Intestinal parasites in Przewalski's horses (Equus ferus przewalskii): a field survey at the Hortobágy National Park, Hungary.

The Pentezug Wildhorse Reserve, located in the Hortobágy National Park, Hungary, has one of the biggest ex situ populations of Przewalski's horses and aims to preserve its landscape and to study this subspecies. Between September and November 2018, 79 faecal samples were collected from Przewalski's horses. The McMaster, Willis flotation, natural sedimentation and coproculture methods were applied to all the samples. Results showed an average level of 1287 eggs per gram (EPG), which is a high faecal egg-shedding level. All the samples were positive for strongyle-type eggs (100%). There were no statistical differences regarding the EPG values between different harems of the population. The same happened when considering sexes, ages, lactating status or when bachelors are compared with harem members. Cyathostominae were dominant, when compared to Strongylinae and Tricostrongylidae, and 15 different morphological infective third-stage larvae types and/or species belonging to the order Strongylida were identified. The subfamily Cyathostominae was prevalent in 100% of the horses. Strongylus vulgaris was the most prevalent strongylin (40.5%). Additionally, 27.8% were positive for Parascaris sp. and 2.5% showed Oxyuris equi in their faeces. This study revealed that there is a higher prevalence of Triodontophorus serratus and Poteriostomum spp. in juveniles. Horses with S. vulgaris showed lower levels of EPG. This was the first study involving this population, showing 100% prevalence of intestinal parasites.

Helminth infections in Italian donkeys: Strongylus vulgaris more common than Dictyocaulus arnfieldi.

Donkeys have been used as working animals for transport and farm activities worldwide. Recently, in European countries, there has been an increasing interest in donkeys due to their use as pets, onotherapy or milk production. During 2014-2016, a countrywide survey was conducted to determine prevalence and risk factors of principal helminth infections in 1775 donkeys in 77 Italian farms. A questionnaire on management and parasite control practices was filled out for each farm. Faecal samples were examined using a modified McMaster technique, a centrifugation/flotation method and a sedimentation technique. Pooled coprocultures were performed for differentiation of strongylid eggs. Strongyles were the most common parasites detected (84.9%), followed by Dictyocaulus arnfieldi (6.9%), Oxyuris equi (5.8%), Parascaris spp. (3.6%), Anoplocephala spp. (1.0%), Strongyloides westeri (0.3%). Coprocultures revealed an omnipresence of cyathostomins (100%), followed by Strongylus vulgaris (31.0%), Poteriostomum spp. (25.0%), Triodontophorus spp. (9.0%), Strongylus edentatus (7.0%), Strongylus equinus (5.0%). Logistic regression analysis identified breed, co-pasture with horses, living area, herd size and number of treatments as significantly associated with strongyles. Sex, age, living area and herd size were significantly associated with Parascaris spp. Dictyocaulus arnfieldi was significantly associated with sex, grass, co-pasture with horses, living area and herd size. Strongylus vulgaris was significantly associated with living area and herd size. The mean number of anthelmintic treatments/year was 1.4; most of the donkeys (71.8%) were dewormed using an ivermectin drug. It is important to design parasite programs to specifically address both D. arnfieldi and S. vulgaris in donkeys, and this is especially important if donkeys co-graze with horses.

Detection of Strongylus vulgaris in equine faecal samples by real-time PCR and larval culture - method comparison and occurrence assessment.

BACKGROUND: Strongylus vulgaris has become a rare parasite in Germany during the past 50 years due to the practice of frequent prophylactic anthelmintic therapy. To date, the emerging development of resistance in Cyathostominae and Parascaris spp. to numerous equine anthelmintics has changed deworming management and the frequency of anthelmintic usage. In this regard, reliable detection of parasitic infections, especially of the highly pathogenic S. vulgaris is essential. In the current study, two diagnostic methods for the detection of infections with S. vulgaris were compared and information on the occurrence of this parasite in German horses was gained. For this purpose, faecal samples of 501 horses were screened for S. vulgaris with real-time PCR and an additional larval culture was performed in samples of 278 horses. A subset of 26 horses underwent multiple follow-up examinations with both methods in order to evaluate both the persistence of S. vulgaris infections and the reproducibility of each diagnostic method. RESULTS: The real-time PCR revealed S. vulgaris-DNA in ten of 501 investigated equine samples (1.9%). The larval culture demonstrated larvae of S. vulgaris in three of the 278 samples (1.1%). A direct comparison of the two methods was possible in 321 samples including 43 follow-up examinations with the result of 11 S. vulgaris-positive samples by real-time PCR and 4 S. vulgaris-positive samples by larval culture. The McNemar's test (p-value = 0.016) revealed a significant difference and the kappa values (0.525) showed a moderate agreement between real-time PCR and larval culture. CONCLUSIONS: The real-time PCR detected a significantly higher proportion of positives of S. vulgaris compared to larval culture and should thus be considered as a routine diagnostic method for the detection of S. vulgaris in equine samples.

Identification of third stage larvae of strongyles and molecular diagnosis of Strongylus vulgaris in the feces of Thoroughbred horses kept in training centers in Rio de Janeiro, Brazil.

The aims of the present study were to identify strongyles in the feces of Thoroughbred horses based on larval morphology; to detect Strongylus vulgaris using molecular diagnosis and compare results to those of feces culture; and to determine the association between the presence of S. vulgaris with corresponding animal information (age range, gender, and anthelmintic use). Feces of horses kept in six Training Centers in Rio de Janeiro State, that showed the presence of ≥500 eggs per gram of feces (EPG) were subjected to strongyle identification. Of the 520 fecal samples collected, 35 had an EPG ≥ 500. After fecal culture for L3 larvae identification, DNA was extracted, subjected to PCR to amplify the ITS2 region DNA fragment of S. vulgaris, and sequenced. A total of 3500 larvae were analyzed. Most were classified as small strong (99.7%), with an emphasis on the type A subfamily of Cyathostominae. Forms of S. vulgaris only corresponded to 0.2%. In all, 25 samples showed amplified S. vulgaris DNA products and 11 showed nucleotide sequences with high sequence identity. Fecal culture and PCR results showed poor agreement (kappa = 0.105) for S. vulgaris diagnosis. Age, gender, anthelmintic use, and anthelmintic administration interval were not statistically significant. The present study showed the presence of S. vulgaris in the feces of horses kept in Rio de Janeiro Training Centers, mainly seen via PCR, which has emerged as the most effective tool for diagnosis. This study made it possible to identify strongyles that infect horses in the region, emphasizing upon the necessity for constant monitoring of the animals.

Selective Anthelmintic Treatment in Horses in Sweden Based on Coprological Analyses: Ten-Year Results.

In Sweden, routine deworming has been used for several decades; however, to slow down the development of anthelmintic resistance, selective treatment is currently recommended. As part of a monitoring programme, equestrian premises submitted faecal samples to the National Veterinary Institute (SVA) twice per year between 2008 and 2017. Analyses for strongyles (small and large), tapeworms and ascarids, followed by premise-specific advice regarding deworming and parasite control strategies, were provided. In total, 43,330 faecal samples, collected from 26,625 horses on 935 premises in springtime (March to June), were analysed by quantitative or semi-quantitative flotation. Moreover, Strongylus vulgaris was detected by larval culture or PCR. Between 4 and 11% of individual horses tested positive for S. vulgaris and 3-10% were shedding tapeworm eggs. There were recurrent high and low egg shedders; 75% of horses with S. vulgaris appeared to have been recently introduced into the herd; the proportion of S. vulgaris-positive premises increased when individual samples rather than pooled samples were used. Based on the results of S. vulgaris diagnostics and strongyle egg-shedding level, 59% of the horses did not need to be dewormed.

Parasite dynamics in untreated horses through one calendar year.

BACKGROUND: Horses are host to a plethora of parasites. Knowledge of the seasonality of parasite egg shedding and transmission is important for constructing parasite control programs. However, studies describing these patterns are sparse, and have largely been conducted only in the United Kingdom. This study evaluated strongylid egg shedding patterns and transmission dynamics of Strongylus vulgaris in naturally infected and untreated mares and foals through one calendar year in Kentucky, USA. The study also investigated the existence of a peri-parturient rise (PPR) in strongylid egg counts in foaling mares and collected information about Strongyloides westeri and Parascaris spp. in the foals. METHODS: This study was conducted from January to December 2018. A herd of 18 mares, one stallion, and 14 foals born in 2018 were followed throughout the year. Sera and feces were collected biweekly from all horses, and worm burdens enumerated in 13 foals at necropsy. An S. vulgaris ELISA antibody test was run on all serum samples. Fecal egg counts were determined for all horses, and coproculture and qPCR assay were employed to test for the presence of S. vulgaris in the mature horses. Data were analyzed using the proc glimmix procedure in the SAS 9.4 software program. RESULTS: We found a general lack of seasonality in strongylid egg shedding throughout the year among the mature horses, and no PPR was demonstrated. Shedding of S. vulgaris eggs displayed a higher abundance during the spring, but findings were variable and not statistically significant. Anti-S. vulgaris antibody concentrations did not display significant fluctuations in the mature horses, but evidence of passive transfer of antibodies to the foals was demonstrated, and foals assumed their own production of antibodies starting at approximately 20 weeks of age. Overall, colts shed higher numbers of strongylid, ascarid, and S. westeri eggs than fillies. CONCLUSIONS: This study demonstrated a lack of seasonality in strongylid egg shedding for the study population, which is in stark contrast to previous studies conducted elsewhere. This strongly suggests that more studies should be done investigating these patterns under different climatic conditions.

Morphomolecular characterization of Strongylus vulgaris isolated from donkeys with special references to histopathological study on the affected organs.

Equine gastrointestinal tract is infected with Strongylus vulgaris (S. vulgaris) which is highly pathogenic parasite for its harmful effect on cranial mesenteric artery during its migration. So, this study was applied for identification of S. vulgaris in donkeys ultramorphologically and molecularly. In addition to, detection of the pathological effect of larval stage of S. vulgaris on the mesenteric arterial system using histopathology and immunohistochemistry. During the period from September to December; 2019, 60 male and 20 female donkeys at the Giza Zoo was postmortem examined. S. vulgaris adults and larvae were collected from the large intestine and cranial mesenteric arteries (CMAs), respectively. Ultramorphological examination of the collected adults was done using scanning electron microscope (SEM). DNA was extracted from 5 larvae and 6 adults for further conventional PCR studies and sequencing of the internal transcriped spacer 2 (ITS2) gene. The ITS2 gene were amplified and showed bands at 148 base pair (bp). The ITS2 gene nucleotide sequences of all isolates were aligned using Basic Local Alignment Search Tool (BLAST). Histological sections of S. vulgaris affected mesenteric arteries exhibited the presence of the parasite larvae either in the lumen with thrombus formation or attached to the intima. Most of the detected inflammatory cell populations were CD68-positive cells. From these results, it can be concluded that the ribosomal spacers genes could be used as markers for Strongylus species identification in eggs collected from equine feces as a beneficial method of diagnosis. Also, it could be important in disease surveillance, improving preventive measures and developing an effective control strategy.

Evaluation of Strategies to Reduce Equine Strongyle Infective Larvae on Pasture and Study of Larval Migration and Overwintering in a Nordic Climate.

Horses, as grazing animals, are inadvertently exposed to intestinal parasites that, if not controlled, may cause disease. However, the indiscriminate use of anthelmintic drugs has led to drug resistance, highlighting the need for pasture-management practices to reduce the level of parasitic exposure and lessen reliance on drugs. The efficacy of such methods depends both on the epidemiology of the parasites and the prevailing weather conditions. The aim of the study was to investigate the effect of faecal removal and harrowing on reducing the number of parasite larvae in herbage. Moreover, the migratory and survival ability of strongyle larvae in a Nordic climate was studied. Faeces from horses naturally infected with strongyle nematodes were used to contaminate pastures and grass samples were collected to harvest larvae. Twice-weekly faecal removal significantly reduced larval yields, whereas harrowing on a single occasion under dry weather conditions in the summer did not. Strongyle larvae were able to migrate 150 cm from the faecal pats, but most larvae were found within 50 cm. Both Cyathostominae and S. vulgaris survived the winter months with larvae harvested up to 17-18 months after faecal placement. Resting of pastures for one year greatly reduced the parasite level, but two years of rest were required for parasite-free pasture.

Helminths and their management in Swiss Army horses: differences between riding horses and pack horses evidence the need of improvement.

INTRODUCTION: Intestinal helminth management in horses has both clinical and epidemiologic relevance, in additional association with anthelmintic resistance. The Swiss Army employs military owned riding horses and privately owned pack horses, which are brought together for service periods up to 12 weeks. We compared husbandry conditions and intestinal helminth management of both groups via questionnaire and analysed faecal samples of 53 riding horses and 130 pack horses using combined sedimentation/flotation, the McMaster method and larval cultures. Riding horses only had cyathostomin infections (prevalence: 60,4 %), while pack horses harboured cyathostomins (71,5 %), Parascaris sp. (6,9 %) and Strongylus vulgaris (1,5 %). Regression models combining faecal sample results with questionnaire data unveiled correlations of husbandry practices with parasite frequencies identifying risk and protective factors. Pasture management, hygiene and deworming practices were highly variable for pack horses, while for riding horses there was an overall concept. This included a selective deworming strategy with faecal egg counts (FECs) of strongyles prior to deworming, applying a threshold of 200 eggs per gram of faeces (epg). Anthelmintic treatments based on FECs, weekly faeces removal on pastures (pastures), the use of macrocyclic lactones and deworming horses regularly were identified as protective factors regarding the 200 epg threshold for strongyle eggs. Accordingly, the mean epg for strongyle eggs between the groups (111 and 539 in riding and pack horses, respectively) was significantly different (p < 0,001). Overall, intestinal helminth management in pack -horses showed room for improvement regarding pasture hygiene, the used anthelmintics and the frequency of deworming, from which all Swiss Army horses would benefit, as they share pastures during their -service, therefore entailing the risk of parasite transmission.

INTRODUCTION: La gestion des helminthes intestinaux chez les chevaux présente une importance clinique et épidémiologique, en association avec la résistance aux anthelminthiques. L'armée suisse emploie des chevaux de selle militaires et des chevaux de bât privés qui sont réunis pour des périodes de service allant jusqu'à 12 semaines. Nous avons comparé la détention animale et la gestion des helminthes intestinaux des deux groupes par le biais d'un questionnaire et analysé les échantillons fécaux de 53 chevaux de selle et 130 chevaux de bât en utilisant la sédimentation/flottation combinée, la méthode McMaster et les cultures larvaires. Les chevaux de selle ne présentaient que des infections à cyathostomes (prévalence: 60,4 %), tandis que les chevaux de bât hébergeaient des cyathostomes (71,5 %), Parascaris sp. (6,9 %) et Strongylus vulgaris (1,5 %). Des modèles de régression combinant les résultats des échantillons fécaux et les données du questionnaire ont révélé des corrélations entre les pratiques de détention animale et la fréquence des parasites, identifiant les facteurs de risque et de protection. La gestion des pâturages, l'hygiène et les pratiques en matière de vermifugation étaient très variables pour les chevaux de bât, tandis que pour les chevaux de selle, il existait un concept unitaire. Ceci comprenait stratégie de vermifugation sélective avec comptage des œufs fécaux (CEF) de strongles avant la vermifugation, en appliquant un seuil de 200 œufs par gramme de fèces (opg). Les traitements anthelminthiques basés sur les CEF, l'enlèvement hebdomadaire des crottins sur le pâturage, l'utilisation de lactones macrocycliques et la vermifugation régulière des chevaux ont été identifiés comme des facteurs de protection concernant le seuil de 200 opg pour les œufs de strongles. En conséquence, l'iog moyen pour les œufs de strongles entre les groupes (111 et 539 chez les chevaux de selle et de bât, respectivement) était significativement différent (p < 0,001). Globalement, la gestion des helminthes intestinaux chez les chevaux de bât a montré qu'il est possible d'améliorer l'hygiène des pâturages, le choix des anthelminthiques et la fréquence des vermifuges, ce dont tous les chevaux de l'armée suisse bénéficieraient, car ils partagent les pâturages pendant leur service, ce qui entraîne un risque de transmission des parasites.

Reviving a tradition: The Development of Strongylus vulgaris in larval culture.

All horses are susceptible to the equine gastrointestinal parasite, Strongylus vulgaris, which is known to cause significant disease and death. The parasite undergoes development from the egg through the first (L1), second (L2) and third (L3) larval stages outside the horse. The L3 is the infective stage. The universally available technique for detection of S. vulgaris larvae is the larval culture method. This requires a 10-14 day culture period to induce development from egg to L3, followed by Baermannization and identification of the L3s to genus and/or species. It is unknown if the culture duration is necessary or ideal for S. vulgaris identification. The purpose of this study was to perform daily examinations of known S. vulgaris positive fecal samples in coproculture. Fresh feces were collected from a horse known to be shedding S. vulgaris eggs. A total of 140 cultures were set up using 10 g of feces. Cultures remained at room temperature and moistened every other day. Every day, 10 samples were examined, and all larvae were identified to stage, genus/species, and enumerated. Throughout the study, L1, L2, and L3 stages were observed, and S. vulgaris, Strongylus edentatus, Triodontophorus spp., and cyathostomin L3s were identified. Third stage larvae were observed on Day 5, and the mean number of L3s significantly increased on Day 10 (P < .001), and declined thereafter. Strongylus vulgaris was first observed on Day 6 with a mean count of 4.1 (95 % CI: 1.1, 7.1) S. vulgaris larvae, accounting for 4.1 % (95 % CI:1.8, 7) of the total L3s observed. The number of S. vulgaris larvae was significantly higher on Day 10 with a mean of 156.8 (95 % CI: 120.7, 192.9) S. vulgaris larvae (P < .001), and the proportion was also significantly higher with S. vulgaris comprising 50 % (95 % CI: 45.9, 54.8) (P = .006) of the total larvae. However, after 10 days, the mean number of S. vulgaris larvae declined, as did the proportion of S. vulgaris larvae compared to the total number of larvae. Using the described methods, it is possible to identify S. vulgaris as early as 6 days, and the optimal period is 10 days to detect the maximum number of S. vulgaris.

Strongylus vulgaris: Infection rate and molecular characterization from naturally infected donkeys at Sadat City, Egypt.

Strongylus vulgaris has high pathogenicity to equines. It causes aneurysm and thrombosis in the arteries particularly an anterior mesenteric artery, that is fatal to equines. In this study, we aimed to diagnose microscopically the natural infection of donkeys with Strongylus vulgaris from Sadat City, Minoufiya Governorate, Egypt. Fecal egg culture was used after the diagnosis of strongyle eggs to identify the species. Hematological and biochemical parameters were assessed. Adult worms were collected after post mortem examination of the infected animal. The sequence of ITS-2 was used to confirm the species of the parasite. The infection rate was 15.85% using the microscopical examination. The larval culture confirmed the infection with strongyle eggs as Strongylus vulgaris larvae. The sequence of ITS-2 was highly identical (about 95%) to sequences from Germany, China, and Turkey and occurred in the same genetic clade with the sequence from Germany. In conclusion, the study presented the diagnosis, the changes in the hematological and biochemical parameters in the infected animals, and the genetic characterization of Strongylus vulgaris from Sadat City, Minoufiya Governorate Egypt for the first time.

Parasite Occurrence and Parasite Management in Swedish Horses Presenting with Gastrointestinal Disease-A Case-Control Study.

All grazing horses are exposed to intestinal parasites, which have the potential to cause gastrointestinal disease. In Sweden, there is a concern about an increase in parasite-related equine gastrointestinal disease, in particular Strongylus vulgaris, since the implementation of prescription-only anthelmintics approximately 10 years ago. In a prospective case-control study, parasitological status, using fecal analyses for strongyle egg counts, the presence of Anoplocephala perfoliata eggs and S. vulgaris Polymerase chain reaction (PCR) as well as serology for S. vulgaris, were compared between horses presenting with or without gastrointestinal disease at a University hospital during a one-year period. Information regarding anthelmintic routines and pasture management was gathered with an owner-filled questionnaire. Although the prevalence of S. vulgaris PCR was 5.5%, 62% of horses were positive in the enzyme-linked immunosorbent assay (ELISA) test and horses with peritonitis showed higher antibody levels for S. vulgaris, as compared to other diagnoses or controls. Overall, 36% of the horse owners used only fecal egg counts (FEC), 32% used FEC combined with specific diagnostics for S. vulgaris or A. perfoliata, and 29% dewormed routinely without prior parasite diagnostics. Effective management methods to reduce the parasitic burden on pastures were rare and considering exposure to S. vulgaris appears high; the study indicates a need for education in specific fecal diagnostics and pasture management.

Comparative Analysis of Intestinal Helminth Infections in Colic and Non-Colic Control Equine Patients.

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.

Prevalence of Strongylus vulgaris in horses after ten years of prescription usage of anthelmintics in Sweden.

Strongylus vulgaris is considered the most pathogenic nematode parasite of the horse. Frequent deworming programs since the 1970s have reduced the prevalence of S. vulgaris to low levels, but to the price of widespread benzimidazole resistance in the small strongyles (cyathostominae) and ivermectin resistance especially in the equine roundworm Parascaris spp. To slow down the progression of anthelmintic resistance in Sweden, horse anthelmintics were made prescription only medicine in 2007 and selective therapy principles were introduced. This means that only individuals with high egg excretion or clinical signs of helminth infection were treated instead of blanket treatment of all horses on a farm. The aims of this study were to investigate the prevalence and risk factors associated with S. vulgaris infection in Sweden ten years after the introduction of a selective therapy regime. A total of 529 faecal samples from 106 farms were collected during March to June in 2016 and 2017. A web-based questionnaire was used to collect information about deworming routines. Strongyle faecal egg counts (FEC) were performed by McMaster and presence of S. vulgaris was demonstrated with a specific PCR on individual larval cultures. Results were analysed by mixed logistic (S. vulgaris prevalence) or linear (FEC) regression models, where farm was included as random factor. The overall prevalence of S. vulgaris at individual and farm levels was 28% and 61%, respectively. We observed a 2.9 increased odds risk of S. vulgaris infection on farms that based their treatment on strongyle FEC only as compared to farms that complemented strongyle FECs with larval cultures or dewormed regularly, 1-4 times per year, without prior diagnosis. We found no association between the prevalence of S. vulgaris and strongyle FEC level, horse age, geographical region or signs of colic. The prevalence of S. vulgaris was 25% in horses shedding ≤150 eggs per gram. Thus horses with low strongyle FECs that are left untreated could be an important source of S. vulgaris infection. This may be an important reason for the approximately three-fold increase in S. vulgaris prevalence since 1999 in Sweden. However, our combined results indicate that selective therapy based on a combination of strongyle FECs and larval cultivation was not associated with an increased risk of S. vulgaris infection. Still, S. vulgaris needs to be monitored continuously and should be taken into careful consideration when the treatment frequency is reduced.

Prevalence of Strongylus vulgaris in horses after ten years of prescription usage of anthelmintics in Sweden.

Strongylus vulgaris is considered the most pathogenic nematode parasite of the horse. Frequent deworming programs since the 1970s have reduced the prevalence of S. vulgaris to low levels, but to the price of widespread benzimidazole resistance in the small strongyles (cyathostominae) and ivermectin resistance especially in the equine roundworm Parascaris spp. To slow down the progression of anthelmintic resistance in Sweden, horse anthelmintics were made prescription only medicine in 2007 and selective therapy principles were introduced. This means that only individuals with high egg excretion or clinical signs of helminth infection were treated instead of blanket treatment of all horses on a farm. The aims of this study were to investigate the prevalence and risk factors associated with S. vulgaris infection in Sweden ten years after the introduction of a selective therapy regime. A total of 529 faecal samples from 106 farms were collected during March to June in 2016 and 2017. A web-based questionnaire was used to collect information about deworming routines. Strongyle faecal egg counts (FEC) were performed by McMaster and presence of S. vulgaris was demonstrated with a specific PCR on individual larval cultures. Results were analysed by mixed logistic (S. vulgaris prevalence) or linear (FEC) regression models, where farm was included as random factor. The overall prevalence of S. vulgaris at individual and farm levels was 28% and 61%, respectively. We observed a 2.9 increased odds risk of S. vulgaris infection on farms that based their treatment on strongyle FEC only as compared to farms that complemented strongyle FECs with larval cultures or dewormed regularly, 1-4 times per year, without prior diagnosis. We found no association between the prevalence of S. vulgaris and strongyle FEC level, horse age, geographical region or signs of colic. The prevalence of S. vulgaris was 25% in horses shedding ≤150 eggs per gram. Thus horses with low strongyle FECs that are left untreated could be an important source of S. vulgaris infection. This may be an important reason for the approximately three-fold increase in S. vulgaris prevalence since 1999 in Sweden. However, our combined results indicate that selective therapy based on a combination of strongyle FECs and larval cultivation was not associated with an increased risk of S. vulgaris infection. Still, S. vulgaris needs to be monitored continuously and should be taken into careful consideration when the treatment frequency is reduced.

Wild horse populations in south-east Australia have a high prevalence of Strongylus vulgaris and may act as a reservoir of infection for domestic horses.

Australia has over 400,000 wild horses, the largest wild equid population in the world, scattered across a range of different habitats. We hypothesised that wild horse populations unexposed to anthelmintics would have a high prevalence of Strongylus vulgaris infections. Verminous endarteritis and colic due to migrating S. vulgaris larvae is now absent or unreported in domestic horses in Australia, yet wild horses may pose a risk for its re-emergence. A total of 289 faecal egg counts (FECs) were performed across six remote wild horse populations in south-east Australia, of varying densities, herd sizes and habitats. Total strongyle egg counts ranged from 50 to 3740 eggs per gram (EPG, mean 1443) and 89% (257/289) of faecal samples had > 500 EPG, classifying them as 'high level shedders'. There were significant differences in mean total strongyle FECs between different locations, habitats and population densities. Occurrence of S. vulgaris was not predictable based on FECs of total strongyle eggs or small (<90 µm) strongyle eggs. A high prevalence of S. vulgaris DNA in faecal samples was demonstrated across all six populations, with an overall predicted prevalence of 96.7%. This finding is important, because of the ample opportunity for transmission to domestic horses. The high prevalence of S. vulgaris suggests vigilance is required when adopting wild horses, or when domestic horses graze in environments inhabited by wild horses. Appropriate veterinary advise is required to minimize disease risk due to S. vulgaris. Monitoring horses for S. vulgaris using larval culture or qPCR remains prudent. Gastrointestinal parasites in wild horse populations may also serve as parasite refugia, thus contributing to integrated parasite management when facing emerging anthelmintic resistance.

Management practices associated with strongylid parasite prevalence on horse farms in rural counties of Kentucky.

Anthelmintic resistance among cyathostomin parasites is a wide-spread problem. The parasite control guidelines written by the American Association of Equine Practitioners (AAEP) encourages the preservation of anthelmintic efficacy by reducing treatment frequency, using targeted deworming, and implementing environmental management practices. While there is knowledge regarding parasite management practices of affluent horse farms in the United States, surveys rarely explore the rural and underserved regions. The purpose of this study was to observe the management practices of horse farms in rural regions Kentucky, including working Amish farms, and determine factors associated with strongyle prevalence. A total of 160 horses among 38 owners from 28 different farms were enrolled in this study. A questionnaire survey regarding equine information, farm management, and deworming history was performed with each owner. Fecal samples were collected to determine fecal egg counts, perform coprocultures for subsequent strongyle larvae identification, and Strongylus vulgaris specific PCR. Serum samples were collected for the S. vulgaris antibody specific ELISA. The mean number of deworming treatments given in the last year was 2.1 with a 95% confidence interval of 1.9-2.3 with ivermectin being the most common active used. Statistical analysis showed horses treated within the last three months with a macrocylic lactone (ML) drug had significantly lower egg counts than horses treated with a ML 7-9 months ago (p = .0005). Despite the AAEP recommendations to reduce the overall number of treatments by using a surveillance-based approach and to no longer rotate treatments, only 17 horses reportedly had a fecal sample submitted for a fecal egg count and 65 horses were dewormed in a rotational manner. Horses whose owners utilized an informative deworming source (i.e., veterinarian, internet, magazine, local feed store) also had significantly lower counts (p = .0026). All coprocultures were negative for S. vulgaris while five horses were PCR positive. Interestingly, 95 horses tested ELISA positive for S. vulgaris. The strongyle egg counts of the working Amish horses were not significantly different from the other horses in this study and deworming practices including the use of efficacious drugs and low treatment frequencies were in accordance with the AAEP guidelines. This study was the first to summarize deworming management practices of rural regions in Kentucky, including a working Amish community. Overall, horse owners employed deworming practices recommended by the AAEP, however rotational deworming is still commonly implemented and fecal egg counts are rarely used.

Correlation between fecal egg count, presence of Strongylus vulgaris, and body score of feral horses on Fort Polk, Louisiana.

Approximately 700 feral horses, dubbed "trespass horses" by the United States Army, occupy Fort Polk, Louisiana and the surrounding Kisatchie National Forest. These horses are considered a nuisance and hazard, and the military is seeking to remove the horses via adoption. The aim of this research was to evaluate the fecal egg count (FEC), body condition score (BCS), and the presence of Strongylus vulgaris within this previously unstudied horse population prior to removal. The feral horse data was compared to domestic horses living on a single farm in the same area. A modified McMaster FEC, Henneke body scoring via photography, and PCR were used to evaluate 10 domestic horses and 28 feral horses. A significantly higher FEC was identified for feral horses when compared to domestic horses (p = 0.004), and 69.2% of feral horses were positive for S. vulgaris while all domestic horses tested negative. Additionally, no correlation was found between FEC and BCS for domestic (p = 0.213) or feral (p = 0.099) horses, and no association was found between FEC and S. vulgaris presence (p = 0.21) or BCS and S. vulgaris presence (p = 0.52). This study provides insight into S. vulgaris and strongyle prevalence in a previously unstudied group of horses and indicates a need for anthelmintic treatment and monitoring of the feral horses once they are adopted.

Nonstrangulating intestinal infarction associated with Strongylus vulgaris in referred Danish equine cases.

REASONS FOR PERFORMING STUDY: Strongylus vulgaris is a pathogenic helminth parasite infecting horses and was once considered to be the primary cause of colic. Migrating larvae cause ischaemia and infarction of intestinal segments. This knowledge is derived from case reports and experimental inoculations of parasite-naïve foals, and it remains unknown to what extent the parasite is associated with different types of colic. OBJECTIVES: To evaluate the role of S. vulgaris as a risk factor for different types of colic in horses. STUDY DESIGN: A retrospective case-control study among horses referred with abdominal pain to the University of Copenhagen Large Animal Teaching Hospital during 2009-2011. METHODS: Each colic case was matched with an equid of the same type (pony, Warmblooded or Coldblooded), age, sex and admitted in the same month and year but for problems unrelated to the gastrointestinal tract. Serum samples were analysed for antibodies to migrating S. vulgaris larvae using a recently developed enzyme-linked immunosorbent assay (ELISA). The following 4 case definitions were used: colic sensu lato, i.e. all horses presenting with colic (n = 274), with further subgroups, i.e. undiagnosed colics (n = 48), strangulating obstructions (n = 76) and nonstrangulating infarctions (n = 20). RESULTS: Strongylus vulgaris antibody levels were similar to control values in colics sensu lato and horses with undiagnosed colic. In contrast, nonstrangulating intestinal infarctions were significantly associated with positive S. vulgaris ELISAs (odds ratio 5.33, 95% confidence interval 1.03-27.76, P = 0.05). Also, horses with nonstrangulating infarctions had a significantly higher occurrence of positive ELISAs than horses with strangulating obstructions (odds ratio 3.79, 95% confidence interval 1.34-10.68, P = 0.01) and the colic sensu lato group (odds ratio 3.09, 95% confidence interval 1.20-8.01, P = 0.02). CONCLUSIONS: Nonstrangulating intestinal infarction was strongly associated with S. vulgaris-specific antibodies, whereas the more broadly defined colic categories were not associated with positive ELISA results. Thus, the ELISA holds potential to become a helpful adjunct in diagnosis and management of horses with colic.

Changes in Serum Strongylus Vulgaris-Specific Antibody Concentrations in Response to Anthelmintic Treatment of Experimentally Infected Foals.

Strongylus vulgaris is the most pathogenic nematode parasite of horses. Its extensive migration in the mesenteric blood vessels can lead to life-threatening intestinal infarctions. Recent work has shown that this parasite is still identified among managed horse populations. A serum enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of migrating larvae of S. vulgaris. Previous work has documented an increase in ELISA values following larvicidal treatment with ivermectin and suggested that the target parasite antigen is primarily produced by the later larval stages. The aim of this study was to experimentally inoculate cohorts of foals with S. vulgaris, and then compare ELISA responses to early or later ivermectin treatments. Fifteen foals were held in confinement and infected orally with ~25 S. vulgaris third-stage larvae on Days 0, 7, 14, and 21. Foals were weaned on Day 43 and turned out to a pasture not previously grazed by horses. Foals remained at pasture continuously until the study was terminated on Day 196. On Day 55, foals were randomly allocated to three treatment groups of five each. Group 1 received ivermectin on Day 56, Group 2 received ivermectin on Day 112, and Group 3 foals served as untreated controls. Serum and fecal samples were collected at 28-day intervals throughout the study. Serum samples were analyzed with the S. vulgaris-specific ELISA and fecal samples were processed for fecal egg counting. The ELISA values of Group 1 foals were significantly lower than Groups 2 or 3 on Days 140-196. Both treated groups exhibited increased ELISA values following ivermectin treatment. Results indicate that the target diagnostic antigen is produced throughout the course of arterial infection with S. vulgaris, but that an early ivermectin treatment can reduce the cumulative antigen produced over the course of an infection.