Performance of three techniques for diagnosing equine tapeworm infection.

Tapeworm infection in horses can cause serious health concerns, and recent data have documented treatment failures in the most common species, Anoplocephala perfoliata. The threat of anthelmintic resistance in A. perfoliata is of particular concern because of poor diagnostic performance of standard egg counting techniques for detecting this parasite. This study compared the performance of three diagnostic techniques 1) Mini-FLOTAC, 2) Cornell-Wisconsin, and 3) Proudman and Edwards used to detect and quantify A. perfoliata eggs in naturally infected horses. Eighteen adult female horses from the University of Kentucky's historic parasitology herd were included in this study. Fecal samples were collected from all horses at five collection time points two weeks apart and analyzed with the three techniques. A total of 90 samples were collected and 270 counts determined in the study. The proportions of positive samples determined by the three techniques were significantly different from each other (p<0.05): Mini-FLOTAC (16%), Cornell-Wisconsin (47%), and Proudman and Edwards (70%). The Proudman and Edwards technique counted consistently higher numbers of tapeworm eggs compared to the other two techniques throughout the study [p < 0.05]. Total raw counts of tapeworm eggs across the study for each technique were 16, 88, and 410 for the Mini-FLOTAC, Cornell-Wisconsin, and Proudman and Edwards, respectively. This study demonstrated that the Proudman and Edwards technique was superior in diagnosing A. perfoliata infection. Future work needs to assess this technique's potential for Fecal Egg Count Reduction Testing (FECRT).

Parascaris spp. eggs shedding patterns in juvenile horses.

Parascaris spp. infect foals worldwide and foals typically shed eggs in the feces from about three to six months of age, upon which natural immunity is incurred. High levels of anthelmintic resistance of Parascaris spp. are a global concern, and further understanding egg shedding patterns and fecal egg counting (FEC) data variability is of high importance. The aims of this study were to monitor Parascaris spp. egg shedding in untreated foals during 12-23 weeks of age, estimate sources of data variability, and assess precision of two ascarid FEC techniques. Fecal samples were collected weekly from 11 foals born in 2022, from May through November (29 weeks). Six subsamples were extracted from each weekly sample to determine 30 FECs between two techniques: a McMaster technique and an Automated Egg Counting System (AECS). Mixed linear modeling was carried out with age, sex, birth month, seasonality, spring- or summer-born foals, and egg counting technique as explanatory variables. Ascarid FECs were associated with age (p < 0.001), seasonality (p < 0.001), and technique (p < 0.001). The McMaster technique was more precise with a mean coefficient of variation (CV) of 34.57% and a 95% confidence interval (CI) of 30.80%- 38.30% compared to the CV for the AECS, which was 42.22% (CI: 37.70%-46.70%). Seasonality accounted for the highest proportion of variance (PV) of all covariates, but differences in PVs for covariates existed between techniques with foal age and subsample contributing more variance to the McMaster, and individual foal and seasonality contributing more to the AECS. Subsamples and replicate counts accounted for less than 1% of the total data variance. The results highlighted substantial differences in PVs between the two techniques at the subsample (AECS: 57.14%; McMaster: 77.51%) and replicate count levels (AECS: 42.86%; McMaster: 22.49%). While differences in precision were observed between the two FEC techniques, they were negligible in the data set, as the overwhelming majority of the data variability in ascarid FECs was attributed to individual foal, seasonality, and foal age.

The microbial community associated with Parascaris spp. infecting juvenile horses.

BACKGROUND: Parasitic nematodes, including large roundworms colloquially known as ascarids, affect the health and well-being of livestock animals worldwide. The equine ascarids, Parascaris spp., are important parasites of juvenile horses and the first ascarids to develop widespread anthelmintic resistance. The microbiota has been shown to be an important factor in the fitness of many organisms, including parasitic nematodes, where endosymbiotic Wolbachia have been exploited for treatment of filariasis in humans. METHODS: This study used short-read 16S rRNA sequences and Illumina sequencing to characterize and compare microbiota of whole worm small intestinal stages and microbiota of male and female intestines and gonads. Diversity metrics including alpha and beta diversity, and the differential abundance analyses DESeq2, ANCOM-BC, corncob, and metagenomeSeq were used for comparisons. RESULTS: Alpha and beta diversity of whole worm microbiota did not differ significantly between groups, but Simpson alpha diversity was significantly different between female intestine (FI) and male gonad (MG) (P= 0.0018), and Shannon alpha diversity was significantly different between female and male gonads (P = 0.0130), FI and horse jejunum (HJ) (P = 0.0383), and FI and MG (P= 0.0001). Beta diversity (Fig. 2B) was significantly different between female and male gonads (P = 0.0006), male intestine (MI) and FG (P = 0.0093), and MG and FI (P = 0.0041). When comparing organs, Veillonella was differentially abundant for DESeq2 and ANCOM-BC (p < 0.0001), corncob (P = 0.0008), and metagenomeSeq (P = 0.0118), and Sarcina was differentially abundant across four methods (P < 0.0001). Finally, the microbiota of all individual Parascaris spp. specimens were compared to establish shared microbiota between groups. CONCLUSIONS: Overall, this study provided important information regarding the Parascaris spp. microbiota and provides a first step towards determining whether the microbiota may be a viable target for future parasite control options.

Equine strongylids: Ivermectin efficacy and fecal egg shedding patterns.

Equine cyathostomins are ubiquitous in grazing horses around the world and a main target in parasite control programs. Anthelmintic resistance has been reported with increasing frequency in these parasites over the past decades, and recent findings of fulminant resistance to the macrocyclic lactone class have raised severe concerns. This study aimed to evaluate ivermectin efficacy in cohorts of yearlings and mares present on four different farms in Central Kentucky. Strongylid egg counts were determined with an automated egg counting system, and the percent fecal egg count reduction (FECR) was calculated using a hierarchical Bayesian analysis. Novel principles were used for classification of groups to either no signs of anthelmintic resistance, evidence of resistance, or inconclusive. Furthermore, an epidemiological analysis was conducted evaluating the possible influence of pregnancy status, time of foaling, and year of arrival on mare strongylid egg shedding levels. A total of 102 yearlings and 247 mares were enrolled in the study. Evidence of ivermectin resistance was documented in one group of yearlings with a mean FECR of 91.2% and a 95% credible interval of 84.0-95.8. The results from one mare group and one additional yearling group were deemed inconclusive, whereas the remaining five groups displayed no evidence of ivermectin resistance. Strongylid shedding in the mares was not positively associated with any of the evaluated factors. This study is the first to demonstrate ivermectin resistance in US bred horses, and the findings emphasize the need for routine monitoring of anthelmintic efficacy on horse farms.