Anthelmintic resistance of nematodes in communally grazed goats in a semi-arid area of South Africa

A survey was conducted on the occurrence of anthelmintic resistance of nematodes in communally grazed goats in a semi-arid area in South Africa. In herds belonging to 10 small-holder goat farmers, the efficacies of fenbendazole, levamisole and rafoxanide were tested by faecal egg count reduction (FECR) tests. Efficacies of 80% were considered a threshold for anthelmintic resistance. The FECR tests showed that all drugs tested more than 80% effective in most instances, but there were notable exceptions. In 1 case, rafoxanide was only 31% effective and in another case fenbendazole was only 47% effective. The occurrence of anthelmintic resistance in this farming sector is of concern. Steps should be taken to prevent its further spread and to avoid the development of a situation as on numerous commercial sheep farms in South Africa where resistance is very common.     

Detecting benzimidazole resistance with faecal egg count reduction tests and in vitro assays

Composite strains of Trichostrongylus colubriformis and Ostertagia spp consisting of 0, 1, 10, 25, 50, 75, 90, and 100% of known resistant strains were prepared and tested for benzimidazole resistance using faecal egg count reduction tests, in vitro egg hatch assays and tubulin binding assays. All tests detected resistance where the proportion of the resistant strain in the composite was 50% or more, whereas none of the tests unequivocally detected resistance below 25%. Egg count reduction tests were no less sensitive than the in vitro tests in detecting low levels of resistance but the egg hatch and tubulin binding assays provided a better quantitative estimate of moderate to high levels of resistance. Faecal egg count reduction therefore, provides a suitable means of detecting resistance in the field but tests, more sensitive to low levels of resistance are required. Results indicate that the use of post-treatment counts alone provides an adequate indication of anthelmintic efficiency.     

A preliminary undifferentiated faecal egg count reduction test survey in the Caledon area

During October 1998 a study was performed in the Caledon area to determine the presence of anthelmintic resistance in the southern Western Cape, which falls within the winter rainfall region of South Africa. The study took the form of an undifferentiated faecal egg count reduction test (FECRt) survey. The predominant worm genera of the region are Teladorsagia and Trichostrongylus, but Haemonchus causes occasional outbreaks in sheep. No resistance data for any area in the Western Cape Province are available at present. Resistance (< 90% FECR) was recorded on 73% of the farms included in the study, with 46% of these involving resistance to 1 drench, 36% to 2 drenches and 18% to 3 drenches. No moxidectin resistance was detected when it was administered at the therapeutic dosage of 0.2 mg/kg. The results of this study show clearly that resistance of nematodes to other anthelmintics occurs in the Caledon area.     

Potential limitations of the undifferentiated faecal egg count reduction test for the detection of anthelmintic resistance in sheep

Larval cultures were used to determine the identities and occurrences of those parasites (excluding Nematodirus) represented by strongylid eggs at the time of anthehnintic administration in ovine faecal egg count reduction tests submitted to the Batchelar Animal Health Laboratory between 1992 and 1994. The numbers of individual nematode genera recorded in pre-treatment samples from these cases ranged from one to five and included infections of Haemonchus, Ostertagia, Ttichostrongylus, Cooperia and Oesophagostomum/Chabertia. Adequate egg representation for testing purposes by all five genera simultaneously was found to occur in only 17 (10%) of the 163 cases examined, with the majority (71%) of them containing between one and three nematode genera. The greatest representation occurred in those tests conducted during the months of February to May. However, even during this period, worm eggs of all five genera were concurrently present on only 16% of occasions. The importance of knowing what nematode genera are adequately represented at the time of routine faecal egg count reduction testing and the relevance of this information to reducing the likelihood of being misled when under- taking assessments of farm resistance status are discussed.     

The detection of resistance to ivermectin by the faecal egg count reduction test

Extract

Perception of, and relief from, pain differs markedly between individual animals⁽¹⁾. In a humane system of animal care, this requires addressing. I have investigated the analgesic effects of different opioid agonists in male and female sheep, and the results suggest a sex-related difference in the efficacy of both kappa (𝛋) and mu (𝛍) opioids.     

A comparison of in vitro tests and a faecal egg count reduction test in detecting anthelmintic resistance in horse strongyles.

This study reports a comparison between faecal egg count reduction test (FECRT), egg hatch assay (EHA) and larval development assay (LDA) for detecting anthelmintic resistance in equine strongyles. Resistance to benzimidazoles was demonstrated in 33 of 42 (79%) farms tested by FECRT and in 32 (62%) of the 52 farms tested by EHA. As the reference strain used was not fully susceptible to benzimidazoles it was not possible to determine the level of resistance by LDA. Pyrantel resistance was indicated on three of 15 farms by faecal egg count reduction. Resistance was also indicated by LDA for one of these farms. In addition resistance was indicated by LDA on two more farms that were not tested by FECRT. Further testing is needed to confirm if these findings are truly indicative of resistance. Generally, correlations between the tests were poor and it was not possible to use the outcome of one test to predict the outcome of another.     

Further potential limitations of the undifferentiated faecal egg count reduction test for the detection of anthelmintic resistance in sheep

Fifteen (36%) of 42 mixed gastro-intestinal nematode infections in sheep, identified as drench-susceptible by the undifferentiated faecal egg count reduction test, were found to harbour anthelmintic-resistant worms when analysed on the basis of changes in the egg counts of individual nematode genera. Most of these cases involved resistance in a single nematode genus, with Ostertagia and Trichostrongylus being implicated most frequently. The possible contributory role of larval culture results in helping to reduce the chances of the faecal egg count reduction test producing these and similar types of errors is discussed.     

A small scale survey of ivermectin resistance in sheep nematodes using the faecal egg count reduction test on samples collected from Scottish sheep

Thirty-eight sheep flocks, predominantly from the south/central Scotland, were examined using a faecal egg count reduction test (FECRT) for the presence of ivermectin (IVM) resistant nematodes. Efficacies of less than 95%, 14-17 days post-treatment, were identified in 6 of 17 naturally grazing flocks where pre-treatment faecal egg counts were in excess of 150 eggs per gram. Efficacies on these IVM resistant farms ranged from 66 to 92%. One other suspected cases of IVM resistance was also identified in returned material. The larvae detected in post-treatment coprocultures from resistant flocks were from the genera Teladorsagia (4 from 6) and Trichostrongylus (2 from 6).     

Benefits of urea-molasses block supplementation and symptomatic and tactical anthelmintic treatments of communally grazed indigenous goats in the Bulwer area, Kwazulu-Natal Province, South Africa

This study was carried out with the cooperation of farmers owning communally grazed indigenous goats in southwestern KwaZulu-Natal Province, South Africa, where farmers had identified poor reproductive performance in their herds as one of their major problems. The aim was to quantify the effects of 3 interventions and the interaction between these interventions on goat productivity and gastrointestinal nematode infection. The interventions were: urea-molasses block supplementation during the dry winter seasons of 2004 and 2005, tactical anthelmintic treatment with ivermectin (400 microg/kg) during the wet summer period (on 3 January 2005) and symptomatic treatment with ivermectin (400 microg/kg) of all goats judged anaemic throughout the entire study period. The FAMACHA system was used as a gauge of anaemia. It was noted that goats considered anaemic tended to remain so throughout the study period. The tactical anthelmintic treatment was effective as it markedly reduced (P = 0.066) the summer peak in faecal egg counts and is therefore recommended. By contrast, while the urea-molasses block supplementation appeared to reduce the faecal egg counts immediately following the 2004 supplementation (P < 0.05), this did not hold true in 2005. Interestingly, in the tactically treated anaemic goats, the improvement in the number of kids suckled per doe year-on-year tended to be greater than in the non-anaemic goats. It is considered that the routine symptomatic treatment of anaemic goats may have been a key factor. More detailed investigations into the routine symptomatic treatment of anaemic goats are therefore recommended.     

A survey of seasonal changes in nematode faecal egg count levels of sheep and goats in Senegal

A survey of the gastro-intestinal nematodes in domestic sheep and goats of the Sahelian zone of Senegal was undertaken. Their prevalence and the egg production were estimated by faecal egg count in 2234 sheep and 577 goats slaughtered at the Dakar slaughterhouse during a 12 month period. The highest prevalence (%) and highest mean egg counts (EPG) were recorded at the end of the wet season. Amongst the species of nematodes encountered there are two of potential importance: Haemonchus contortus and Oesophagostomum columbianum. It is suggested that the most effective control can be achieved by a critical treatment at the end of dry season (May–June) when the EPG starts to increase, and when hypobiosis is supposed to be terminating. This would insure the greatest proportion of the existing worm population to be exposed to the anthelmintic.     

Preserving new anthelmintics: a simple method for estimating faecal egg count reduction test (FECRT) confidence limits when efficacy and/or nematode aggregation is high

As it has been 30 years since a new anthelmintic class was released, it is appropriate to review management practices aimed at slowing the development of anthelmintic resistance to all drug classes. Recommendations to delay anthelmintic resistance, provide refugia and the use of a simulation model were reviewed to find optimum treatment strategies that maintain nematode control. Simulated Australian conditions indicated that a common successful low-risk treatment program was a rapid rotation between a "triple-combination" product (benzimidazole+levamisole+abamectin) and a new high-efficacy drug (monepantel). Where Haemonchus contortus was a threat, moxidectin was required at critical times because of its persistent activity against this parasite. Leaving up to 4% of adult sheep untreated provided sufficient "refugia" for non-selected worms to reduce the risk of selecting for anthelmintic resistance without compromising nematode control. For a new anthelmintic, efficacy estimated by faecal egg count reduction (FECR) is likely to be at or close to 100%, however using current methods the 95% confidence limits (CL) for 100% are incorrectly determined as 100%. The fewer eggs counted pre-treatment, the more likely an estimate of 100% will occur, particularly if the true efficacy is >90%. A novel way to determine the lower-CL (LCL) for 100% efficacy is to reframe FECR as a binomial proportion, i.e. define: n and x as the total number of eggs counted (rather than eggs per gram of faeces) for all pre-treatment and post-treatment animals, respectively; p the proportion of resistant eggs is p = x/n and percent efficacy is 100 ×(1-p) (assuming equal treatment group sizes and detection levels, pre- and post-treatment). The LCL is approximated from the cumulative inverse beta distribution by: 95%LCL=100 ×(1-(BETAINV(0.975, x+1, n-x+1))). This method is simpler than the current method, independent of the number of animals tested, and demonstrates that for 100% efficacy at least 37 eggs (not eggs per gram) need to be counted pre-treatment before the LCL can exceed 90%. When nematode aggregation is high, this method can be usefully applied to efficacy estimates lower than 100%, and in this case the 95% upper-CL (UCL) can be estimated by: 95% UCL = 100 ×(1((BETAINV(0.025, x+1, n-x+1))), with the LCL approximated as described above. A simulation study to estimate the precision and accuracy of this method found that the more conservative 99%CL was optimum; in this case 0.975 and 0.025 are replaced by 0.995 and 0.005 to estimate the LCL and UCL, respectively.     

Periparturient rise in faecal helminth egg counts of Udah sheep in the Zaria area of Nigeria

The trichostrongylid egg output in the faeces of seven lambing and seven barren Udah ewes was compared during the beginning of the rainy season in 1974. Both groups showed an increase in egg counts which reached a considerably higher level in the lambing ewes. The highest level, a mean count of 2 650 eggs/g, was reached 2 weeks after lambing.It is suggested that the increase was due to helminths, mainly Haemonchus sp., which had been inhibited during the dry season and resumed their development at the beginning of the rainy season.The rapid decline in egg counts recorded 5–6 weeks after lambing was considered to be due to self cure associated with high rainfall.     

Comparison of two assays, a faecal egg count reduction test (FECRT) and a coproantigen reduction test (CRT), for the diagnosis of resistance to triclabendazole in Fasciola hepatica in sheep

A sheep trial was performed to evaluate two diagnostic assays, a faecal egg count reduction test (FECRT) and a coproantigen reduction test (CRT), for the diagnosis of resistance of Fasciola hepatica to triclabendazole (TCBZ). The FECRT defines successful TCBZ treatment as a 95% or greater reduction in fluke faecal egg counts (FECs) at 14 days post-treatment (dpt). The CRT defines effective TCBZ treatment as faeces negative for Fasciola coproantigens at 14dpt, as measured by the commercial BIO K201 coproantigen ELISA (Bio-X Diagnostics, Jemelle, Belgium). Forty-nine indoor-reared sheep were split into four trial groups and each sheep was infected with 200 metacercariae of 1 of 4 F. hepatica isolates, previously described as susceptible (Cullompton and Fairhurst) and resistant (Leon and Oberon) to TCBZ action, respectively. TCBZ treatment was administered at 12 weeks post-infection (wpi) to one sub-group in each infected sheep group, and these sheep were culled at 4 weeks post-treatment (wpt). Untreated sheep sub-groups, were culled at a parallel time-point, that is, at 16wpi. Necropsy was performed to confirm treatment efficacy. Individual faecal samples were collected twice-weekly throughout the trial period, sub-sampled and examined by a standardised egg sedimentation protocol and by the BIO K201 ELISA. Results supported the use of both the FECRT and the CRT for the diagnosis of resistance of F. hepatica to TCBZ. In addition, the study confirmed the TCBZ susceptibility of the Cullompton and Fairhurst F. hepatica isolates and the TCBZ resistance of the Oberon F. hepatica isolate. However, the Leon F. hepatica isolate was found to be susceptible, rather than resistant, to TCBZ action.     

Effect of nematode burden as assessed by means of faecal egg counts on body condition in goats farmed under resource-poor conditions in South Africa

A longitudinal study was conducted of the nematode faecal egg counts (FECs) and body condition scores (BCSs) of goats of resource-poor farmers at Rust de Winter, Gauteng Province, Impendle, KwaZulu-Natal Province, and Kraaipan, North-West Province, South Africa. Periods of higher FECs occurred from December/January to March/April at Rust de Winter and at Impendle and from January to March at Kraaipan. Seasonal variations in body condition were evident in the goats at Impendle with the animals showing lower BCSs from June to October. The goats at Rust de Winter and at Kraaipan did not show clear seasonal variations, although the goats at Rust de Winter showed lower BCSs from mid-July to early December. The BCSs for Rust de Winter where the animals were grazed on a private farm were generally higher than those of the other sites, where communal grazing is practised.     

Estimates of genetic parameters for faecal egg count of Haemonchus contortus infection and relationship with growth traits in Avikalin sheep

Genetic parameters for faecal egg count were estimated in naturally challenged Avikalin sheep developed and maintained at Central Sheep & Wool Research Institute, Avikanagar, India, over a period of 4 years (2004–2007). The data on faecal egg count for 433 animals descended from 41 sires, and 151 dams were used for the study. Genetic analyses were carried out using restricted maximum likelihood, fitting an animal model and ignoring or including maternal genetic or permanent environmental effects. Direct heritability for the trait was 0.149 ± 0.096 when maternal effects were ignored. In the model which takes in to account direct genetic, maternal genetic and maternal permanent environment effect together, it was observed that maternal heritability (m²) accounts for 0.6% of total variation whereas maternal permanent environmental effect (c²) accounts for 6.14% of total phenotypic variation. Effect of faecal egg count on the growth characteristics was observed to be significant. It was seen that wherever FEC was high, body weight or average daily gain declined in active infective stage. After termination of the infection, these effects were found to be non-significant. Result suggests that direct genetic and maternal permanent environmental effects were important for this trait; thus, they need to be considered for improvement in the trait.     

Novel insights in the faecal egg count reduction test for monitoring drug efficacy against gastrointestinal nematodes of veterinary importance

The faecal egg count reduction test (FECRT) is the method of choice to monitor anthelmintic efficacy against gastro-intestinal nematodes in livestock. Guidelines on how to conduct a FECRT are made available by the World Association for the Advancement of Veterinary Parasitology (WAAVP). Since the publication of these guidelines in the early 1990 s, some limitations have been noted, including (i) the ignorance of host-parasite interactions that depend on animal and parasite species, (ii) their feasibility under field conditions, (iii) appropriateness of study design, and (iv) the high detection limit of the recommended faecal egg count (FEC) method. Therefore, the objective of the present study was to empirically assess the impact of the level of excretion and aggregation of FEC, sample size and detection limit of the FEC method on the sensitivity and specificity of the FECRT to detect reduced efficacy (<90% or <95%) and to develop recommendations for surveys on anthelmintic resistance. A simulation study was performed in which the FECRT (based on the arithmetic mean of grouped FEC of the same animals before and after drug administration) was conducted under varying conditions of mean FEC, aggregation of FEC (inversely correlated with k), sample size, detection limit and 'true' drug efficacies. Classification trees were built to explore the impact of the above factors on the sensitivity and specificity of detecting a truly reduced efficacy. For a reduced-efficacy threshold of 90%, most combinations resulted in a reliable detection of reduced and normal efficacy. For the reduced-efficacy threshold of 95% however, unreliable FECRT results were found when sample sizes <15 were combined with highly aggregated FEC (k=0.25) and detection limits ≥ 5 EPG or when combined with detection limits ≥ 15 EPG. Overall, an increase in sample size and mean preDA FEC, and a decrease in detection limit improved the diagnostic accuracy. FECRT remained inconclusive under any evaluated condition for drug efficacies ranging from 87.5% to 92.5% for a reduced-efficacy-threshold of 90% and from 92.5% to 97.5% for a threshold of 95%. The results highlight that (i) the interpretation of this FECRT is affected by a complex interplay of factors, including the level of excretion and aggregation of FEC and (ii) the diagnostic value of FECRT to detect small reductions in efficacy is limited. This study, therefore, provides a framework allowing researchers to adapt their study design according to a wide range of field conditions, while ensuring a good diagnostic performance of the FECRT.     

Natural infection of land-snails by protostrongylids on a pasture grazed by sheep in the Rabat area of Morocco

The output by sheep of first-stage protostrongylid larvae, and the population density and level of infection of terrestrial molluscs were assessed monthly during a 1-year study. Limax maximus was the most infected mollusc. Juveniles were less infected than adults, for Cernuella virgata and Theba pisana; the reverse was observed for Otala lactea. Banded individuals had a lower intensity of infection than the non-banded ones for C. virgata and T. pisana. Sixty-one percent of protostrongylid larvae in molluscs on the pastures studied were found in C. virgata, and 25% in O. lactea. The infectivity of the pasture for sheep was related to rainfall.