Minimising the development of anthelmintic resistance, and optimising the use of the novel anthelmintic monepantel, for the sustainable control of nematode parasites in Australian sheep grazing systems

Objective To compare the risk of different treatment scenarios on selecting for anthelmintic resistance on Australian sheep farms. Design A computer simulation model predicted populations of Trichostrongylus colubriformis, Haemonchus contortus or Teladorsagia (Ostertagia) circumcincta, and the frequency of anthelmintic resistance genes. Method Nematode populations and the progression of drug resistance for a variety of treatment options and management practices in sheep‐rearing areas of Western Australia (WA), Victoria (VIC) and New South Wales (NSW) were simulated. A scoring system was devised to measure the success of each option in delaying resistance to each anthelmintic and in controlling nematode populations. Results The best option at all sites was combining the new anthelmintic (monepantel) with a triple mixture of benzimidazole, levamisole and abamectin (COM). The next best option was: in NSW, rotation at each treatment between monepantel, moxidectin and COM; in VIC, rotation at each treatment between monepantel and COM; and in WA, rotation at each treatment between monepantel (used in winter) and COM or moxidectin (used in summer–autumn). In WA, rapid selection for resistance occurred as a consequence of summer–autumn treatments; however, if a small percentage of adult stock were left untreated then this selection could be greatly reduced. Despite purposely assuming relatively high resistance to benzimidazole and levamisole, COM was still effective in controlling worms and delaying resistance. Conclusions Because of cost constraints, it may not be feasible or profitable for producers to always use the combination of all drugs. However, the second‐ and third‐best options still considerably slowed the development of anthelmintic resistance.     

Multiple anthelmintic resistance and the possible contributory factors in Beetal goats in an irrigated area (Pakistan)

This paper presents the first report of multiple anthelmintic resistance in the gastrointestinal nematodes of goats and its possible contributory factors in an irrigated area (Pakistan). A total of 18 privately owned Beetal goat flocks were selected in order to determine the anthelmintic resistance against commonly used anthelmintics. Forty to 48 animals from each flock were selected according to their weight and egg count. The three anthelmintics viz., oxfendazole, levamisole and ivermectin, were given to three groups at manufacturer’s recommended dose while one group was kept as untreated control. Anthelmintic resistance was determined through faecal egg count reduction and egg hatch tests while assessment of the contributory factors of anthelmintic resistance was measured through the rural participatory approach. Faecal egg count reduction test revealed high prevalence of anthelmintic resistance (83.3%) and it was either single (levamisole) or multiple (oxfendazole and levamisole). Egg hatch test confirmed the resistance against oxfendazole as detected with faecal egg count reduction test. None of the goat flocks was resistant to ivermectin. Copro-cultures revealed that Haemonchus contortus, Trichostrongylus colubriformis and Teladorsagia circumcincta were the most common species exhibiting resistance to levamisole and oxfendazole. Step-wise logistic regression of the data on worm control practices revealed significant role of under-dosing, low-protein diets, healthcare supervision by the traditional healers and mass treatments.     

Drench-and-shift is a high-risk practice in the absence of refugia

Abstract

AIM: To examine the effect of an anthelmintic treatment to lambs, followed immediately by a shift onto pastures with differing levels of larval contamination, on the development of anthelmintic resistance, in order to support recommendations to farmers regarding drench-and-shift practices for sustainable worm control.

METHODS: Newly weaned Romney lambs (n=72) were dosed with third-stage infective larvae (L3) of two nematode parasite species, Teladorsagia (=Ostertagia) circumcincta and Trichostrongylus colubriformis, comprising benzimidazole-resistant and -susceptible isolates, calculated to yield, after treatment with albendazole, a 95% reduction in faecal nematode egg count (FEC). Once infections became patent (Day 0), lambs were randomised into nine groups of eight animals, treated with albendazole at the manufacturer's recommended dose rate, and moved to individual pastures each previously prepared to have one of three different levels of parasite larval infestation (Treatment 1 = low contamination, Treatment 2 = medium contamination, and Treatment 3 = high contamination), and grazed on those pastures before receiving a second treatment with albendazole at Day 47. Anthelmintic resistance status in each group of lambs was measured using FEC reduction (FECR) and egghatch assays (EHA) after the first anthelmintic treatment, and FECR after the second treatment.

RESULTS: Egg-hatch assays demonstrated significant differences between treatments. The concentration of anthelmintic required to kill 50% of the eggs (LC₅₀) for Treatment 1, comprising the least contaminated pastures, was significantly higher than for Treatments 2 and 3 on Days 33 and 40. Treatment 1 also had a significantly lower FECR at the final anthelmintic treatment, and significantly lower FEC than the other two treatments from Days 26 to 47.

CONCLUSIONS: The results demonstrated that the populations of T. circumcincta and T. colubriformis in lambs treated with anthelmintic had significantly higher levels of albendazole resistance at the end of the grazing period in lambs moved onto pastures with relatively low levels of parasite contamination than those moved onto pastures with relatively higher contamination, confirming drench-and-shift onto ‘clean’ pasture as a high-risk practice for the selection for anthelmintic resistance. While this does not necessarily preclude the use of this practice it does emphasise the importance of taking appropriate remedial action to minimise the risk.     

Dynamics of the free-living stages of sheep intestinal parasites on pasture in the North Island of New Zealand. 2. Weather variables associated with development

Abstract

AIM: To identify weather variables associated with the development of eggs of Teladorsagia (=Ostertagia) circumcincta, Trichostrongylus colubriformis and Haemonchus contortus to third-stage infective larvae (L3) under a range of climatic conditions on pasture in the North Island of New Zealand.

METHODS: Sheep faeces containing known numbers of eggs of all three nematode species were deposited on, or buried in, pasture plots at three sites, viz coastal Manawatu, Upper Hutt Valley, and East Cape hill country. Development was measured by recovering L3 from faeces, herbage and soil 28–31 days after deposition, on 13–18 occasions between January 2005 until July 2006. Weather data were recorded at each site, and the association between weather variables and number of L3 recovered was analysed using subsets regression to select best-fitting models from several candidate variables, after adjustment for efficiency of recovery of L3. A multiple linear regression model was then developed for each species, to select weather variables that had both significant and substantive effects on the number of L3 recovered.

RESULTS: For all species, mean daily temperature was the bestpredictor of the number of L3 recovered (p=0.001). For T.circumcincta the final model included mean daily temperature and soil temperature (R²=51%), and for T. colubriformis the model only included mean daily temperature (R²=55%). For development of H. contortus, mean daily temperature was the most significant variable, but moisture in the form of rainfall entropy, i.e. the temporal distribution of rainfall, overthe first 14 days was also significant in the final model (R²=34%).

CONCLUSIONS: Temperature was the most important determinant of developmental success of free-living nematodes on pasture at the study sites, and probably also for other parts ofNew Zealand with similar climates. Moisture was not significant in the development of T. circumcincta or T.colubriformis, implying that under the generally moist temperate climate in New Zealand, moisture is seldom limiting for development of these species. Haemonchus contortusappeared to be sensitive to moisture availability and precipitation in the first 14 days after deposition of faeces. The results of this study will be further developed as part of species-specific climate-driven models of parasite development in New Zealand.     

Dynamics of the free-living stages of sheep intestinal parasites on pasture in the North Island of New Zealand. 1. Patterns of seasonal development

Abstract

AIM: To describe the seasonal pattern of development of third-stage infective larvae (L3) from eggs of Teladorsagia (=Ostertagia) circumcincta, Trichostrongylus colubriformis and Haemonchus contortus on pasture in the North Island of New Zealand.

METHODS: Sheep faeces containing known numbers of eggs of all three nematode species were deposited on, or buried in, pasture plots at three sites, viz coastal Manawatu, Upper Hutt Valley, and East Cape hill country. Development was measured by recovering L3 from faeces, herbage and soil 28–31 days after deposition on 13–18 occasions, between January 2005 and July 2006. Analysis of the number of larvae recovered used a mixed model including number of eggs deposited, weight of faeces recovered (an assumed indicator of earthworm activity), site, contamination date, and position of deposited faeces, i.e. on the surface or buried.

RESULTS: There was a significant effect of contamination date on development of all three species, with maximum numbers ofL3 developing between late spring (November) and early autumn (March), and minimum numbers in June and July. There were large differences between species, with H. contortus exhibiting a long period (April to October) where development was close to zero, whereas T. circumcincta developed to some extent all year round. Development of T. colubriformis was intermediate between the other two species.

Burying faeces containing nematode eggs increased the number of L3 recovered compared with surface deposition (p≤0.001), although there were a small number of exceptions involving only T. colubriformis. The weight of faeces recovered at harvest, which was assumed to be an indication of earthworm activity, was correlated with the number of L3 recovered for all species (p<0.001). In a separate analysis, earthworms were assumed tohave been active if <5 g faeces remained at harvest. Where this occurred, the number of L3 of T. colubriformis and T.circumcincta recovered was reduced by 56% and 58%, respectively (p<0.001).

CONCLUSIONS: A marked seasonal pattern of development was observed for all three species, with the most larvae developing in spring-early autumn and the least in winter. This seasonal pattern was most pronounced in H. contortus and least obvious in T. circumcincta. Burying faeces containing eggs generally resulted in more L3 being recovered, whilst the apparent activity of earthworms resulted in fewer larvae being recovered.     

Morphological changes in trichostrongylid eggs after treatment with fenbendazole

The influence of daily doses of 0.05–0.1 mg fenbendazole/kg body weight on egg development was tested in twenty-seven lambs artificially infected with Haemonchus contortus plus Trichostrongylus colubriformis or Ostertagia circumcincta, as well as in two additional sheep naturally infected with Nematodirus spp.Atypical eggs were detected before a reduction in the number of eggs. Atypical eggs had blastomeres of different sizes and shapes; knot-like, crater-like or bubble-like. In isolated cases eggs with atypical first larvae were recognizable. Morphologically changed eggs occurred in the reproductive organs of female O. circumcincta worms.None, or only a very small number of third stage larvae, could be cultured from the faeces of treated sheep.     

Pharmacokinetic behaviour and anthelmintic efficacy of 1-n-butyl carbamoyl oxfendazole given by intramuscular injection

Oxfendazole (OFZ) was chemically modified to 1-n-butyl carbamoyl OFZ (C₄-OFZ) in an attempt to improve the solubility of OFZ and enable it to be administered by injection. After intramuscular injection to sheep and cattle, C₄-OFZ was metabolised to OFZ which resulted in higher plasma OFZ concentrations that persisted for a considerably longer period than those observed following administration of OFZ orally. The anthelmintic efficacy of injected C₄-OFZ was tested, in sheep, against strains ofTrichostrongylus colubriformis,Haemonchus contortus andOstertagia circumcincta, which were highly resistant to benzimidazoles. In all cases, the C₄-OFZ treatment showed a significant improvement in efficacy over the conventional oral OFZ drench.     

Duration of Anthelmintic Effect of Three Formulations of Ivermectin (Oral,Injectable and Pour-on) Against Multiple Anthelmintic-Resistant <Emphasis Type="Italic">Haemonchus contortus</Emphasis> in Sheep

We report the results of investigations that were conducted in a sheep flock in Uttaranchal, India where repeated failure of anthelmintic medication was noted. The study revealed that Haemonchus contortus in sheep had developed resistance to benzimidazoles (fenbendazole, mebendazole and albendazole), imidazothiazole (levamisole) and salicylanide (rafoxanide), while it was fully susceptible to avermectins (ivermectin). Further, the suppression of nematode egg output in faeces of sheep naturally infected with multiple anthelmintic-resistant H. contortus following treatment with ivermectin tablet (0.4 mg/kg body weight (bw), orally), ivermectin injection (1% w/v, 0.2 mg/kg bw, subcutaneously) and ivermectin pour-on (0.5 w/v, 0.5 mg/kg bw) was also studied over a period of 10 weeks post treatment. It was noted that ivermectin tablet after initial clearance of infection (faecal egg count reduction 100%), could not prevent establishment of new patent natural infection for even a single day, while ivermectin pour-on and injection prevented the establishment of new infection for 7 and 14 days post treatment, respectively. Maximum protection period (duration for which mean faecal egg count of sheep reaches 500 eggs per gram of faeces or more) of 68 days was recorded in sheep treated with injectable ivermectin, followed by pour-on (60 days) and oral (53 days) preparations.     

Farm management practices associated with macrocyclic lactone resistance on sheep farms in New Zealand

AIM: To identify farm practices associated with the presence of resistance to a macrocyclic lactone (ML) anthelmintic on sheep farms in New Zealand.

METHODS: A cross-sectional study was conducted to test for associations between the presence of resistance to an ML anthelmintic (ivermectin) and management practices on sheep farms in New Zealand. Selection of farms was both random (n=80) and purposive (n=32; being farms with a history of suspected ML resistance). Resistance was inferred from faecal nematode egg count (FEC) reduction (FECR) tests (FECRTs) when there was <95% reduction in FEC 7–10 days after treatment with a half dose of ivermectin (0.1 mg/kg). A logistic regression model was built to identify farm-level factors that were associated with the presence or absence of ML resistance.

RESULTS: Of the 112 flock managers that were approached for interview, 103 (92%) returned useable questionnaires. The odds of ML resistance were increased: on farms that had used long-acting ML products in ewes as a pre-lambing treatment for ≥3 of the previous 5 years (odds ratio (OR) = 7.2; 95% confidence interval (CI) = 1.7–30.3); on farms where <70% of the total stock units mid-winter were from sheep (OR=6.5; 95% CI=1.6–25.6); on farms which over the year purchased ≥10% of the number of sheep present mid-winter (OR=7.1; 95% CI=1.5–34.7); and on farms where the average wool diameter of the main flock was <37 (OR=4.1; 95% CI=1.1–14.7) microns. The model provided a good fit to the data (pseudo R²=0.64; Hosmer-Lemeshow statistic = 0.38).

CONCLUSIONS: Explanatory factors identified as associated with the presence of ML (ivermectin) resistance on farms included the use of long-acting anthelmintic formulations in ewes pre-lambing, sources of refugia of unselected parasites on the farm, breed of sheep and their requirements for anthelmintic treatments, and the importing of resistant parasites with purchased stock. The study provides support for controls that aim to provide refugia of susceptible worms and that minimise the risk of introduction of resistance through effective quarantine-drenching.     

Resistance to moxidectin and abamectin in naturally acquired Ostertagia circumcincta infections in sheep

AIM: To investigate the occurrence of resistance to macrocylic lactone (ML) anthelmintics by Ostertagia circumcincta in lambs on a sheep and cattle property in the North Island of New Zea- land.

METHODS: Thirty lambs were randomly allocated to one of five equal-sized groups, consisting of an untreated control and four treatment groups. The treatments, which were administered at the manufacturer's recommended dose rates, included oral moxidectin, oral abamectin (both at 0.2 mg/kg), an albendazole-levamisole combination, and an albendazole-levamisole-ivermectin combination. Post mortem worm counts were undertaken 7 days after treatment to determine the efficacy of each anthelmintic.

RESULTS: The albendazole-levamisole and albendazole-levam-isole-ivermectin combinations both reduced O. circumcincta burdens to zero whereas for moxidectin and abamectin efficacies of only 72% and 29%, respectively, were recorded.

CONCLUSIONS: These results clearly demonstrated the occurrence of resistance to MLs by O. circumcincta. Although this is not the first occasion where resistance to this anthelmintic family has been detected in this parasite in sheep in New Zealand, it is the first instance that resistance to either moxidectin or abamectin has been reported.     

Observations on the emergence of multiple anthelmintic resistance in sheep flocks in the south-east of Scotland

Multiple resistance to benzimidazole, imidazothiazole and macrocyclic lactone anthelmintics is an emerging problem in the south-east of Scotland. The general management and nematode control strategies employed in four affected flocks (flocks 1-4) were investigated in an attempt to identify the risk factors which might have led to the appearance of production limiting disease associated with anthelmintic resistance. The important risk factors for multiple anthelmintic resistance could not be confirmed and it proved easier to criticise nematode control practices on theoretical grounds, than to propose practical solutions. It seems likely that different risk factors were involved in the four flocks. Lambs in flocks 1 and 2 had been treated with an anthelmintic at 3-4 weekly intervals with the aim of achieving suppressive nematode control, while sheep in flock 1 had been treated with an anthelmintic after they were moved onto clean grazing. Recently lambed ewes had been treated with moxidectin in three of the four flocks, with the aim of controlling their periparturient rise in faecal nematode egg output. All of these factors might have contributed to the emergence of multiple anthelmintic resistance, because they could have led to anthelmintic treatments at times when the nematode population in refugia was small, compared to that in the sheep. Annual rotation of the anthelmintic group was compromised by the emergence of benzimidazole resistance and did not prevent the emergence of multiple resistance in any of the flocks described, although the practice may have slowed the development of resistance. Underdosing may have selected for benzimidazole and imidazothiazole resistance in flock 2, associated with inaccurate estimation of the weights of terminal sire lambs. These investigations also highlighted problems associated with the diagnosis of anthelmintic resistance, in particular the confounding effects of the onset of host immunity to nematode parasites, the possible influence of the age of the adult nematode population, and the insensitivity of the undifferentiated faecal egg count reduction test in situations where resistance is emerging.     

Benzimidazole resistance of sheep nematodes in Norway confirmed through controlled efficacy test

Background

Resistance against benzimidazoles (BZ) has recently been detected in Norwegian sheep flocks through a large scale prevalence survey based on the faecal egg count reduction test (FECRT). The use of this test in combination with bulk larval culture only gives an indication of which gastrointestinal nematodes genera that are involved and these results have to be confirmed by a controlled efficacy test (CET) to get accurate information about resistant nematodes populations at species level. A CET was therefore performed with larvae from two flocks where BZ resistance was previously detected through FECRT.

Results

The latter test confirmed the previous results in both flocks. In flock A, the BZ resistant nematode population consisted solely of Haemonchus contortus, whereas H. contortus and Teladorsagia circumcincta comprised the resistant worm population in flock B.

Conclusions

Some discrepancies that have been recorded between FECRT and CET results regarding time for post-treatment coproscopical examination and a temporary suppression of faecal egg excretion are discussed.     

Survey of anthelmintic resistance in Western Australian sheep flocks 2. Relationship with sheep management and parasite control practices

Owners of 116 farms, whose flocks had been tested for anthelmintic resistance, were interviewed to determine their use of various sheep management and parasite control practices and their knowledge and adoption of recommended procedures for the prevention and control of resistance. Farmers knowledge of current recommendations related mainly to changing drenches and drench groups. Other aspects of the recommended program including reduction of drenching frequency and the use of alternative management strategies were not considered as important by farmers. For most questions a high proportion of farmers (greater than 20%) had no opinion. Associations between various strategies for nematode control and resistance of Trichostrongylus and Teladorsagia to thiabendazole and levamisole were examined. These relationships differed between anthelmintics and nematode genera. A number of factors were related to resistance of one or both nematode genera to one or both anthelmintic groups. These factors included flock size, percentage of ewes in the flock, cattle number, main sheep production activity, grazing strategy, frequency of drenching, changes in the frequency of drenching, number of summer drenches and the method of estimating dose rates. It was concluded that the methods employed to control anthelmintic resistance may vary with the nematode, its resistance status and the anthelmintic to which it is exposed. Modifications to the previously recommended program have been proposed which incorporate selection of the anthelmintic to be used following a test for anthelmintic resistance.     

Efficacy of Copper Oxide Needles for the Control of Nematode Parasites in Dairy Goats

The spread of benzimidazole-resistant nematodes in dairy goat farms is of a great concern as probably more than 70% of the flocks are involved. As there are very few other anthelmintic options during the lactating period, we have evaluated the efficacy of copper oxide needles (CON, Copinox, Bayer, UK) in both experimental and natural infections in goats. The curative effect of CON (2–4 g) on existing worm burdens was assessed in goats experimentally infected with Teladorsagia circumcincta, Haemonchus contortus and Trichostrongylus colubriformis, compared to controls. The preventive effect of CON (4 g) on worm establishment was monitored for 2 months in animals experimentally infected with H. contortus and for 3 months in naturally infected animals on a farm exhibiting predominant infections with T. circumcincta and Oesophagostomum venulosum. In both experimental and natural conditions, the efficacy of CON was nil against Teladorsagia, Trichostrongylus and Oesophagostomum infections. In contrast, the efficacy of CON against Haemonchus was clearly established in reducing the worm burden (75%) as well as in lowering the egg output (37–95%) in relation to the establishment of new infections over several weeks. Copper oxide needles may represent an alternative to conventional anthelmintics in the control of Haemonchus infection in some goat farms.     

Traumatic myositis ossificans in a dog

AIM: To determine the distribution of the three common Trichostrongylus spp. infecting sheep and their resistance status on farms throughout New Zealand, using PCR.

METHODS: Cultures were prepared from faecal samples from 70 farms while conducting faecal egg count reduction tests (FECRT) in lambs between 2010 and 2012. Trichostrongylus-type infective stage larvae (L3) were recovered from cultures, derived from untreated control and albendazole-, levamisole- and ivermectin-treated groups of lambs on each of the farms involved, and these were identified to species using PCR analysis of the second internal transcribed spacer region of ribosomal DNA. The species composition of the larvae present in cultures from the untreated control groups was examined across all farms to assess any potential differences in geographical distribution. In addition, the species composition of larvae cultured from the untreated and anthelmintic-treated lamb groups were compared to determine which species exhibited resistance to each of the anthelmintics used in the FECRT.

RESULTS: Of 67 farms with Trichostrongylus spp. present, 42 (63%) cultures from the untreated control groups contained all three Trichostrongylus spp. and no significant geographical patterns in their distribution were detected. Seven samples contained only one species. Irrespective of the anthelmintic efficacy levels, Trichostrongylus colubriformis dominated cultures prepared from lambs following treatment with albendazole (99.1 (95%CI?=?97?100)% of larvae) or levamisole (81.6 (95%CI?=?75.3?87.9)% of larvae), indicating the presence of widespread resistance in this species. In cultures prepared from levamisole-treated lambs, small numbers of T. axei larvae were also frequently present (5.4 (95% CI?=?1.3?12.4)% of larvae). Resistance to ivermectin was not found in any of the three Trichostrongylus spp. after PCR identification. Although larvae were identified, based on length, as being Trichostrongylus spp., for 24 of the 48 samples cultured following treatment with ivermectin, 100% of the larvae present were identified as Teladorsagia circumcincta.

CONCLUSIONS: As in previous surveys, all three Trichostrongylus spp. were common throughout New Zealand and no geographical patterns were detected in the current study. On all farms where resistance to albendazole and/or levamisole was indicated (i.e. efficacy <95%), the species identified as being resistant was T. colubriformis. Even where efficacies were >95%, T. colubriformis still tended to dominate in post-treatment cultures. While this could reflect a lower susceptibility of T. colubriformis to these anthelmintics, it seems more likely to indicate the presence of resistant genotypes in these populations. Similarly, T. axei also tended to be present after treatment with levamisole, which likely reflects a known lower susceptibility of this species to these anthelmintics.     

Fractional protein synthesis rate and polyamine concentrations in tissues of Merino sheep selected for gastrointestinal nematode resistance

Selection of sheep for resistance to gastrointestinal nematodes has been considered as one option to control nematode infection. It is essential to understand the influence of selection on the diversification of protein metabolism in individual tissues. In this experiment we examined fractional protein synthesis rate (FSR) and polyamine (putrescine, spermidine and spermine) and histamine concentrations in tissues of Merino sheep selected for low faecal worm egg count (parasite resistance) and control sheep. Sixty 18-month-old Merino rams, 40 from the resistant flock and 20 from the control line, were kept in individual pens of an animal house and fed ad libitum. Half of each genotype group were dosed orally with infective larvae of Trichostrongylus colubriformis and Ostertagia circumcincta thrice weekly to deliver 10,000 larvae from each species per week for 18 weeks. At the end of week 18, FSR was measured using a flooding dose of l-[ring-d₅]phenylalanine, and polyamine concentrations were determined in all the sheep. Regardless of sheep genotype, the nematode infection significantly increased FSR in the mesenteric lymph nodes (37.0% vs. 32.6%/day, P < 0.01) and abomasal lymph nodes (28.8% vs. 16.0%/day, P < 0.001) and the bone marrow (5.12% vs. 3.96%/day, P < 0.01), but reduced FSR in skin (17.4% vs. 20.6%/day, P < 0.001), muscle (2.46% vs. 2.73%/day, P < 0.05) and liver (24.1% vs. 25.8%/day, P < 0.05). Nematode infection did not affect FSR in the jejunum. Polyamine concentrations in the tissues were generally increased by the infection (P < 0.05) except for a decline in spermine concentration in the muscle (P < 0.01). Overall there was no significant effect of selection for nematode resistance on tissue FSR or the concentration of amines. It was concluded that nematode infection resulted in a divergence of protein synthesis between tissues, and selection for nematode resistance in Merino sheep did not result in significant change in tissue protein synthesis compared with control sheep.     

Development and spatial distribution of the free-living stages of Teladorsagia circumcincta and Trichostrongylus colubriformis on pasture: A pilot study

Abstract

AIMS: To measure the development of Teladorsagia (=Ostertagia) circumcincta and Trichostrongylus colubriformis eggs to third-stage infective larvae (L3) at different times of the year. Also, to measure the spatial distribution of L3 across herbage, soil and faeces, in order to assess whether spatial issues could be important in larval dynamics on pasture.

METHODS: Field plots were contaminated with sheep faeces containing approximately 20,000 eggs of each of T. circumcincta and T. colubriformis on five separate occasions, viz 01 December 1996 (summer), 18 March 1998 (autumn), 17 June 1998 (winter), 15 October 1998 (spring), and 23 July 1999 (winter). Replicate plots (n=10) were harvested at intervals for up to 12 months after deposition of faeces, and the number and distribution of L3 were measured. Larvae were sampled from faeces (where these remained), herbage, and three soil zones to a depth of 145 mm.

RESULTS: There were large differences between contamination dates in the percentage of eggs that developed to L3. For both species the highest percentage development was for eggs deposited in December (7.8% and 25.9% for T. circumcincta and T. colubriformis, respectively) and the lowest for June (0.4% and 0.03% T. circumcincta and T. colubriformis, respectively). Development in winter was often delayed, and this was always associated with a low yield of larvae, probably due to compounding mortalities associated with long periods of exposure to low temperatures.

The relative distribution of L3 present on herbage, in faeces or in the soil varied between sampling times. However, overall the most L3 were recovered from soil (74% and 66% for T.circumcincta and T. colubriformis, respectively, averaged over all samples), and the lowest recoveries were from the herbage.

CONCLUSIONS: Although the data are limited, the results indicated that the highest percentage of eggs developed to infective larvae in summer and only minimal development occurred in winter. The data do not support the view that substantial contamination of pastures with sheep parasites occurs over winter. Large numbers of larvae were recovered from soil, which indicates that, assuming they can subsequently migrate onto herbage, soil is a potentially important reservoir ofinfective larvae in New Zealand. Therefore, the spatial distribution of L3 on pasture may affect both the dynamics and transmission of parasite populations. Further work on both these issues is warranted.     

Managing anthelmintic resistance: untreated adult ewes as a source of unselected parasites, and their role in reducing parasite populations

AIMS: To test the hypotheses that when untreated adult ewes are rotationally grazed (follow behind) on pastures after lambs receiving routine anthelmintic treatments, the ewes can function as a source of unselected parasites in refugia, capable of slowing the development of anthelmintic resistance, and suppress the build-up of parasites resulting from the development of anthelmintic resistance. METHODS: Firstly, the potential of untreated adult ewes to slow the development of anthelmintic resistance, and to suppress parasite populations under differing levels of anthelmintic efficacy, was investigated using a simulation model. Secondly, a field trial with three replicates of each treatment compared two grazing systems (lambs only vs lambs followed by ewes) and two types of anthelmintic, viz albendazole (ALB), to which resistance was present (faecal nematode egg count reduction (FECR)=57-59%) and ivermectin plus levamisole (IL), to which resistance was absent (FECR=97-99%), in a factorial treatment structure. Parasite populations were monitored using faecal nematode egg counts (FEC), faecal larval cultures, pasture larval sampling, and slaughter of tracer lambs. Animal performance was measured using liveweight, dag score, body condition score, and fleece weights. RESULTS: Model simulations indicated that parasites cycling in the untreated ewes could slow the development of resistance being selected for by the anthelmintic treatments given to lambs and this could occur without a nett increase in larval numbers on pasture. Further, as worm control in the lambs declined with increasing levels of anthelmintic resistance the ewes increasingly functioned as nett removers of parasite larvae, effectively reducing parasite population size. In the field trial, untreated adult ewes contributed to pasture infestations of most parasite species, but not Nematodirus spp. Parasite species on pasture and infecting lambs changed when ewes were present, but larval populations on pasture in the autumn were no greater than when lambs grazed alone. In the presence of anthelmintic resistance, parasite populations were reduced when ewes grazed in rotation with lambs, implicating the ewes as nett removers of parasite challenge. CONCLUSIONS: Untreated adult ewes were a source of unselected genotypes, capable of slowing the development of anthelmintic resistance in most, but not all, parasite species. Further, the potential of adult ewes to remove from pasture more parasite larvae than they contribute through faecal contamination indicates a potentially useful role in suppressing parasite populations, particularly when worm control in lambs is less effective as a result of anthelmintic resistance.     

Anthelmintic resistance survey in New Zealand

The prevalence and level of thiabendazole resistance of sheep nematodes in the North Island of New Zealand was investigated by means of an in vitro egg-hatch technique; samples from sheep on 52 properties in six country areas were examined. Resistance of Haemonchus contortus and Trichostrongylus spp. was identified on the basis of egg hatch in 0.1 ppm thiabendazole. Evidence of resistance was obtained on 11 (21.2%) of the properties. LD 50's and resistance ratios were calculated for the resistant nematode populations found.     

Anthelmintic resistance of nematode parasites of small ruminants in eastern Ethiopia: exploitation of refugia to restore anthelmintic efficacy

Faecal egg count reduction tests (FECRT) were conducted in May 2003 to determine the efficacy of anthelmintics used for treatment against nematode parasites in separately managed sheep and goat flocks at Alemaya University in eastern Ethiopia. These tests revealed high levels of anthelmintic resistance to albendazole, tetramisole, the combination of these two drugs, and to ivermectin in the goat flock (predominantly infected by Haemonchus contortus and Trichostrongylus spp.), whereas all drugs were highly efficacious in the sheep flock. A second FECRT confirmed these observations. Following this, a new management system was implemented on the goat flock for a period of 9 months (January-September 2004) in an attempt to restore the anthelmintic efficacy. This involved a combination of measures: eliminating the existing parasite infections in the goats, exclusion from the traditional goat pastures, and introducing communal grazing of the goats with the university sheep flock and livestock owned by neighbouring small-holder farmers. A second series of FECRTs (Tests 3 and 4) conducted 7 months after this change in management, showed high levels of efficacy to all three drugs (albendazole, tetramisole and ivermectin) in the goat flock. This is the first field study to demonstrate that anthelmintic efficacy in the control of nematode parasites of small ruminants can be restored by exploiting refugia.