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Abstract Background Current methods for obtaining fecal egg counts in horses are often inaccurate and variable depending on the analyst’s skill and experience. Automated digital scanning of fecal sample slides integrated with analysis by an artificial intelligence (AI) algorithm is a viable, emerging alternative that can mitigate operator variation compared to conventional methods in companion animal fecal parasite diagnostics. Vetscan Imagyst is a novel fecal parasite detection system that uploads the scanned image to the cloud where proprietary software analyzes captured images for diagnostic recognition by a deep learning, object detection AI algorithm. The study describes the use and validation of Vetscan Imagyst in equine parasitology. Methods The primary objective of the study was to evaluate the performance of the Vetscan Imagyst system in terms of diagnostic sensitivity and specificity in testing equine fecal samples (n = 108) for ova from two parasites that commonly infect horses, strongyles and Parascaris spp., compared to reference assays performed by expert parasitologists using a Mini-FLOTAC technique. Two different fecal flotation solutions were used to prepare the sample slides, NaNO3 and Sheather’s sugar solution. Results Diagnostic sensitivity of the Vetscan Imagyst algorithm for strongyles versus the manual reference test was 99.2% for samples prepared with NaNO3 solution and 100.0% for samples prepared with Sheather’s sugar solution. Sensitivity for Parascaris spp. was 88.9% and 99.9%, respectively, for samples prepared with NaNO3 and Sheather’s sugar solutions. Diagnostic specificity for strongyles was 91.4% and 99.9%, respectively, for samples prepared with NaNO3 and Sheather’s sugar solutions. Specificity for Parascaris spp. was 93.6% and 99.9%, respectively, for samples prepared with NaNO3 and Sheather’s sugar solutions. Lin’s concordance correlation coefficients for VETSCAN IMAGYST eggs per gram counts versus those determined by the expert parasitologist were 0.924–0.978 for strongyles and 0.944–0.955 for Parascaris spp., depending on the flotation solution. Conclusions Sensitivity and specificity results for detecting strongyles and Parascaris spp. in equine fecal samples showed that Vetscan Imagyst can consistently provide diagnostic accuracy equivalent to manual evaluations by skilled parasitologists. As an automated method driven by a deep learning AI algorithm, VETSCAN IMAGYST has the potential to avoid variations in analyst characteristics, thus providing more consistent results in a timely manner, in either clinical or laboratory settings. Graphical Abstract

Abstract Background Parascaris spp. represent a significant threat to equine health worldwide, particularly in foals. The long-term survival of parasites in the host necessitates persistent modulation of the host immune response. Intercellular communication achieved through the exchange of molecules via extracellular vesicles (EVs) released from the parasite could be a crucial factor in this regard. This study aimed to isolate and characterize EVs released by adult male and female Parascaris worms and conduct a proteomic analysis to identify sex-specific proteins and potential immunomodulatory factors. Methods Live adult Parascaris worms were collected, and EVs were isolated from spent culture media using differential ultracentrifugation. Nanoparticle tracking analysis and transmission electron microscopy confirmed the size, concentration, and morphology of the isolated EVs. Proteins within the isolated EVs were analyzed using mass spectrometry-based proteomics (LC–MS/MS). Results Proteomic analysis revealed a total of 113 proteins in Parascaris EVs, with several proteins showing homology to known helminth exosome proteins and exhibiting immunomodulatory functions. Sex-specific differences in EV protein composition were observed, with a distinct abundance of C-type lectins in female EVs, suggesting potential sex-specific roles or regulation. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses revealed metabolic pathways shared between male and female Parascaris EVs, as well as differences in signal transduction, and cell growth and death pathways, indicating sex-specific variations. Conclusions These findings imply that Parascaris EVs and their protein cargo are complex. This data potentially opens avenues for discovering innovative approaches to managing and understanding helminth infection. Graphical Abstract

Background: Current methods for obtaining fecal egg counts in horses are often inaccurate and variable depending on the analyst's skill and experience. Automated digital scanning of fecal sample slides integrated with analysis by an artificial intelligence (AI) algorithm is a viable, emerging alternative that can mitigate operator variation compared to conventional methods in companion animal fecal parasite diagnostics. Vetscan Imagyst is a novel fecal parasite detection system that uploads the scanned image to the cloud where proprietary software analyzes captured images for diagnostic recognition by a deep learning, object detection AI algorithm. The study describes the use and validation of Vetscan Imagyst in equine parasitology. Methods: The primary objective of the study was to evaluate the performance of the Vetscan Imagyst system in terms of diagnostic sensitivity and specificity in testing equine fecal samples (n = 108) for ova from two parasites that commonly infect horses, strongyles and Parascaris spp., compared to reference assays performed by expert parasitologists using a Mini-FLOTAC technique. Two different fecal flotation solutions were used to prepare the sample slides, NaNO3 and Sheather's sugar solution. Results: Diagnostic sensitivity of the Vetscan Imagyst algorithm for strongyles versus the manual reference test was 99.2% for samples prepared with NaNO3 solution and 100.0% for samples prepared with Sheather's sugar solution. Sensitivity for Parascaris spp. was 88.9% and 99.9%, respectively, for samples prepared with NaNO3 and Sheather's sugar solutions. Diagnostic specificity for strongyles was 91.4% and 99.9%, respectively, for samples prepared with NaNO3 and Sheather's sugar solutions. Specificity for Parascaris spp. was 93.6% and 99.9%, respectively, for samples prepared with NaNO3 and Sheather's sugar solutions. Lin's concordance correlation coefficients for VETSCAN IMAGYST eggs per gram counts versus those determined by the expert parasitologist were 0.924–0.978 for strongyles and 0.944–0.955 for Parascaris spp., depending on the flotation solution. Conclusions: Sensitivity and specificity results for detecting strongyles and Parascaris spp. in equine fecal samples showed that Vetscan Imagyst can consistently provide diagnostic accuracy equivalent to manual evaluations by skilled parasitologists. As an automated method driven by a deep learning AI algorithm, VETSCAN IMAGYST has the potential to avoid variations in analyst characteristics, thus providing more consistent results in a timely manner, in either clinical or laboratory settings. [ABSTRACT FROM AUTHOR]

Background: Parascaris spp. represent a significant threat to equine health worldwide, particularly in foals. The long-term survival of parasites in the host necessitates persistent modulation of the host immune response. Intercellular communication achieved through the exchange of molecules via extracellular vesicles (EVs) released from the parasite could be a crucial factor in this regard. This study aimed to isolate and characterize EVs released by adult male and female Parascaris worms and conduct a proteomic analysis to identify sex-specific proteins and potential immunomodulatory factors. Methods: Live adult Parascaris worms were collected, and EVs were isolated from spent culture media using differential ultracentrifugation. Nanoparticle tracking analysis and transmission electron microscopy confirmed the size, concentration, and morphology of the isolated EVs. Proteins within the isolated EVs were analyzed using mass spectrometry-based proteomics (LC–MS/MS). Results: Proteomic analysis revealed a total of 113 proteins in Parascaris EVs, with several proteins showing homology to known helminth exosome proteins and exhibiting immunomodulatory functions. Sex-specific differences in EV protein composition were observed, with a distinct abundance of C-type lectins in female EVs, suggesting potential sex-specific roles or regulation. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses revealed metabolic pathways shared between male and female Parascaris EVs, as well as differences in signal transduction, and cell growth and death pathways, indicating sex-specific variations. Conclusions: These findings imply that Parascaris EVs and their protein cargo are complex. This data potentially opens avenues for discovering innovative approaches to managing and understanding helminth infection. [ABSTRACT FROM AUTHOR]

Abstract Migration of Ascaris lumbricoides through the papilla of Vater in humans, and entry into the biliary tree, is well‐recognised. Ascaris suum and Toxocara vitulorum have been recovered from the liver of swine and buffalo. We necropsied a Persian Kurdish filly at age 6 months, weighing ∼100 kg. Death evidently was caused by oleander (Nerium oleander) intoxication. An 8‐cm adult male Parascaris was found at the lobar‐left hepatic bile duct junction. We suggest that the nematode entered anteriorly into the hepatic tree, via the duodenum, major duodenal papilla, bile duct, left hepatic duct and finally the lobar duct. Considering the brief 4‐h elapsed time between death and necropsy, and the 18‐cm distance from the major duodenal papilla to the location of the parasite, we conclude that entry into the biliary tree likely occurred ante‐mortem. We advise consideration of Parascaris infection in differential diagnosis of equine hepatic and pancreatic dysfunction.

Anthelmintic resistance is reported in equine nematodes with increasing frequency in recent years, and no new anthelmintic classes have been introduced during the past 40 years. This manuscript reviews published literature describing anthelmintic resistance in cyathostomins, Parascaris spp., and Oxyuris equi with special emphasis on larvicidal efficacy against encysted cyathostomin larvae and strongylid egg reappearance periods (ERP). Resistance to benzimidazoles and pyrimidines is highly prevalent in cyathostomin populations around the world, and macrocyclic lactone resistance has been documented in cyathostomins in recent years as well. Two recent studies have documented resistance to the larvicidal regimen of fenbendazole, whereas the larvicidal efficacy of moxidectin is variable, but with no evidence of a reduction from historic levels. In the 1990s, ERP estimates were 8–10 and 12–16 weeks for ivermectin and moxidectin, respectively, while several studies published after year 2000 found ERPs to be 5 weeks for both compounds. This is a clear change in anthelmintic performance, but it remains unclear if this is due to development of anthelmintic resistance or selection for other biological traits leading to a quicker resumption of strongylid egg shedding following anthelmintic treatment. Macrocyclic lactone resistance is common in Parascaris spp. around the world, but recent reports suggests that resistance to the two other classes should be monitored as well. Finally, O. equi has been reported resistant to ivermectin and moxidectin in countries representing four continents. In conclusion, multi-drug resistance is becoming the norm in managed cyathostomin populations around the world, and a similar pattern may be emerging in Parascaris spp. More work is required to understand the mechanisms behind the shortened ERPs, and researchers and veterinarians around the world are encouraged to routinely monitor anthelmintic efficacy against equine nematodes.

Abstract 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

Migration of Ascaris lumbricoides through the papilla of Vater in humans, and entry into the biliary tree, is well‐recognised. Ascaris suum and Toxocara vitulorum have been recovered from the liver of swine and buffalo. We necropsied a Persian Kurdish filly at age 6 months, weighing ∼100 kg. Death evidently was caused by oleander (Nerium oleander) intoxication. An 8‐cm adult male Parascaris was found at the lobar‐left hepatic bile duct junction. We suggest that the nematode entered anteriorly into the hepatic tree, via the duodenum, major duodenal papilla, bile duct, left hepatic duct and finally the lobar duct. Considering the brief 4‐h elapsed time between death and necropsy, and the 18‐cm distance from the major duodenal papilla to the location of the parasite, we conclude that entry into the biliary tree likely occurred ante‐mortem. We advise consideration of Parascaris infection in differential diagnosis of equine hepatic and pancreatic dysfunction. [ABSTRACT FROM AUTHOR]

Summary: Background: Pulmonary oedema is an uncommon but life‐threatening condition in horses. It is defined as an abnormal accumulation of liquid and solute in the extravascular tissues and spaces of the lungs, which causes oxygen diffusion impairment and a reduction in lung compliance. Diagnosis is determined by typical clinical signs, including frothy, haemorrhagic nasal discharge and respiratory distress, and can be supported by thoracic imaging and arterial blood gas analysis. In horses, post‐anaesthetic pulmonary oedema (PAPOE) is poorly reported, and there is a lack of information regarding risk factors and prognosis. Objectives: The objective of the study was to determine risk factors and short‐term outcome for PAPOE during the recovery period in horses. Study design: A retrospective case‐control study including 17 cases and 53 controls. Methods: Medical records of clinical cases and controls over a period of four and a half years were reviewed. Short‐term outcomes and risk factors for PAPOE were investigated using conditional multivariable logistic regression analysis. Results: Ascarid impaction prevalence in PAPOE cases was significantly higher than controls (p = 0.03). In a univariable analysis, the following variables were significantly associated with PAPOE (p ≤ 0.05): gastric reflux on admission, haemorrhage during surgery, intravenous crystalloid treatment prior to admission, fresh frozen plasma and hydroxyethyl starch administration during anaesthesia. On multivariable analysis, gastric reflux on admission (OR = 7.9, 95% CI 1.1, 57.2) and fresh frozen plasma treatment during anaesthesia (OR = 9.6, 95% CI 1.2, 79.1) were found as risk factors for PAPOE. There was no significant difference in outcome between cases and controls. Main limitations: The low number of cases, the lack of gross and histological post‐mortem examinations of the horses that did not survive and the clinical definition of PAPOE. Conclusions: Gastric reflux and fresh frozen plasma administration during general anaesthesia are associated with PAPOE. [ABSTRACT FROM AUTHOR]

Anthelmintic resistance is reported in equine nematodes with increasing frequency in recent years, and no new anthelmintic classes have been introduced during the past 40 years. This manuscript reviews published literature describing anthelmintic resistance in cyathostomins, Parascaris spp., and Oxyuris equi with special emphasis on larvicidal efficacy against encysted cyathostomin larvae and strongylid egg reappearance periods (ERP). Resistance to benzimidazoles and pyrimidines is highly prevalent in cyathostomin populations around the world, and macrocyclic lactone resistance has been documented in cyathostomins in recent years as well. Two recent studies have documented resistance to the larvicidal regimen of fenbendazole, whereas the larvicidal efficacy of moxidectin is variable, but with no evidence of a reduction from historic levels. In the 1990s, ERP estimates were 8–10 and 12–16 weeks for ivermectin and moxidectin, respectively, while several studies published after year 2000 found ERPs to be 5 weeks for both compounds. This is a clear change in anthelmintic performance, but it remains unclear if this is due to development of anthelmintic resistance or selection for other biological traits leading to a quicker resumption of strongylid egg shedding following anthelmintic treatment. Macrocyclic lactone resistance is common in Parascaris spp. around the world, but recent reports suggests that resistance to the two other classes should be monitored as well. Finally, O. equi has been reported resistant to ivermectin and moxidectin in countries representing four continents. In conclusion, multi-drug resistance is becoming the norm in managed cyathostomin populations around the world, and a similar pattern may be emerging in Parascaris spp. More work is required to understand the mechanisms behind the shortened ERPs, and researchers and veterinarians around the world are encouraged to routinely monitor anthelmintic efficacy against equine nematodes. [Display omitted] • Cyathostomins are widely resistant to pyrimidines and benzimidazoles. • Cyathostomins have been reported resistant to macrocyclic lactones in recent years. • The interpretation of shortened egg reappearance periods is not clear. • Parascaris spp. are widely resistant to macrocyclic lactones. • Oxyuris equi has been documented resistant to macrocyclic lactones. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The equine ascarids, Parascaris spp., are important nematode parasites of juvenile horses and were historically model organisms in the field of cell biology, leading to many important discoveries, and are used for the study of chromatin diminution. In veterinary parasitology, Parascaris spp. are important not only because they can cause clinical disease in young horses but also because they are the only ascarid parasites to have developed widespread anthelmintic resistance. Despite this, much of the general biology and mechanisms of anthelmintic resistance are poorly understood. This review condenses known basic biological information and knowledge on the mechanisms of anthelmintic resistance in Parascaris spp., highlighting the importance of foundational research programs. Although two variants of this parasite were recognized based on the number of chromosomes in the 1870s and suggested to be two species in 1890, one of these, P. univalens, appears to have been largely forgotten in the veterinary scientific literature over the past 100 years. We describe how this omission has had a century-long effect on nomenclature and data analysis in the field, highlighting the importance of proper specimen identification in public repositories. A summary of important basic biology, including life cycle, in vitro maintenance, and immunology, is given, and areas of future research for the improvement of knowledge and development of new systems are given. Finally, the limited knowledge regarding anthelmintic resistance in Parascaris spp. is summarized, along with caution regarding assumptions that resistance mechanisms can be applied across clades. [ABSTRACT FROM AUTHOR]

The presence of digestive parasites can alter behaviour, fertility, body condition, juvenile development, decreased resistance to other pathogens, or the performance for which animals are bred. The aim of this study was to monitor gastrointestinal parasite infestation in equines in Mehedinți County. The horses, sport and working, various breeds and half-breeds, came from different localities from Mehedinți County. Samples were collected in plastic bags and examined by flotation, sedimentation, and Baermann methods. Horses were divided into several categories: sex, age, breed, and service. Strongylidae and Parascaris spp. represented parasitism in recreational and working horses studied in Mehedinți county, no other parasitic elements were identified. The overall prevalence was 43.5% for Strongylidae and 21.17% for Parascaris spp. The simultaneous presence of at least two parasite species was observed in 17.64% of the horses examined. [ABSTRACT FROM AUTHOR]

Abstract Background Equine ascariosis, caused by Parascaris spp., is a worldwide endoparasitic disease affecting young horses in particular. Despite the great number of horses reared in Italy, large-scale epidemiological surveys dealing with ascariosis prevalence in the country are not reported in the current literature. For this reason, the present survey aims to describe, for the first time, the spread and infestation of Parascaris spp. in a large population of Italian horses (6896 animals) using faecal egg counts, and further to identify risk factors associated with ascarid egg shedding. Methods Individual rectal faecal samples collected during routine veterinary examinations were used and Parascaris spp. prevalence was tested against the animal’s age, sex, housing conditions, geographic provenance as well as the respective sampling season. Results Among the examined stables, 35.8% showed at least one horse to be positive for Parascaris spp. eggs and an overall prevalence of 6.3% was found. Ascariosis rates tended to decrease significantly with age and, proportionally, 80.0% of the recorded Parascaris spp. eggs were found in 0.7% of the examined animals. Statistically significant differences among prevalence rates were found between the different geographic areas of provenance and prevalence was found to be higher in horses reared outdoors compared to those raised indoors. Analysis of data based on sex and season did not show any significant differences. Despite the lower prevalence found compared to other European countries, ascariosis was concluded to represent a significant health challenge for horses reared in Italy, especially foals. Age (foals and yearlings) and outdoor rearing were identified to be significant risk factors for Parascaris spp. egg shedding. Furthermore, the relevance of the infected horses over 6 years of age should not be underestimated as these represent a significant source of contamination for younger animals. Conclusions The development of improved treatment protocols based on regular faecal examination combined with follow-up assessment of the efficacy of integrated action plans would prove beneficial in regard to animal health and anthelmintic resistance reduction in the field. Graphic Abstract

Benzimidazoles resistance is widespread in strongyle parasitic nematodes and associated with polym orphisms in the codons 167, 198 and 200 of isotype 1 β-tubulin (tbb-1). In ascarids, benzimidazole (BZ) resistance has rarely been reported and in none of these cases were any of these polymorphisms detected. Here, available genome and transcriptome data from WormBase ParaSite were used to compare the complete β-tubulin reservoirs of Parascaris univalens, Ascaris suum and Ascaris lumbricoides. Adult Parascaris spp. specimens collected in Australia from horses after BZ treatment (susceptible, n = 13) or surviving BZ treatment and collected after ivermectin treatment (resistant, n = 10) were genotyped regarding codons 167, 198 and 200 using Sanger sequencing. Phylogenetic analyses clearly showed that there are no one-to-one ascarid orthologs of strongyle tbb-1 genes. In the reference genomes, as well as phenotypically susceptible and resistant Parascaris spp. from Australia, six out of seven β-tubulin genes showed a BZ-susceptible genotype (F167, E198, F200). The only exception were the testis-specific β-tubulin D genes from all three ascarid species that encode tyrosine at codon 200. This was observed independently of the BZ-susceptibility phenotype of Parascaris spp. These data suggest that different mechanisms lead to BZ resistance in ascarid and strongyle nematodes. [ABSTRACT FROM AUTHOR]

Background: Equine ascariosis, caused by Parascaris spp., is a worldwide endoparasitic disease affecting young horses in particular. Despite the great number of horses reared in Italy, large-scale epidemiological surveys dealing with ascariosis prevalence in the country are not reported in the current literature. For this reason, the present survey aims to describe, for the first time, the spread and infestation of Parascaris spp. in a large population of Italian horses (6896 animals) using faecal egg counts, and further to identify risk factors associated with ascarid egg shedding. Methods: Individual rectal faecal samples collected during routine veterinary examinations were used and Parascaris spp. prevalence was tested against the animal's age, sex, housing conditions, geographic provenance as well as the respective sampling season. Results: Among the examined stables, 35.8% showed at least one horse to be positive for Parascaris spp. eggs and an overall prevalence of 6.3% was found. Ascariosis rates tended to decrease significantly with age and, proportionally, 80.0% of the recorded Parascaris spp. eggs were found in 0.7% of the examined animals. Statistically significant differences among prevalence rates were found between the different geographic areas of provenance and prevalence was found to be higher in horses reared outdoors compared to those raised indoors. Analysis of data based on sex and season did not show any significant differences. Despite the lower prevalence found compared to other European countries, ascariosis was concluded to represent a significant health challenge for horses reared in Italy, especially foals. Age (foals and yearlings) and outdoor rearing were identified to be significant risk factors for Parascaris spp. egg shedding. Furthermore, the relevance of the infected horses over 6 years of age should not be underestimated as these represent a significant source of contamination for younger animals. Conclusions: The development of improved treatment protocols based on regular faecal examination combined with follow-up assessment of the efficacy of integrated action plans would prove beneficial in regard to animal health and anthelmintic resistance reduction in the field. [ABSTRACT FROM AUTHOR]

Ascaris and Parascaris are important parasites in the family Ascarididae, large, ubiquitous intestinal-dwelling nematodes infecting all classes of vertebrates. Parasitic nematode drug resistance in veterinary medicine and drug recalcitrance in human medicine are increasing worldwide, with few if any new therapeutic classes on the horizon. Some of these parasites are zoonotic, e.g., Ascaris is passed from humans to pigs and vice versa. The development of new therapies against this family of parasites would have major implications for both human and livestock health. Here we tested the therapeutic ability of a paraprobiotic or dead probiotic that expresses the Bacillus thuringiensis Cry5B protein with known anthelmintic properties, against zoonotic Ascaris suum and Parascaris spp. This paraprobiotic, known as IBaCC, intoxicated A. suum larvae in vitro and was highly effective in vivo against intestinal A. suum infections in a new mouse model for this parasite. Fermentation was scaled up to 350 l to treat pigs and horses. Single dose Cry5B IBaCC nearly completely cleared A. suum infections in pigs. Furthermore, single dose Cry5B IBaCC drove fecal egg counts in Parascaris-infected foals to zero, showing at least parity with, and potential superiority to, current efficacy of anthelmintics used against this parasite. Cry5B IBaCC therefore represents a new, paraprobiotic One Health approach towards targeting Ascarididae that is safe, effective, massively scalable, stable, and useful in human and veterinary medicine in both the developed and developing regions of the world.

Benzimidazoles resistance is widespread in strongyle parasitic nematodes and associated with polym orphisms in the codons 167, 198 and 200 of isotype 1 β-tubulin (tbb-1). In ascarids, benzimidazole (BZ) resistance has rarely been reported and in none of these cases were any of these polymorphisms detected. Here, available genome and transcriptome data from WormBase ParaSite were used to compare the complete β-tubulin reservoirs of Parascaris univalens, Ascaris suum and Ascaris lumbricoides. Adult Parascaris spp. specimens collected in Australia from horses after BZ treatment (susceptible, n = 13) or surviving BZ treatment and collected after ivermectin treatment (resistant, n = 10) were genotyped regarding codons 167, 198 and 200 using Sanger sequencing. Phylogenetic analyses clearly showed that there are no one-to-one ascarid orthologs of strongyle tbb-1 genes. In the reference genomes, as well as phenotypically susceptible and resistant Parascaris spp. from Australia, six out of seven β-tubulin genes showed a BZ-susceptible genotype (F167, E198, F200). The only exception were the testis-specific β-tubulin D genes from all three ascarid species that encode tyrosine at codon 200. This was observed independently of the BZ-susceptibility phenotype of Parascaris spp. These data suggest that different mechanisms lead to BZ resistance in ascarid and strongyle nematodes.

Horses in Iceland have been isolated for more than 1,000 yr but still harbor a similar range of gastrointestinal parasites as do horses across the world. The long isolation of the horses and their parasites presumably means that no resistance genes have been introduced into the Parascaris spp. population. It is therefore of particular interest to investigate the efficacy of ivermectin on Parascaris spp. infecting Icelandic foals. Potential treatment failure of ivermectin in Iceland will add substantial new information on how resistance can arise independently. This study aimed to determine the efficacy of subcutaneous injection of ivermectin for the treatment of Parascaris spp. infection in foals and to identify the Parascaris species present in the west and north of Iceland. A fecal egg count reduction (FECR) test (FECRT) was performed on 50 foals from 8 farms, including an untreated control group of 6 foals, from September to November 2019. The foals were between 3 and 5 mo of age at the start of the study and had not previously been treated with anthelmintic drugs. Each foal was treated subcutaneously with off-label use of Ivomec® injection 10 mg/ml or Noromectin® 1% at a dose of 0.2 mg/kg. The FECR for each farm was calculated in 2 ways, by the eggCounts package in R and by the Presidente formula (FECRT). Both calculation methods resulted in efficacy levels between 0% and 80.78%, indicating ivermectin resistance on all farms. We also confirmed, by karyotyping, that the species of equine ascarid present in the west and north of Iceland is Parascaris univalens. This study provides evidence for treatment failure of ivermectin against P. univalens infection in foals. Since Icelandic horses have been isolated on the island for more than 1,000 yr, this implies that resistance alleles have developed independently in the Icelandic Parascaris population. The actual clinical impact of ivermectin resistance is unknown but another drug of choice should be considered to treat Parascaris infection in foals in Iceland. [ABSTRACT FROM AUTHOR]

An increasing number of metazoans undergo programmed DNA elimination (PDE), where a significant amount of DNA is selectively lost from the somatic genome during development. In some nematodes, PDE leads to the removal and remodeling of the ends of all germline chromosomes. In several species, PDE also generates internal breaks that lead to sequence loss and increased numbers of somatic chromosomes. The biological significance of these karyotype changes associated with PDE and the origin and evolution of nematode PDE remain largely unknown. Here, we assembled the single germline chromosome of the nematode Parascaris univalens and compared the karyotypes, chromosomal gene organization, and PDE features among other nematodes. We show that PDE in Parascaris converts an XX/XY sex-determination system in the germline into an XX/XO system in the somatic cells. Comparisons of Ascaris , Parascaris , and Baylisascaris ascarid chromosomes suggest that PDE existed in the ancestor of these nematodes, and their current distinct germline karyotypes were derived from fusion events of smaller ancestral chromosomes. The DNA breaks involved in PDE resolve these fused germline chromosomes into their pre-fusion karyotypes. These karyotype changes may lead to alterations in genome architecture and gene expression in the somatic cells. Cytological and genomic analyses further suggest that satellite DNA and the heterochromatic chromosome arms are dynamic and may play a role during meiosis. Overall, our results show that chromosome fusion and PDE have been harnessed in these ascarids to sculpt their karyotypes, altering the genome organization and serving specific functions in the germline and somatic cells. [Display omitted] • Complete genomes reveal karyotype differences in the Parascaris germline and soma • Current ascarid germline chromosomes are derived from fusion of smaller chromosomes • PDE converts distinct germline karyotypes to the same somatic karyotype in ascarids • PDE provides a means for organisms to benefit from both fused and split chromosomes Chromosome fusion and fission change karyotypes; they are prevalent in nematodes with holocentric chromosomes. Simmons et al. show some parasitic nematodes, including the unichromosomal Parascaris univalens , use programmed DNA elimination (PDE) to split evolutionarily fused germline chromosomes to restore their ancestral karyotype in somatic cells. [ABSTRACT FROM AUTHOR]

• Parasite loads of 1,673 young horses examined at necropsy over 29 years were analysed. • Parasite burden varied across season and was reduced in race horses. • Fatal Parascaris spp. and cyathostomin infections have been on the rise since 2000. • Available deworming history data was compatible with lower drug efficacy after 2000. Horses are infected by a wide range of parasite species that form complex communities. Parasite control imposes significant constraints on parasite communities whose monitoring remains, however, difficult to track through time. Postmortem examination is a reliable method to quantify parasite communities. Here, we compiled 1,673 necropsy reports accumulated over 29 years, in the reference necropsy centre from Normandy (France). The burden of non-strongylid species was quantified and the presence of strongylid species was noted. Details of horse deworming history and the cause of death were registered. Building on these data, we investigated the temporal trend in non-strongylid epidemiology and we determined the contribution of parasites to the deaths of horses throughout the study period. Data analyses revealed the seasonal variations of non-strongylid parasite abundance and reduced worm burden in race horses. Beyond these observations, we found a shift in the species responsible for fatal parasitic infection from the year 2000 onward, whereby fatal cyathostominosis and Parascaris spp. infection have replaced cases of death caused by Strongylus vulgaris and tapeworms. A concomitant break in the temporal trend of parasite species prevalence was also found within a 10 year window (1998–2007) that has seen the rise of Parascaris spp. and the decline of both Gasterophilus spp. and tapeworms. A few cases of parasite persistence following deworming were identified, which all occurred after 2000. Altogether, these findings provide insights into major shifts in non-strongylid parasite prevalence and abundance over the last 29 years. They also underscore the critical importance of Parascaris spp. in young equids. [ABSTRACT FROM AUTHOR]

• High horse movement frequency is a risk factor for Parascaris sp. egg shedding. • Horses originating from large breeding farms are more likely to shed ascarid eggs. • Reduced efficacy of pyrantel against Parascaris sp. was observed. • Proportion of horses shedding strongylid eggs was lowest during the spring time. Gastrointestinal parasites, Parascaris sp. and strongyles, are common in young horses worldwide and control of these parasites is challenged by increasing anthelmintic resistance. Our aim was to identify risk factors for these infections as well as to assess the efficacy of fenbendazole (dose 7.5 mg/kg) and pyrantel embonate (dose 19 mg/kg) against Parascaris sp. We also evaluated association between owner observed symptoms and patent infections with these parasites. Fecal samples were collected from 367 young horses in Finland and a questionnaire study was conducted. Fecal egg counts were performed by Mini-FLOTAC® method. Univariable logistic regression models using patent infection status (Yes/No), separately for Parascaris sp. and strongyle infections as an outcome were run initially to screen potential risk factors collected by the questionnaire. After the initial screening, multiple logistic regression models were constructed and run to account for correlated data structure, risk factors and potential confounders simultaneously. Two significant risk factors for a patent Parascaris sp. infection were found: breeding farm size (p = 0.028) and frequency of horse movements (p = 0.010). Horses originating from large breeding farms were more likely (OR = 2.47, 95% confidence interval (CI) 1.10–5.51) to shed Parascaris sp. eggs upon relocation to training stables compared to horses originating from small breeding farms. Horses living in farms with frequent horse movements to other premises had higher odds (OR = 3.56, 95% CI: 1.35–9.39) of a patent Parascaris sp. infection compared to farms with less frequent horse movements. Risk factors for patent strongyle infection included age (p < 0.001) and season (p = 0.017). Horses were less likely (OR = 0.27, 95% CI: 0.10 - 0.66) to shed strongylid eggs during the spring compared to the winter. Horses excreting over 200 ascarid eggs per gram were included in the anthelmintic efficacy trial. A mean FECR less than 90% was interpreted as presence of anthelmintic resistance. The mean FECR was 98.5% (95% CI: 95.8–100) and 68.0% (95% CI: 52.7–83.3) in the fenbendazole (n = 31) and pyrantel (n = 26) treatment groups, respectively. In conclusion, we identified two new risk factors for patent Parascaris sp. infection; breeding farm size and frequency of horse movements. Reduced efficacy of pyrantel against Parascaris sp. was observed for the second time in Europe. A relatively high Parascaris sp. prevalence in yearlings (34%) and two-year-olds (20%) was observed, which has not been reported earlier. An association between symptoms and a patent Parascaris sp. infection was observed in foals. [ABSTRACT FROM AUTHOR]

• The variability in faecal egg count results can influence clinical decision making. • Egg density in faeces is inhomogeneous and follows a negative binomial distribution. • A single or mean egg count does not account for variation in egg density in faeces. • The negative predictive value and sensitivity of a single egg count of zero is low. • Confidence intervals should be used to estimate the distribution mean egg count. This study investigated the impact of variability in Parascaris spp. and strongyle faecal egg counts (FEC) from foals on treatment decision-making and detection of a patent infection. A single faecal sample was collected once daily for three days from 53 foals and a FEC was performed on three separate portions of each sample (total of nine egg counts per foal). Differences in the decision to administer an anthelmintic using the results of a single count (C 1), the mean of three (X ¯ 1–3) or nine counts (X ¯ 1–9) and the upper 5% confidence limit of the gamma confidence interval (CI) of the estimate of the distribution mean (μ) from three (UCL 1–3) and nine counts (UCL 1–9) were determined for a range of egg count thresholds. The UCL 1–9 was used as the best estimate of μ, hypothesis testing for treatment and the comparison of treatment decision-making using C 1 , X ¯ 1–3 , X ¯ 1–9 and UCL 1–3. The results of this study demonstrated that a point estimate (C 1 or X ¯ 1–3) was of limited value for estimating the distribution mean of egg counts in faeces and there was overall poor agreement in treatment decision-making for individual foals using C 1 compared with UCL 1–9. Of the foals with C 1 of zero eggs per gram, 54% and 47% had Parascaris and strongyle eggs in subsequent counts, respectively. The egg density in faeces is inhomogeneous, resulting in considerable variability in egg count results for an individual foal: between faecal piles, different portions of a faecal pile and days. The use of the negative binomial distribution CI for μ takes this variability into account and is recommended for use when interpreting FEC data from horses. [ABSTRACT FROM AUTHOR]

Parascaris sp. is the only ascarid parasitic nematode in equids and one of the most threatening infectious organisms in horses. Only a limited number of compounds are available for treatment of horse helminthiasis, and Parascaris sp. worms have developed resistance to the three major anthelmintic families. In order to overcome the appearance of resistance, there is an urgent need for new therapeutic strategies. The active ingredients of herbal essential oils are potentially effective antiparasitic drugs. Carvacrol is one of the principal chemicals of essential oil from Origanum, Thymus, Coridothymus, Thymbra, Satureja and Lippia herbs. However, the antiparasitic mode of action of carvacrol is poorly understood. Here, the objective of the work was to characterize the activity of carvacrol on Parascaris sp. nicotinic acetylcholine receptor (nAChR) function both in vivo with the use of worm neuromuscular flap preparations and in vitro with two-electrode voltage-clamp electrophysiology on nAChRs expressed in Xenopus oocytes. We developed a neuromuscular contraction assay for Parascaris body flaps and obtained acetylcholine concentration-dependent contraction responses. Strikingly, we observed that 300 µM carvacrol fully and irreversibly abolished Parascaris sp. muscle contractions elicited by acetylcholine. Similarly, carvacrol antagonized acetylcholine-induced currents from both the nicotine-sensitive AChR and the morantel-sensitive AChR subtypes. Thus, we show for the first time that body muscle flap preparation is a tractable approach to investigating the pharmacology of Parascaris sp. neuromuscular system. Our results suggest an intriguing mode of action for carvacrol, being a potent antagonist of muscle nAChRs of Parascaris sp. worms, which may account for its antiparasitic potency.

All around the world, intestinal helminths constitute one of the most prevalent life-long occurring infections and re-infections affecting all horse age groups. A range of parasite species among strongyles, ascarids, and tapeworms is known to have the potential to cause colic in horses. However, there is a lack of current scientific evidence on the actual relevance of helminth infection levels in the context of colic in horses kept during prevailing epidemiological conditions. Thus, a prospective case-control study on the occurrence of intestinal helminths in a total of 620 mainly adult equine clinic patients was conducted to investigate the association between colic and helminth infection. For each horse, a range of copromicroscopic, serological, and clinical data was obtained, in addition to a questionnaire on relevant anamnestic data, including previous anthelmintic treatment and husbandry. Using a FLOTAC-based copromicroscopic diagnosis, the highest infection rates were seen for strongyles (41.8%), followed by Anoplocephala perfoliata and Parascaris spp. (both 0.8%), with no significant difference between the two study groups. Employing a real-time PCR a 1.1% S. vulgaris DNA prevalence was found. Considerably higher seroprevalences were observed using S. vulgaris and A. perfoliata ELISAs, with 32.3% and 10.7%, respectively. It was noteworthy that no association concerning either serologic status was encountered with colic status. The shedding of strongyle eggs was associated with a 1.8-times increased risk of S. vulgaris seropositivity. Recent anthelmintic treatment was associated with the onset of colic, as animals who had received an anthelmintic during the previous week had a 2.4-times higher risk of signs of colic compared to those who had been treated at least eight weeks prior. Another noteworthy observation was that ponies were significantly less often affected by colic than warmbloods. The high S. vulgaris and considerable A. perfoliata seroprevalences encountered in this investigation should prompt veterinarians, farm managers, and horse owners to maintain consequent and effective worm control measures.

Highlights • Transcripts of P-glycoprotein were detected by a novel multiple nucleic acid hybridization method. • Multiple nucleic acid hybridization can detect mRNA transcripts within delicate nematode tissues. • P-glycoproteins are expressed in a variety of tissues in Parascaris. Graphical abstract Abstract Macrocyclic lactone-resistant Parascaris have been reported throughout the world. In part, the drug resistant phenotype is hypothesized to be associated with ATP-binding cassette transporters known as P-glycoproteins. In many systems, P-glycoproteins efflux drugs out of cells thereby precluding drug binding to target receptors. Parascaris may evade macrocyclic lactone-mediated death by effluxing drugs away from target receptors in the nervous system. Alternatively, P-glycoprotein expression in the gut or body wall could prevent penetration of drugs into the body of the parasite altogether. In the present study, we evaluate expression of Peq-pgp-11 and Peq-pgp-16 using a novel multiple nucleic acid hybridization method. This method allowed for visualization of individual mRNA transcripts within fixed tissue sections of Parascaris adults. Our investigation revealed expression of Peq-pgp-11 and Peq-pgp-16 in the intestine, body wall, nerves, lateral cords, and reproductive tissues of male and female parasites. These results suggest that Pgp could efflux drugs locally at the level of parasite neuronal tissue as well as at sites of entry for drugs such as the hypodermis and intestine. The multiple nucleic acid hybridization method could be useful for providing tissue context for gene expression in a variety of nematode parasites. [ABSTRACT FROM AUTHOR]

A previously described model for the dynamics of the parasitic stages of Parascaris spp. was modified to include eggs outside the host and the genetics of anthelmintic resistance before being used to address questions regarding the development of resistance. Three broad questions were addressed; i) How sustainable is the current common practice of treating foals monthly for their first year of life (i.e. 12 treatments/year)? ii) Does the timing of treatments have an effect on resistance development? (i.e. do certain treatments select for resistance more strongly than others?), and iii) How sustainable is the currently recommended strategy of targeting ascarid infections in foals with two treatments applied during the first five months of life? A range of variations within these broad questions were considered, such as the value in rotational deworming, whether larvicidal treatments are more selective for resistance, and whether combination anthelmintics should be introduced. Twelve anthelmintic treatments at monthly intervals resulted in the development of resistance to all the anthelmintics used, regardless of how they were used, indicating that such intensive treatment frequency is unlikely to be sustainable. The timing of a single annual treatment influenced resistance development with treatments at 3 and 4 months of age being more selective than treatments at other times. Treatments administered to foals older than 6 months of age did not select for resistance within the timeframe of these simulations. Treatments with activity against migrating third stage larvae (ivermectin and a programme of 5 daily treatments with fenbendazole) were more selective for resistance than those which only killed worms in the intestine. Restricting the number of treatments to young foals to two, administered at 2 and 5 months of age slowed the development of resistance by allowing a small contribution from susceptible genotype worms to subsequent generations. If the interval between treatments was reduced, resistance developed more rapidly demonstrating the importance of allowing some susceptible worms to reach patency before the second treatment is administered. Under a reduced treatment schedule with a clearly defined ‘refugium’ of susceptibility, the use of effective actives in combination appears to offer advantages for delaying resistance development. The model offers insights into more sustainable drug use strategies and has identified some priority questions for future research. [ABSTRACT FROM AUTHOR]

Parasite control in juvenile horses remains a substantial challenge for veterinary practitioners and their clients. This age group is at risk for various types of parasitic disease because their worm burdens tend to be larger and more diverse than in mature horses. This contrast is largely attributable to marked differences in relative levels of acquired immunity. Frequent, perennial anthelmintic treatment is not a sustainable approach for parasite control in this age group, but the simple, surveillance-based parasite control strategies recommended for mature horses are not appropriate for juveniles either. Maneuvering through the early years of a horse's life requires up-to-date knowledge about biology and epidemiology of the parasite species in play as well as local evidence regarding the anthelmintic resistance status of each herd. This article provides an overview of the most important helminth parasites to target in a control programme for juvenile horses, and highlights how the target species change as horses age. Young foals (<2 months) are exposed to Strongyloides westeri and Parascaris spp., whereas the primary target parasite in foals between age 2 months and weaning is Parascaris spp. Beyond weaning, the focus shifts away from ascarids, and the targets instead are strongyles and tapeworms. Gasterophilus spp. and Oxyuris equi are also discussed. [ABSTRACT FROM AUTHOR]

Literature documenting the growth and development of Parascaris spp. infections was used to develop a model describing worm dynamics in the young horse. The model incorporates four main variables; the rate at which larvae migrate through host tissues to return to the small intestine, the proportion of migrating larvae which succeed in returning to the small intestine, the rate of growth in size of maturing and adult worms and the survival rate of maturing and adult worms. In addition, the number of eggs laid each day by adult female worms is calculated as a function of worm size (length) and is used to calculate faecal egg output of the foal. Published data describing the rate of migration through host tissues, and the growth of worms following their return to the small intestine, was used to derive relationships describing these processes. However, only limited data exists relating the survival of migrating larvae and mature worms in the intestine to host age and experience of infection. Therefore, relationships and coefficients describing these variables were modified so that output aligned with published experimental results. As a consequence, the model has not yet been evaluated against an independent data set, and so remains as the best ‘current hypothesis’ for the dynamics of this parasite. Hopefully, future experiments designed to test specific assumptions and outputs of the model will lead to a better understanding of the biology of this important parasite. For example, the most influential variable in determining model output is the survival rate of worms in the small intestine. In the model, worm survival declines in response to both the increasing age of the horse and the increasing cumulative length of worms in the intestine (used as a proxy for crowding). Given the importance of this variable to model behaviour and the paucity of experimental data on this topic this would seem a priority for future study. Initial experiments using the model suggest that a single anthelmintic treatment, administered soon after patency of initial infection, may effectively control environmental contamination with Parascaris spp. eggs while allowing a small ‘refugia’ of susceptibility to delay the emergence of anthelmintic resistance. Further evaluations of the practicality of this approach may be worthwhile. [ABSTRACT FROM AUTHOR]

Reasons for performing study: Parascaris spp. infections can lead to life-threatening small intestinal impactions in foals. Currently available diagnostic techniques cannot estimate the magnitude of an ascarid burden, and hence identify foals potentially at risk of developing impactions. Objectives: To describe and evaluate an ultrasonographic transabdominal scoring technique for monitoring of ascarid burdens in foals and to perform a cost-benefit analysis of the application of this technique. Study design: A transabdominal ultrasonographic technique was validated against ascarid worm counts from 10 foals aged 162–294 days. In a treatment trial, 15 foals were randomly allocated to 3 treatment groups: ivermectin, oxibendazole and no treatment. Blinded ultrasound examinations were performed daily for 5 consecutive days following treatment. Foals were examined ultrasonographically twice by the same investigator, and by different investigators for intra- and interobserver agreement evaluation. Cost-benefit analyses identified threshold values for the probability of ascarid impactions above which the screening method becomes cost-effective. Methods: The ultrasound technique used 3 locations along the ventral midline. An ascarid scoring system was established that assessed the magnitude of ascarid burden ranging from 1–4. The method was validated against worm burdens of 10 worms and above with calculation of diagnostic specificity, sensitivity, and predictive values. Treatment trial data were evaluated statistically using mixed model analysis. Kappa values were generated for intra- and interobserver agreement. Results: Two consecutive examinations were found to detect worm burdens >10 ascarids reliably. Ascarid scores declined in response to both anthelmintic treatments, although differences were not statistically significant. Kappa values indicated fair to moderate intra- and interobserver agreements. The majority of cost-benefit analyses indicated that ultrasound examinations are cost effective when the probability of ascarid impactions is above a range of 0.0001–0.0082 (i.e. 1 in 10,000 to 8 in 1000 foals). Conclusions: The ultrasonographic screening techniques can be a useful tool for monitoring ascarid burdens in foals. [ABSTRACT FROM AUTHOR]

This article reports the results of a faecal egg count reduction test on 4 farms in France, as an integrated part of the routine deworming strategy against horse cyathostomins and Parascaris equorum. Treatment with fenbendazole ( FBZ) or ivermectin ( IVM) was evaluated in yearlings on Farms 1 and 2 and treatment with pyrantel embonate ( PYR) was tested on Farms 3 and 4. Calculation of the arithmetic mean faecal egg count reduction and the 95% confidence intervals (95% CI) around the mean was performed using bootstrap analysis. For equine cyathostomins, resistance to FBZ was found with an arithmetic mean reduction of 48.8% (95% CI: 1.9-69.3%). On Farms 1 and 2, horses with reduced efficacy were identified. PYR was found to be effective against cyathostomins, with an arithmetic mean reduction of 95.3% (95% CI: 84.6-99.8%), as well as IVM (100%). For P. equorum, both FBZ and PYR were effective (100% reduction). The efficacy of IVM, however, was low (45.5%; 95% CI: 0-96.3%). These results confirm that FBZ resistance in equine cyathostomins is present in France and that anthelmintic resistance to IVM is present in P. equorum. This study underlines the necessity to evaluate the efficacy of horse deworming strategies on a regular basis under field conditions. [ABSTRACT FROM AUTHOR]

Abstract: A study was undertaken at the Texas A&M Horse Center to evaluate and compare the effectiveness of three anthelmintics—ivermectin, fenbendazole, and a combination of ivermectin and pyrantel pamoate—on fecal egg count reductions of cyathostomes and Parascaris equorum in 30 naturally infected foals. The foals were randomized into three treatment groups, with individuals being rerandomized after each 8-week observation period. The treatments of ivermectin and fenbendazole were given at the manufacturer''s recommended doses, and the pyrantel treatment was given at two times the manufacturer''s recommended dose. Fecal egg counts were performed at the time of treatment and at 2-week intervals after treatment for a total of 8 weeks. Each foal received a total of three treatments during the course of the study. Fecal egg counts were performed by a modified McMaster''s test, with a sensitivity of 25 eggs per gram of feces, and by the modified Wisconsin double centrifugal flotation technique, with a sensitivity of 0.2 eggs per gram of feces. Fecal egg reduction percentages were calculated. Analysis of the results showed that ivermectin, either used alone or with pyrantel, was a more effective anthelmintic for cyathostome (small strongyle) control than fenbendazole. Fenbendazole and pyrantel showed a higher initial reduction in Parascaris egg counts when compared with the ivermectin-only-treated group, but this difference lessened over time. The use of the combination treatment showed the best results for controlling both parasites, indicating that a combination of anthelmintics may be necessary to control parasites on some equine farms. [Copyright &y& Elsevier]

Abstract: The expanding prevalence of Parascaris equorum populations that are resistant to macrocyclic lactone (ML) anthelmintics makes it desirable to identify dewormers which remain effective. The objective was to evaluate the efficacy of pyrantel pamoate in 14 suckling foals that had been infected orally with ∼600 larvated eggs of a P. equorum isolate selected for ML resistance (ML-R). Seventy days after inoculation, foals were weaned, housed individually, and fecal samples were examined frequently to detect the onset of patency. Between 73 and 80 days post-inoculation, all 14 foals developed P. equorum egg counts ≥150 eggs per gram (EPG). An initial cohort of eight foals was treated orally with ivermectin paste (200μg/kg) 84–91 days post-inoculation. Egg counts were reduced by only 47% at 2 weeks after ivermectin treatment, confirming the ML-R status of the isolate. A second cohort of six foals was not treated with ivermectin. Within each cohort, eligible foals were allocated randomly to treated (pyrantel pamoate; n =7) or untreated control (n =7) groups. Treated foals were dosed orally on Day 0 with a paste formulation of pyrantel pamoate at 13.2mg/kg. Mean ascarid egg counts of treated foals were reduced by 96.0% and 98.8% at 1 and 2 weeks post-treatment, respectively. On Day 14, foals were euthanatized and specimens of P. equorum were recovered from the gut contents, preserved in 10% formalin, and counted. Mean numbers of P. equorum adults recovered postmortem were significantly lower (P =0.0031) in foals treated with pyrantel pamoate (; range 0–16) compared to control foals (; range 0–320). A paste formulation of pyrantel pamoate, at a dosage of 13.2mg/kg, was 97.3% effective against a ML-R isolate of P. equorum. [Copyright &y& Elsevier]

In Parascaris developmental commitment to the germ line and somatic lineages is indicated by the orientation of the mitotic spindle in blastomeres, the topology of cells in the embryo, and chromatin diminution in presomatic blastomeres. Using three different experimental techniques: transient pressure treatment, application of cytochalasin B, and isolation of blastomeres, we have succeeded in uncoupling several developmental processes during cleavage of P. univalens. The following results were obtained: (1) Following mitotic nondisjunction we observed identical behavior of all chromatids in each blastomere. Thus chromosome differentiation by differential replication does not occur. (2) Chromosome fragments obtained by pressure treatment of egg cells underwent chromatin diminution. Thus this process does not require an intact germ-line chromosome. However, chromosomes immobilized on a monopolar spindle did not undergo chromatin diminution. Thus diminution appears to require segregation of chromatids. (3) Blastomeres that completely lacked chromosomes as a result of mitotic nondisjunction underwent normal early cleavage divisions. (4) Pressure treatment or prolonged treatment with cytochalasin B caused egg cells or germ line blastomeres to lose their germ line quality, as deduced from the coincident occurrence of symmetrical (presomatic-like) cleavage and chromatin diminution. (5) Isolated blastomeres from 2-cell embryos, i.e. 1/2 blastomeres, usually cleaved according to their prospective fates in the whole embryo. However, in some partial embryos derived from such blastomeres, chromatin diminution was delayed for either one or two cleavage mitoses. An 'activation' model as an alternative to a prelocalization model is presented, which can account for early blastomere topogenesis and chromatin diminution. [ABSTRACT FROM AUTHOR]

Fifteen bare paddocks, which consisted of soil only and were located on 12 different studs, were examined for their content of Parascaris equorum eggs in the upper 15 cm of the soil profile. The paddocks were classified into three different groups according to the type of soil: clayey soil (A), morainic soil (B) and gravel or gravel-like sand (C). Soil profiles were collected down to a depth of 15 cm and were divided into three layers: 0-5 cm (D1), 5-10 cm (D2) and 10-15 cm (D3). The eggs in each layer were counted and identified as infective or noninfective eggs. The paddocks in group C, which had good drainage conditions, had significantly lower numbers of eggs in the whole profile and in D1 and D2 than the paddocks in groups A or B. Furthermore, there was a significantly higher proportion of the total egg count present in D3 in the group C paddocks. This may have been due to a higher degree of passive transportation of eggs down the profile in the gravel or gravel-like sand. Even though there was a significantly higher proportion of infective eggs in the soil from the group C paddocks, the lower total numbers of eggs resulted in a lower total number of infective eggs in those paddocks. The study showed that the soil type was an important factor in determining the content of P. equorum eggs in the upper layer of the soil profile in bare paddocks and consequently for the potential infestation of horses with P. equorum. [ABSTRACT FROM AUTHOR]

Parascaris sp. is the only ascarid parasitic nematode in equids and one of the most threatening infectious organisms in horses. Only a limited number of compounds are available for treatment of horse helminthiasis, and Parascaris sp. worms have developed resistance to the three major anthelmintic families. In order to overcome the appearance of resistance, there is an urgent need for new therapeutic strategies. The active ingredients of herbal essential oils are potentially effective antiparasitic drugs. Carvacrol is one of the principal chemicals of essential oil from Origanum, Thymus, Coridothymus, Thymbra, Satureja and Lippia herbs. However, the antiparasitic mode of action of carvacrol is poorly understood. Here, the objective of the work was to characterize the activity of carvacrol on Parascaris sp. nicotinic acetylcholine receptor (nAChR) function both in vivo with the use of worm neuromuscular flap preparations and in vitro with two-electrode voltage-clamp electrophysiology on nAChRs expressed in Xenopus oocytes. We developed a neuromuscular contraction assay for Parascaris body flaps and obtained acetylcholine concentration-dependent contraction responses. Strikingly, we observed that 300 µM carvacrol fully and irreversibly abolished Parascaris sp. muscle contractions elicited by acetylcholine. Similarly, carvacrol antagonized acetylcholine-induced currents from both the nicotine-sensitive AChR and the morantel-sensitive AChR subtypes. Thus, we show for the first time that body muscle flap preparation is a tractable approach to investigating the pharmacology of Parascaris sp. neuromuscular system. Our results suggest an intriguing mode of action for carvacrol, being a potent antagonist of muscle nAChRs of Parascaris sp. worms, which may account for its antiparasitic potency. [ABSTRACT FROM AUTHOR]

Simple Summary: The most important equine intestinal worm species include the roundworms Strongylus vulgaris and Parascaris spp. as well as tapeworms such as Anoplocephala perfoliata. These parasites reside in the small and large intestine and may cause various signs of disease, such as diarrhea, unthriftiness, or colic. However, following decades of routine anti-worm treatments, it is currently unclear what relevance these infections have in the context of colic under the prevailing situation. Therefore, we examined the signs of the presence of the infection concerning the above-mentioned parasites in 620 equine clinic patients, half of these admitted after being diseased with colic and the other admitted due to non-intestinal diseases. With approximately on third of all horses being positive for antibodies directed against S. vulgaris, we detected an unexpectedly high infection rate. With every tenth horse showing respective antibodies, also tapeworm infections were encountered in a considerable proportion of the examined horses. Somewhat unexpectedly, no association between worm infection and colic was detected. However, recent—i.e., during the last seven days—anthelmintic treatment was 2.4 times more often seen in horses showing signs of colic. Overall, the considerable S. vulgaris and tapeworm infection rates mean we should stay alert in continuing worm monitoring and control. All around the world, intestinal helminths constitute one of the most prevalent life-long occurring infections and re-infections affecting all horse age groups. A range of parasite species among strongyles, ascarids, and tapeworms is known to have the potential to cause colic in horses. However, there is a lack of current scientific evidence on the actual relevance of helminth infection levels in the context of colic in horses kept during prevailing epidemiological conditions. Thus, a prospective case-control study on the occurrence of intestinal helminths in a total of 620 mainly adult equine clinic patients was conducted to investigate the association between colic and helminth infection. For each horse, a range of copromicroscopic, serological, and clinical data was obtained, in addition to a questionnaire on relevant anamnestic data, including previous anthelmintic treatment and husbandry. Using a FLOTAC-based copromicroscopic diagnosis, the highest infection rates were seen for strongyles (41.8%), followed by Anoplocephala perfoliata and Parascaris spp. (both 0.8%), with no significant difference between the two study groups. Employing a real-time PCR a 1.1% S. vulgaris DNA prevalence was found. Considerably higher seroprevalences were observed using S. vulgaris and A. perfoliata ELISAs, with 32.3% and 10.7%, respectively. It was noteworthy that no association concerning either serologic status was encountered with colic status. The shedding of strongyle eggs was associated with a 1.8-times increased risk of S. vulgaris seropositivity. Recent anthelmintic treatment was associated with the onset of colic, as animals who had received an anthelmintic during the previous week had a 2.4-times higher risk of signs of colic compared to those who had been treated at least eight weeks prior. Another noteworthy observation was that ponies were significantly less often affected by colic than warmbloods. The high S. vulgaris and considerable A. perfoliata seroprevalences encountered in this investigation should prompt veterinarians, farm managers, and horse owners to maintain consequent and effective worm control measures. [ABSTRACT FROM AUTHOR]