This study quantified the extent of anthelmintic resistance (AR) in ascarid and strongylid nematodes against commonly used anthelmintics in Australian Thoroughbred horses. Faecal egg count reduction tests (FECRTs, n = 86) and egg reappearance period (ERP) tests were conducted on 22 farms across Australia. Faecal egg counts (FECs) were determined using the modified McMaster technique, and percent faecal egg count reduction (%FECR) was calculated using the Bayesian hierarchical model and hybrid Frequentist/Bayesian analysis method. The results were interpreted using old (published in 1992) and new (2023) research guidelines of the World Association for the Advancement of Veterinary Parasitology (WAAVP). The species composition of strongylid nematodes was detected utilising a DNA-metabarcoding method using pre- and post-treatment samples. Resistance was observed in strongylid nematodes to commonly used single-active and combination anthelmintics, including ivermectin (IVM %FECR range: 82%-92%; 95% lower credible interval (LCI) range: 80%-90%), abamectin (ABM: 73%-92%; 65%-88%), moxidectin (MOX: 89%-91%; 84%-89%), oxfendazole (OFZ: 0%-56%; 0%-31%) and its combination with pyrantel (OFZ + PYR: 0%-82%; 0%-78%). Resistance in Parascaris spp. was observed to IVM (10%-43%; 0%-36%), ABM (0%; 0%) and MOX (0%; 0%). When the new thresholds recommended by the WAAVP were used, AR was detected in six additional FECRTs for strongylids and three more tests for Parascaris spp., introducing resistance to OFZ and OFZ + PYR in the latter. Shortened ERPs (4-6 weeks) of strongylids were observed in 31 FECRTs in which AR was not detected at 2 weeks post-treatment for all the anthelmintics tested. Among cyathostomins, Cylicocyclus nassatus, Cylicostephanus longibursatus and Coronocyclus coronatus were the most prevalent species at 2 weeks post-treatment, whereas the main species appearing at five weeks following treatments with macrocyclic lactones were Cylicocyclus nassatus, Cylicostephanus longibursatus and Cylicocyclus ashworthi. After treatment with OFZ + PYR, the latter three, plus Coronocyclus coronatus and Cyathostomum catinatum, were detected at 5 weeks post-treatment. Overall, the study highlights the prevalence of AR in both ascarids and strongylid nematodes against commonly used anthelmintic products to control worms in Australian horses. The results indicate that ML combination products provided acceptable efficacy at 2 weeks. However, ERP calculations suggest that products work less effectively than previously measured. It is suggested to regularly monitor the efficacy of the anthelmintics and consider changing the worm control practices to better manage worms and AR in Australian horses.

BACKGROUND: The roundworms, Parascaris spp., are important nematode parasites of foals and were historically model organisms in the field of cell biology, leading to many important discoveries. According to karyotype, ascarids in Equus are commonly divided into Parascaris univalens (2n = 2) and Parascaris equorum (2n = 4).
METHODS: Here, we performed morphological identification, karyotyping and sequencing of roundworms from three different hosts (horses, zebras and donkeys). Phylogenetic analysis was performed to study the divergence of these ascarids based on cytochrome c oxidase subunit I (COI) and internal transcribed spacer (ITS) sequences.
RESULTS: Karyotyping, performed on eggs recovered from worms of three different Equus hosts in China, showed two different karyotypes (2n = 2 in P. univalens collected from horses and zebras; 2n = 6 in Parascaris sp. collected from donkeys). There are some differences in the terminal part of the spicula between P. univalens (concave) and Parascaris sp. (rounded). Additionally, it was found that the egg\'s chitinous layer was significantly thicker in Parascaris sp. (> 5 μm) than P. univalens (< 5 μm) (F(2537) = 1967, P < 0.01). Phylogenetic trees showed that the sequences of Parascaris from Equus hosts were divided into two distinct lineages based on sequences of the COI and ITS.
CONCLUSIONS: Comparing the differences in roundworms collected from three different Equus hosts, this study describes a Parascaris species (Parascaris sp.) with six chromosomes in donkeys. It is worth noting that the thickness of the chitinous layer in the Parascaris egg may serve as a diagnostic indicator to distinguish the two roundworms (P. univalens and Parascaris sp.). The Parascaris sp. with six chromosomes in donkeys in the present study may be a species of P. trivalens described in 1934, but the possibility that it is a new Parascaris species cannot be ruled out. Both karyotyping and molecular analysis are necessary to solve the taxonomic problems in Parascaris species.

Parascaris spp. are major gastro-intestinal nematodes that infect foals and can lead to respiratory symptoms, poor growth, and in some cases obstruction of the small intestine and death. Ivermectin resistance has been reported for Parascaris spp. in many countries. In Poland, the knowledge of the level of resistance against ivermectin in Parascaris spp. is limited. The aim of this study was to examine the efficacy of ivermectin against Parascaris spp. in foals from south-eastern Poland. Foals (n = 225 = reared in 7 stud farms) were treated orally with ivermectin paste. Faecal samples were collected from the rectum of each foal or from the environment straight after defaecation on 1 day prior and 2 weeks after deworming. A faecal egg count (FEC) was performed using the McMaster method with a minimum detection limit of 50 eggs/g. FEC reduction (FECR) was calculated using the Faecal Egg Count Reduction Test. The statistical analysis was limited to foals excreting more than 150 eggs/g before treatment and to stud farms with at least 6 foals excreting at or above this level. Confidence intervals were determined by 1000 bootstraps at farm level and the contribution of sex and age to FECR was quantified using a generalized equation estimation procedure. Parascaris spp. eggs were found in 40% of the foals. Following ivermectin treatment, Parascaris spp. eggs were identified in 28.4% of the foals. The mean estimated FECR ranged from 44% to 97% and average efficacy was 49.3%. FECR was more pronounced in older foals (P-values = 0. 003). The FECR was more pronounced in males than in females (P value = 0.028). This study is the first to indicate a reduced efficacy of ivermectin against Parascaris spp. in foals in Poland.

The equine Parascaris spp. is large, parasitic nematodes, and predominantly focuses on the intestine of foals and young weanlings. There are two roundworms, Parascaris equorum and Parascaris univalens, recognized among equine hosts. In this study, all fifty-nine Parascaris worms were harvested from three different equine hosts (twenty specimens from Equus zebra, twenty specimens from E. caballus, and nineteen specimens from E. asinus). The ribosomal gene (ITS) and mitochondrial genes (cox1 and nadh1) were amplified to identify and genetically characterize these worms. Analysis of ITS sequences revealed five genotypes among the fifty-nine worms, and the sequence similarity among the worms from E. zebra and E. caballus was at a high level (99.87%), while the one of E. asinus worms showed an apparent difference from the worms either from the E. zebra or from the E. caballus (sequence similarity ranging from 93.04 to 93.42%). Analysis of mitochondrial genes revealed that twenty-one (cox1 gene) and thirteen (nadh1 gene) unique haplotypes were defined among the fifty-nine worms. The shared haplotypes (four cox1 haplotypes and one nadh1 haplotype) only occurred between the worm populations from E. zebra and E. caballus. The cox1 and nadh1 haplotype sequences were respectively applied to construct phylogenetic trees. Although the topologies showed that E. asinus worm population had an obvious boundary with the worm populations of the E. zebra and the E. caballus, however, no noticeable boundary was found within the two later worm populations. Meanwhile, the E. asinus worm population showed an obvious genetic differentiation and an extremely low gene flow (close to zero) with the worm populations from E. zebra and E. caballus, indicating that the genetic characteristics of the worms from the E. asinus have an obvious difference with the one from E. zebra and E. caballus.

Parasite control in foals is complicated by the concurrent presence of biologically diverse parasites with differing levels of anthelmintic resistance. Several combination anthelmintic products are available for use in horses, but information on their efficacies against important equine parasites is scarce. Two trials were performed in New Zealand during 2008 and 2011 on four different farms with substantially different anthelmintic treatment histories. The first trial evaluated the efficacy of an ivermectin/praziquantel/oxibendazole combination, a single active oxibendazole, and a single-active macrocyclic lactone (ML) in 49 foals located on three farms. The second trial evaluated two combination anthelmintic products and three single-active ML products and enrolled a total of 110 foals on three farms. Foals in the second trial were allocated to one of six anthelmintic treatment groups; oxfendazole/pyrantel embonate, pyrantel embonate/ivermectin/praziquantel, ivermectin/praziquantel, abamectin/praziquantel, moxidectin/praziquantel, and a placebo-treated control. In both trials, foals were monitored monthly prior to treatment, and fecal egg counts (FECs) of Parascaris spp., strongylid, and Strongyloides westeri were determined. A \"rolling enrolment\" process was implemented whereby foals were systematically allocated to a treatment group and treated with the corresponding anthelmintic following the first appearance of Parascaris spp. eggs in the faeces. A generalised linear model was used to evaluate the effect of farm and treatment on Day14 FEC (ln) for each parasite. Three different FECR calculation methods were employed as follows; i) FECR(T) pre and post treatment ii) FECR (C) in the treated group compared with control, and iii) FECR (P) pre- and post- treatment in the treated and control groups. Across both trials, treatment with ML single active products failed to achieve >95% reduction in Parascaris spp. FEC on two of three farms. The pyrantel embonate/oxfendazole and ivermectin/ praziquantel/oxibendazole combinations demonstrated full efficacy against Parascaris spp. This is in contrast to the anti-strongylid efficacies determined, where the pyrantel embonate/oxfendazole combination and single active oxibendazole had reduced efficacy on one farm, while the macrocyclic lactones generally had good efficacy. Strongyloides egg counts were sporadic in both trials, and allowed limited insight into anthelmintic efficacy. The study illustrated the importance of keeping an untreated or placebo-treated control group in studies evaluating anti-Parascaris efficacy and it demonstrated the utility of a rolling enrolment procedure, where foals are enrolled over the course of a defined period of time. Furthermore, the study demonstrated the value of a farm specific FECR monitoring programme and the complexity of parasite control in foals, where combination anthelmintic products can be employed to target multiple species of parasites.

The aims of this study were to determine the species of Parascaris present in foals in Sweden and to establish whether anthelmintic resistance to pyrantel and fenbendazole is present on Swedish stud farms. Ascarid eggs collected from different regions in Sweden were karyotyped and were all identified as Parascaris univalens, characterized by one chromosomal pair. Faecal egg count reduction tests were performed on a total of 142 foals on 9 farms between September 2016 and May 2017. Healthy foals with at least 150 eggs per gram faeces (EPG) were included in the study and treated with oral pastes of pyrantel embonate or fenbendazole according to manufacturer instructions. The efficacy of the drugs was calculated by a Bayesian model using the R package \"eggCounts\". In accordance with the American Association of Equine Practitioners, parasites were classified as resistant to pyrantel if the reduction in EPG was ≤ 85% and to fenbendazole if the observed efficacy was ≤ 90%. Four of eleven groups treated with pyrantel had an observed efficacy of ≤ 85%, and as many as 43% of the foals treated with pyrantel excreted eggs 10-16 days after treatment. In contrast, one of the six groups treated with fenbendazole had an observed efficacy of ≤ 90%, and only 6% of all foals were excreting eggs 10-16 days after treatment. Since resistance to ivermectin has earlier been shown to be widespread in Parascaris spp. in Sweden it is likely that multiresistant populations are present on Swedish stud farms. This is the first study showing the existence of pyrantel-resistant Parascaris spp. in Europe, and the first ever study where anthelmintic resistance has been shown in P. univalens.

BACKGROUND: In the last decade, Parascaris spp. resistance to anthelmintics has been recorded in many countries. In Saudi Arabia, there are limited data available on Parascaris spp. resistance to anthelmintics.
OBJECTIVE: To determine the current status of ivermectin, abamectin and praziquantel combined, and fenbendazole resistance to Parascaris spp. in horses in Saudi Arabia.
METHODS: Three hundred and forty-one foals from eleven different farms were examined by faecal egg count (FEC). The foals were all Arab horses aged 17.2 ± 4.5 (SD) months. Ivermectin (n = 46 foals), abamectin and praziquantel combined (n = 46), and fenbendazole (n = 46) were administered on day 0 and faeces were collected on day 14. The study comprised 41 untreated foals as controls. Animals that have FEC of ≥100 eggs per gram (EPG) were used to measure anthelmintic efficacy. Parascaris spp. populations were considered susceptible when faecal egg count reduction (FECR) was ≥95% associated with a lower 95% confidence limit (LCL) >90%, suspected resistant when FECR ≤90% or LCL <90% and resistant when FECR <90% and LCL <90%.
RESULTS: Prevalence of Parascaris spp. infection was 53% (179/341 horses). Anthelmintic resistance to Parascaris spp. were highest following fenbendazole (55% of farms and 65% of foals) and to a lower extent following ivermectin or the combination of abamectin and praziquantel which comprised 27% of farms (and 46% of foals) and 18% of farms (and 10% of foals), respectively.
CONCLUSIONS: These data indicate that anthelmintics-resistant Parascaris spp. populations are present on horse farms in Saudi Arabia.

Parascaris spp. infects foals worldwide and may cause airway inflammation in addition to small intestinal impaction and rupture. It is observed that acquired immunity eliminates ascarid burdens beginning at about 6 months of age, and current evidence suggests that a single parasite generation propagates in each foal crop. The purpose of this study was to monitor natural parasitic infections in untreated mixed breed horse foals over the course of 0-300 days of age. Fecal samples were collected monthly from all foals born in 2014 (n=13), beginning July 2014 through February 2015. Fecal egg counts (FECs) were performed in triplicates using the Mini-FLOTAC method. The foals were necropsied between 154 and 298 days of age and all intestinal ascarid were collected and identified to stage. Ascarid FECs exhibited a biphasic distribution with an initial peak at 91-120 days of age and, after a steady decline, a second, smaller peak at 241-300 days of age. Numbers of corticated and decorticated ascarid eggs were compared, with decorticated FECs remaining consistently low with a slight increase directly after the first corticated FEC peak. Overall, 4.36% of the total ascarid eggs counted were decorticated. Ascarid FECs showed a sharp peak in September, declined, and then steadily increased beginning in December and continuing through February. Upon necropsy, moderate to high number of ascarid specimens were recovered from foals between 8 and 10 months of age, coinciding with the second peak for the FECs. Eleven of the 13 foals harbored immature ascarid stages indicating a recent reinfection. However, these data demonstrates that apparently a second, smaller wave of infection is present in 8-10 month old foals. It may be of value to monitor egg counts in this age group to make sure that all parasite categories are well controlled.