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The aim of this study was to assess the changes in strongylid communities of domestic horses after two decades of regular anthelmintic treatments; the changes in prevalence and relative abundance of individual strongylid species and their contribution to the observed alterations were estimated. The study was conducted in 2023; data collected in 2004 were used for comparison; 39 horses from two farms in Ukraine (22 horses in 2004 and 17 in 2023) were examined. In total, 18,999 strongylid specimens were collected by in vivo diagnostic deworming method before (in 2004, 9119 specimens) and after (in 2023, 9880 specimens) frequent application of anthelmintics. Strongylids were identified morphologically. Thirteen strongylid species were found in horses in 2023; only small strongylids (Cyathostominae) were recorded. In 2004, 21 species were found: 6 species of large strongylids (Strongylinae) and 15 of cyathostomins. Species richness (Margalef’s index) and species diversity (Shannon’s and Simpson’s indexes) decreased over two decades on both farms; the Berger-Parker dominance index for C. nassatus dramatically increased up to 74.4. The dissimilarity of strongylid communities of 2023 and 2004 was primarily connected with the disappearance of large strongylids and rare cyathostomin species after two decades of regular anthelmintic treatments. SIMPER analysis revealed that C. nassatus and C. catinatum mostly contributed to this dissimilarity. A trend of gradual transformation of the strongylid community structure from multimodal (in 2004) to bimodal (in 2023) was observed on both farms. [ABSTRACT FROM AUTHOR]

This article aims to report an unusual case of equine colic associated with cecal lesions induced by parasite compatible with strongyles. A ten-year-old mare, Mangalarga Marchador breed, was attended with history of severe abdominal pain. Upon admission to the Veterinary Hospital, altered circulatory status and blackened feces were observed, in addition to the presence of cylindrical, reddish parasites less than 1 cm in length, consistent with cyathostomins. The patient had no history of deworming, and fecal egg count (FEC) resulted in 1,500 eggs per gram. Due to pain unresponsive to analgesia, the mare underwent exploratory celiotomy, revealing diffuse areas of infarction in the cecum and small colon, leading to euthanasia due to the severity and extent of the lesions. Histopathological analysis of tissue samples collected from the affected segments revealed multifocal tissue necrosis associated with hemorrhage, congestion, and inflammation, along with tubular filamentous structures compatible with cylindrical parasites in the cecum, leading to the diagnosis of parasitic necrohemorrhagic typhlitis. Studies suggest various possibilities regarding the pathogenesis of non-strangulating cecal infarction associated with strongyle parasitism, considered the main gastrointestinal endoparasites of horses and potential agents associated with colic, thus underscoring the need for effective strategies for parasite control in horses. [ABSTRACT FROM AUTHOR]

Abstract Cyathostomins are the largest group of parasites in horses that can be controlled by ivermectin (IVM). This study aimed to run a four-dose titration trial of IVM in 28 naturally infected Thoroughbred yearlings. The local Strongyle population had been recorded to be resistant to IVM (200 µg/kg). The parasite fecal egg count (FEC) was performed to investigate the egg reappearance period (ERP) of two and five weeks (w2pt and w5pt) after IVM treatment. FEC was > 1000 on day zero for all groups. Although 100% FEC reduction was reported at w2pt for all concentrations, the FEC at w5pt revealed < 83% efficacy. This study reports the reduction of ERP using the label dose as well as 300, and 400 µg/kg (double dose) of IVM. The protocol allowed IVM to significantly suppress FEC w2pt although not eliminating adult worms, failing to guarantee an extension of its protection period over 8 weeks. Moreover, the FEC at w5pt possibly means the infection was not cleared, and worms reestablished egg laying. We raised the possibility of withdrawing IVM of control programs when the drug has less than 80% FEC reduction at w5pt.

Consisting of approximately 50 different species, the cyathostomin parasites are ubiquitous in grazing horses. Co-infection with several species is common, and large burdens can cause the fatal disease of larval cyathostominosis. Due to intense anthelmintic drug use, cyathostomin resistance has developed to all available anthelmintic drug groups. Resistance to the anthelmintic drug pyrantel (PYR) has been documented in over 90% of studies published over the past two decades. In Sweden, a study performed in the early 2000s only confirmed resistance in 4.5% of farms. Further, prescription-only administration of equine anthelmintic drugs was enforced in Sweden in 2007. However, it is unknown if this conservative drug use has maintained PYR efficacy in cyathostomins. The aim of the present study was to investigate the effect of PYR on cyathostomin infection in Sweden using fecal egg count reduction tests (FECRTs). Further, the effect of PYR treatment on cyathostomin species composition was studied using metabarcoding.Sixteen farms with at least six horses excreting a minimum of 100 eggs per gram feces were included. Using the current World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines, PYR resistance was demonstrated in nine of farms, with seven farms showing full susceptibility. Farms with low biosecurity measures had significantly lower efficacy of PYR treatment. The most common cyathostomin species were Cylicocyclus nassatus, Cyathostomum catinatum, Cylicostephanus longibursatus, Cys. calicatus, Cys. goldi, Cys. minutus, Coronocyclus coronatus and Cya. pateratum, accounting for 97% of all sequence reads prior to treatment. Of these, Cyc. nassatus and Cya. catinatum had the highest occurrence, accounting for 68% of all sequence reads prior to PYR treatment. Treatment did not significantly affect the species composition. The results highlight the importance of drug efficacy testing when using PYR to treat cyathostomin infection, even when selective anthelmintic treatment and thus low treatment intensity, is used on the farm.

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.

Consisting of approximately 50 different species, the cyathostomin parasites are ubiquitous in grazing horses. Co-infection with several species is common, and large burdens can cause the fatal disease of larval cyathostominosis. Due to intense anthelmintic drug use, cyathostomin resistance has developed to all available anthelmintic drug groups. Resistance to the anthelmintic drug pyrantel (PYR) has been documented in over 90% of studies published over the past two decades. In Sweden, a study performed in the early 2000s only confirmed resistance in 4.5% of farms. Further, prescription-only administration of equine anthelmintic drugs was enforced in Sweden in 2007. However, it is unknown if this conservative drug use has maintained PYR efficacy in cyathostomins. The aim of the present study was to investigate the effect of PYR on cyathostomin infection in Sweden using fecal egg count reduction tests (FECRTs). Further, the effect of PYR treatment on cyathostomin species composition was studied using metabarcoding. Sixteen farms with at least six horses excreting a minimum of 100 eggs per gram feces were included. Using the current World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines, PYR resistance was demonstrated in nine of farms, with seven farms showing full susceptibility. Farms with low biosecurity measures had significantly lower efficacy of PYR treatment. The most common cyathostomin species were Cylicocyclus nassatus, Cyathostomum catinatum, Cylicostephanus longibursatus, Cys. calicatus, Cys. goldi, Cys. minutus, Coronocyclus coronatus and Cya. pateratum , accounting for 97% of all sequence reads prior to treatment. Of these, Cyc. nassatus and Cya. catinatum had the highest occurrence, accounting for 68% of all sequence reads prior to PYR treatment. Treatment did not significantly affect the species composition. The results highlight the importance of drug efficacy testing when using PYR to treat cyathostomin infection, even when selective anthelmintic treatment and thus low treatment intensity, is used on the farm. [Display omitted] • Pyrantel resistance in cyathostomins despite selective anthelmintic strategy. • Poor biosecurity measures associated with lower pyrantel drug efficacy. • Efficacy testing of pyrantel vital, regardless anthelmintic treatment regime. • No change in cyathostomin species composition after pyrantel treatment. [ABSTRACT FROM AUTHOR]

Helminths are commonly found in grazing equids, with cyathostomin nematodes and the cestode Anoplocephala perfoliata being the most prevalent. Most horses harbour low burdens of these parasites and do not develop signs of infection; however, in a small number of animals, high burdens can accumulate and cause disease. Cyathostomins are associated with a syndrome known as larval cyathostominosis. This occurs when large numbers of larvae emerge from the large intestinal wall. This disease has a case fatality rate of up to 50%. A. perfoliata infection has been associated with various types of colic, with burdens of >20 worms associated with pathogenicity. Anthelmintic resistance is a serious problem in cyathostomins and is emerging in A. perfoliata. Control methods that reduce reliance on anthelmintics now need to be applied, especially as no new dewormer compounds are on the horizon. Sustainable control methods must employ diagnostics to identify horses that require treatment. Coprological tests (faecal egg counts, FECs) have been used for several decades to inform treatment decisions to reduce helminth egg shedding. These tests cannot be used to assess host burdens as FECs do not correlate with cyathostomin or A. perfoliata burdens. In the last decade, new tests have become available that measure parasite-specific antibodies, the levels of which have been shown to correlate with parasite burden. These tests measure antigen-specific IgG(T) and are available in serum (cyathostomin, A. perfoliata) or saliva (A. perfoliata) formats. Tests for other helminths have been developed as research tools and need to be translated to support equine clinicians in practice. A key element of sustainable control strategies is that diagnostics must be used in combination with management approaches to reduce environmental transmission of helminths; this will help limit the proportion of horses harbouring parasite burdens that need to be targeted by treatment. This manuscript provides a review of the development, performance and general utility of various diagnostic methods for informing equine helminth management decisions. [ABSTRACT FROM AUTHOR]

Ivermectin (IVM) is an anthelmintic compound commonly used off-label in mules due to its broad-spectrum of activity. Despite the general use of IVM in mules with the same dose and route of administration licensed for horses, significant pharmacokinetic differences might exist between horses and mules, as already observed for donkeys. The aim of the present study was to evaluate the pharmacokinetic profile and anthelmintic efficacy of an oral paste of IVM in mules naturally infected with cyathostomins. Fifteen adult mules with fecal egg counts (FEC) ≥200 eggs per gram (EPG), with exclusive presence of cyathostomins, were included in the study. All mules were orally treated with IVM according to the manufacturers recommended horse dosage (200 µg/kg body weight). FECs were performed before (day-10 and day-3) and after treatment at days 14 and 28 by using a modified McMaster method. The FEC reduction (FECR%) was also calculated. Blood samples were collected from five animals at various times between 0.5 h up to 30 days post treatment to determine pharmacokinetic parameters. The maximum IVM serum concentration (Cmax) was 42.31 ± 10.20 ng/mL and was achieved at 16.80 ± 9.96 hours post-treatment (Tmax), area under the curve (AUC) was 135.56 ± 43.71 ng × day/mL. FECR% remained high (>95%) until the 28th day.

Cyathostomins are the most common and highly prevalent parasites of horses worldwide. Historically, the control of cyathostomins has mainly relied on the routine use of anthelmintic products. Increasing reports on anthelmintic resistance (AR) in cyathostomins are concerning. A potential method proposed for detecting emerging AR in cyathostomins has been estimating the egg reappearance period (ERP). This paper reviews the data available for the ERP of cyathostomins against the three major classes of anthelmintics, macrocyclic lactones, tetrahydropyrimidines, and benzimidazoles. Published peer-reviewed original research articles were obtained from three databases (PubMed, CAB Direct and Web of Science) and were evaluated for their inclusion in a systematic review. Subsets of articles were then subjected to a review of ERP data. A total of 54 (of 134) studies published between 1972 and 2022 met the criteria for inclusion in the systematic review. Until the beginning of 2022, there was no agreed definition of the ERP; eight definitions of ERP were identified in the literature, complicating the comparison between studies. Additionally, potential risk factors for the shortening of the ERP, including previous anthelmintic use and climate, were frequently not described. Reports of shortened ERP for moxidectin and ivermectin are frequent: 20 studies that used comparable ERP definitions reported shortened moxidectin and ivermectin ERPs of 35 and 28 days, respectively. It is unclear whether the ERPs of these anthelmintics reduced to such levels are due to the development of AR or some biological factors related to horses, cyathostomin species, and/or the environment. The ERPs for other anthelmintics, such as fenbendazole and pyrantel, were frequently not reported due to established resistance against these drugs. Future research in horses is required to understand the mechanism(s) behind the shortening of ERP for cyathostomins. Based on this systematic review, we propose recommendations for future ERP studies.

In recent years, resistance to the benzimidazole (BZ) and tetrahydropyrimidine (PYR) anthelmintics in global cyathostomin populations, has led to reliance on the macrocyclic lactone drugs (ML-of which ivermectin and moxidectin are licensed in horses) to control these parasites. Recently, the first confirmed case of resistance to both ivermectin (IVM) and moxidectin (MOX) was reported in the USA in yearlings imported from Ireland. This suggests that ML resistance in cyathostomins has emerged, and raises the possibility that regular movement of horses may result in rapid spread of ML resistant cyathostomins. Resistance may go undetected due to a lack of surveillance for ML efficacy. Here, we report anthelmintic efficacies in cyathostomins infecting UK Thoroughbreds on four studs. Faecal egg count reduction tests (FECRT) were performed to define resistance (resistance = FECR

Context. Deworming all horses with commercial anthelmintics on a rotational basis every 90 days is still popular worldwide. Aims. To investigate anthelmintic resistance and therapeutic requirements of adult horses on the basis of faecal examinations after a rotation between commercial anthelmintics commonly used in Brazil. Methods. We followed up faecal egg counts and coprocultures of 29 horses for 7 months, after pre-established anthelmintic treatments, every 3 months (August and November 2019 and February 2020). The following commercial anthelmintics were used respectively: moxidectin (1), ivermectin + praziquantel (2) and trichlorfon + mebendazole (3). Faecal egg count (FEC), FEC reduction tests (FECRT) and coproculture were performed before (Day 0) and after 14 days for each of the three treatments. Wilcoxon test (95% confidence level, P < 0.05) was used to test the efficacy of the treatment on the FECRT and the occurrence of resistance to the treatments used. Results. The FEC results and percentage of individual FECRT indicated parasite resistance, and 82 of 87 (94%) deworming treatments were unnecessary. The efficacy rate was 78%, 34% and 53% for Treatments 1, 2 and 3 respectively. In the coprocultures before deworming, 98.5% were small strongyles, 1.1% Strongylus equinus and 0.4% Gyalocephalus capitatus. After deworming, 98.7% of parasites were small strongyles and 1.3% Dictyocaulus arnfieldi. FEC values at Day 0 and 14 days after anthelminthics were different (lower) only for the moxidectin treatment (U = 271.5; P-value <0.01). For both ivermectin + praziquantel (U = 373; P-value >0.05) and mebendazole + trichlorfon (U = 391; P-value >0.05) treatments, there was no difference in FEC between Day 0 and Day 14 at 95% confidence level. Conclusions. The anthelmintics used in the deworming protocol of this study had reduced efficacy against the parasites identified in the coprocultures and the pre-established treatments were probably unnecessary in 93% of the horses with a low level of infection. Implications. This study has highlighted not only the issue of the indiscriminate use of anthelmintics regarding parasitic resistance, but also the exposure of horses to unnecessary treatments. Such widespread use is a costly waste of resources and further reinforces the discontinuation of schedule rather than strategic deworming protocols. [ABSTRACT FROM AUTHOR]

The present paper describes a new strongylid nematode belonging to the genus Cylicostephanus Ihle, 1922 (Strongyloidea, Cyathostominae), from Asiatic wild ass (Khulan) and domestic horses in Mongolia. The external leaf-crown (ELC) of Cylicostephanus scharchui sp. n. consists of approximately 12 elements and the internal leaf-crown (ILC) of approximately 36 elements. The buccal capsule is cylindrical, circular in cross-section, and slightly deeper than wide. A long dorsal gutter extends from the floor of the buccal capsule closely to base of ILC. The duct of the dorsal esophageal gland opens at the tip of the dorsal gutter. The tail of females is short, and the vulva is close to the anus. The buccal capsule and dorsal gutter are most similar to those of Cylicostephanus calicatus, but differ significantly in size of dorsal gutter and relative width and depth of the buccal capsule as well as characteristics of the leaf-crowns. [ABSTRACT FROM AUTHOR]

In recent years, resistance to the benzimidazole (BZ) and tetrahydropyrimidine (PYR) anthelmintics in global cyathostomin populations, has led to reliance on the macrocyclic lactone drugs (ML-of which ivermectin and moxidectin are licensed in horses) to control these parasites. Recently, the first confirmed case of resistance to both ivermectin (IVM) and moxidectin (MOX) was reported in the USA in yearlings imported from Ireland. This suggests that ML resistance in cyathostomins has emerged, and raises the possibility that regular movement of horses may result in rapid spread of ML resistant cyathostomins. Resistance may go undetected due to a lack of surveillance for ML efficacy. Here, we report anthelmintic efficacies in cyathostomins infecting UK Thoroughbreds on four studs. Faecal egg count reduction tests (FECRT) were performed to define resistance (resistance = FECR <95% lower credible interval (LCI) < 90%). Stud A yearlings had FECRs of 36.4–78.6% (CI:15.7–86.3) after three IVM treatments, 72.6% (CI: 50.8–85.2) after MOX, and 80.8% (CI: 61.9–90.0) after PYR. Mares on stud A had a FECR of 97.8% (CI: 93.3–99.9) and 98% (95.1–99.4) after IVM and MOX treatment, respectively. Resistance to MLs was not found in yearlings or mares on studs B, C or D with FECR after MOX OR IVM treatment ranging from 99.8 to 99.9% (95.4–100); although yearlings on studs B, C and D all had an egg reappearance period (ERP) of six weeks for MOX and stud C had a four-week ERP for IVM. This study describes the first confirmed case of resistance to both licensed ML drugs on a UK Thoroughbred stud and highlights the urgent need for a) increased awareness of the threat of ML resistant parasites infecting horses, and b) extensive surveillance of ML efficacy against cyathostomin populations in the UK, to gauge the extent of the problem. [Display omitted] • Increasing anthelmintic resistance in cyathostomins is a global concern. • Four thoroughbred studs in England were sampled using faecal egg count reduction tests. • Anthelmintic resistance to ivermectin, moxidectin and pyrantel was detected in yearlings. • Egg reappearance periods were lower than the original label claims in yearlings. • Extensive surveillance of ML efficacy against cyathostomin populations is required. [ABSTRACT FROM AUTHOR]

Cyathostomins are the most common and highly prevalent parasites of horses worldwide. Historically, the control of cyathostomins has mainly relied on the routine use of anthelmintic products. Increasing reports on anthelmintic resistance (AR) in cyathostomins are concerning. A potential method proposed for detecting emerging AR in cyathostomins has been estimating the egg reappearance period (ERP). This paper reviews the data available for the ERP of cyathostomins against the three major classes of anthelmintics, macrocyclic lactones, tetrahydropyrimidines, and benzimidazoles. Published peer-reviewed original research articles were obtained from three databases (PubMed, CAB Direct and Web of Science) and were evaluated for their inclusion in a systematic review. Subsets of articles were then subjected to a review of ERP data. A total of 54 (of 134) studies published between 1972 and 2022 met the criteria for inclusion in the systematic review. Until the beginning of 2022, there was no agreed definition of the ERP; eight definitions of ERP were identified in the literature, complicating the comparison between studies. Additionally, potential risk factors for the shortening of the ERP, including previous anthelmintic use and climate, were frequently not described. Reports of shortened ERP for moxidectin and ivermectin are frequent: 20 studies that used comparable ERP definitions reported shortened moxidectin and ivermectin ERPs of 35 and 28 days, respectively. It is unclear whether the ERPs of these anthelmintics reduced to such levels are due to the development of AR or some biological factors related to horses, cyathostomin species, and/or the environment. The ERPs for other anthelmintics, such as fenbendazole and pyrantel, were frequently not reported due to established resistance against these drugs. Future research in horses is required to understand the mechanism(s) behind the shortening of ERP for cyathostomins. Based on this systematic review, we propose recommendations for future ERP studies. [Display omitted] • Is the egg reappearance period (ERP) an early indicator of anthelmintic resistance? • A summary of cyathostomins ERP for equine registered anthelmintics from 1972 to 2022. • Recent developments on the definition of ERP. • Proposed recommendations for future ERP studies. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the effectiveness of oral ivermectin and moxidectin against natural cyathostomin infection in four horse farms located in the central regions of Mexico. Of the 445 horses: Warmblood (145), Thoroughbreds (100), and Quarter Horses (200) breeds, aged between 6 months and 27 years, were used. Data on horses and parasite control methods were collected through interviews with farm owners and veterinarians. Using the McMaster technique, fecal samples were processed from all 445 horses, 180 of which were positive for cyathostomins. On each farm, a selection was made of 45 animals meeting the criteria of a Faecal Egg Count Reduction Test yielding results exceeding 150 eggs per gram of strongylid-type nematodes. Subsequently, three separate experimental groups were formed for each farm, each consisting of 15 horses The first group was treated with oral ivermectin 1.87 %; the second group with oral moxidectin 2 %; and the third was the non-treatment control group. Coprocultures were also performed to identify the presence of nematode species. The data obtained were analyzed with RESO.exe©️. Three of the four farms achieved a 100 % reduction in eggs per gram with both macrocyclic lactones. One farm achieved 93 % reduction with ivermectin and 87 % with moxidectin. This study demonstrates that macrocyclic lactones effectively reduce cyathostomins in three of the four farms studied. The results suggest potential cyathostomin resistance to macrocyclic lactones, particularly moxidectin, on one farm. Given these findings, sustainable parasite management is required on horse breeding farms in Mexico. [ABSTRACT FROM AUTHOR]

Helminths are commonly found in grazing equids, with cyathostomin nematodes and the cestode Anoplocephala perfoliata being the most prevalent. Most horses harbour low burdens of these parasites and do not develop signs of infection; however, in a small number of animals, high burdens can accumulate and cause disease. Cyathostomins are associated with a syndrome known as larval cyathostominosis. This occurs when large numbers of larvae emerge from the large intestinal wall. This disease has a case fatality rate of up to 50%. A. perfoliata infection has been associated with various types of colic, with burdens of >20 worms associated with pathogenicity. Anthelmintic resistance is a serious problem in cyathostomins and is emerging in A. perfoliata. Control methods that reduce reliance on anthelmintics now need to be applied, especially as no new dewormer compounds are on the horizon. Sustainable control methods must employ diagnostics to identify horses that require treatment. Coprological tests (faecal egg counts, FECs) have been used for several decades to inform treatment decisions to reduce helminth egg shedding. These tests cannot be used to assess host burdens as FECs do not correlate with cyathostomin or A. perfoliata burdens. In the last decade, new tests have become available that measure parasite-specific antibodies, the levels of which have been shown to correlate with parasite burden. These tests measure antigen-specific IgG(T) and are available in serum (cyathostomin, A. perfoliata) or saliva (A. perfoliata) formats. Tests for other helminths have been developed as research tools and need to be translated to support equine clinicians in practice. A key element of sustainable control strategies is that diagnostics must be used in combination with management approaches to reduce environmental transmission of helminths; this will help limit the proportion of horses harbouring parasite burdens that need to be targeted by treatment. This manuscript provides a review of the development, performance and general utility of various diagnostic methods for informing equine helminth management decisions.

This study aimed to collect information on local and systemic inflammatory responses, and goblet cell‐associated components, following anthelmintic treatment with moxidectin and ivermectin in horses naturally infected with cyathostomin parasites. Thirty‐six horses aged 2–5 years of age were randomly allocated to three groups. Group 1 received ivermectin/praziquantel (0.2 mg/kg), Group 2 received moxidectin/praziquantel (0.4 mg/kg) and Group 3 were untreated controls. Tissue samples from the Cecum, Dorsal and Ventral Colons were used for histopathological evaluation and preserved for RNA isolation and gene expression analysis. Whole blood was collected weekly for gene expression analysis as well. The control group had significantly higher inflammation associated with higher larval scores. The treatment groups displayed no differences in larval counts and inflammatory cell populations (p >.05). Mucosal larval counts were positively correlated with goblet cell hyperplasia scores (p =.047). The moxidectin‐treated group had a significantly lower expression of IFN‐γ (p <.05). The data suggest that removal of cyathostomins reduced the pro‐inflammatory response associated with cyathostomin infections. Pro‐inflammatory reactions associated with anthelmintic treatment were minimal, but lowest for moxidectin‐treated horses. Results suggested that cecum, ventral and dorsal colons responded differently to cyathostomin larvae, which may have implications in the disease process. [ABSTRACT FROM AUTHOR]

Abstract Background Cyathostomins are the most important and common parasitic nematodes of horses, with > 50 species known to occur worldwide. The frequent and indiscriminate use of anthelmintics has resulted in the development of anthelmintic resistance (AR) in horse nematodes. In this study we assessed the efficacy of commonly used anthelmintics against cyathostomins in Australian thoroughbred horses. Methods Two drug efficacy trials per farm were conducted on two thoroughbred horse farms in the state of Victoria, Australia. In the first trial, the horses on Farm A were treated with single and combinations of anthelmintics, including oxfendazole (OFZ), abamectin (ABM), abamectin and morantel (ABM + MOR), moxidectin (MOX) and oxfendazole and pyrantel (OFZ + PYR), at the recommended doses, whereas the horses on Farm B only received MOX, at the recommended dose. The faecal egg count reduction test (FECRT) was used to determine the efficacy and egg reappearance period (ERP) of anthelmintics. Based on the results of the first trial, the efficacies of MOX and a combination of ABM + MOR were reassessed to confirm their activities against cyathostomins. Results Of the five anthelmintic products tested on Farm A, resistance against OFZ, ABM and OFZ + PYR was found, with efficacies of − 41% (− 195% lower confidence limit [LCL]), 73% (60% LCL) and 82% (66% LCL) at 2 weeks post-treatment, respectively. The FECRT showed high efficacies of MOX and ABM + MOR (100%) at 2 week post-treatment and shortened ERPs for these anthelmintics (ABM + MOR: 4 weeks; MOX: 5 weeks). Resistance to MOX was found on Farm B, with a reduced efficacy of 90% (70% LCL) and 89% (82% LCL) at 2 weeks post-treatment in trials one and two, respectively. Conclusions This study provides the first evidence of MOX- and multidrug-resistant (ABM and combinations of anthelmintics) cyathostomins in Australia and indicates the need for continuous surveillance of the efficacy of currently effective anthelmintics and large-scale investigations to assess the ERP for various anthelmintics. Graphical Abstract

Anthelmintic resistance in equine cyathostomins is both widespread and highly prevalent in the benzimidazole and tetrahydropyrimidine classes; however, reports of resistance to macrocyclic lactone (ML) drugs are sparse and sporadic. This study reports a case of clear ML resistance in a group of Thoroughbred yearlings imported from Ireland to the US in 2019. Fecal egg count reduction (FECR) following ivermectin administered in February 2020 demonstrated 100% reduction in the US bred yearlings, but 93.5%, 70.5%, and 74.5% reduction in three groups of the imported yearlings. The two former groups were then retreated with ivermectin, yielding FECRs of 33.8% and 23.5%, respectively. Horses from these two groups were then assigned randomly to two possible treatments; moxidectin or a triple combination of moxidectin, oxibendazole, and pyrantel pamoate. The groups treated with moxidectin had FECRs of 90.2%, 57.3%, and 50.0%, while the triple combination had a 100% FECR in all treated groups. Subsequently, the efficacy of ivermectin was reassessed in June 2020 yielding FECRs of 99.8%, 87.7%, and 62.0% in the three imported groups. The FECRs of the US bred yearlings all remained in the 99–100% range. This is the first study to clearly demonstrate ML resistance in cyathostomins and to confirm the suspicion through reassessment. These data demonstrate that ML-resistant cyathostomins were imported from Ireland and serve to illustrate that the global movement of horses has the potential to quickly spread ML-resistant parasite isolates around the world. The equine industry is strongly encouraged to routinely monitor anthelmintic efficacy, so occurrence of ML resistant cyathostomins can be detected and appropriate interventions implemented as early as possible.

Abstract Background Anthelmintic treatment is a risk factor for intestinal disease in the horse, known as colic. However the mechanisms involved in the onset of disease post anthelmintic treatment are unknown. The interaction between anthelmintic drugs and the gut microbiota may be associated with this observed increase in risk of colic. Little is known about the interaction between gut microbiota and anthelmintics and how treatment may alter microbiome function. The objectives of this study were: To characterise (1) faecal microbiota, (2) feed fermentation kinetics in vitro and (3) metabolic profiles following moxidectin administration to horses with very low (0 epg) adult strongyle burdens. Hypothesis: Moxidectin will not alter (1) faecal microbiota, (2) feed fermentation in vitro, or, (3) host metabolome. Results Moxidectin increased the relative abundance of Deferribacter spp. and Spirochaetes spp. observed after 160 h in moxidectin treated horses. Reduced in vitro fibre fermentation was observed 16 h following moxidectin administration in vivo (P = 0.001), along with lower pH in the in vitro fermentations from the moxidectin treated group. Metabolic profiles from urine samples did not differ between the treatment groups. However metabolic profiles from in vitro fermentations differed between moxidectin and control groups 16 h after treatment (R 2 = 0.69, Q2Y = 0.48), and within the moxidectin group between 16 h and 160 h post moxidectin treatment (R 2 = 0.79, Q2Y = 0.77). Metabolic profiles from in vitro fermentations and fermentation kinetics both indicated altered carbohydrate metabolism following in vivo treatment with moxidectin. Conclusions These data suggest that in horses with low parasite burdens moxidectin had a small but measurable effect on both the community structure and the function of the gut microbiome.

Cyathostomins are the most abundant equid endoparasites globally. There are approximately fifty cyathostomin species and, whilst they occupy distinct niches within the large intestine, they are generally considered to share similar characteristics in terms of pathogenicity and response to drug treatment. There are three classes of anthelmintic licensed in the UK to treat cyathostomins (benzimidazoles, tetrahydropyrimidines and macrocyclic lactones) and cases of resistance have been documented for all classes. Previously, faecal egg count reduction tests (FECRT) on four UK Thoroughbred studs revealed multidrug resistant cyathostomins on one stud (A), with evidence of resistance to the macrocyclic lactones (MLs) ivermectin (IVM) and moxidectin (MOX), and to pyrantel (PYR). The remaining three studs (B-D) lacked resistance to IVM and MOX but had a shortened egg reappearance period post treatment. To determine whether specific species could be associated with the observed resistance and shortened egg reappearance period, strongyle eggs collected from between six and 15 individual horses per stud were copro-cultured to third larval stage (L3), before and after anthelmintic treatment, over a three-year timeframe (2021-2023). Quantitative DNA metabarcoding of the ITS-2 region was carried out on all samples. On stud A, single but differing species were found to be responsible for ML and pyrantel resistance in yearlings, Cyathostomum catinatum and Cylicocyclus nassatus, respectively. On studs B-D, with shortened egg reappearance periods, species composition remained largely unchanged post treatment. This study is the first to quantitatively profile cyathostomin species composition pre- and post-treatment in a multidrug resistant population in the UK, revealing that resistance in cyathostomins was species specific. This raises the question of whether these species may be responsible for ML and PYR resistance more widely and indicates that anthelmintic resistance in cyathostomins may not be a multi-species phenomenon., Competing Interests: Declaration of competing interest None., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Cyathostomins are the most important and common parasitic nematodes of horses, with > 50 species known to occur worldwide. The frequent and indiscriminate use of anthelmintics has resulted in the development of anthelmintic resistance (AR) in horse nematodes. In this study we assessed the efficacy of commonly used anthelmintics against cyathostomins in Australian thoroughbred horses. Methods: Two drug efficacy trials per farm were conducted on two thoroughbred horse farms in the state of Victoria, Australia. In the first trial, the horses on Farm A were treated with single and combinations of anthelmintics, including oxfendazole (OFZ), abamectin (ABM), abamectin and morantel (ABM + MOR), moxidectin (MOX) and oxfendazole and pyrantel (OFZ + PYR), at the recommended doses, whereas the horses on Farm B only received MOX, at the recommended dose. The faecal egg count reduction test (FECRT) was used to determine the efficacy and egg reappearance period (ERP) of anthelmintics. Based on the results of the first trial, the efficacies of MOX and a combination of ABM + MOR were reassessed to confirm their activities against cyathostomins. Results: Of the five anthelmintic products tested on Farm A, resistance against OFZ, ABM and OFZ + PYR was found, with efficacies of − 41% (− 195% lower confidence limit [LCL]), 73% (60% LCL) and 82% (66% LCL) at 2 weeks post-treatment, respectively. The FECRT showed high efficacies of MOX and ABM + MOR (100%) at 2 week post-treatment and shortened ERPs for these anthelmintics (ABM + MOR: 4 weeks; MOX: 5 weeks). Resistance to MOX was found on Farm B, with a reduced efficacy of 90% (70% LCL) and 89% (82% LCL) at 2 weeks post-treatment in trials one and two, respectively. Conclusions: This study provides the first evidence of MOX- and multidrug-resistant (ABM and combinations of anthelmintics) cyathostomins in Australia and indicates the need for continuous surveillance of the efficacy of currently effective anthelmintics and large-scale investigations to assess the ERP for various anthelmintics. [ABSTRACT FROM AUTHOR]

Since the introduction of the last equine broad-spectrum anthelmintic group in the 1980’s, the investment in new drugs to control horse’s parasites did not result in new advancements. These drugs allowed a very effective and extensive control of equine nematodes through successful interval dosing programs, firstly introduced in the 1960’s. However, the widespread and indiscriminate use of anthelmintics in these intensive treatments have led to increasing resistance in the major equine nematodes. Reports of reduced effectiveness are virtually worldwide and repercussions in livestock production farms have already been seen. Based on recent questionnaires about horse farm practices, preventive measures and international recommendations, it is clear that most of them are still not being widely implemented. It is also clear that these recommendations are outdated and new approaches must be considered to correctly tackle this rapidly evolving issue in horse management, as more accurate diagnostic methods are currently available, such as Mini-FLOTAC. This article intends to do a general review of the history and current situation of anthelmintic resistance in horses, with emphasis in Europe, as well as, how to diagnose and delay or even prevent its further development, mentioning new methods of diagnostic and directions in which to develop research.

The objective of this study was to assess the efficacy of commercial anthelmintics in horses naturally infected with gastrointestinal nematodes from the southern Rio Grande do Sul state, Brazil. The anthelmintic efficacy was assessed by fecal egg count reduction test (FECRT) and coproculture. Fecal samples were collected from 110 animals in seven farms located in the southern region of the state of Rio Grande do Sul, in the south of Brazil. Criteria used to suspect anthelmintic resistance included percentages of efficacy lower than 95% and a lower limit of the confidence interval below 90%. Anthelmintic resistance was detected in five of the seven farms studied. Coproculture results show that, even after horses were treated with anthelmintics, infection by nematodes of the subfamily Cyathostominae was still present in animals from five farms.

The avermectin and pyrantel resistance of equine parasites was monitored on four farms in the Czech Republic in 2014. The aim of the testing was to investigate the anthelmintic resistance of cyathostomins and ascarids. One hundred and eighty-six horses were included in the test, 54 of which were infected by ascarids and 174 by cyathostomins. Faecal egg count reduction tests were used to assess anthelmintic resistance. The faecal egg count reduction for ascarids was within the range of 87.9% to 100% for avermectins and 100% for pyrantel embonate. Faecal egg count reduction for cyathostomins ranged from 99% to 100% for avermectins and 89.3% to 98.2% for pyrantel embonate.

Anthelmintic resistance in equine cyathostomins is both widespread and highly prevalent in the benzimidazole and tetrahydropyrimidine classes; however, reports of resistance to macrocyclic lactone (ML) drugs are sparse and sporadic. This study reports a case of clear ML resistance in a group of Thoroughbred yearlings imported from Ireland to the US in 2019. Fecal egg count reduction (FECR) following ivermectin administered in February 2020 demonstrated 100% reduction in the US bred yearlings, but 93.5%, 70.5%, and 74.5% reduction in three groups of the imported yearlings. The two former groups were then retreated with ivermectin, yielding FECRs of 33.8% and 23.5%, respectively. Horses from these two groups were then assigned randomly to two possible treatments; moxidectin or a triple combination of moxidectin, oxibendazole, and pyrantel pamoate. The groups treated with moxidectin had FECRs of 90.2%, 57.3%, and 50.0%, while the triple combination had a 100% FECR in all treated groups. Subsequently, the efficacy of ivermectin was reassessed in June 2020 yielding FECRs of 99.8%, 87.7%, and 62.0% in the three imported groups. The FECRs of the US bred yearlings all remained in the 99–100% range. This is the first study to clearly demonstrate ML resistance in cyathostomins and to confirm the suspicion through reassessment. These data demonstrate that ML-resistant cyathostomins were imported from Ireland and serve to illustrate that the global movement of horses has the potential to quickly spread ML-resistant parasite isolates around the world. The equine industry is strongly encouraged to routinely monitor anthelmintic efficacy, so occurrence of ML resistant cyathostomins can be detected and appropriate interventions implemented as early as possible. Image 1 • Reduced macrocyclic lactone efficacy in yearlings imported from Ireland. • Ivermectin efficacy was retested twice and was reduced on all occasions. • 100% strongyle fecal egg count reduction in US bred yearlings on all occasions. • A combination of moxidectin, oxibendazole and pyrantel pamoate was 100% effective. • Global movement of horses likely to spread macrocyclic lactone resistance quickly. [ABSTRACT FROM AUTHOR]

Intestinal strongyles are common helminths of donkeys, in which they may be responsible for disease and poor performance. This study aimed to identify intestinal strongyle genera in 55 naturally infected donkeys from three different farm typologies in Tuscany, central Italy, using morphological and metrical analysis of third stage larvae (L3) obtained from faecal cultures. Larvae were identified using two previous reported morphological identification keys. Moreover, eggs per gram (EPG) data were also evaluated to assess differences, if any, according to the farm typology, sex, and age of the examined donkeys. The results showed that small strongyles were prevalent in all donkey farms. In all examined farms, most (92-100%) of L3 were identified as cyathostomin species of the genera Cylicocyclus spp. and Cylicostephanus spp. Large strongyles of the genera Strongylus spp. and Triodontophorus spp., were identified at low percentage (8%), only in the single organic farm included in the study. A high agreement was observed between the two different morphometric keys used. No significant differences were found for EPG according to farm typology, and sex and age from the examined donkeys. This is the first report about genera identification of intestinal strongyles infecting donkeys in Tuscany, Italy. [ABSTRACT FROM AUTHOR]

Background: Information regarding parasite control practices currently used on Thoroughbred and Standardbred studs in Australia is lacking. Anthelmintic resistance (AR) is a global problem which has implications for equine health and welfare. Objectives: To identify parasite control practices currently used on horse studs in Australia and investigate the frequency of use of management factors that have been associated with the likelihood of promoting or delaying AR. Study design: Questionnaire study of equine parasite control on Thoroughbred and Standardbred studs in Australia. Methods: An online questionnaire was emailed to 300 studs in Australia. Information obtained included property details, grazing management, anthelmintic use, non‐chemotherapeutic parasite control practices, use of faecal egg counts (FECs) and perceptions of AR. Results: Seventy‐five completed questionnaires were received (25% response rate). Macrocyclic lactones (MLs) were the most commonly administered anthelmintics in mares and foals and less than 5% of respondents used targeted treatment regimens. The implementation of pasture hygiene practices was variable. The majority of respondents (97%) considered AR to be important; however, few respondents were aware of the use of FEC reduction tests for monitoring of drug efficacy. Main limitations: The potential for nonresponse bias was the main limitation of this study. Conclusions: Parasite control strategies on Australian stud farms remain over‐reliant on anthelmintic use. The frequent use of MLs is of concern for the increased selection pressure for AR. There is a lack of awareness of the importance of non‐chemotherapeutic strategies in integrated approaches to parasite control aimed at delaying the development of AR. This study highlights the need for greater veterinary involvement in the implementation of more sustainable parasite control practices with greater emphasis placed on surveillance through FEC testing. [ABSTRACT FROM AUTHOR]

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. [Display omitted] [ABSTRACT FROM AUTHOR]

• Encysted larval counts were significantly higher in the cecum. • Evidence of spatial variation was observed within intestinal organs. • Refrigeration of tissue samples did not affect encysted larval counts. Cyathostomins are pervasive equine parasites in horses across the world, and larval stages are known to cause the deadly disease larval cyathostominosis. The mucosal digestion technique is widely used for enumeration of encysted larval stages. Previous studies have investigated the spatial variation of encysted larvae, however current protocols lack a description of a standardized area from which to take the tissue sample. This study sought to evaluate spatial variation in encysted cyathostomin larval counts among the large intestinal organs and their subsections. Following humane euthanasia, ceca, ventral, and dorsal colons were harvested from 8 foals (aged 4–8 months) raised in an anthelmintic naïve parasitology research herd. Each organ was weighed and separated into 3 equal sections by length: the orad, intermediate, and aborad portions. From each of those sections, two 5% weight tissue samples were collected and digested to quantify the early third stage larvae (EL3) and late third stage larvae/fourth stage larvae (LL3/L4). A mixed model statistical analysis was carried out to evaluate for differences of larval counts among the different organs, sections, and the interaction term between the organs and sections. There were significant differences among organs (P < 0.0001), with the ceca having higher counts than the ventral and dorsal colons. However, there were no significant differences among the three defined organ sections (P = 0.1076). Coefficients of variation (CV) were all calculated to be greater than 1, suggesting a high level of variability among the samples; the least amount of variation can be found in the cecal data with a CV of 1.4024 compared with the ventral colon's 1.529845 and dorsal colon's 3.339135 within the respective organ. The following sections had the highest mean counts of encysted larvae: intermediate cecum, orad ventral colon, and aborad dorsal colon. Though only a portion of the results were significant, trends were observed and these should be investigated further in future studies and potentially employed in larvicidal efficacy evaluations. [ABSTRACT FROM AUTHOR]

Since the introduction of the last equine broad-spectrum anthelmintic group in the 1980's, the investment in new drugs to control horse's parasites did not result in new advancements. These drugs allowed a very effective and extensive control of equine nematodes through successful interval dosing programs, firstly introduced in the 1960's. However, the widespread and indiscriminate use of anthelmintics in these intensive treatments have led to increasing resistance in the major equine nematodes. Reports of reduced effectiveness are virtually worldwide and repercussions in livestock production farms have already been seen. Based on recent questionnaires about horse farm practices, preventive measures and international recommendations, it is clear that most of them are still not being widely implemented. It is also clear that these recommendations are outdated and new approaches must be considered to correctly tackle this rapidly evolving issue in horse management, as more accurate diagnostic methods are currently available, such as Mini-FLOTAC. This article intends to do a general review of the history and current situation of anthelmintic resistance in horses, with emphasis in Europe, as well as, how to diagnose and delay or even prevent its further development, mentioning new methods of diagnostic and directions in which to develop research. [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]

• Changes in the equine faecal microbiota occur after anthelmintic treatment. • These changes are accompanied by increases in faecal albumin and serum fibrinogen. • Gut dysbiosis may be involved in the pathogenesis of acute larval cyathostominosis. • Manipulation of the equine "helminthome" could be used to probe gut ecosystems. The interactions between parasitic helminths and gut microbiota are considered to be an important, although as yet incompletely understood, factor in the regulation of immunity, inflammation and a range of diseases. Infection with intestinal helminths is ubiquitous in grazing horses, with cyathostomins (about 50 species of which are recorded) predominating. Consequences of infection include both chronic effects, and an acute inflammatory syndrome, acute larval cyathostominosis, which sometimes follows removal of adult helminths by administration of anthelmintic drugs. The presence of cyathostomins as a resident helminth population of the equine gut (the "helminthome") provides an opportunity to investigate the effect helminth infection, and its perturbation, has on both the immune system and bacterial microbiome of the gut, as well as to determine the specific mechanisms of pathophysiology involved in equine acute larval cyathostominosis. We studied changes in the faecal microbiota of two groups of horses following treatment with anthelmintics (fenbendazole or moxidectin). We found decreases in both alpha diversity and beta diversity of the faecal microbiota at Day 7 post-treatment, which were reversed by Day 14. These changes were accompanied by increases in inflammatory biomarkers. The general pattern of faecal microbiota detected was similar to that seen in the relatively few equine gut microbiome studies reported to date. We conclude that interplay between resident cyathostomin populations and the bacterial microbiota of the equine large intestine is important in maintaining homeostasis and that disturbance of this ecology can lead to gut dysbiosis and play a role in the aetiology of inflammatory conditions in the horse, including acute larval cyathostominosis. [ABSTRACT FROM AUTHOR]

• A model for the complete cyathostomin lifecycle was constructed. • Multiple mechanisms for the genetics of anthelmintic resistance were incorporated. • Treatment strategies were compared for development of anthelmintic resistance. • The genetics of resistance had a large effect on the rate at which resistance developed. • Different strategies had similar relative ranking regardless of genetic mechanism assumed. Previously described models for the free-living and parasitic phases of the cyathostomin life-cycle were combined into a single model for the complete life-cycle. The model simulates a single free-living population on pasture utilising parasite egg output from the horses and localised temperature and rainfall data to estimate infective larval density on herbage. Multiple horses of different ages are possible, each with an individualised anthelmintic treatment programme. Genotypes for anthelmintic resistance are included allowing for up to three resistance genes with 2 alleles each. Because little is known of the genetics of resistance to anthelmintics in cyathostomins, the first use of this model was to compare the effect of different assumptions regarding the inheritance of resistance on model outputs. Comparisons were made between single and two-gene inheritance, where the heterozygote survival was dominant, intermediate or recessive under treatment, and with or without a fitness disadvantage associated with the resistance mechanism. Resistance developed fastest when the heterozygotes survived anthelmintic treatment (i.e. , were dominant) and slowest when they did not (i.e. , were recessive). Resistance was slower to develop when inheritance was poly-genic compared to a single gene, and when there was a fitness cost associated with the resistance mechanism, although the latter variable was the least influential. Importantly, while these genetic factors sometimes had a large influence on the rate at which resistant genotypes built up in the model populations, their order of ranking was always the same, when different anthelmintic use strategies were compared. Therefore, the described model is a useful tool for evaluating different treatment and management strategies on their potential to select for resistance. [ABSTRACT FROM AUTHOR]

Grazing equids are constantly exposed to three clinically important gastrointestinal parasites (small strongyles/cyathostomins, Anoplocephala spp. and Parascaris spp.). Knowledge of the local seasonal dynamic of these parasitic infections is important for constructing a sustainable parasite control program with a rational number of anthelmintic treatments. However, studies describing these patterns are sparse in France. In this context, a two-year study was carried out to assess i) the seasonal dynamic and variability of strongyle faecal egg counts (FEC) and infective larvae (L3) counts on pastures, and ii) the prevalence of Anoplocephala spp. and Parascaris spp. and the dynamic evolution of their presence. During 2021 and 2022 grazing seasons, monthly individual faecal egg counts (FEC) and diarrhea scores (DS) were determined on 428 equids divided into 33 groups. A monthly body condition score (BCS) was also attributed to animals ≥3 years old and a monthly bodyweight was estimated for each animal <3 years old. At the group level, the strongyle L3 counts on grazed pastures were carried out at least in spring, summer and autumn. Eggs of strongyles were observed in 97% of equids. In 64% of the groups, the peaks of FEC were noted in September and October. At the individual level, the maximum strongyle FEC was related to age, group of breeds, number of grazed plots and number of anthelmintic treatments. No negative association was observed between strongyle FEC and BCS or average daily weight gain. In the pastures, cyathostomin larvae were found almost exclusively. Over the two years, the peaks of cyathostomin L3 counts occurred in 87% of the groups between September and November and ranged from 635 to 87,500 L3 kg-1 dry herbage. The variability of the maximum cyathostomin L3 count in each group was explained by the year and the number of grazed plots. Eggs of Anoplocephala spp. were observed in 12% of equids. Eggs of Parascaris spp. were noted in 34% of one year-old animals, 9% of two years-olds and 2% of olders. Anoplocephala spp. and Parascaris spp. eggs were observed every month with a peak in the percentage of shedders in groups in October for Anoplocephala spp. and May-June for Parascaris spp.This study highlights the prevalence of each parasite, the variability in cyathostomin egg excretion and L3 counts amongst groups and individuals and the factors involved in this variation These local epidemiological data will help us to re-think a newer strategy against these parasites. [ABSTRACT FROM AUTHOR]

Intestinal strongyles are common helminths of donkeys, in which they may be responsible for disease and poor performance. This study aimed to identify intestinal strongyle genera in 55 naturally infected donkeys from three different farm typologies in Tuscany, central Italy, using morphological and metrical analysis of third stage larvae (L3) obtained from faecal cultures. Larvae were identified using two previous reported morphological identification keys. Moreover, eggs per gram (EPG) data were also evaluated to assess differences, if any, according to the farm typology, sex, and age of the examined donkeys. The results showed that small strongyles were prevalent in all donkey farms. In all examined farms, most (92–100%) of L3 were identified as cyathostomin species of the genera Cylicocyclus spp. and Cylicostephanus spp. Large strongyles of the genera Strongylus spp. and Triodontophorus spp., were identified at low percentage (8%), only in the single organic farm included in the study. A high agreement was observed between the two different morphometric keys used. No significant differences were found for EPG according to farm typology, and sex and age from the examined donkeys. This is the first report about genera identification of intestinal strongyles infecting donkeys in Tuscany, Italy.

Ivermectin (IVM) is an anthelmintic compound commonly used off-label in mules due to its broad-spectrum of activity. Despite the general use of IVM in mules with the same dose and route of administration licensed for horses, significant pharmacokinetic differences might exist between horses and mules, as already observed for donkeys. The aim of the present study was to evaluate the pharmacokinetic profile and anthelmintic efficacy of an oral paste of IVM in mules naturally infected with cyathostomins. Fifteen adult mules with fecal egg counts (FEC) ≥ 200 eggs per gram (EPG), with exclusive presence of cyathostomins, were included in the study. All mules were orally treated with IVM according to the manufacturer's recommended horse dosage (200 µg/kg body weight). FECs were performed before (day-10 and day-3) and after treatment at days 14 and 28 by using a modified McMaster method. The FEC reduction (FECR%) was also calculated. Blood samples were collected from five animals at various times between 0.5 h up to 30 days post treatment to determine pharmacokinetic parameters. The maximum IVM serum concentration (Cmax) was 42.31 ± 10.20 ng/mL and was achieved at 16.80 ± 9.96 h post-treatment (Tmax), area under the curve (AUC) was 135.56 ± 43.71 ng × day/mL. FECR% remained high (>95%) until the 28th day.

Horse cyathostominosis is a large intestine helminthosis caused by parasites belonging to the family Strongylidae, subfamily Cyathostominae. The cyathostomins (small strongyles) represent a challenge for the parasitologists and animal owners due to the different ontogenesis, the high number of parasite species and their ability to develop anthelmintic resistance. The faeces were examined by flotation (Willis) method and the infestation level was determined by McMaster method in day 0, 7 and 14 post treatments. The product Vanbendazol (30% fenbendazole) had a 97.7% efficacy in the treated horses from Şofronea, Arad County, using the Faecal egg count reduction test (FECRT). Also were performed Presidente and Borgsteede relations and the anthelmintic efficacy was 98.2% for the both relations.

Highlights • Improved identification of infective larvae with a morphological key is proposed. • Intestinal cell characteristics displayed high differences between L3 types. • Classification of larval types reduces the probable species in equine faecal samples. • Identification key will increase the diagnostic of classical parasitology techniques. Abstract Cyathostomins comprise around 50 parasite species of equids, offering a great challenge regarding their individual identification. The objective of our work was to improve identification of infective third stage larvae (L3) with a morphological key supplemented with detailed scientific illustrations based on our research and available literature. The highlighted features were; the number, arrangement, and shape of intestinal cells (IC), general features and the total body length of the eight different Cyathostomin sensu latum types (Type A, B, C, D, E, F, G, H), Gyalocephalus capitatus , and Posteriostomum spp. Due to variability, we were unable to define final body length measurements to differentiate L3 of cyathostomins. However, IC characteristics displayed a higher difference between L3 types than total body length. Through the number and arrangement of IC, 14 species were classified within three larval types. The classification of L3 into distinct larval types sensu latum gives us the advantage of reducing the number of probable species presented in equine faecal samples using a low-cost technique when monitoring the parasite fauna present in individual horses or on the farm level. The present improved identification key shall increase the diagnostic capabilities of classical equine parasitology techniques, using general L3 morphology thereby pragmatically improving regional and transnational epidemiological and biodiversity studies. The present key may also assist in defining the cyathostomin community in cyathostominosis clinical cases and within drug resistant populations across different management systems and geographical locations. [ABSTRACT FROM AUTHOR]

The avermectin and pyrantel resistance of equine parasites was monitored on four farms in the Czech Republic in 2014. The aim of the testing was to investigate the anthelmintic resistance of cyathostomins and ascarids. One hundred and eighty-six horses were included in the test, 54 of which were infected by ascarids and 174 by cyathostomins. Faecal egg count reduction tests were used to assess anthelmintic resistance. The faecal egg count reduction for ascarids was within the range of 87.9% to 100% for avermectins and 100% for pyrantel embonate. Faecal egg count reduction for cyathostomins ranged from 99% to 100% for avermectins and 89.3% to 98.2% for pyrantel embonate. [ABSTRACT FROM AUTHOR]

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A growing body of evidence, particularly in humans and rodents, supports the existence of a complex network of interactions occurring between gastrointestinal (GI) helminth parasites and the gut commensal bacteria, with substantial effects on both host immunity and metabolic potential. However, little is known of the fundamental biology of such interactions in other animal species; nonetheless, given the considerable economic losses associated with GI parasites, particularly in livestock and equines, as well as the global threat of emerging anthelmintic resistance, further explorations of the complexities of host-helminth-microbiota interactions in these species are needed. This study characterises the composition of the equine gut commensal flora associated with the presence, in faecal samples, of low (C low ) and high (C high ) numbers of eggs of an important group of GI parasites (i.e. the cyathostomins), prior to and following anthelmintic treatment. High-throughput sequencing of bacterial 16S rRNA amplicons and associated bioinformatics and statistical analyses of sequence data revealed strong clustering according to faecal egg counts ( P  = 0.003). A trend towards increased populations of Methanomicrobia (class) and Dehalobacterium (genus) was observed in C low in comparison with C high . Anthelmintic treatment in C high was associated with a significant reduction of the bacterial Phylum TM7 14 days post-ivermectin administration, as well as a transient expansion of Adlercreutzia spp. at 2 days post-treatment. This study provides a first known insight into the discovery of the intimate mechanisms governing host-parasite-microbiota interactions in equines, and sets a basis for the development of novel, biology-based intervention strategies against equine GI helminths based on the manipulation of the commensal gut flora. [ABSTRACT FROM AUTHOR]

The study presents the results of a cross-sectional survey to describe the epidemiology of ascarid and strongylid nematodes in horses, the impact of diverse climatic conditions on parasite diversity and the levels of faecal egg shedding in different age groups of managed Thoroughbred horses. Individual faecal samples (n = 1377) collected from 62 Thoroughbred farms across four climatic zones in Australia were analysed using the modified McMaster technique for faecal egg counts (FECs) and strongylid nematodes were identified utilising PCR-directed next-generation sequencing (NGS) of the second internal transcribed spacer of the nuclear ribosomal DNA (ITS-2). Across all age groups, the prevalence of ascarid and strongylid nematodes was 12% (95% confidence interval 10–14%) and 72% (70–74%), respectively. Based on strongylid FECs, yearlings had the highest prevalence (89%) followed by weanlings (83%), foals (79%), wet mares (61%), dry mares (59%) and stallions (54%). However, for Parascaris spp., foals had the highest prevalence (46%) followed by weanlings (32%) and yearlings (13%). The highest mean FECs for Parascaris spp. were observed in foals (418 eggs per gram [EPG] of faeces) while those for strongylids were in yearlings (1002 EPG). Of the adult horses (mares and stallions), 67% (489 of 729) and 11% (77 of 729) were low (i.e., ≤250 EPG) and moderate (i.e., 251–500 EPG) strongylid egg-shedders, respectively. Strongylid egg shedding varied across climatic zones, with the highest mean FECs in the summer rainfall (723 EPG) followed by non-seasonal rainfall (629 EPG), winter rainfall (613 EPG), and Mediterranean (606 EPG) rainfall zones. Twenty-three nematode species were detected using NGS, with Cylicostephanus longibursatus (28%), Cylicocyclus nassatus (23%) and Coronocyclus coronatus (23%), being the most abundant species. Three species of Strongylus (i.e., S. vulgaris, S. equinus and S. edentatus) were also detected. The nemabiome composition, species richness and relative abundance varied within horse age and between climatic zones. These empirical findings provide a comprehensive understanding of the prevalence of parasites within horse populations and the multifaceted factors that influence their occurrence, thereby allowing for the formulation of tailored strategies aimed at parasite control in domestic horses. [Display omitted] • A cross-sectional study of 62 Thoroughbred farms revealed a high prevalence of ascarid and strongylid nematodes. • Majority of the adult horses were low strongylid egg shedders (i.e., < 250 eggs per gram). • Horse age and diverse climatic conditions impacted the parasite diversity and egg shedding patterns. • Cylicostephanus longibursatus, Coronocyclus coronatus and Cylicocyclus nassatus were the most abundant species. • Study provided insights for tailoring parasite control strategies in horses. [ABSTRACT FROM AUTHOR]

Cyathostominae are ubiquitous to grazing horses and regarded the most prevalent internal parasite in the horse. Unfortunately, decades of indiscriminate use of anthelmintic drugs have resulted in the development of resistance in cyathostomins to all currently available drug groups, the most recent being a documented lack of efficacy to the macrocyclic lactones (ML). In vivo determination of anthelmintic resistance in horses most often utilises the faecal egg count reduction test (FECRT). Further, a shortened egg reappearance period (ERP) can indicate a change in response to the applied treatment and suggest an upcoming reduction of efficacy. Although both true resistance as demonstrated by the FECRT and shorter ERPs after ML treatment have now been shown in cyathostomins worldwide, the efficacy of ML as regards to cyathostomins in Sweden is currently unknown. The aim of the present study was therefore to determine FECRTs and ERPs after ivermectin (IVM) treatment in Swedish horses. Sixteen equestrian establishments with a minimum of six horses excreting at least 150 eggs per gram faeces (EPG) at screening were selected. For each establishment, FECRTs and ERPs were determined by collecting faecal samples prior to and 14 days after IVM treatment (200 µg/kg), and thereafter at weekly intervals for a total of eight weeks. All participants responded to a questionnaire detailing pasture management methods and anthelmintic routines.Questionnaire results showed that the majority of establishments (69%) only treated horses with anthelmintic drugs if indicated by faecal diagnostics and all of the establishments had a mean FECRT exceeding 99.0% and ERPs ranging from six to over eight weeks. The ERP was shown to increase with age as young individuals were shown to excrete cyathostomin eggs earlier after treatment compared with older horses (R = 0.21, p = 0.015). Riding schools, stud farms and those declaring not to use separate summer and winter paddocks had significantly shorter ERPs (p <0.01).In conclusion, retained ERPs and no confirmed resistance to IVM were found in Swedish equine establishments practising selective anthelmintic treatment, and supports the use of selective deworming regimens as a means of reducing the risk of anthelmintic resistance development. [Display omitted] • Routine anthelmintic treatment of horses without prior diagnostics is rare in Sweden. • Ivermectin resulted in FECRs > 99% in all studied horse establishments. • ERPs after ivermectin treatment were 6–8 weeks in all studied horse establishments. • No evidence of resistance in cyathostomins to ivermectin shown in Swedish horses. • Selective treatment regimens can reduce anthelmintic resistance development. [ABSTRACT FROM AUTHOR]

Human and animal health is globally affected by a variety of parasitic helminths. The impact of co-infections and development of anthelmintic resistance requires improved diagnostic tools, especially for parasitic nematodes e.g., to identify resistant species or attribute pathological effects to individual species or particular species combinations. In horses, co-infection with cyathostomins is rather a rule than an exception with typically 5 to 15 species (out of more than 40 described) per individual host. In cyathostomins, reliable morphological species differentiation is currently limited to adults and requires highly specialized expertize while precise morphological identification of eggs and early stage larvae is impossible. The situation is further complicated by a questionable validity of some cyathostomins while others might actually represent cryptic species complexes. Several molecular methods using different target sequences were established to overcome these limitations. For adult worms, PCR followed by sequencing of mitochondrial genes or external or internal ribosomal RNA spacers is suitable to genetically confirm morphological identifications. The most commonly used method to differentiate eggs or larvae is the reverse-line-blot hybridization assay. However, both methods suffer from the fact that target sequences are not available for many species or even that GenBank® entries are unreliable regarding the cyathostomin species. Recent advances in proteomic tools for identification of metazoans including insects and nematodes of the genus Trichinella will be evaluated for suitability to diagnose cyathostomins. Future research should focus on the comparative analysis of morphological, molecular and proteomic data from the same cyathostomin specimen to optimize tools for species-specific identification.

Methanol extracts of plant structures are promising alternatives to traditional pharmaceutical anthelminthic treatments. An in vitro evaluation was done of how methanol extracts of Diospyros anisandra bark and leaves, and Petiveria alliacea stems and leaves, collected during the rainy and dry seasons, effected cyathostomin larval development and egg hatching. Seven concentrations (600, 300, 150, 75, 37.5, 18.7 and 9.3 μg/ml) were tested using the egg hatch assay. An ANOVA was applied to identify differences between the concentrations and the controls. Fifty percent lethal concentration (LC 50 ) and the 95% confidence interval were calculated with a probit analysis. At and above 37.5 μg/ml, the D. anisandra bark extracts from both seasons exhibited ≥95% egg hatch inhibition (EHI), while the D. anisandra leaf extracts had >90% EHI at and above 75 μg/ml. For P. alliacea , the extracts from leaves and stems from either season exhibited >97% EHI at and above 300 μg/ml, although similar efficacy was also observed at lower concentrations with the rainy season stems (75 μg/ml) and leaves (150 μg/ml). Values for LC 50 were lowest for the rainy season D. anisandra bark (10.2 μg/ml) and leaf extracts (18.4 μg/ml), followed by the rainy season P. alliacea stems extract (28.2 μg/ml). In the D. anisandra extracts, EHI was largely due to its ovicidal activity (≥96% beginning at 37.5 μg/ml), whereas in the P. alliacea extracts it was due to L 1 larval hatch failure (≥90% beginning at 75 μg/ml). Overall, the rainy season D. anisandra bark extracts had a strong in vitro anthelminthic effect against cyathostomins by inhibiting larval development, and the rainy season P. alliacea stem extracts had a strong effect by preventing egg hatching. Both are possible control alternatives for these nematodes. [ABSTRACT FROM AUTHOR]

Anthelmintic resistance is a global problem that threatens sustainable control of the equine gastrointestinal cyathostomins (Phylum Nematoda; Superfamily Strongyloidea). Of the three novel anthelmintic classes that have reached the veterinary market in the last decade, none are currently licenced in horses, hence current control regimens focus on prolonging the useful lifespan of licenced anthelmintics. This approach would be facilitated by knowledge of the resistance mechanisms to the most widely used anthelmintics, the macrocyclic lactones (ML). There are no data regarding resistance mechanisms to MLs in cyathostomins, although in other parasitic nematodes, the ABC transporters, P-glycoproteins (P-gps), have been implicated in playing an important role. Here, we tested the hypothesis that P-gps are, at least in part, responsible for reduced sensitivity to the ML ivermectin (IVM) in cyathostomins; first, by measuring transcript levels of pgp-9 in IVM resistant versus IVM sensitive third stage larvae (L3) pre-and post-IVM exposure in vitro . We then tested the effect of a range of P-gp inhibitors on the effect of IVM against the same populations of L3 using the in vitro larval development test (LDT) and larval migration inhibition test (LMIT). We demonstrated that, not only was pgp-9 transcription significantly increased in IVM resistant compared to IVM sensitive L3 after anthelmintic exposure (p < 0.001), but inhibition of P-gp activity significantly increased sensitivity of the larvae to IVM in vitro, an effect only observed in the IVM resistant larvae in the LMIT. These data strongly implicate a role for P-gps in IVM resistance in cyathostomins. Importantly, this raises the possibility that P-gp inhibitor-IVM combination treatments might be used in vivo to increase the effectiveness of IVM against cyathostomins in Equidae. [ABSTRACT FROM AUTHOR]