L.F. Le Jambre

THE GENETICS OF RESISTANCE AND RESILIENCE TO HAEMONCHUS CONTORTUS INFECTION IN YOUNG MERINO SHEEP

Abstract Young Merino sheep in 60 half-sib groups, totalling over 1000 lambs, were given a single dose of 11,000 Haemonchus contortus larvae. Resistance was measured, together with productivity during infection and when uninfected. One half-sib group exhibited an extremely high level of resistance; its sire was postulated to be the carrier of a major resistance gene. Genetic parameters were estimated for productivity during infection and in the absence of infection, for resistance as measured by faecal egg count and haematocrit, and for resilience as measured by the depression of productivity due to infection. The heritability of faecal egg count was estimated at 0.3, that for haematocrit after infection at 0.4. The heritability of resilience was too low to allow substantial progress by direct selection for this trait. Genetic correlations between resistance to infection and production traits when uninfected were not significantly different from zero. It was concluded that selection for polygenically controlled resistance would lead to substantial progress for this trait and would also increase productivity of infected animals.

Egg hatch as an in vitro assay of thiabendazole resistance in nematodes

Abstract A technique is described in which thiabendazole resistance is assayed by hatching nematode eggs in solutions of thiabendazole. Eggs of Haemonchus contortus and Ostertagia circumcincta were hatched in a series of concentrations of thiabendazole in a 0.1% NaCl solution. Thiabendazole resistant strains hatched in higher concentrations of thiabendazole than non-resistant strains. The dose response thus obtained for a strain of worms can be tested statistically against strains which are known to be either resistant or non-resistant.

Regulation of Haemonchus contortus populations in sheep exposed to continuous infection

Abstract Groups of sheep were infected three times weekly for 15 weeks with infective larvae of Haemonchus contortus at four rates ranging from 600 to 4800 larvae per week. At 3-weekly intervals, sheep from each group were killed for total worm counts after receiving a dose of radiolabelled larvae which enabled a direct measurement of establishment of incoming larvae during the continuing infection. Peak H. contortus burdens were reached between 6 and 9 weeks of infection, and were related to infection rate. In the groups receiving 2400 and 4800 larvae per week, worm numbers then declined rapidly and by the end of the experiment were lower than those in the groups given 600 or 1200 larvae per week. Establishment of incoming larvae was not influenced by infection rate, and declined from 45% in the first 4 weeks of infection to insignificant levels during the final 6 weeks. The proportion of incoming larvae arrested in their development increased as the infection progressed. It was concluded that H. contortus numbers were regulated by development of resistance to infection and by a loss of established worms which was related to the current rate of larval intake, and to the hosts previous experience of infection.

Rotational grazing for control of gastrointestinal nematodes of goats in a wet tropical environment

A preliminary experiment involving contamination of pasture plots with eggs of Haemonchus contortus, Trichostronglyus colubriformis and Oesophagostomum columbianum every month for a year established that in the tropical environment of the Pacific island of Tongatapu, hatching and development of all species was rapid and continuous, with a short survival on pasture (3-7 weeks) of the resulting infective larvae. These results indicated that a rotational grazing system consisting of ten paddocks grazed in sequence for 3.5 days at a time may permit a reduction in the frequency of anthelmintic treatment of goats. In comparison with an adjacent set-stocked flock which required treatment on three occasions during the year when mean flock egg counts exceeded 2000 eggs per gram (EPG), rotationally grazed goats generally maintained mean egg counts of less than 1000 EPG. Anthelmintic treatment was only given to rotationally grazed goats individually as they kidded, and there were indications that even this precaution was unnecessary. Because of the expense of frequent anthelmintic treatment and the resulting selection of strains of anthelmintic-resistant nematodes, rotational grazing of small ruminants through fencing, tethering or herding deserves further investigation as a nematode control option in wet tropical environments.

The impact of refugia on the development of thiabendazole resistance in Haemonchus contortus

Abstract Martin P. J. , Le Jambre L. F. and Claxton J. H. 1981. The impact of refugia on the development of thiabendazole resistance in Haemonchus contortus . International Journal for Parasitology 11 : 35–41. Beginning with a mixture of 5% thiabendazole-resistant and 95% non thiabendazole-resistant Haemonchus contortus larvae, worm free sheep were artificially infected with either 10,000, 9000, 7000, 2500 or zero larvae. Four weeks later, the infected sheep were treated with 44 mg/kg of thiabendazole. Seven days after anthelmintic treatment the same sheep were infected with zero, 1000, 3000, 7500 or 10,000 larvae respectively so that each sheep was given a total of 10,000 larvae. The larvae given after thiabendazole treatment are referred to as being in refugia (defined as an area where individual members of a population can escape exposure to a drug). This was repeated for six generations of parasites, maintaining the same proportion of larvae in refugia in each generation. The egg hatch assay for resistance indicated that refugia delayed the development of resistance in Haemonchus contortus . Where none or a small proportion of larvae were in refugia, a rapid increase in resistance occurred. With an increased proportion of larvae in refugia, resistance was slower to develop. Population size, as estimated by faecal egg counts done after anthelmintic treatment and corrected for the number of larvae given in each treatment, was lower at the higher levels of refugia. This was due to a lower level of resistance in the higher refugia lines and consequently a greater effect of the drug. However, the estimate of population size, from egg counts done after the larvae introduced from refugia had matured, were similar in all lines.

Principles for the use of macrocyclic lactones to minimise selection for resistance

OBJECTIVE To provide principles for the appropriate use of avermectin/milbemycin or macrocyclic lactone (ML) anthelmintics in sheep, to ensure effective worm control and to minimise selection for ML resistance. STRATEGY The principles were based on an assessment of the information currently available. The MLs were categorised into three groups (ivermectin [IVM], abamectin [ABA] and moxidectin [MOX]) based on structural differences, persistence and efficacy against ML resistant strains. The reported order of activity or efficacy against ML resistant worm strains was IVM<ABA<MOX. General treatment schemes were considered for Australian conditions and were divided into the following situations: 1. quarantine treatment, 2. treatment on/to clean pasture, 3. treatment on/to safe pasture, 4. treatment on/to moderate/heavily contaminated pasture. For each of these situations a strategy was considered for farms where ML resistance was present or absent. It was assumed that resistance commonly occurs in some or all other broad spectrum anthelmintics, and even where ML resistance has been detected, the ML group remains the most effective. The guidelines provided are general and it is expected that state agencies and sheep/veterinary advisers would give specific advice to suit their environments and drug resistance/worm problems. CONCLUSIONS The primary recommendation is to use a mixture of effective drugs when treating sheep. However, unless the combination treatment is highly effective it is unlikely to delay selection for ML resistance if sheep are being treated and moved to a clean or safe pasture. Where possible, reliance on the ML anthelmintics should be reduced by not using them every year, not using them in low risk stock or by using narrow spectrum and low efficacy drugs such as naphthalophos when appropriate. Anthelmintic treatment should be given as part of a strategic worm control program. It is suggested that IVM-oral and IVM-capsules should not be used when ML resistance is present. In this situation MOX or ABA should be used in combination with other drugs, provided that the chosen ML is effective against the resistant parasite. It is essential to monitor the efficacy of ML and drug combinations by post-treatment worm egg counts, particularly when ML resistance has been detected.

Development and survival of infective larvae of Haemonchus contortus and Trichostrongylus colubriformis on pasture in a tropical environment.

A trial to determine the seasonal pattern of egg hatching and larval survival on pasture was carried out in representative wet and dry zones of Fiji. Fourteen plots were established on parasite-free pasture at each of two sites. One plot at each site was contaminated every month with faeces from naturally infected goats containing a known proportion of Haemonchus contortus and Trichostrongylus colubriformis eggs. Pasture was sampled at regular intervals after contamination and infective larvae identified and counted. Larvae of both species developed throughout the year in the wet zone but development was more sporadic in the dry zone. Larval counts generally declined to below detectable levels within 9 weeks of contamination between September and March but longevity increased during the cooler weather from April to August. The comparatively short larval survival times noted in this experiment may present opportunities for manipulation of parasite population dynamics in the wet tropics.

Larval paralysis as an in vitro assay of levamisole and morantel tartrate resistance inOstertagia

The paper describes a method for detecting levamisole and morantel tartrate resistance by determining the percentage of paralysed third stage larvae in serial dilutions of anthelmintic. Levamisole resistantOstertagia spp. had a smaller proportion of larvae undergoing tonic paralysis in either levamisole or morantel tartrate than did a non-resistant strain. The dose response thus obtained for a strain of worms can be tested statistically against strains which are known to be resistant or non-resistant.

The inheritance of thiabendazole resistance in Haemonchus contortus.

Haemonchus contortus worm populations isolated from naturally infected sheep at the Pastoral Research Laboratory, Armidale, N.S.W., were found to contain approximately 20% of worms resistant to a 50 mg/kg dose of thiabendazole. Following 3 generations of selection with 50 mg/kg thiabendazole the number of worms removed by the anthelmintic was too small to detect differences between treated and control groups. After more than 15 generations of selection, matings between males from the selected strain and non-resistant females produced resistant males and females in equal numbers. Thus, thiabendazole resistance does not appear to be sex-linked. A dose--response assay on the F2 adults indicated that worms from female resistant x male non-resistant crosses were more resistant than F2 adults of the reciprocal cross. An in vitro technique that identified thiabendazole-resistant eggs by their ability to hatch in a solution containing thiabendazole and 0.1% NaCl solution was also used to study the inheritance of resistance. F1 eggs had similar LC50s to the resistant parents. F2 and back-cross eggs from an original mating of thiabendazole-resistant females x non-resistant males had a higher LC50 than F2 and back-cross eggs from the reciprocal mating, indicating a degree of matroclinous inheritance of resistance. However, the resistant parents had tolerances to thiabendazole exceeding those of F2. F3 eggs had a resistance distribution that ranged from that of the resistant to the non-resistant parent. No significant deviation from linearity was observed in any of the dose--response lines. These results indicate that thiabendazole resistance in H. contortus worms is inherited as an autosomal and semi-dominant trait.

Parasitological and immunological responses of genetically resistant Merino sheep on pastures contaminated with parasitic nematodes

One hundred and twenty lambs were grazed continuously from weaning until 9 months of age on 12 plots contaminated with larvae of three nematode species (Haemonchus contortus, Trichostrongylus colubriformis and Ostertagia circumcincta). The lambs were sired by either a genetically resistant ram or susceptible rams (determined by the response of previous progeny to artificial H. contortus infection). Half the resistant and half the susceptible lambs were given strategic anthelmintic treatment and the remainder remained untreated. Faecal egg counts and blood packed cell volume were measured frequently in all animals. One and 5 months after weaning, two lambs from each plot were slaughtered, and worm burdens and larval establishment rates of the three species of nematode were estimated. At the second slaughter, leukotriene levels and larval migration inhibitory (LMI) activity were measured in mucus collected from the small intestine. The dominant species in all faecal samples and the gastrointestinal tract was T. colubriformis. Lambs of the resistant genotype had lower faecal worm egg counts, lower worm burdens and higher levels of resistance to larval establishment. There were no differences in larval migration inhibition (LMI) activity, but resistant lambs had higher levels of the leukotriene LTC4/D4/E4. Further, the resistant genotype, identified on responsiveness to artificial infections with H. contortus, was more resistant to infections of three important species acquired naturally from contaminated pastures. All these genetic differences were maintained while the lambs were subject to strategic anthelmintic treatment.