chemical control.

Trichostrongylus colubriformis and Ostertagia circumcincta resistant to levamisole, morantel tartrate and thiabendazole: occurrence of field strains.

Field strains of Trichostrongylus colubriformis and Ostertagia circumcincta, designated PF4 and PF5 respectively, were recovered from a farm on which the sole use of levamisole over a preceding 12 year period led to the development of anthelmintic resistance. The results of field observations and preliminary critical trials in both Merino and crossbred sheep showed that both species have varying degrees of resistance to three major anthelmintics; levamisole, morantel tartrate and thiabendazole. Mean worm count reductions for adult T colubriformis (PF4) for therapeutic doses of morantel tartrate, thiabendazole and levamisole in crossbreds were 45.7 per cent, 97.3 per cent and zero respectively, and for Merinos 80.7 per cent, 88.3 per cent and 92.0 per cent respectively. Against O circumcincta the corresponding reductions for crossbreds were 51.4 percent, 95.4 per cent and 20.3 per cent and for Merinos, 52.5 per cent, 73.1 per cent and 29.8 per cent. There was no statistically significant difference in the responses of both parasite species to either levamisole or morantel. This result suggests that resistance to the two chemically unrelated drugs may be co-inherited.

The efficacy of fenbendazole and albendazole against immature and adult stages of benzimidazole-resistant sheep trichostrongylids.

The efficacy of two recently introduced benzimidazole anthelmintics, albendazole and fenbendazole, was determined for six-day, 10-day and adult stages of resistant strains of Haemonchus contortus and Trichostrongylus colubriformis. Albendazole, at 3.8 mg/kg reduced H contortus worm counts by 92.4, 70.8 and 67.1 per cent while fenbendazole, at 5.0 mg/kg, reduced worm burdens by 51.7, 95.5 and 93.4 per cent against six-, 10- and 25-day-old parasites respectively. For T colubriformis, the corresponding reductions with albendazole were 97.7, 95.8 and 64.9 per cent and for fenbendazole 29.0, 66.3 and 33.4 per cent. Compared with susceptible strains of H contortus and T colubriformis, for which therapeutic doses of benzimidazole anthelmintics are generally highly active against all stages of development, the present results show that these drugs do not have a uniform level of activity against all developmental stages of resistant strains.

The effect of single and divided dose administration on the efficacy of fenbendazole against adult stages of benzimidazole resistant sheep trichostrongylids.

Sheep infected with benzimidazole resistant strains of Haemonchus contortus and Trichostrongylus colubriformis were used to compare the anthelmintic efficacy of fenbendazole given as a single dose or administered in a divided dose regime over five days. Statistical analysis showed no significant difference between the two methods of administration for H contortus. On the other hand, divided dose fenbendazole was significantly less effective than single doses against adult T colubriformis at dose rates of 5 and 7.5 mg/kg. In the case of H contortus a highly significant correlation coefficient between post treatment egg counts and worm counts (r = 0.789) was obtained. This suggests that reduction in faecal egg output following drug treatment would provide a useful field indication of anthelmintic performance of fenbendazole (and possibly related compounds) against benzimidazole resistant strains of this parasite.

Physiological characteristics of free-living and parasitic stages of strains of Haemonchus contortus, susceptible or resistant to benzimidazole anthelmintics.

The effect of changes in genetic constitution (associated with developing benzimidazole resistance) on the physiological characteristics of Haemonchus contortus was investigated. Benzimidazole resistant H contortus were significantly more infective for sheep (20 per cent) than benzimidazole susceptible strains. Pathological changes were more severe in sheep infected with resistant H contortus as measured by packed cell volume, plasma protein concentration and haemoglobin levels. Faecal egg output was significantly higher for resistant strains as was the development and survival of eggs and free-living stages on pasture. Infective third stage larvae of benzimidazole resistant H contortus had a significantly faster exsheathment rate in rumenal fluid than did susceptible strains. These adaptive changes in response to continuing anthelmintic selection of resistant worm populations are discussed and the implications of these findings for worm control programmes indicated.

The dose response of several benzimidazole anthelmintics against resistant strains of Haemonchus contortus and Trichostrongylus colubriformis selected with thiabendazole.

Dose response lines for eight benzimidazole anthelmintics and thiophanate were determined, using standardised strains of thiabendazole selected and resistant Haemonchus contortus and Trichostrongylus colubriformis. Against H contortus, thiophanate, thiabendazole, parbendazole and oxibendazole were inactive. Mebendazole was inactive at dose rates of 6.26 and 12.5 mg/kg, although significant activity occurred at 25 mg/kg. Fenbendazole, cambendazole, oxfendazole and albendazole demonstrated significant activity at dose rates equal to or greater than the recommended therapeutic level. Thiophanate was inactive against resistant T colubriformis. The remaining compounds only showed significant activity when used at dose rates in excess of the recommended therapeutic level. These results show that a side resistance exists among the benzimidazole anthelmintics and suggests that changes in dose response lines could be expected to occur if resistant strains are selected with benzimidazoles other than thiabendazole.

The effect of route of administration on the anthelmintic efficacy of benzimidazole anthelmintics in sheep infected with strains of Haemonchus contortus and Trichostrongylus colubriformis resistant or susceptible to thiabendazole.

Observations of erratic anthelmintic activity of fenbendazole against known standardised thiabendazole-resistant strains of Haemonchus contortus and Trichostrongylus colubriformis in sheep were investigated. Fenbendazole at a dose rate of 10 mg/kg body weight was administered by oral, intra-ruminal or intra-abomasal routes, and was most effective against both resistant strains following intra-ruminal administration. In addition thiabendazole, oxibendazole, fenbendazole, parbendazole and mebendazole plus two unrelated compounds, levamisole and morantel tartrate, were used at one and a half times their suggested or recommended therapeutic dose rate against thiabendazole-resistant strains of H contortus and T colubriformis in sheep; each drug being administered by the intra-ruminal or intra-abomasal routes. Fenbendazole was more effective against both strains following intra-ruminal administration. Parbendazole was more effective against the resistant strain of T colubriformis following intra-ruminal administration. At the dose rate chosen for the other benzimidazoles used against these resistant strains, there was no difference in anthelmintic efficacy due to route of administration. Levamisole was highly effective against both resistant strains, irrespective of the route of administration. In the groups treated with morantel tartrate, the results obtained were difficult to interpret due to mortalities and a highly variable response in the surviving sheep. Fenbendazole, thiabendazole and mebendazole when used at their suggested or recommended therapeutic dose rate in sheep, were highly effective against known thiabendazole-susceptible strains of H contortus and T colubriformis following both intra-ruminal or intra-abomasal administration.

The anthelmintic efficacy of fenbendazole against thiabendazole-resistant strains of Haemonchus contortus and Trichostrongylus colubriformis in sheep.

The anthelmintic efficacy of fenbendazole (methyl 5-(phenylthio)-2-benzimidazole carbamate) was tested in sheep against standardised strains of Hcaemonchus contortus and Trichostrongylus colubriformis, known to be resistant to thiabendazole (LD90 for thiabendazole against H ontortus was 200 mg/kg bodyweight and against T colubriformis was 150 mg/kg). Fenbendazole at dose rates of 5, 10 and 20 mg/kg per os reduced total worm counts in H contortus infected sheep by 66, 90 and 100 per cent respectively, with similar reductions recorded for worm egg outputs. For the thiabendazole resistant strain of T colubriformis, fenbendazole reduced total worm counts in infected sheep by 4, 44 (40-48), 79 (75-83), 96 and 100 per cent at dose rates of 5, 10, 20, 40 and 80 mg/kg per os. Significant suppression of worm-egg production by thiabendazole resistant T colubriformis was obtained with fenbendazole at dose levels of 5 mg/kg and above. The implications of these results are discussed in the light of the increasing occurrence of strains of trichostrongylid nematodes resistant to currently available benzimidazole anthelmintics.

Infectivity of Ancylostoma caninum larvae after freezing over liquid nitrogen.

Third-stage larvae of Ancyclostoma caninum were exsheathed either by carbon dioxide or sodium hypochlorite treatment. Exsheathment per se reduced the infectivity of third-stage larvae by 45% to 59% without affecting larval motility. Both ensheathed and exsheathed larvae were frozen over liquid nitrogen for 90 days, and thawed. Frozen ensheathed larvae were all killed; but frozen exsheathed larvae were 40-54% as infective as non-frozen exsheathed larvae, and the addition of 10% dimethyl sulphoxide raised the infectivity to 89%, relative to non-frozen exsheathed larvae. Taken together, the deleterious effects of exsheathment and freezing resulted in an infectivity of 16-37%, relative to normal larvae.

The anthelmintic efficacy of fenbendazole against a mixed nematode infection in sheep.

Fenbendazole (methyl-5-(phenylthio)-2-benzimidazole carbamate) at dose rates of 5 mg/kg and above was 100 per cent effective in eliminating a naturally acquired Dictyocaulus filaria infection in sheep. The drug was 100 per cent effective in eliminating concurrent infections of adult Trichostrongylus axei, Haemonchus contortus, Haemonchus placei, Ostertagia circumcincta, Ostertagia ostertagii, Cooperia oncophora, Cooperia mcmasterii, Nematodirus spathiger, Neumatodirus filcollis, Oesophagostomum venulosum and Chabertia ovina. Fenbendazole was 93 per cent and 97 per cent effective at doses of 5 and 10 mg/kg respectively in removing infection with adult T colubriformis, and post-treatment worm-egg production was completely suppressed in surviving female worms. No adverse side-effects were observed in treated sheep at either of the two dose rates used.