Faecal Egg Counts Research Articles

Successful treatment of Trypanoxyuris sp. infection in naturally infected southern brown-howlers (Alouatta guaribaclamitans)

Southern brown-howler monkeys (Alouatta guariba clamitans) may harbor Trypanoxyuris sp., a pinworm parasite with documented fatal consequences in this species. Despite this risk, effective treatment protocols remain unclear. Therefore, the present study aimed to evaluate the efficacy of two anthelmintic protocols against natural infections in two brown-howler monkeys received at the Wild Animal Care and Rehabilitation Sector (SARAS-CAV-UDESC). The protocols utilized pyrantel pamoate & praziquantel (600.0 mg, PO, single dose) and albendazole (20.0 mg, PO, daily for 5 days). Fecal egg counts were carried out daily at the Laboratory of Parasitology and Parasitic Diseases (LAPAR-CAV-UDESC) before and after drug administration. Both treatments successfully eliminated Trypanoxyuris sp. infections. The animal treated with pyrantel pamoate & praziquantel achieved egg clearance by day 6 (144 h), demonstrating effectiveness with a single administration. Albendazole cleared the infection within 2 days of treatment, indicating its potential as a fast-acting treatment. No adverse effect were observed in the treated monkeys. These findings contribute to the development of evidence-based treatment protocols for Trypanoxyuris sp. in primates, enhancing animal health and welfare of captive and wild populations.

First evaluation and detection of ivermectin resistance in gastrointestinal nematodes of sheep and goats in South Darfur, Sudan.

In Sudan, resistance to benzimidazoles has been reported recently in cattle and goats from South Darfur. Herein, ivermectin efficacy against gastrointestinal nematodes (GINs) was evaluated in sheep and goats in three study areas in South Darfur. The faecal egg count reduction test (FECRT) was used to evaluate the efficacy of ivermectin in sheep and goats naturally infected with GINs in the region of Bulbul (goats: n = 106), Kass (goats: n = 40) and Nyala (Domaia (sheep: n = 47, goats: n = 77) and the University farm (goats: n = 52)), using different treatment plans, and the efficacy was evaluated 12 days after treatment. Ivermectin efficacy was also evaluated in goats experimentally infected using local Haemonchus contortus isolates from Kass and Nyala. Nematodes surviving ivermectin treatment in goats in Bulbul and Nyala were harvested and larvae used to infect worm-free male sheep (n = 6, ≤6 months old). Infected sheep were dosed subcutaneously with ivermectin every eight days with increasing doses from 0.2 mg/kg to 1.6 mg/kg bodyweight (bw). Reduced ivermectin efficacy was identified in sheep and goats in the four study locations. Using a paired statistic, the efficacy of a therapeutic dose in sheep was 75.6% (90% upper credible limit (UCrL): 77.5%), while twice the recommended dose led to a reduction of 92.6% (90% UCrL: 93.3%). In goats, the FECRs of a therapeutic dose were 72.9-95.3% (90% UCrL range: 73.6-95.7%) in Bulbul, Nyala Domaia, Nyala University farm and Kass. Twice the dose recommended for goats in Bulbul revealed a 90% UCrL of 87.6%. All post-treatment faecal cultures contained only Haemonchus spp. larvae. The experimental infection trials in sheep and goats supported our findings from field trials and calculated upper 90% CrL of below 98.9%. For the first time highly ivermectin resistant H. contortus populations have been identified in sheep and goats in Sudan, and resistance was experimentally confirmed.

Patterns of gastrointestinal parasite infections in bighorn sheep, Ovis canadensis, with respect to host sex and seasonality

Males and females in sexually dimorphic species show differences in their physiology and behaviour due to differences in energetic investment into reproduction and soma. This means that the two sexes may show different patterns of parasitism at different times of the year. In this study, we evaluate the abundance of fecal eggs and larvae of 5 parasite types (Strongyles, Nematodirus spp., Marshallagia marshalli., Protostrongylus spp. lungworms, and Eimeria spp.) in relation to season and sex in Rocky Mountain bighorn sheep (Ovis canadensis). We use fecal egg counts (FEC) as a proxy for infection intensity. Parasite FECs differed between male and female bighorn sheep and varied with season. We found pronounced fluctuations in fecal egg counts of various parasite species in males and females across different seasons and reproductive stages. Strongyle counts were significantly higher during late gestation and lactation/summer, and particularly pronounced in males. Nematodirus counts were highest during late gestation in females and during the rut in males. Marshallagia counts peaked during late gestation in females and during the rut in males. Protostrongylus spp. lungworm counts were highest during late gestation in females and in males during lactation/summer and the rut. Eimeria oocyst counts varied across seasons, with higher counts in males during the rut and in females during winter and late gestation. Additionally, significant differences in Strongyle counts were observed between coursing and tending rams, with tending rams exhibiting higher counts. We discuss why the sexes might differ in FECs and suggest that differences between FECs of the parasites across seasons may be due to different life cycles and cold tolerance of the parasites themselves.

Behavioural response to gastrointestinal parasites of yearling dairy calves at pasture

ABSTRACT Aims To investigate the association between gastrointestinal parasites (GIP) and animal behaviour in dairy calves under New Zealand pastoral conditions, using animal-mounted, accelerometer-based sensors. Methods Thirty-six, 5–6-month-old, Friesian-Jersey, heifer calves fitted with animal activity sensors to track behaviour were randomly allocated to one of two treatment groups. Half the animals were challenged with an oral dose of 20,000 larvae of Ostertagia ostertagi and Cooperia oncophera once a week for 3 weeks and half were unchallenged. Five weeks after the last dose, seven infected and nine uninfected animals were treated with an oral anthelmintic (AHC) and data collected for a further week. Accelerometer data were classified into minutes per day eating, ruminating, in moderate–high activity or in low activity. Live weight and faecal egg counts (FEC) were recorded weekly over the study period. All animals co-grazed a newly sown pasture not previously grazed by ruminants and were moved every week to fresh grazing. Treatment status was blinded to those managing the animals which were otherwise treated identically. Results Complete behavioural records were available from 30/36 calves, (13 challenged and 17 unchallenged). Before treatment with AHC, FEC increased in infected and un-treated calves over the study, while uninfected animals maintained a near zero FEC. There was no difference in live weight gain between the two groups over the study period. Bayesian, multinomial regression predicted differences in animal behaviour between infected and uninfected animals that were not treated with AHC over the 7 weeks following initial infection. Parasitised calves not treated with AHC were less active and spent up to 6 (95% highest density interval (HDI) = 1–11) minutes/day less in low level activity and up to 15 (95% HDI = 7–20) minutes/day less in moderate to high level activity. They ruminated up to 9 (95% HDI = 2–15) minutes/day more and ate up to 10 (95% HDI = 2–19) minutes/day more than control calves that were not treated with AHC. The effect of AHC on time spent in each behaviour differed between infected and uninfected calves and increased the coefficient of dispersion of the behavioural data. Conclusions and clinical relevance Small differences in animal behaviour can be measured in calves with GIP. However, to use this to target treatment, further validation studies are required to confirm the accuracy of behavioural classification and understand the complex drivers of animal behaviour in a dynamic and variable pasture-parasite–host environment.

Fecal egg counts and individual milk production in temperate pastoral dairy systems of Australia

Anthelmintic use in lactating dairy cattle, has been shown to result in a milk production response in some previous studies. If individual animals within a herd could be identified that would most benefit from anthelmintic treatment, this may reduce anthelmintic resistance. Australian dairy systems are predominantly pasture-based, allowing sustained exposure and immune stimulation of cattle to gastrointestinal nematodes. We assessed the relationship between milk production and early lactation fecal egg counts (FEC) to determine whether cows with higher FEC produced less milk. Ten pasture-based dairy farms in south-west Victoria, Australia, entered an observational study. Individual FECs at a minimum detectable count of 2.5 eggs per gram of feces (epg) were recorded for recently calved primiparous and multiparous cohorts on each farm. All animals were calved ≤30 d at sampling. Body condition scores were assessed at sampling and milk production data was collected from daily milk meters and herd tests to give first 100-d milk production. When separated by primiparous or multiparous status, no difference in the milk production between cows with FEC = 0 and FEC ≥2.5 epg was identified. Between farm variation was large for FEC and milk production. Fecal egg count at a minimum detectable count of 2.5 epg detected parasitism in primiparous cows post calving but the presence and magnitude of parasitism measurable by FEC was not related to milk production in pasture based Australian dairy systems. In multiparous cows, the rate detection of worm eggs at this analytical sensitivity was lower and the significance of a positive FEC at this analytical sensitivity requires further assessment to ascertain the effect on milk production. Based on our study, it seems unlikely that individual FEC results would be useful as a basis to select individual cows in southwest Victorian dairy herds for anthelmintic treatment.

Anthelmintic effects of Podang mango (Mangifera indica) fruit peel waste extract through in vivo application on Indonesian Etawa goat production and health.

The continuous use of anthelmintic drugs has led to global issues of resistance. One breakthrough to address this problem is the utilization of bio-anthelmintics derived from active compounds in agro-industrial waste. This in vivo study investigated the effectiveness of Podang mango (Mangifera indica L.) fruit peel waste extract for anthelmintic purposes, using concentrations up to 5%. This study included 28 Etawa crossbred goats aged 17 months. Goats were randomly assigned to four groups: A negative control, an aqueous fruit peel extract (AFPE) group at 2.5%, another AFPE group at 5%, and a positive control receiving ivermectin. Goats chosen had egg per gram (EPG) counts surpassing 1000 before exposure to Haemonchus contortus. For 7 days within a 9-week study, AFPE from Podang mangoes was given. On the 7th day, the positive control group was administered ivermectin. AFPE dosage relied on the average abomasum fluid per kilogram of animal weight. The feeding regimen consisted of concentrate and Pennisetum purpureum cv. Mott is customized for the nutritional needs of livestock. Data on feed consumption, digestibility, average daily gain, percentage reduction in fecal egg count, body condition score, and clinical parameters were collected throughout the study. At higher treatment levels (AFPE), there was a greater reduction in both EPG and fecal egg counts. The expected and normal ranges were maintained for consumption and digestibility. While body weight increased, FAMACHA parameters showed a decrease. Compared to the negative control group, substantial disparities (p < 0.05) existed for hemoglobin, red blood cells, and hematocrit in both the positive control and the treatment groups. Blood urea nitrogen and creatinine, indicative of liver and kidney health, were within normal ranges. At a concentration of up to 5%, Podang mango waste extract (AFPE) can function as a substitute for traditional helminth medicines or bio-anthelmintics in goats, enhancing their production.

Genetic and environmental influences on faecal worm egg counts following natural infection in Barbari goats

Genetic and environmental influences on faecal worm egg counts following natural infection in Barbari goats

Comparison of traditional copromicroscopy with image analysis devices for detection of gastrointestinal nematode infection in sheep

Sustainable parasite control practices are necessary to combat the negative effects of gastrointestinal nematodes on animal health and production while reducing the selection pressure for anthelmintic resistance. Parasite diagnostic tests can inform treatment decisions, the timing and effectiveness of treatment and enable livestock breeding programmes. In recent years new diagnostic methods have been developed, some incorporating machine learning (ML), to facilitate the detection and enumeration of parasite eggs. It is important to understand the technical characteristics and performance of such new methods compared to long standing and commonly utilised methods before they are widely implemented. The aim of the present study was to trial three new diagnostic tools relying on image analysis (FECPAKG2, Micron and OvaCyte) and to compare them to traditional manual devices (McMaster and Mini-FLOTAC). Faecal samples were obtained from 41 lambs naturally infected with gastrointestinal nematodes. Samples were mixed and separated into 2 aliquots for examination by each of the 5 methods: McMaster, Mini-FLOTAC, FECPAKG2, Micron and OvaCyte. The techniques were performed according to their respective standard protocols and results were collected by trained staff (McMaster and Mini-FLOTAC) or by the device (FECPAKG2, Micron and OvaCyte). Regarding strongyle worm egg count, McMaster values varied from 0 to 9,000 eggs per gram (EPG). When comparing replicate aliquots, both the Mini-FLOTAC and Micron methods displayed similar repeatability to McMaster. However, we found FECPAKG2 and OvaCyte significantly less precise than McMaster. When comparing parasite egg enumeration, significant positive linear correlations were established between McMaster and all other methods. No difference was observed in EPG between McMaster and Mini-FLOTAC or FECPAKG2; however, Micron and OvaCyte returned significantly higher and lower EPG, respectively, compared to McMaster. The number of eggs ascribed to other parasite species was not sufficient for performing a robust statistical comparison between all methods. However, it was noted that FECPAKG2 generally did not detect Strongyloides papillosus eggs, despite these being detected by other methods. In addition, Moniezia spp and Trichuris spp eggs were detected by OvaCyte and Mini-FLOTAC, respectively, but not by other methods. The observed variation between traditional and new methods for parasite diagnostics highlights the need for continued training and enhancing of ML models used and the importance of developing clear guidelines for validation of newly developed methods.

First report of ivermectin resistance in cyathostomins (small strongyles) of horses in Argentina

In Argentina, as in the rest of the world, cyathostomins are the most common nematodes parasitizing horses. Control is based almost exclusively on the administration of benzimidazoles, pyrimidines, and macrocyclic lactones. However, intensive use of these drugs is resulting in the development of anthelmintic resistance (AR). For example, AR to benzimidazoles is currently distributed throughout Argentina, while incipient AR to pyrimidines (pyrantel embonate) is appearing in areas where this drug is used. Macrocyclic lactones and especially ivermectin, are by far the most used drugs by the vast majority of equine premises in the country. Although ivermectin has been used since 1982, its efficacy against equine strongylid parasites has remained very high until the present. In this study we report for the first time, the presence of a cyathostomin population with resistance to ivermectin in adult horses belonging to an equine premise located in central Argentina. Fecal egg count reduction tests (FECRT) were performed following the most recent guidelines of the World Association for the Advancement of Veterinary Parasitology (WAAVP) for the diagnosis of anthelmintic resistance (research protocol) and resistance was considered when the Upper 90% Credible Interval fell below the expected efficacy threshold of 99.9%. Calculations were carried out using two different online calculation interfaces suggested by WAAVP. For the 14-day post-treatment interval, ivermectin efficacy was 79.5% (90% Credible Interval: 68.1–88.8) and 79.3% (74.2–83.6.3%) with the two methods, respectively. At 19 days post treatment, fecal egg count reductions were 68.6% (50.5–83.1) and 68.4% (61.9–74.1), respectively. At both intervals, this cyathostomin population fullfilled the criteria for AR. These findings suggest dispersion of ivermectin resistant cyathostomins in Argentina. Given the widespread use of macrocyclic lactones, it is important that veterinarians and the equine industry promote a more selective and evidence-based use of these drugs and establish routine monitoring to determine anthelmintic field efficacy to detect treatment failures as early as possible and avoid potential health problems as well as further spread of resistant genes.

Occurrence of gastrointestinal nematodes in lambs in Norway, as assessed by copromicroscopy and droplet digital polymerase chain reaction

BackgroundGastrointestinal nematodes (GINs) have a major impact on sheep production, health, and welfare worldwide. Norway is no exception, but there are only a few studies on the prevalence of GINs in Norwegian sheep. The aim of this study was to investigate the current occurrence of the most important nematodes in sheep flocks in Norway.Faecal samples were collected from flocks in 2021/2022, mainly from three geographical regions in Norway, i.e., northern, eastern, and western. In each of 134 flocks included, individual samples from 10 lambs (autumn) were pooled. Third stage larvae (L3) were cultivated and harvested (Baermann method) from the pooled samples. The DNA was then extracted and further analysed using droplet digital PCR (ddPCR). This enables assessment of the proportions of the three most important nematode species/genera, i.e., H. contortus, T. circumcincta, and Trichostrongylus. The fractional abundance/relative proportion of each species/genus was assessed by performing duplex assays with universal strongyle and species/genus-specific primers and probe sets. In addition, the occurrence of Nematodirus eggs was assessed by standard faecal egg counts (i.e., McMaster method).ResultsOf the 134 flocks sampled, 24 were from the northern region, 31 from eastern, and 71 from western Norway. In addition, some flocks from central (n = 7), and southern (n = 1) Norway were included. Among the sampled flocks, T. circumcincta occurred most commonly (94%), followed by H. contortus (60%) and Trichostrongylus (55%), and Nematodirus (51%). In general, mixed infections were observed, with 38% and 18% of flocks infected with three or all four genera, respectively.ConclusionsThe results of this study indicate that GINs are widespread in Norway. Teladorsagia circumcincta seems to be present in most flocks based on this screening. Moreover, the results show that Nematodirus spp. infect lambs throughout the country, predominantly N. battus, and indicate that this nematode has become more abundant, which could lead to an increase in nematodirosis.

Strongylids of Domestic Horses in Eastern Slovakia: Species Diversity and Evaluation of Particular Factors Affecting Strongylid Communities

PurposeThis study aimed to examine the species diversity and structure of the strongylid community in domestic horses in Eastern Slovakia. Also, an analysis of the impact of age, sex, and collection location factors on the strongyid communities was performed.MethodsTwenty-seven horses 1.5–21 years old from two farms in eastern Slovakia with different horse-management conditions were studied. Strongylids were collected after horse treatments with Noromectin (0.2 mg ivermectin); 66,170 specimens were collected and identified. Faecal egg count reduction test (FECRT) was performed following fenbendazole (FBZ) and ivermectin (IVM) treatments.ResultsTwenty-four strongylid species were found; horses were infected with 6 to 16 (average = 11.7) species. Six cyathostomin species (Cylicocyclus nassatus, Cyathostomum catinatum, C. pateratum, Cylicostephanus longibursatus, C. goldi, C. calicatus) were the most prevalent; C. catinatum was the dominant species in both farms (Berger-Parker index 0.34 and 0.42). The structure of the strongylid community was multimodal with dominant, subdominant, background, and rare species. The Mantel test showed that horse age and sex did not significantly affect the nematode infracommunity composition (p > 0.05), while the impact of the collection location (farm) was significant (p = 0.03). Additionally, C. longibursatus was identified as the species contributing significantly to the observed farm differences. Strong resistance to FBZ was documented on both farms (FECRT was 36.4% and 22.7%); IVM resistance was not observed (FECRT = 100%).ConclusionThis study presents the first report on the strongylids parasitizing domestic horses in Eastern Slovakia and gives basic information for further studies of horse parasites and their control in the region.

Evaluation of anthelmintic efficacy of ethanolic leaf extract of Juglans regia L. on Ascaridia galli: a comprehensive in vitro and in vivo study.

Anthelmintic resistance in livestock animals has been spreading across the world in prevalence and severity. As a result, researchers are exploring alternative strategies to combat this issue, and one promising avenue is the utilization of medicinal plants. This study aims to investigate the anthelmintic efficacy of the crude ethanolic extract (CEE) derived from the leaves of Juglans regia against one of the most detrimental nematode parasites affecting poultry, namely Ascaridia galli (A. galli). For the in vitro studies, adult A. galli worms were collected from the naturally infected chickens and the efficacy of CEE was measured at the concentration of 25, 50, and 100mg/ml using adult worm motility inhibition (WMI) assay. In addition, levamisole (0.55mg/ml) was used as the positive control. Likewise, Phosphate buffered saline (PBS) was used as the negative control. For the in vivo studies, CEE of J.regia at the doses of 500, 1000, and 2000mg/kg were evaluated in chickens experimentally infected with A. galli. The anthelmintic efficacy was monitored using faecal egg count reduction (FECR) and worm count reduction (WCR) assays. In vitro studies revealed significant (P < 0.001) anthelmintic effects of CEE of J.regia on the motility of A. galli worms at different hours post-exposure. At the concentration of 100mg/ml, CEE resulted in 96.5% inhibition of worm motility at 24h post-exposure. While the synthetic anthelmintic drug, levamisole caused the highest inhibition of worm motility (100%) at the same time period. The in vivo anthelmintic activity of CEE of J. regia demonstrated a maximum effect on day 14 post-treatment by inducing 67.28% FECR and 65.03% WCR. We observed no significant difference (P > 0.05) in worm counts between the negative control group and the chickens treated with CEE at the dosage of 500mg/kg. Together, the results of the present study suggest that CEE of J. regia leaves possess anthelmintic properties and could be a potential source of novel anthelmintic compounds for controlling helminth parasites.

Single-Cell Transcriptomic Profiling Unveils Dynamic Immune Cell Responses during Haemonchus contortus Infection.

Haemonchus contortus is a parasite widely distributed in tropical, subtropical, and warm temperate regions, causing significant economic losses in the livestock industry worldwide. However, little is known about the genetics of H. contortus resistance in livestock. In this study, we monitor the dynamic immune cell responses in diverse peripheral blood mononuclear cells (PBMCs) during H. contortus infection in goats through single-cell RNA sequencing (scRNA-Seq) analysis. A total of four Boer goats, two goats with oral infection with the L3 larvae of H. contortus and two healthy goats as controls, were used in the animal test. The infection model in goats was established and validated by the fecal egg count (FEC) test and qPCR analysis of the gene expression of IL-5 and IL-6. Using scRNA-Seq, we identified seven cell types, including T cells, monocytes, natural killer cells, B cells, and dendritic cells with distinct gene expression signatures. After identifying cell subpopulations of differentially expressed genes (DEGs) in the case and control groups, we observed the upregulation of multiple inflammation-associated genes, including NFKBIA and NFKBID. Kyoto Encyclopedia of the Genome (KEGG) enrichment analysis revealed significant enrichment of NOD-like receptor pathways and Th1/Th2 cell differentiation signaling pathways in CD4 T cells DEGs. Furthermore, the analysis of ligand-receptor interaction networks showed a more active state of cellular communication in the PBMCs from the case group, and the inflammatory response associated MIF-(CD74 + CXCR4) ligand receptor complex was significantly more activated in the case group, suggesting a potential inflammatory response. Our study preliminarily revealed transcriptomic profiling characterizing the cell type specific mechanisms in host PBMCs at the single-cell level during H. contortus infection.

Association of SERPING1 gene single nucleotide polymorphism with resistance to natural strongyle infection among native goats in humid tropical Kerala

Strongylosis poses a significant threat to goat populations, particularly in humid tropical regions like Kerala, a southern peninsular state in India. This study aimed to compare breed differences in host resistance and resilience to natural strongyle infection between Attappady Black and Malabari goats reared under semi-intensive systems. Additionally, we analyzed the association of a potential single nucleotide polymorphism (SNP) in the Serine Proteinase Inhibitor Clade G member 1 (SERPING1) gene with these traits. Faecal egg count (FEC) and volume of packed red cells (VPRC) were employed as indicators of host resistance and resilience, respectively. Over the course of one year, fortnightly phenotypic data on FEC and VPRC were collected from 109 native goats, including 58 Malabari and 51 Attappady Black goats. The Malabari goats exhibited greater resistance and resilience to strongylosis, evidenced by significantly lower FEC (p≤0.01) and higher VPRC (p≤0.05) compared to Attappady Black goats. Pearson correlation analysis revealed a highly significant negative correlation between FEC and VPRC (r= −0.41, p<0.01). Genotyping of the −312-C>T SNP in SERPING1 gene was conducted using modified bidirectional PCR amplification of specific alleles (Bi-PASA). While the association between SERPING1 genotypes and FEC was not significant, a significant association was observed between SERPING1 genotypes and VPRC (p<0.05). These findings underscore the importance of breed-specific differences in goat resilience to strongyle infection and highlight the potential role of SERPING1 gene polymorphism in modulating host resilience traits.

Surveillance of Ancylostoma caninum in naturally infected dogs in Quebec, Canada, and assessment of benzimidazole anthelmintics reveal a variable efficacy with the presence of a resistant isolate in imported dogs.

Ancylostoma caninum is a widely prevalent parasitic nematode in dogs across the world. There has been a notable increase in reports of anthelmintic resistance in A. caninum within the United States of America in recent years, which has led us to investigate the potential of this scenario in Canada. The study objectives were to assess the prevalence of A. caninum in two different groups, including a colony of rescued dogs in Canada and three imported Greyhound dogs from USA, and to evaluate the efficacy of two benzimidazole (BZ) anthelmintics against A. caninum, complemented with a molecular genetic analysis adapted to low prevalence. Fecal samples were collected at pre- and post-treatment with fenbendazole for the native shelters-origin group, and a combination of anthelmintic formulations, including the pro-BZ febantel for the USA-origin group. The coprology analyses found several genera of internal parasites. Canine ancylostomiasis was the most prevalent parasitosis with 30.77% in the native group and 100% in the USA group, but with overall low average of A. caninum eggs per gram. Through the fecal egg count reduction test (FECRT), applying a cut-off at 90% as baseline of egg reduction for successful efficacy, BZ showed variable efficacy. Furthermore, molecular analysis confirmed the presence of A. caninum in both groups of dogs and found differences in the genetics linked to BZ resistance on the A. caninum β-tubulin isotype 1 gene. In the isolate from the native group, both codons 167 and 200 were homozygous without the presence of single nucleotide polymorphism (SNP). In contrast, the selected isolate from the USA group, showed a homozygous allele at position 200 and a heterozygous SNP at position 167. The latter was congruent with the low efficacy in FECRT and agrees with the recent findings of USA A. caninum isolate resistant phenotype to the BZ anthelmintics. The limitations of the study include an overall low eggs-per-gram in both canine groups, and the shortage of additional fecal samples from the USA group, restraining the molecular analysis only to one out of the three Greyhounds. This study provided some insights on the efficacy of BZs against A. caninum and revealed the presence of BZ resistant isolates in imported dogs in Quebec, Canada. All this information should be considered, for choosing the best strategy in the control of A. caninum using anthelmintic drugs.

Tuft cell acetylcholine is released into the gut lumen to promote anti-helminth immunity

Upon parasitic helminth infection, activated intestinal tuft cells secrete interleukin-25 (IL-25), which initiates a type 2 immune response during which lamina propria type 2 innate lymphoid cells (ILC2s) produce IL-13. This causes epithelial remodeling, including tuft cell hyperplasia, the function of which is unknown. We identified a cholinergic effector function of tuft cells, which are the only epithelial cells that expressed choline acetyltransferase (ChAT). During parasite infection, mice with epithelial-specific deletion of ChAT had increased worm burden, fitness, and fecal egg counts, even though type 2 immune responses were comparable. Mechanistically, IL-13-amplified tuft cells release acetylcholine (ACh) into the gut lumen. Finally, we demonstrated a direct effect of ACh on worms, which reduced their fecundity via helminth-expressed muscarinic ACh receptors. Thus, tuft cells are sentinels in naive mice, and their amplification upon helminth infection provides an additional type 2 immune response effector function.

In vitro and in vivo anthelmintic effect of essential oil obtained from Thymus capitatus flowers against Haemonchus contortus and Heligmosomoides polygyrus

Sheep haemonchosis is a disease that causes serious losses in livestock production, particularly with the increase of cases of anthelmintic resistance around the world. This justifies the urgent need of alternative solutions. The aim of this study was to determine the chemical profile, in vitro, and, in vivo, anthelmintic properties of Thymus capitatus essential oil. To evaluate the, in vitro, anthelmintic activity of the T. capitatus EO on Haemonchus contortus, two tests were used: egg hatch assay (EHA) and adult worm motility (AWM) assay. The nematicidal effect of this oil was evaluated, in vivo, in mice infected artificially with Heligmosomoides polygyrus using faecal egg count reduction (FECR) and total worm count reduction (TWCR). Chromatographic characterization of T.capitatus composition using gas chromatography coupled to mass spectrometry (GC-MS) demonstrated the presence of carvacrol (81.16%), as the major constituents. The IC50 values obtained was 1.9 mg/mL in the EHT. In the AWM assay; T. capitatus essential oil achieved 70.8% inhibition at 1 mg/mL after 8 h incubation. The in vivo, evaluation on H. polygyrus revealed a significant nematicidal effect 7 days post-treatment by inducing 49.5% FECR and 64.5% TWCR, using the highest dose (1600 mg/kg). The results of present study, demonstrate that T.capitatus EO possess a significant anthelmintic properties. Furthermore, it could be an alternative source of anthelmintic agents against gastrointestinal infections caused by H. contortus.

Faecal egg count reduction tests and nemabiome analysis reveal high frequency of multi-resistant parasites on sheep farms in north-east Germany involving multiple strongyle parasite species

Anthelmintic resistance in sheep parasitic gastrointestinal nematodes is widespread and a severe health and economic issue but prevalence of resistance and involved parasite species are unknown in Germany. Here, the faecal egg count reduction test (FECRT) was performed on eight farms using fenbendazole, ivermectin and moxidectin and on four farms using only moxidectin. A questionnaire was used to obtain data on management practices to potentially identify risk factors for presence of resistance. All requirements of the recently revised WAAVP guideline for diagnosing anthelmintic resistance using the FECRT were applied. Nematode species composition in pre- and post-treatment samples was analysed with the nemabiome approach. Using the eggCounts statistic package, resistance against fenbendazole, ivermectin and moxidectin was found on 7/8, 8/8 and 8/12 farms, respectively. No formal risk factor analysis was conducted since resistance was present on most farms. Comparison with the bayescount R package results revealed substantial agreement between methods (Cohen's κ = 0.774). In contrast, interpretation of data comparing revised and original WAAVP guidelines resulted in moderate agreement (Cohen's κ = 0.444). The FECR for moxidectin was significantly higher than for ivermectin and fenbendazole. Nemabiome data identified 4 to 12 species in pre-treatment samples and treatments caused a small but significant decrease in species diversity (inverse Simpson index). Non-metric multidimensional scaling and k-means clustering were used to identify common patterns in pre- and post-treatment samples. However, post-treatment samples were scattered among the pre-treatment samples. Resistant parasite species differed between farms. In conclusion, the revised FECRT guideline allows robust detection of anthelmintic resistance. Resistance was widespread and involved multiple parasite species. Resistance against both drug classes on the same farm was common. Further studies including additional drugs (levamisole, monepantel, closantel) should combine sensitive FECRTs with nemabiome data to comprehensively characterise the anthelmintic susceptibility status of sheep nematodes in Germany.

93 Evaluating the efficacy of anthelmintics in an upper Midwest horse herd

Abstract Cyathostomins in the gastrointestinal tract are ubiquitous in horses around the world and remain a target in parasite control programs. Resistance to anthelmintics has risen in recent years with widespread resistance to benzimidazoles and pyrantels and emerging resistance to macrocyclic lactones. Thus, investigating the efficacy of anthelmintics in different locations is important to direct future parasite control programs. This 2-yr study evaluated the efficacy of three anthelmintic drugs in a university-owned equine herd in the upper Midwest using fecal egg count reduction tests (FECRT). Additionally, the effect of age group and body condition score (BCS) on fecal egg shedding was evaluated. We hypothesized that drug resistance is present in the parasite population of this equine herd. Fecal egg counts (FEC) of strongelid eggs were determined using the McMaster method. The efficacy of anthelmintics was determined by calculating the percentage of reduction in FEC between d 0 and d 14 post-treatment. Eighteen horses of various breeds were used in the study. The horses were grouped by age (&amp;lt; 10, 10-16, and &amp;gt; 16 yr; n = 6 in each group) and BCS using the Henneke system (5.0-5.5, n = 9; 6.0-6.5., n = 6; and 7.0-8.0, n = 3). In yr 1, horses within age group were randomly assigned to anthelmintic treatment: pyrantel pamoate, fenbendazole, or ivermectin. In yr 2, all horses received ivermectin. Individual FEC were performed on the day of anthelmintic treatment (d 0) using freshly voided feces. FEC were repeated 14 d after treatment. FEC were log transformed before analysis. Efficacy of the anthelmintics were compared using a mixed model ANOVA with repeated measures (SAS 9.4). Results were considered significant when P &amp;lt; 0.05. The FECRT was greater when treated with ivermectin and pyrantel pamoate (94.8 ± 11.6 % and 96.6 ± 14.3 %, respectively) compared with fenbendazole (54.4 ± 10.8 %; P &amp;lt; 0.05). The FECRT increased from 54.4% when treated with fenbendazole to 100% when the same horses were treated with ivermectin (P &amp;lt; 0.05). Age tended to affect egg shedding in horses (P = 0.0548). Young horses (&amp;lt; 10 yr) had a greater FEC compared with older horses (&amp;gt;16 yr; P &amp;lt; 0.05) but were not different from horses aged 10 to 16 yr (P &amp;gt; 0.05). There was no effect of BCS on strongelid egg shedding in this study (P = 0.9468). These results indicate the efficacy was within the expected range with no signs of resistance to pyrantel pamoate (93.3 to 100% reduction). There was evidence of emerging resistance to ivermectin (70.4 to 100% reduction) and significant resistance to fenbendazole (0 to 71.5% reduction) in the cyathostomin population in this horse herd. The findings of this study agree with previous research demonstrating resistance to anthelmintics. These results highlight the importance of designing improved parasite control programs to limit the spread of drug resistance with the ultimate goal of improving overall health of the horse.

98 Adding parasite resistance to the breeding objective in hair sheep

Abstract Economic selection indexes provide the best method to achieve genetic gain for multiple traits simultaneously. Hair sheep enterprises, particularly in environments with substantial gastrointestinal nematode challenge, may benefit from incorporating parasite resistance in a selection index. However, obtaining the relative economic value (REV) for parasite resistance is challenging; information on its impact on productivity is incomplete, as are the costs of treatment given anthelmintic resistance. The National Sheep Improvement Program (NSIP) currently provides producers with an index designed to increase total weight of lamb weaned per ewe lambing (TW). Our objective was to assess the effects of including fecal egg count, an indicator of parasite resistance, as an additional goal trait. Including the benchmark scenario with TW as its sole objective, 10 indexes were constructed using standard index theory. Post-weaning fecal egg count (PFEC) was added to this breeding objective with a REV of -1 and the REV for TW was varied from +1 to +5 at +0.5 increments. Weaning weight, number of lambs born and weaned, and PFEC were the selection criteria, modelled as either phenotypes or estimated breeding values (EBV). Phenotypes and EBV were simulated for 1,000 lambs by Cholesky decomposition and used to generate index scores. Ten replicates were run and the top 3% of males and 26% of females were retained on index scores. Annual responses in the goal traits were estimated. Responses in PFEC were expressed as a percentage, assuming a mean PFEC of 2,089 eggs/g as in NSIP. Concordances with those selected in the benchmark scenario were determined. Using EBV as selection criteria, TW increased by 1.7 kg/yr in the benchmark scenario (Table 1). However, unfavorably, PFEC increased by 2.3%/yr. When phenotypic criteria were used, TW increased by 0.5 kg/yr, and PFEC by 0.3%/yr. Once added to the breeding objective, PFEC decreased regardless of the REV of TW. However, response in TW also reduced, although less so as its REV increased. With EBV as the criteria, the REV of +3 for TW appeared preferable: PFEC reduced by 7.1%/yr with 98% of the response in TW retained. With phenotypic selection criteria, PFEC reduced by 5.2%/yr with 94% of the response in TW retained. With +3 REV for TW and EBV as the selection criteria, on average 65% and 82% of males and females, respectively, were chosen in common with the benchmark. When phenotypic criteria were used, those percentages were 60% and 80%, respectively. Implementing a more holistic index with REV of +3 for TW and -1 for PFEC would result in reasonable responses in both traits. However, producers will need to decide if reduced gains in TW is acceptable to improve parasite resistance in a hair sheep breeding program.