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Quantitative studies of ovine haemonchosis. 3. The interpretation and diagnostic significance of the changes in serial egg counts of Haemonchus contortus in a sheep flock.

An examination of the level and distribution of egg counts of H. contortus within an extensively managed flock of Merino sheep in south-east Queensland was carried out on three occasions over a ten week period. During this time the geometric mean egg count increased from 527 to 1331 eggs per gram, and although 26.3% of the flock were responsible for this increase, clinical evidence of haemonchosis was not observed. Examination of the distribution of counts supported the hypothesis that counts followed a negative binomial distribution. The values obtained for "k" (the index of over-dispersion), ranged from 0.5 to 1.4. However, on the two latter examinations, counts showed a divergence from the expected pattern (P = 0.015; P = 0.014). Further examinations of flocks at higher mean levels of count are required to establish if the trend towards a more normal distribution were maintained. The evidence from the present study and from outbreaks previously reported, suggested that in extensively managed sheep, the level of variance remained high as the mean egg count increased. It therefore appeared likely that in outbreaks of acute haemonchosis under grazing conditions, only a small proportion of the flock may become seriously affected.

Animals↗

Relationship among particle size distribution, dissolution profile, plasma values, and anthelmintic efficacy of oxfendazole.

Three mean particle sizes of oxfendazole raw material (1.65 micron, lot A; 3.2 micron, 10t B; 12.0 micron, lot C) were prepared and identically formulated as corresponding (A, B, and C) suspensions at 2.26% (W/V) concentration. Studies involving microscopic examination, scanning electron microscope analysis, particle size distribution, and surface area measurement were carried out on raw materials. In vitro dissolution profiles were obtained for the suspensions. A comparative bioavailability study of these 3 suspensions was performed in 12 sheep with each sheep given each formulation in a Latin square crossover study design; oxfendazole was dosed at rate of 5 mg/kg of body weight. Plasma-value measurements were made followed by an analysis of various bioavailability studies. Plasma area values indicated that suspension C (dw = 12.0 micron) was significantly (P less than 0.05) less bioavailable than was suspension A (dw = 1.65 micron); there was no difference between suspension A and suspension B. Significant differences were not seen in biological half-life and maximum plasma concentrations. The term dw refers to that particle diameter (determined by Coulter counting) at which 50% of the oxfendazole mass was in the form of particles having a lesser diameter and 50% was in the form of particles having a greater diameter. In a separate study involving 20 Merino weaner sheep infected with benzimidazole-resistant Haemonchus contortus larvae, oxfendazole's anthelmintic efficacy was demonstrated in the 2.26% suspension dosage form (90% particles less than 10 micron) at a dosing rate of 5 mg/kg. A correlation was found between its anthelmintic activity and plasma area values when compared in individual sheep. Data demonstrated that substantial differences in particle size distribution of oxfendazole could influence its dissolution rate, plasma concentrations, and absorption characteristics, thus indicating that oxfendazole's absorption could be dissolution-rate limited.

Administration, Oral↗

Thiabendazole-resistant strains of Haemonchus and Ostertagia in California lambs.

Ten parasite-free experimental lambs were inoculated intraruminally with a mixed-larvae inoculum which contained approximately 5,000 Haemonchus contortus infective larvae. Five lambs were treated with thiabendazole (66 mg/kg of body weight) and 5 were untreated controls. All lambs were killed 35 days after inoculations were done, and parasites present were recovered. Reduction in numbers of recovered Haemonchus contortus (25.7%) and Ostertagia spp (26.8%) were not significant (P greater than 0.05).

Animals↗

Experiments on inhibited development of Haemonchus contortus and Ostertagia circumcincta in sheep in The Netherlands.

Inhibited development of Haemonchus contortus and Ostertagia spp was studied in sheep. Exposure of infective Haemonchus contortus larvae for five weeks to 15 degrees C or 16 degrees C very effectively induced inhibited development but exposure to lower temperatures was much less effective. Infection with fresh larvae cultured for seven or 12 days at 25 degrees C in the dark resulted only in very low rates of inhibited development. This was not much influenced by the age of the host or the month of infection. Exposure of infective Ostertagia spp larvae for five weeks to 4 degrees C was fairly effective in inducing inhibited development while exposure to 16 degrees C was not effective. On two occasions inhibited development of Ostertagia spp was observed in lambs after a primary infection with fresh infective larvae cultured for seven days at 25 degrees C in the dark. This implies that a conditioning for inhibited development of Ostertagia spp is possible within this seven day culture period.

Animals↗

Resistance of Haemonchus contortus to thiophanate.

The occurrence of a field strain of Haemonchus contortus which was resistant to thiophanate and susceptible to thiabendazole and levamisole is reported. The LD95 of thiophanate for the strain was found to be 156.1 mg/kg and its resistance factor was 6.7. In possessing specific thiophanate resistance it differed from previously reported thiophanate resistant strains of H contortus which have been benzimidazole resistant worms that have possessed cross resistance to thiophanate.

Animals↗

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.

Animals↗

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.

Animals↗

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.

Animals↗

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.

Administration, Oral↗

Haemonchus contortus infections in Targhee and Targhee-Barbados Black-Belly cross lambs.

Targhee and Targhee-Barbados Black-Belly cross lambs that were raised helminth-free were experimentally infected with 10,000 or 25,000 infective larvae of Haemonchus contortus. There were no significant (P = less than 0.05) differences in results of weight gain, packed cell volumes, numbers of eggs per gram of feces, or numbers of helminths between the 2 groups of lambs.

Animals↗

Inheritance of cambendazole resistance in Haemonchus contortus.

Fourth-molt larvae (M4) of cambendazole-resistant (CBZ-R) and cambendazole-susceptible (BPL) strains of Haemonchus contortus were recovered from donor lambs and inoculated orally into source lambs to effect mating in 2nd lamb. Eggs produced by these parasitic matings were collected, cultured, and inoculated into lambs to compare the anthelmintic activity of cambendazole (CBZ), given at a dose rate of 20 mg/kg, against the F1 adults with its activity against the 2 parenteral strains (CBZ-R and PBL). In 2 replications, CBZ was 94% and 99% efficacious against the progeny of the cross matings and 91% and 95% against the BPL strain; CBZ was only 39% and 42% efficacious against the CBZ-R strain. In 1 test of F2 progeny of the reciprocal matings, CBZ was 81.2% efficacious against the F2 adults, whereas CBZ was 97.5% efficacious against the BPL strain and 37% against the CBZ-R strain. The results indicated that heredity of resistance to CBZ in H contortus is not sex-linked and is probably a result of a heterozygous recessive allele. The infective larvae of the CBZ-R groups in these 3 trials were from the 20th and 24th passages through lambs without exposure to any anthelmintics, yet the anthelmintic activity of CBZ ranged from 37% to 42%, indicating that there had been no reversion to susceptibility.

Alleles↗

A residual anthelmintic 2,6-diiodo-4-nitrophenol (Disophenol). Methods of testing its anthelmintic efficacy.

A subcutaneous injection of 2,6-diiodo-4-nitrophenol (Disophenol) at 10 mg/kg sheep leaves a residue bound to serum albumin which is lethal to Haemonchus contortus for 3 months after treatment. In the larval anthelmintic test, susceptible worm free sheep are dosed so that either third stage larvae (L3), or fourth stage larvae (L4) or 5th and adult stages are present on the day of treatment but slaughter is delayed to allow these larval stages to develop to adults because the larger worms are more easily seen, identified and counted. The larval anthelmintic test in sheep had to be altered and sheep killed within a few days of treatment, because the residues may be more effective against subsequent stages of development. Disophenol was greater than 60% effective against L3 of Oesophagostomum columbianum and L4 of H. contortus in greater than 60% of sheep (Class B). It rose to greater than 80% effective against adult H. contortus in greater than 80% of sheep (Class A). Against H. contortus it maintained Class A for 32 days, fell to Class B from 45--76 days and Class C (greater than 50% effective in greater than 50% of sheep) at 91 days after treatment respectively. In the RSA a treatment in December followed by another in March would protect sheep adequately against H. contortus for the entire season.

Animals↗

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.

Animals↗

Efficiency of oxfendazole administered as a single dose or in a controlled release capsule against benzimidazole-resistant haemonchus contortus, Ostertagia circumcincta and Trichostrongylus colubriformis.

Laboratory strains of Haemonchus contortus and Ostertagia circumcincta selected for thiabendazole resistance were found to have a strong side resistance to a single dose of oxfendazole. The LD50 and LD95 in mg of drug per host liveweight were respectively 4.28 and 18.46 mg/kg for H contortus and 3.61 and 11.20 mg/kg for O circumcincta. A field strain of Trichostrongylus colubriformis that had not been selected with thiabendazole for seven years also had a strong side resistance to oxfendazole with approximately 50 per cent of its population resistant to the recommended dose rate of 5 mg/kg. Prolonged administration of oxfendazole by intraruminal controlled release capsules was found to be effective against both susceptible and resistant strains of the above parasites. The first observed effect of oxfendazole, from controlled release capsules, on resistant worms was a decrease in the percentage of eggs developing to third stage larvae. This was followed by a decrease in egg count and in worm numbers.

Animals↗

Haemonchus contortus third stage larvae in sheep: kinetics of arrival into the abomasum and transformation during rumino-omasal transit.

The kinetics of passage of Haemonchus contortus third stage larvae through the omasoabomasal orifice were analysed in sheep for six hours after their administration by the nasooesophageal route. The larvae moved into the abomasum more slowly than the liquid phase of the ruminal contents. They arrived in three different forms: sheathed, exsheathed or coiled. A very small number of exsheathed larvae seemed to be necessary for inducing early modifications in permeability of the abomasal mucosa.

Abomasum↗

Circulating antigens in ovine haemonchosis.

We have developed a sandwich enzyme linked immunosorbent assay (sandwich ELISA), which allows the sensitive assay of Haemonchus contortus antigens. Using this test we have attempted to detect H. contortus circulating antigens in the serum, urine and milk of experimentally infected ewes. The results of this study indicate that circulating antigens may not be a general feature in ovine haemonchosis.

Animals↗

Immune response of sheep to Haemonchus contortus: serum antibodies against cross reacting antigens between parasites.

Normal and H. contortus infected sera were studied by ELISA technique against different stages of the parasites. In all cases antibody activity was detected. This activity in serum is partially eliminated after absorption with an adult worm extract of N brasiliensis. N. brasiliensis and H. contortus antigens were analysed by TCIEP with a rabbit anti-N. brasiliensis serum to examine shared antigens of H. contortus. A minimum of seven cross reacting antigens were detected. H. contortus adult worm extract was absorbed by the rabbit anti-N. brasiliensis serum. After absorption all cross reacting antigens were removed but at least one antigen reacting with a rabbit serum anti-H. Contortus is maintained. When this antigen is tested in elisa technique only a weak antibody activity is found in normal serum.

Animals↗