An alternative control regimen for drug-resistant parasites is combination deworming, where two drugs with different modes of action are administered simultaneously to target the same parasite. Few studies have investigated this in equine cyathostomins. We previously reported that an oxibendazole (OBZ) and pyrantel pamoate (PYR) combination was not sustainable against a cyathostomin population with high levels of OBZ and PYR resistance. This study consisted of a field study and two computer simulations to evaluate the efficacy of a moxidectin-oxibendazole (MOX-OBZ) combination against the same cyathostomin population. In the field study, anthelmintic treatments occurred when ten horses exceeded 100 eggs per gram. Fecal egg counts and efficacy evaluations were performed every two weeks. The two simulations utilized weather data as well as equine and parasite population parameters from the field study. The first simulation repeated the treatment schedule used in the field study over a 40 year period. The second evaluated efficacies of combination treatments using selective therapy over 40 years. In the field study, efficacies of MOX and both combination treatments were 100%. The egg reappearance period for MOX was 16 weeks, and the two combination treatments were 12 and 18 weeks. The first (46.7%) and last (40.1%) OBZ efficacies were not significantly different from each other. In the simulation study, the combination treatment delayed MOX resistance development compared to when MOX was used as a single active. This occurred despite the low efficacy of OBZ. The second set of simulations identified combination treatments used with selective therapy to be the most effective at delaying MOX resistance. Overall, this study supports the use of combination treatment against drug-resistant cyathostomins, when one of the actives exhibits high efficacy, and demonstrates benefits of this approach despite substantially lowered efficacy of the other active ingredient.

A parasitological survey of 651 northern fur seals Callorhinus ursinus L. from five subpopulations was conducted on St. Paul Island, Alaska, during July–August 2012–2014. Digenean trematodes were found in 210 of 651 fur seals with a total prevalence of 32.3%. Intensity of infection varied from 1 to 1540 parasites with mean intensity 18.4 ± 111.1 SD and median intensity of 2 specimens per host. Significant differences in prevalence and intensity of infection in northern fur seals between separate rookeries was not observed (Mann–Whitney test; p > 0.05). Four species of digeneans belonging to the families Heterophyidae (Apophallus zalophi Price, 1932, Phocitrema fusiforme Goto and Ozaki, 1930, and Galactosomum ubelakeri (Dailey, 1969)) and Troglotrematidae (Nanophyetus salmincola (Chapin, 1926)) were found. Nanophyetus salmincola is reported from C. ursinus for the first time. We obtained partial 28S rDNA sequences for all digenean species and conducted molecular phylogenetic analysis to demonstrate their phylogenetic relationships.

Equine cyathostomin are pervasive gastrointestinal parasites with wide-spread resistance to the benzimidazole and tetrahydropyrimidine drug classes worldwide. Combination deworming has been proposed as a more sustainable parasite control strategy. Simulation studies have found combination deworming to be effective in controlling drug resistant ovine trichostrongylid parasites. One equine study demonstrated an additive effect of a combination of oxibendazole and pyrantel pamoate against cyathostomins. However, this is the only equine study evaluating combination therapy, and the effects of repeated combination treatments administered over time remain unknown. The purpose of this study was to observe the efficacy of repeated oxibendazole/pyrantel pamoate combination therapy administered over one year against a cyathostomin population with resistance to benzimidazole and pyrantel products. Fecal egg counts were determined for the entire herd (N = 21) at the day of anthelmintic treatment and at two-week intervals for eight weeks post treatment. Starting efficacies of oxibendazole (OBZ, 10 mg/kg) and pyrantel pamoate (PYR, 6.6 mg base/kg) were 66.7% and 63.3%, respectively. Hereafter, the herd was treated four times with an oxibendazole/pyrantel pamoate combination, eight weeks apart, followed by repeating the single active treatments before concluding the study. While the first combination treatment exhibited an additive effect of the two active ingredients, this efficacy was not sustained over the course of the study. Mean fecal egg count reduction (FECR) was significantly greater for the first combination treatment (76.6%) than the second (42.6%, p = 0.0454), third (41.6%, p = 0.0318), and fourth (40.7%, p = 0.0372) combination treatments. The final single active mean FECRs were 42.3% for oxibendazole, and 42.7% for pyrantel pamoate. These efficacies were not significantly different from the initial single active efficacies (OBZ, p = 0.4421; PYR, p = 0.8361). These results suggest that combination therapy against double resistant equine cyathostomin populations is not sustainable, when using actives with markedly decreased starting efficacies.

Parasite infections are more quantifiable postmortem than antemortem in horses. Thus a study was carried out examining dead horses for specific parasite species. Most of the weanling and older horses submitted to the University of Kentucky Veterinary Diagnostic Laboratory (UKVDL) for postmortem examination between November 22, 2016 and March 23, 2017 were examined for certain species of internal parasites. The stomach and duodenum from 69 horses were examined for bots (Gasterophilus spp.). Combined data for both Thoroughbred and non-Thoroughbred (16 other than Thoroughbred breeds/mixed breeds) horses revealed that the prevalence of Gasterophilus intestinalis was 19% (n=12) with 2nd instars (x̄ 8.5) and 39% (n=27) with 3rd instars (x̄ 90). The prevalence of Gasterophilus nasalis was 1.5% (n=1) for 2nd instars (x̄ 1) and 7% (n=5) for 3rd instars (x̄ 25). A few third instar G. intestinalis placed in 10% formalin showed slight movement at over two hundred hours later. The cecum and about 25cm of the terminal part of the ileum were examined from 139 horses for tapeworms (Anoplocephala spp.) and large strongyles (Strongylus spp.). The prevalence of A. perfoliata was 44% (n=62) and the average number of specimens per infected horse was 92.5. Strongylus vulgaris and Strongylus edentatus were not found in the gut of any horse.

Strongyloides westeri is a threadworm parasite found in the small intestine mainly in foals at up to about 4 months of age. This parasite is associated with small intestinal enteritis and diarrhea as well as skin irritation and a clinical syndrome called "frenzied behavior" due to percutaneous invasion. The aim of this study was to investigate S. westeri parasite burdens in age groups older than the initial lactogenic transmission phase. Secondary aims were to examine the impact of host age and gender on worm burden and egg output in naturally infected foals. Thirteen foals were enrolled in the study, exposed to natural infection with S. westeri and were humanely euthanized at ages ranging from 89 to 221days old. Egg counts per gram of feces were determined using the Mini-FLOTAC method. Worms were retrieved from the mucosa of the anterior half of the small intestine using a digestion method and enumerated. The foals were all observed to be healthy with no signs of parasitic disease. Foals at 4-5 months of age had significantly higher S. westeri worm and egg counts compared to those aged 6-8 months (p<0.05). Fecal egg counts were significantly associated with worm counts (p=0.0045). While egg and worm counts declined markedly in the 6-8 month age range, they were not eliminated completely. This could either reflect a new infection acquired orally or transcutaneously, or a subset of adult intestinal parasites acquired via the lactogenic route and surviving for longer time periods. This study provided new information about S. westeri infection patterns in young horses at 4-8 months of age.

Field tests were performed on Thoroughbred yearlings (n=143), evaluating efficacy of moxidectin (MOX) against ascarids and strongyles and the efficacy of an ivermectin (IVM)-praziquantel (PRAZ) combination against those nematodes and tapeworms on a farm in Central Kentucky. The study was started on March 1, 2016 and completed on August 23, 2016. Fecal samples were collected from yearlings every two weeks for counts of ascarid and strongyle eggs per gram of feces (EPGs) and for determining the presence of tapeworm eggs. MOX was given to 88 yearlings on March 1; 55 yearlings were nontreated controls on this date but were treated two weeks later with MOX. On June 15, 2016, all yearlings (n=87) remaining on the farm were treated with IVM-PRAZ combination. Ascarids were present in low numbers for MOX-treated horses, and the drug had virtually no effect. The low number of ascarid infected horses before any treatment during the study was most likely due to age related immunity. Efficacy against strongyles was alike for both MOX and the IVM- PRAZ combination. At two weeks post-treatment, strongyle EPG values were reduced over 90%, started increasing at four weeks post-treatment, and returned to near pre-treatment values at six weeks post-treatment. Tapeworm efficacy of the IVM-PRAZ treatment was 96% at two weeks and was maintained for the entire ten-week post-treatment examination period.

Parasite control in foals is of utmost importance due to the high susceptibility to parasitic infection and disease in this age group. Foals are commonly co-infected with strongyle and ascarid parasites, which complicate parasite control strategies. The present study retrospectively investigated necropsy records of foals born into a university herd kept without anthelmintic treatment since 1979. The aims were to statistically analyze the relationship between fecal egg counts, worm burdens, foal age, sex, and season with specific focus on Parascaris and Strongylus spp. A total of 83 foals born between 1999 and 2015 were included. Foals were born between January and September within the given year and age at necropsy ranged between 27 and 563 days of age with a mean and median of 202 and 204 days, respectively. One set of multivariate mixed linear models was constructed analyzing strongyle and ascarid fecal egg counts as outcome variables, and another set of analyses investigated the following worm counts as outcome variables: Intestinal Parascaris spp. counts (immatures and adults), S. vulgaris (migrating and intestinal stages), S. edentatus (migrating and intestinal stages). Both ascarid and strongyle egg counts were influenced significantly by differences between study years (p < 0.05). In addition, total ascarid egg counts were statistically influenced by age (p = 0.020) exhibiting a peak at four months of age and fillies had significantly higher ascarid worm burdens (p = 0.043). Foal age had significant influences on intestinal counts of immature Parascaris spp. (p = 0.034) and adult S. edentatus counts (p = 0.028). Larval counts of S. edentatus were significantly associated with birth month (p = 0.023), whereas counts of migrating S. vulgaris larvae were not statistically associated with any of the investigated covariates. This study provides novel information on the dynamics of important parasites in naturally infected foals.

Summary Necropsy and extensive parasitological examination of dead northern elephant seal (NES) pups was done on San Miguel Island, California, in February, 2015. The main interest in the current study was to determine if hookworms were present in NESs on San Miguel Island where two hookworm species of the genus Uncinaria are known to be present - Uncinaria lyonsi in California sea lions and Uncinaria lucasi in northern fur seals. Hookworms were not detected in any of the NESs examined: stomachs or intestines of 16 pups, blubber of 13 pups and blubber of one bull. The results obtained in the present study of NESs on San Miguel Island plus similar finding on Año Nuevo State Reserve and The Marine Mammal Center provide strong indication that NES are not appropriate hosts for Uncinaria spp. Hookworm free-living third stage larvae, developed from eggs of California sea lions and northern fur seals, were recovered from sand. It seems that at this time, further search for hookworms in NESs would be nonproductive.

Anisakid nematodes parasitize the alimentary tracts of aquatic vertebrates, including northern fur seals (Callorhinus ursinus) (NFS). The main purpose of this study was to estimate the prevalence, intensity, and species composition of anisakids in NFSs on St. Paul Island, Alaska, and to analyze changes in NFS infection with these nematodes during the last decades. The study was carried out on St. Paul Island, Alaska, in July-August 2011-2013. Stomachs of 443 humanely harvested NFS males were collected from five separate haul-out areas. A total of 4,460 anisakid specimens were collected and identified by morphological criteria. Gross and histological examinations of stomachs were also performed. The overall prevalence of NFS infection was 91.2 %; overallmean intensity 10.9, and overall median intensity 6. Five species, Anisakis simplex s. l., Contracaecum osculatum s. l., Pseudoterranova decipiens s. l., P. azarazi and Phocascaris cystophorae, were found. The prevalence of Anisakis was 46.5%, its intensity 7.5. The prevalence and intensity of Contracaecum, Pseudoterranova, and Phocascaris were 33.6 % and 3.5, 81.9 % and 6.5, and 5.2 % and 1.7, respectively. Significant differences in the prevalence of NFS infection were observed between separate haul-outs. Comparison of the present data with the results of studies performed in the North Pacific in the 1960s, showed a significant decrease in the intensity of NFS infection with anisakids during the last decades. The prevalence of mucosal lesions in NFS stomachs caused by anisakids also decreased from 92 to 20.9 %. Possible reasons for the changes in NFS infection with anisakids are discussed.

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

The present research is a continuation of studies conducted periodically over 40 years on transmission of natural infections of internal parasites in the same horse herd on pasture (Field 10) on a farm in Central Kentucky. It included 12 mixed light horse foals born in 2013 and euthanatized between July, 2013 and April, 2014 for collection of internal parasites. Parasites found: Gasterophilus intestinalis, Strongyloides westeri, Parascaris equorum, Anoplocephala perfoliata, small strongyles (cyathostomes), Strongylus vulgaris, Strongylus edentatus and Thelazia lacrymalis. Prevalence generally was related to age of the foals. Overall prevalence and number of specimens were lower than in earlier studies except for P. equorum. There were 15 species (much fewer than previously) of small strongyles found and recorded by location in the large intestines. All stages of small strongyles encysted in the mucosa of the large intestine were recovered by artificial digestion and in significantly lower numbers in older foals.

Summary Review of several aspects of the biology of Strongyloides westeri, especially on the life cycle, are presented. The main reason is because of the potential renewed importance of these parasites since the recently recognized increase in their prevalence in horse foals. Indications are that the first and main source of enteric infections in foals maturing to adults in their small intestines is from parasitic third stage larvae (L3) passing to them in milk of their dams. It appears that a lesser source of infection is from free-living L3 which, when they are ingested with food or penetrate the skin, can mature in foals. In conclusion, the source of enteric infections of adult S. westeri in foals is mainly from parasitic L3 that they ingest in milk of their dams.

The prevalence of patent Strongyloides westeri infections was determined by examination for eggs in fecal samples collected from 513 Thoroughbred (TB) foals between February 25 and June 3, 2014. The study was conducted with 244 colts and 269 fillies from 11 well-managed farms in Central Kentucky. Foals ranged from 17 to 117 days of age and had never been dewormed. The mean prevalence of patent S. westeri infection was 30 % (156/513 foals). This prevalence was substantially greater than historical reports from the same sampling area during the late 1990s and early 2000s. It is hypothesized that this change is attributable to diminished use of ivermectin in foals due to concerns about macrocyclic lactone resistance in Parascaris equorum.

Abstract Prevalence of Hookworms, Uncinaria lucasi (Ancylostomatidae), in Northern Fur Seals (Callorhinus ursinus) on St. Paul Island, Alaska. Lyons, E. T., Kuzmina, T. A., Carie, J. L., Tolliver, S. C., Spraker, T. R. — Review of main studies on biology and ecology of the hookworm Uncinaria lucasi Stiles, 1901 performed on St. Paul Island, Alaska, is presented. Current data on prevalence of adult hookworms parasitizing northern fur seals (NFS), Callorhinus ursinus Linnaeus, 1758, were obtained based on the examination of the intestines of dead NFS pups and subadult 3-4 year-old males in July and August of 2011-2013. In addition, blubber samples collected from subadult NFS males were examined for parasitic third stage hookworm larvae (L3). All current data were compared with previously published studies performed in 1950s-1960s. Current prevalence of U. lucasi in dead pups collected from Reef Rookery was 4.9 % in 2011, 0 % in 2012 and 10.5 % in 2013. This rookery has a rocky substrate. On sandy rookeries prevalence was up to 75 % on Morjovi Rookery and 50 % on Vostochni Rookery. Parasitic L3 were recovered in 2.5 % of subadult males examined in 2013. Decreasing prevalence of hookworm infection of dead pups and subadult males during the last several years follows the tremendous decline in the number of fur seals in the herd on St. Paul Island during last several decades.

Strongylus vulgaris is regarded as the most pathogenic helminth parasite infecting horses. Migrating larvae cause pronounced endarteritis and thrombosis in the cranial mesenteric artery and adjacent branches, and thromboembolism can lead to ischemia and infarction of large intestinal segments. A recently developed serum ELISA allows detection of S. vulgaris-specific antibodies during the six-month-long prepatent period. A population of horses has been maintained at the University of Kentucky without anthelmintic intervention since 1979, and S. vulgaris has been documented to be highly prevalent. In 2012, 12 foals were born in this population, and were studied during a 12-month period (March–March). Weekly serum samples were collected to monitor S. vulgaris specific antibodies with the ELISA. Nine colts underwent necropsy at different time points between 90 and 300 days of age. At necropsy, Strongylus spp. and Parascaris equorum were identified to species and stage and enumerated. Initial statistical findings indicate a significant interaction between foal age and ELISA results (p<0.042). All foals had initial evidence of S. vulgaris-directed maternal antibodies transferred in the colostrum, but then remained ELISA negative during their first three months of life. Foals born in February and March became ELISA positive at about 12 weeks of age, while those born in April and May went positive at about 15 and 21 weeks, respectively. Foal date of birth was significantly associated with ELISA results (p<0.0001). This could be explained by birth date-dependent differences in parasite exposure. One foal remained ELISA-negative throughout the course of 30 weeks during the study. A significant association was found between ELISA values and larval S. vulgaris burdens (p<0.0001) as well as a three-way interaction between S. vulgaris, S. edentatus, and P. equorum burdens (p<0.001). A plateau with a subsequent decline in ELISA values corresponded with S. vulgaris larvae leaving the bloodstream and migrating back to the intestine.

Observations were made on the intestinal threadworm (Strongyloides westeri) and ascarid (Parascaris equorum) in field studies in 373 Thoroughbred foals on nine farms in Central Kentucky (USA) in 2013. Qualitative examination was made of feces of the foals for presence of S. westeri and P. equorum eggs. The main purpose of the prevalence study was to obtain current data on S. westeri in Thoroughbred foals. This was done to compare these findings with earlier studies in this geographical area where the prevalence of this parasite has been very low the last several decades. All the foals except on one farm had been treated one or more times with a parasiticide before the study. Prevalence of S. westeri in foals was 0 to 3 % on two farms, 6 to 9 % on three farms, and 20 to 51 % on four farms. The prevalence of P. equorum in foals was 0 to 14 % on three farms, 27 to 38 % on three farms, and 46 to 51 % on three farms. Effect of drugs given before the current study, on the prevalence of the two parasitic nematode species, is discussed. Also, three field tests were done by the present authors to evaluate activity of three individual or combination of drugs against ascarids. Evaluation was somewhat limited because, on two of the farms, a relatively small number of foals was passing ascarid eggs before treatment. However, oxibendazole and oxibendazole/piperazine combination were much more efficacious than fenbendazole.

Studies of northern fur seal (Callorhinus ursinus Linnaeus, 1758) infection by the filariid nematode Acanthocheilonema odendhali were carried out in 2011–2012 on St. Paul Island, Pribilof Archipelago, Alaska. Skins of 502 humanely harvested northern fur seals from haul-out areas of five rookeries, Polovina (n = 122), Morjovi (n = 54), Zapadni (n = 72), Lukanin (n = 109), and Gorbatch (n = 145), were examined. A. odendhali was found in 18 % of northern fur seals. The prevalence of infection ranged from 12.5 % up to 22.9 % on different haul-out areas on the island. The mean intensity of infection was 1.3 (range 1–7). Detailed morphological examination of collected specimens was performed using light microscopy. Several characters were added to the morphological description of the species, among them lateral thickening of the body cuticle, especially prominent in males, variations in number and position of genital papillae in males, transverse striation of the cuticle, and terminal dilation on tail end in microfilariae. The adult specimens studied had a shorter esophagus than type specimens from the California sea lion described by Perry (1967). Comparison of partial sequences of the mitochondrial cox1 gene from specimens collected from five sampling sites on St. Paul Island and a specimen from the type host and territory in California showed no significant differences and strongly supported conspecificity of the material from Alaska with A. odendhali.

Parascaris equorum generally infects horses less than 18 months old and its pathological effects can be severe. Infection occurs when larvated eggs, present in pastures, paddocks, stalls, and on feeding and watering equipment are ingested. The purpose of this study was to examine the effects of windrow composting on the viability of P. equorum eggs at a cooperating central Kentucky horse farm. Three grams of feces containing 2216 P. equorum eggs per gram were sealed in filter bag sentinel chambers. Chambers were exposed to 1 of 3 treatments: constant exposure or intermittent exposure to the interior of the windrow; controls were stored at 4 °C. At day 0, all chambers in the experimental treatments were placed in the center of 10 locations of the windrow. On subsequent days when the windrow was turned, chambers in the constant exposure treatment were returned to the interior of the windrow and chambers in the intermittent exposure treatment were alternated between resting on top of, or inside, the windrow. Chambers from each treatment and control chambers were removed at days 2, 4, 6, 8, 10, 12, 14, and 18; and incubated for 21 days at room temperature (24 °C). After incubation, eggs were recovered from the chambers using double centrifugation flotation. Eggs were evaluated microscopically, staged according to development and classified as viable or nonviable based on whether embryonation to the larval stage had occurred. Results were reported as the mean percent viable eggs for each treatment and time point. A mixed linear model with repeated measures was used to evaluate the influence of experimental day and treatment on the percent viability of P. equorum eggs. Chambers treated with constant exposure contained 10.73% (SD = 0.29) viable eggs on day 2 and declined to an average of 0.00% by day 8. Chambers exposed to the intermittent treatment contained 16.08% (SD = 0.26) viable eggs on day 2 and decreased to 0.00% by day 6. Control chambers for days 2, 4, 6, 8, 10, 12, 14, and 18 all had viabilities above 79.00%. A significant fixed effect of experimental day (p < 0.0001) and compost treatment (p < 0.0001) was observed. There was no significant interaction between experimental day and compost treatment (p > 0.7459). The results of this study demonstrate that windrow composting was effective at rendering P. equorum eggs nonviable when it was tested under the conditions at a working horse farm.

The aims of the study were to determine fecundity of several strongylid species parasitizing domestic horses and analyze possible relations between numbers of eggs in female uteri and size of both the eggs and the nematodes as well as the influence of fecundity on proportion of species in the strongylid community. Twenty-five specimens from each of 15 strongylid species (Strongylus vulgaris, Strongylus edentatus, Triodontophorus serratus, Triodontophorus brevicauda, Triodontophorus tenuicollis, Cyathostomum catinatum, Coronocyclus coronatus, Cylicocyclus nassatus, Cylicocyclus insigne, Cylicocyclus leptostomus, Cylicostephanus calicatus, Cylicostephanus goldi, Cylicostephanus longibursatus, Cylicostephanus minutus, and Poteriostomum imparidentatum) collected after necropsy were studied. The reproductive system was extracted from the female body; all eggs were removed, counted, and measured under a light microscope. Significant differences in number of eggs in female uteri of various strongylid species were observed (Kruskal–Wallis test, p < 0.001); the least numbers of eggs were registered in C. longibursatus (average = 49) and C. leptostomus (63) and the largest number in S. edentatus (5,918). Significant correlation between nematode body size and number of eggs was observed (p < 0.001). Correlation between size of eggs and body size was insignificant (Spearman R = 0.11, p = 0.70). Negative correlation was observed between number of eggs in female uteri and proportion of these species in strongylid community (Spearman R = −0.78, p < 0.001). Multiple linear regression of species proportion in the community on three predictors (number of eggs, body size, and egg size) was not significant (p > 0.05). However, the question on influence of fecundity on proportion of species in strongylid community needs further studies.