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Background: Veterinary diagnostics aid intervention strategies, track zoonoses, and direct selective breeding programs in livestock. In ruminants, gastrointestinal nematode (GIN) parasites are a major cause of production losses, but morphologically similar species limit our understanding of how specific GIN co-infections impact health in resource-limited settings. To estimate the presence and relative abundance of GINs and other helminths at the species level, we sought to develop a low-cost and low-resource molecular toolkit applied to goats from rural Malawi smallholdings. Methods: Goats were subjected to health scoring and faecal sampling on smallholdings in Lilongwe district, Malawi. Infection intensities were estimated by faecal nematode egg counts with a faecal subsample desiccated for DNA analysis. Two DNA extraction methods were tested (low-resource magbead kit vs high-resource spin-column kit), with resulting DNA screened by endpoint polymerase chain reaction (PCR), semi-quantitative PCR, quantitative PCR (qPCR), high-resolution melt curve analysis (HRMC), and ‘nemabiome’ internal transcribed spacer 2 (ITS-2) amplicon sequencing. Results: Both DNA isolation methods yielded comparable results despite poorer DNA purity and faecal contaminant carryover from the low-resource magbead method. GINs were detected in 100% of samples regardless of infection intensity. Co-infections with GINs and coccidia (Eimeria spp.) were present in most goats, with GIN populations dominated by Haemonchus contortus, Trichostrongylus colubriformis, Trichostrongylus axei, and Oesophagostomum columbianum. Both multiplex PCR and qPCR were highly predictive of GIN species proportions obtained using nemabiome amplicon sequencing; however, HRMC was less reliable than PCR in predicting the presence of particular species. Conclusions: These data represent the first ‘nemabiome’ sequencing of GINs from naturally infected smallholder goats in Africa and show the variable nature of GIN co-infections between individual animals. A similar level of granularity was detected by semi-quantitative PCR methods, which provided an accurate summary of species composition. Assessing GIN co-infections is therefore possible using cost-efficient low-resource DNA extraction and PCR approaches that can increase the capacity of molecular resources in areas where sequencing platforms are not available; and also open the door to affordable molecular GIN diagnostics. Given the diverse nature of infections in livestock and wildlife, these approaches have potential for disease surveillance in other areas.

Additional file 8. Figures S1–S5. Figure S1: Assessing quality of DNA extraction methods. Figure S2: Assessing GIN species detection likelihoods by DNA extraction method and infection intensity. Figure S3: High-resolution melt curves with pan-nematode ITS-2 primers. Figure S4: Nemabiome ITS-2 amplicon with the DADA2 IDTAXA pipeline. Figure S5: Cross-validation of PCR methods with nemabiome using the DADA2 IDTAXA pipeline.

Increasing anthelmintic resistance (AR) in livestock has stimulated growing efforts to monitor anthelmintic effectiveness (AE) on livestock farms. On-farm assessment of AE relies on measuring the reduction in faecal egg count (FEC) following treatment; and if conducted rigorously, qualifies as a formal FEC reduction test (FECRT) for AR. Substantial research effort has been devoted to designing robust protocols for the FECRT and its statistical interpretation; however, a wide range of factors other than AR can affect FEC reduction on farms. These are not always possible to control, and can affect the outcome and repeatability of AE measurements and confound the on-farm classification of AR using FECRT. This review considers confounders of FEC reduction, focusing on gastrointestinal nematodes of ruminants, including host and parasite physiology and demography; pharmacokinetic variation between drugs, parasites and hosts; and technical performance. Drug formulation and delivery, host condition and diet, and seasonal variation in parasite species composition, can all affect AE and hence observed FEC reduction. Causes of variation in FEC reduction should be attenuated, but this is not always possible. Regular monitoring of AE can indicate a need to improve anthelmintic administration practices, and detect AR early in its progression. Careful interpretation of FEC reduction, however, taking into account possible confounders, is essential before attributing reduced FEC reduction to AR. Understanding of confounders of FEC reduction will complement advances in FECRT design and interpretation to provide measures of anthelmintic efficacy that are both rigorous and accessible.Facteurs de confusion affectant la réduction du nombre d’œufs fécaux comme mesure de l’efficacité anthelminthique.L’augmentation de la résistance aux anthelminthiques (RA) chez le bétail a stimulé des efforts croissants pour mesurer l’efficacité des anthelminthiques (EA) dans les élevages. L’évaluation à la ferme de l’EA repose sur la mesure de la réduction du nombre d’œufs dans les selles (NOS) après le traitement ; si effectué rigoureusement, celui-ci est qualifié de test formel de réduction du NOS (TFRNOS) pour la RA. Des efforts de recherche substantiels ont été consacrés à la conception de protocoles robustes pour le TFRNOS et son interprétation statistique, mais un large éventail de facteurs autres que la RA peuvent affecter la réduction du NOS dans les exploitations. Ces facteurs ne sont pas toujours contrôlables et peuvent affecter le résultat et la répétabilité des mesures de l’EA et introduire de la confusion dans la classification à la ferme de la RA à l’aide de la TFRNOS. Cette revue examine les facteurs de confusion de la réduction du NOS, en se concentrant sur les nématodes gastro-intestinaux des ruminants, et considère la physiologie et la démographie de l’hôte et du parasite, la variation pharmacocinétique entre les médicaments, les parasites et les hôtes, et les performances techniques. La formulation et l’administration de médicaments, la condition et le régime alimentaire de l’hôte, ainsi que les variations saisonnières de la composition des espèces de parasites, peuvent tous affecter l’EA et donc la réduction du NOS observé. Les causes de variation de la réduction du NOS doivent être atténuées, mais cela n’est pas toujours possible. Une surveillance régulière de l’EA peut indiquer la nécessité d’améliorer les pratiques d’administration des anthelminthiques et de détecter la RA au début de sa progression. Une interprétation prudente de la réduction du NOS, cependant, en tenant compte des facteurs de confusion possibles, est essentielle avant d’attribuer la réduction du NOS à la RA. La compréhension des facteurs de confusion d’une moindre réduction du NOS complétera les progrès sur la conception et l’interprétation du TFRNOS, pour fournir des mesures de l’efficacité des anthelminthiques à la fois rigoureuses et accessibles.

Angiostrongylusvasorum is a helminth parasite of domestic dogs that is increasing in range and prevalence. Its lifecycle requires terrestrial gastropod mollusc (“gastropod”) intermediate hosts, but research is lacking regarding contact risk in situ. We studied co-occurrence between dogs and gastropods in dog-walking spaces in an A. vasorum hotspot in southern England, United Kingdom, with the aim of quantifying environmental and spatio-temporal overlap. We surveyed 390 quadrats and 180 point-counts along 3 km transects at seven sites, yielding 1672 gastropod and 763 dog observations. Common gastropods comprised Arion, Cornu, Monacha, Deroceras, Tandonia, Cochlicella, and Trochulus species. Habitat was the most important factor structuring both gastropod and dog presence and abundance. Likelihood ratio comparisons from conditional probability trees revealed that dogs were 15× more likely to be present on hardstanding surfaces than other habitats but were also present on natural and amenity grassland. Presence of gastropod species associated with high A. vasorum prevalence was 65.12× more likely in woodland/scrub and 62.17× more likely in amenity grassland than other habitats. For gastropods overall, high abundance was 5.82× more likely in woodland/scrub and natural grassland. The findings suggest co-occurrence is highest in amenity and natural grassland, but infection risk is greatest in amenity grassland and woodland/scrub.

Additional file 2: Text S1. Validation of grass L3 recovery and fecal L3 recovery protocols. Recovery rates were calculated by spiking parasite-free grass and fecal samples with around 1000 L3s.

Additional file 1: Table S1. Results of S. mansoni and S. haematobium qPCR experiments, including Cq scores and eDNA copies for all replicates in the experiment.

Helminth infections are ubiquitous in grazing ruminant production systems, and are responsible for significant costs and production losses. Anthelmintic Resistance (AR) in parasites is now widespread throughout Europe, although there are still gaps in our knowledge in some regions and countries. AR is a major threat to the sustainability of modern ruminant livestock production, resulting in reduced productivity, compromised animal health and welfare, and increased greenhouse gas emissions through increased parasitism and farm inputs. A better understanding of the extent of AR in Europe is needed to develop and advocate more sustainable parasite control approaches. A database of European published and unpublished AR research on gastrointestinal nematodes (GIN) and liver fluke (Fasciola hepatica) was collated by members of the European COST Action "COMBAR" (Combatting Anthelmintic Resistance in Ruminants), and combined with data from a previous systematic review of AR in GIN. A total of 197 publications on AR in GIN were available for analysis, representing 535 studies in 22 countries and spanning the period 1980-2020. Reports of AR were present throughout the European continent and some reports indicated high within-country prevalence. Heuristic sample size-weighted estimates of European AR prevalence over the whole study period, stratified by anthelmintic class, varied between 0 and 48%. Estimated regional (country) prevalence was highly heterogeneous, ranging between 0% and 100% depending on livestock sector and anthelmintic class, and generally increased with increasing research effort in a country. In the few countries with adequate longitudinal data, there was a tendency towards increasing AR over time for all anthelmintic classes in GIN: aggregated results in sheep and goats since 2010 reveal an average prevalence of resistance to benzimidazoles (BZ) of 86%, macrocyclic lactones except moxidectin (ML) 52%, levamisole (LEV) 48%, and moxidectin (MOX) 21%. All major GIN genera survived treatment in various studies. In cattle, prevalence of AR varied between anthelmintic classes from 0-100% (BZ and ML), 0-17% (LEV) and 0-73% (MOX), and both Cooperia and Ostertagia survived treatment. Suspected AR in F. hepatica was reported in 21 studies spanning 6 countries. For GIN and particularly F. hepatica, there was a bias towards preferential sampling of individual farms with suspected AR, and research effort was biased towards Western Europe and particularly the United Kingdom. Ongoing capture of future results in the live database, efforts to avoid bias in farm recruitment, more accurate tests for AR, and stronger appreciation of the importance of AR among the agricultural industry and policy makers, will support more sophisticated analyses of factors contributing to AR and effective strategies to slow its spread.Importance croissante de la résistance aux anthelminthiques chez les ruminants européens : création et méta-analyse d’une base de données ouverte.Les helminthes sont omniprésents dans les systèmes de production de ruminants au pâturage et sont responsables de coûts et de pertes de production importants. La résistance aux anthelminthiques (RA) des parasites est maintenant répandue dans toute l’Europe, bien qu’il existe encore des lacunes dans nos connaissances dans certaines régions et certains pays. La RA est une menace majeure pour la pérennité de la production moderne de ruminants, en diminuant la productivité, en compromettant la santé et le bien-être des animaux, et en augmentant les émissions de gaz à effet de serre au travers d’une augmentation du parasitisme et des intrants agricoles. Une meilleure compréhension de l’étendue de la RA en Europe est nécessaire pour développer et préconiser des approches de lutte antiparasitaire plus durables. Une base de données intégrant des informations publiées et non publiées en Europe concernant la RA des nématodes gastro-intestinaux (NGI) et de la douve du foie (Fasciola hepatica) a été compilée par les membres de l’action européenne COST « COMBAR » (« Combattre la résistance aux anthelminthiques chez les ruminants ») et combinée avec les données d’une précédente étude systématique concernant la RA des NGI. Au total, 197 publications sur la RA des NGI étaient disponibles pour analyse, représentant 535 études dans 22 pays et couvrant la période 1980–2020. Des signalements de RA étaient présents sur tout le continent européen et certains rapports indiquaient une forte prévalence nationale. Les estimations heuristiques pondérées par la taille de l’échantillon de la prévalence de la RA en Europe sur toute la période d’étude, stratifiées par classe d’anthelminthiques, variaient de 0 à 48 %. La prévalence régionale (nationale) estimée était très hétérogène, variant entre 0 % et 100 % selon le secteur de l’élevage et la classe d’anthelminthique, et augmentait généralement avec les efforts de recherche dans le pays. Dans les quelques pays disposant de données longitudinales adéquates, il y avait une tendance à l’augmentation de la RA au fil du temps pour toutes les classes d’anthelminthiques des NGI : les résultats agrégés chez les ovins et caprins depuis 2010 révèlent une prévalence moyenne de résistance aux benzimidazoles (BZ) de 86 %, aux lactones macrocycliques sauf moxidectine (ML) de 52 %, au lévamisole (LEV) de 48 % et à la moxidectine (MOX) de 21 %. Tous les genres principaux de NGI ont survécu au traitement dans diverses études. Chez les bovins, la prévalence de la RA variait selon les classes d’anthelminthiques de 0 à 100 % (BZ et ML), 0 à 17 % (LEV) et 0 à 73 % (MOX), et Cooperia et Ostertagia ont survécu aux traitements. Une RA suspectée chez F. hepatica a été signalée dans 21 études portant sur 6 pays. Pour les NGI, et encore plus pour F. hepatica, il y avait un biais d’échantillonnage en faveur des exploitations individuelles suspectées de RA, et l’effort de recherche était biaisé vers l’Europe occidentale et en particulier le Royaume-Uni. La capture continue des résultats futurs dans la base de données, en direct, les efforts pour éviter les biais dans le recrutement des exploitations, des tests plus précis pour la RA et une meilleure appréciation de l’importance de la RA parmi l’industrie agricole et les décideurs politiques, soutiendront des analyses plus sophistiquées des facteurs contribuant à la RA, et des stratégies efficaces pour ralentir sa propagation.

Helminth infections, mainly by gastrointestinal nematodes (GIN), are one of the main concerns for animal health, welfare and productivity in grazing ruminant livestock worldwide. The use of a sensitive, precise, accurate, low-cost, and easy-to-perform copromicroscopic technique is of pivotal importance to perform reliable fecal egg count (FEC) and fecal egg count reduction test (FECRT), in order to determine the need of anthelmintic treatment, but also anthelmintic efficacy or resistance. This approach is fundamental to a correct and efficient control of GIN. Unfortunately, in worldwide ruminant farm practice, repeated anthelmintic treatments are carried out, without prior diagnosis of infection, contributing to the spread of Anthelmintic Resistance (AR). Tackling this phenomenon, improving mainly the GIN diagnosis and AR status in farm animals, is a priority of the European COST Action "COMBAR-COMBatting Anthelmintic Resistance in Ruminants" and of the STAR-IDAZ International Research Consortium on Animal Health. One of the specific objectives of the COMBAR Working Group 1 (WG1) is to conduct an European market analysis of new diagnostics and develop a business plan for commercial test introduction, leveraging technical know-how of participants. Since the Mini-FLOTAC in combination with the Fill-FLOTAC may be considered a good candidate for a standardized FEC and FECRT in the laboratory, as well as directly in the field, the aim of this study was to conduct SWOT (Strength-Weaknesses-Opportunities-Threats) and PESTEL (Political, Economic, Social, Technological, Environmental, and Legal) analyses of these tools in 20 European countries involved in the COMBAR WG1, in order to identify the opportunities, barriers, and challenges that might affect the Mini-FLOTAC and Fill-FLOTAC commercialization in Europe.

Graphical abstract, Highlights • 47 farmers in Botswana were trained to use targeted selective treatment for worms of small ruminants. • Goats from treated herds showed greater improvements in health than those from untreated herds. • The system was adopted successfully by farmers with a range of levels of literacy and education. • Mixed methods were used for evaluation and interpretation of the system within a cultural context., Due to the threat of anthelmintic resistance, livestock farmers worldwide are encouraged to selectively apply treatments against gastrointestinal nematodes (GINs). Targeted selective treatment (TST) of individual animals would be especially useful for smallholder farmers in low-income economies, where cost-effective and sustainable intervention strategies will improve livestock productivity and food security. Supporting research has focused mainly on refining technical indicators for treatment, and much less on factors influencing uptake and effectiveness. We used a mixed method approach, whereby qualitative and quantitative approaches are combined, to develop, implement and validate a TST system for GINs in small ruminants, most commonly goats, among smallholder farmers in the Makgadikgadi Pans region of Botswana, and to seek better understanding of system performance within a cultural context. After the first six months of the study, 42 out of 47 enrolled farmers were followed up; 52% had monitored their animals using the taught inspection criteria and 26% applied TST during this phase. Uptake level showed little correlation with farmer characteristics, such as literacy and size of farm. Herd health significantly improved in those herds where anthelmintic treatment was applied: anaemia, as assessed using the five-point FAMACHA© scale, was 0.44–0.69 points better (95% confidence interval) and body condition score was 0.18–0.36 points better (95% C.I., five-point scale) in treated compared with untreated herds. Only targeting individuals in greatest need led to similar health improvements compared to treating the entire herd, leading to dose savings ranging from 36% to 97%. This study demonstrates that TST against nematodes can be implemented effectively by resource-poor farmers using a community-led approach. The use of mixed methods provides a promising system to integrate technical and social aspects of TST programmes for maximum uptake and effect.

For many parasites, the full set of hosts that are susceptible to infection is not known, and this could lead to a bias in estimates of transmission. We used counts of individual adult parasites from historical parasitology studies in southern Africa to map a bipartite network of the nematode parasites of herbivore hosts that occur in Botswana. Bipartite networks are used in community ecology to represent interactions across trophic levels. We used a Bayesian hierarchical model to predict the full set of host–parasite interactions from existing data on parasitic gastrointestinal nematodes of wild and domestic ungulates given assumptions about the distribution of parasite counts within hosts, while accounting for the relative uncertainty of less sampled species. We used network metrics to assess the difference between the observed and predicted networks, and to explore the connections between hosts via their shared parasites using a host–host unipartite network projected from the bipartite network. The model predicts a large number of missing links and identifies red hartebeest, giraffe and steenbok as the hosts that have the most uncertainty in parasite diversity. Further, the unipartite network reveals clusters of herbivores that have a high degree of parasite sharing, and these clusters correspond closely with phylogenetic distance rather than with the wild/domestic boundary. These results provide a basis for predicting the risk of cross-species transmission of nematode parasites in areas where livestock and wildlife share grazing land. This article is part of the themed issue ‘Opening the black box: re-examining the ecology and evolution of parasite transmission’.

We describe and discuss the global properties of 45 gamma-ray bursts (GRBs) observed by HETE-2 during the first three years of its mission, focusing on the properties of X-Ray Flashes (XRFs) and X-ray-rich GRBs (XRRs). We find that the numbers of XRFs, XRRs, and GRBs are comparable. We find that the durations and the sky distributions of XRFs and XRRs are similar to those of GRBs. We also find that the spectral properties of XRFs and XRRs are similar to those of GRBs, except that the values of the peak energy $E^{\rm obs}_{\rm peak}$ of the burst spectrum in $\nu F_\nu$, the peak energy flux $\Fp$, and the energy fluence $S_E$ of XRFs are much smaller -- and those of XRRs are smaller -- than those of GRBs. Finally, we find that the distributions of all three kinds of bursts form a continuum in the [$S_E$(2-30 keV),$S_E$(30-400) keV]-plane, the [$S_E$(2-400 keV), $E_{\rm peak}$]-plane, and the [$F_{\rm peak}$(50-300 keV), $E_{\rm peak}$]-plane. These results provide strong evidence that all three kinds of bursts arise from the same phenomenon., Comment: 33 pages, 15 figures, submitted to ApJ

The majority of existing models for predicting disease risk in response to climate change are empirical. These models exploit correlations between historical data, rather than explicitly describing relationships between cause and response variables. Therefore, they are unsuitable for capturing impacts beyond historically observed variability and have limited ability to guide interventions. In this study, we integrate environmental and epidemiological processes into a new mechanistic model, taking the widespread parasitic disease of fasciolosis as an example. The model simulates environmental suitability for disease transmission at a daily time step and 25 m resolution, explicitly linking the parasite life cycle to key weather-water-environment conditions. Using epidemiological data, we show that the model can reproduce observed infection levels in time and space for two case studies in the UK. To overcome data limitations, we propose a calibration approach combining Monte Carlo sampling and expert opinion, which allows constraint of the model in a process-based way, including a quantification of uncertainty. The simulated disease dynamics agree with information from the literature, and comparison with a widely-used empirical risk index shows that the new model provides better insight into the time-space patterns of infection, which will be valuable for decision support.

Helminth infections, mainly by gastrointestinal nematodes (GIN), are one of the main concerns for animal health, welfare and productivity in grazing ruminant livestock worldwide. The use of a sensitive, precise, accurate, low-cost, and easy-to-perform copromicroscopic technique is of pivotal importance to perform reliable fecal egg count (FEC) and fecal egg count reduction test (FECRT), in order to determine the need of anthelmintic treatment, but also anthelmintic efficacy or resistance. This approach is fundamental to a correct and efficient control of GIN. Unfortunately, in worldwide ruminant farm practice, repeated anthelmintic treatments are carried out, without prior diagnosis of infection, contributing to the spread of Anthelmintic Resistance (AR). Tackling this phenomenon, improving mainly the GIN diagnosis and AR status in farm animals, is a priority of the European COST Action "COMBAR-COMBatting Anthelmintic Resistance in Ruminants" and of the STAR-IDAZ International Research Consortium on Animal Health. One of the specific objectives of the COMBAR Working Group 1 (WG1) is to conduct an European market analysis of new diagnostics and develop a business plan for commercial test introduction, leveraging technical know-how of participants. Since the Mini-FLOTAC in combination with the Fill-FLOTAC may be considered a good candidate for a standardized FEC and FECRT in the laboratory, as well as directly in the field, the aim of this study was to conduct SWOT (Strength-Weaknesses-Opportunities-Threats) and PESTEL (Political, Economic, Social, Technological, Environmental, and Legal) analyses of these tools in 20 European countries involved in the COMBAR WG1, in order to identify the opportunities, barriers, and challenges that might affect the Mini-FLOTAC and Fill-FLOTAC commercialization in Europe.

The majority of existing models for predicting disease risk in response to climate change are empirical. These models exploit correlations between historical data, rather than explicitly describing relationships between cause and response variables. Therefore, they are unsuitable for capturing impacts beyond historically observed variability and have limited ability to guide interventions. In this study, we integrate environmental and epidemiological processes into a new mechanistic model, taking the widespread parasitic disease of fasciolosis as an example. The model simulates environmental suitability for disease transmission at a daily time step and 25 m resolution, explicitly linking the parasite life cycle to key weather-water-environment conditions. Using epidemiological data, we show that the model can reproduce observed infection levels in time and space for two case studies in the UK. To overcome data limitations, we propose a calibration approach combining Monte Carlo sampling and expert opinion, which allows constraint of the model in a process-based way, including a quantification of uncertainty. The simulated disease dynamics agree with information from the literature, and comparison with a widely-used empirical risk index shows that the new model provides better insight into the time-space patterns of infection, which will be valuable for decision support.

19 páginas., An elicitation exercise was conducted to collect and identify pressing questions concerning the study of helminths in livestock, to help guide research priorities. Questions were invited from the research community in an inclusive way. Of 385 questions submitted, 100 were chosen by online vote, with priority given to open questions in important areas that are specific enough to permit investigation within a focused project or programme of research. The final list of questions was divided into ten themes. We present the questions and set them briefly in the context of the current state of knowledge. Although subjective, the results provide a snapshot of current concerns and perceived priorities in the field of livestock helminthology, and we hope that they will stimulate ongoing or new research efforts., We thank the officers and members of the Livestock Helminth Research Alliance (LiHRA) for encouraging this initiative and giving space to it in their annual meetings, to the World Association for the Advancement of Veterinary Parasitology for permitting elicitation of questions as part of their 26th biennial conference in Kuala Lumpur, Malaysia, and to the editors of Trends in Parasitology for commissioning this article. The authors credit the EU for funding leading to this work through FP7 STREP GLOWORM. We gratefully acknowledge Hassan Azaizeh, Sarah Beynon, Jacques Cabaret, Gerald Coles, Tina Alstrup Hansen, Alison Howell, Hamadi Karembe, Alvaro Martinez-Moreno, Francisco A. Rojo, Guillaume Sallé, Jože Starič, Eurion Thomas, and numerous anonymous participants for contributing generously to the exercise with questions and votes. This article is based in part upon work from COST Action COMBAR CA16230, supported by COST (European Cooperation in Science and Technology). MMV is funded by the Spanish “Ramón y Cajal” Programme, Ministry of Economy and Competitiveness (RYC-2015-18368), and ERM and HRV by UK BBSRC grant BB/M003949/1.

Haemonchus contortus is a gastrointestinal nematode parasite of small ruminants, which feeds on blood and causes significant disease and production loss in sheep and goats, especially in warmer parts of the world. The life cycle includes free-living immature stages, which are subject to climatic influences on development, survival and availability, and this species therefore exhibits spatio-temporal heterogeneity in its infection pressure based on the prevailing climate. Models that better explain this heterogeneity could predict future epidemiological changes. The basic reproduction quotient (Q0) was used as a simple process-based model to predict climate-driven changes in the potential transmission of H. contortus across widely different geo-climatic zones, and showed good agreement with the observed frequency of this species in the gastrointestinal nematode fauna of sheep (r = 0.81, P