Haemonchus contortus and Ostertagia circumcincta: fenbendazole treatment and abomasal permeability changes in sheep

Effects of fenbendazole (7.5 mg/kg of body weight infused during 90 minutes) on abomasal transmural potential difference (PD), pH, and ionic concentrations were determined in 8 lambs fitted with permanent abomasal cannulae. Four sheep were treated before inoculation and on day 28 after inoculation, with 200,000 3rd-stage Ostertagia circumcincta larvae and 4 others were similarly inoculated with 15,000 3rd-stage Haemonchus contortus larvae. In worm-free lambs, the unique effect was an increase in Na+ concentration of the abomasal contents. The O circumcincta or H contortus-inoculated lambs had an increase in PD, pH, and Na+ concentrations. Treatment further increased these concentrations, but the effects on PD occurred and ceased earlier in sheep inoculated with O circumcincta than in those inoculated with H contortus. The results indicated that part of the changes were linked to substances produced by the parasites or by the host. The increase in PD and pH was associated with a decrease in K+ concentration and indicated that these substances stimulated HCO3- transfer.     

Resistance to macrocyclic lactone anthelmintics by Haemonchus contortus and Ostertagia circumcincta in sheep in New Zealand

AIM: To establish the efficacy of oral formulations of ivermectin and moxidectin against naturally acquired abomasal nematode infections on a North Island sheep farm. METHODS: Two controlled slaughter trials were undertaken. In the first, 30 sheep on pasture were randomly allocated on the basis of faecal egg count to 1 of 3 groups, comprising an untreated control group and 2 treatment groups. One treatment group was given a single oral dose of ivermectin and the other a single oral dose of moxidectin, both at the manufacturer's recommended dose rates of 0.2 mg/kg liveweight. Six days after treatment, all animals were slaughtered and their abomasa recovered for worm counting. The second trial, which involved 47 animals, was essentially the same as the first except that, as well as involving the slaughter of 30 sheep from all 3 groups, 6 days after treatment, it also included a further 8 untreated control animals and 9 moxidectin treated animals which were slaughtered 27 days after treatment. RESULTS: At 6 days after treatment, moxidectin was highly effective against all 3 of the abomasal nematodes present. While ivermectin was similarly effective against Trichostrongylus axei 6 days after treatment, it was not effective against either Ostertagia circumcinta or Haemonchus contortus, against which average efficacies of only 63.6% and 61.6%, respectively, were recorded. At 27 days after treatment, moxidectin, was also highly effective against T. axei (97.3% reduction) but not against either H. contortus (71.4% reduction) or O. circumcinta (61.0% reduction). CONCLUSIONS: These results provide the first record of macrocyclic lactone resistance in H. contortus in sheep or in any other host in New Zealand, and the first case where such resistance has been exhibited in more than one parasite species at a time. Although the therapeutic efficacy of moxidectin was high against these resistant H. contortus and O. circumcincta strains, resistance to moxidectin was indicated by its diminished prophylactic activity against them. It is suggested that this reduction in the prophylactic activity of moxidectin is also likely to reduce its apparent current high therapeutic efficacy. CLINICAL RELEVANCE: As well as providing further evidence that it can no longer be automatically assumed that macrocylic lactone anthelmintics will be effective on sheep farms in this country, these findings also present a warning that increasingly complex parasite control options may have to be faced in the future.     

Hypergastrinaemia of sheep infected with Haemonchus contortus

Radioimmunoassay of plasma gastrin has shown that a hypergastrinaemia occurs in yearling sheep, after infection with 10(6) infective third stage larvae of Haemonchus contortus. Plasma gastrin concentrations began to rise two to four days after infection, that is, at about the same time as, or slightly before, the abomasal pH was observed to increase.     

Enhancement of resistance of lambs to Haemonchus contortus by previous infection with Ostertagia circumcincta

Two experiments were performed to determine whether previous infection of young lambs with Ostertagia circumcincta increased their resistance to the development of Haemonchus contortus and to explore alterations in the microenvironment of the abomasum related to the interaction. The parasitic infections were monitored with periodic fecal egg counts and by recovery and enumeration of parasites at necropsy of the lambs. Alteration of the microenvironment was examined by measurement of serum gastrin concentrations at critical times during both experiments. The results indicated that the numbers of H contortus were significantly (P less than 0.05) less in lambs which had O circumcincta infections before their exposure to H contortus. There was less reduction in lambs from which O circumcincta was removed (anthelmintic therapy) before their exposure to H contortus. Fecal egg counts also were significantly (P less than 0.05) less in the former group of lambs (exposed to both parasites) than those in lambs exposed only to H contortus. Serum gastrin values were significantly (P less than 0.05) increased in lambs infected with O circumcincta and indicated that the interaction observed could be due, in part, to alteration of abomasal pH.     

The distribution of pepsinogen within the abomasa of cattle and sheep infected with Ostertagia spp. and sheep infected with Haemonchus contortus.

The effect of nematode infections on the production of pepsinogen by ruminants was investigated immunohistochemically and biochemically. Abomasal tissues were collected from parasite-naive cattle and sheep, from sheep infected with predominantly Ostertagia circumcincta, sheep infected experimentally with Haemonchus contortus and cattle infected with Ostertagia ostertagi. Pepsinogen was also assayed biochemically in homogenates of fundic mucosae from sheep infected with predominantly O. circumcincta. Infection with Ostertagia spp. parasites was associated mainly with nodular hyperplasia, resulting in increased numbers of cells that produce both pepsinogen and mucus. Measured biochemically, nodules contained more pepsinogen than adjacent more normal mucosa (p < 0.05), and this effect was largely attributable to the greater mass of nodules. Infection of sheep with H. contortus was associated with generalised hyperplasia, characterised by increased numbers of mucopeptic cells and in at least one animal with reductions in parietal cell numbers. At the same time, the zymogen granule content of chief cells was reduced. Similar changes were occasionally seen in sheep infected predominantly with O. circumcincta. Generalised hyperplasia is likely to be indicative of the presence of ambulatory parasitic stages as opposed to those confined to nodules. The potential for the enhanced production of pepsinogen by increased numbers of cells with a joint mucous cell and zymogenic cell phenotype may offset decreases in the numbers of chief cells or reductions in chief cell activity.     

Wool production of sheep chronically infected with Haemonchus contortus

A chronic infection was established in a group of twenty 18-month-old non-reproductive Merino ewes by oral administration of 3000 infective H. contortus larvae twice weekly for 12 weeks. Their live-weights and wool production were compared with those of 20 uninfected ewes grazing the same pasture. In the infected sheep, faecal egg counts increased over a period of 9 weeks to reach a mean of 5000 eggs per gram, accompanied by small, but significant effects on packed-cell volume and live-weight. The effects of infection on wool growth were also small, and not statistically significant, although there was evidence of a faster seasonal decline in wool growth in infected sheep. It was concluded that mortality induced by acute infection is the most important economic effect of this parasite.     

The abomasal histology of worm-free sheep given primary and challenge infections of Haemonchus contortus

The histological changes in the abomasal mucosa of pairs of worm-free ewes and lambs were compared at various stages after infection and reinfection with 350 Haemonchus contortus L3 per kg bodyweight. In terms of worm burdens the ewes were relatively resistant to reinfection compared with the lambs and the histological changes in the abomasum at most stages after infection were more marked in the adult animals. These included increases in the numbers of mast cells, globule leucocytes, eosinophils and IgA plasma cells in the mucosa.     

Cross-reactive antigens of Haemonchus contortus adult worms in Teladorsagia circumcincta infected goats.

The adult Haemonchus contortus somatic antigens responsible for cross-reactivity have been analysed using serum samples from goat kids infected and reinfected with Teladorsagia circumcincta. Goat kids infected with T. circumcincta had similar serum ELISA values against somatic antigens of H. contortus as goats infected with H. contortus itself. Immunoblotting confirmed this extensive cross-reactivity particularly in the molecular weight range 105-29 kDa. However, peptides with high (195, 152 and 119 kDa) or low (23 kDa) molecular weight were only faintly recognized by heterologous sera.     

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.     

Effect of an antimuscarinic drug on the plasma pepsinogen activity of sheep infected with Ostertagia circumcincta

The antimuscarinic drug atropine caused a marked fall in plasma pepsinogen values of sheep with burdens of Ostertagia circumcincta and this response was greater in animals which had higher plasma pepsinogen values before administration of the drug. The response was shown to occur whether the elevated plasma pepsinogen values were a consequence of a larval infection in previously naive or exposed animals or of adult parasites directly transplanted into the abomasum of naive lambs.     

Effect of parasitism with Ostertagia circumcincta on pharmacokinetics of fenbendazole in sheep

Plasma and abomasal fluid concentrations of fenbendazole and its two major metabolites in sheep experimentally infected with Ostertagia circumcincta were compared with those in the same sheep when non-parasitised. Bio-availability of the drug was reduced in the parasitised state. There was also a reduction in the proportion of drug present in the form of metabolites in parasitised as compared with non-parasitised animals.     

Development of protection and of self-cure in lambs infected daily with Ostertagia circumcincta

Lambs were infected daily with 1000 Ostertagia circumcincta larvae. At intervals of 4 weeks groups of lambs were each challenged with a dose of 28,000 larvae, and their worm burdens compared with those of unchallenged controls. The measure of their ability to resist a challenge infection, which would have determined their degree of "protection", was obscured by the separate phenomenon of self-cure. Evidence suggested that the different manifestations of self-cure were determined by the state of resistance of the host and followed a set chronological sequence.     

Variability of resistance in goats infected with Haemonchus contortus in Brazil

The variability between and within breeds with respect to nematode egg counts (EPG), packed cell volume (PCV) and hemoglobin (Hb) was studied in 36 yearling female goats of the Caninde (15), Bhuj (6) and Anglo-Nubian (15) breeds, exposed to Haemonchus contortus. Nematode-free goats were turned to a contaminated paddock in late February. From then on, fecal egg per gram counts (EPG), packed cell volume (PCV) and hemoglobin (Hb) were determined at 2-week intervals up to Week 18. The EPG, transformed as [log(EPG + 75)], varied (P < 0.01) between goats within breeds and between weeks of exposure, but not between goat breeds (P > 0.05). PCV and Hb were affected by goat breeds (P < 0.05), by goats within breeds (P < 0.01) and by weeks of exposure (P < 0.01). Anglo-Nubians had higher (P < 0.01) PCV and Hb than Caninde; Bhuj had intermediate values. There were two EPG rises; one between Weeks 6 and 10 and the other between Weeks 14 and 16. The within breed variability was marked during the EPG rise on Week 6, when individual egg counts ranged from 130 to 2500. The EPG rises coincided with drops in Hb. PCV presented a similar trend, though not as marked. Haemonchus was responsible for more than 95% of nematode eggs counted. Considering the goat as experimental unit, the correlation coefficients (r) were: -0.45, P = 0.0064, between log(EPG + 75) and PCV; and -0.53, P = 0.0009, between log(EPG + 75) and Hb. The negative correlation between egg counts and blood values suggested breed differences in PCV and Hb were related to resistance to H. contortus infection and/or to its effects.