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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., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2019. Published by Elsevier Ltd.)

Species composition and distribution of hindgut ciliates were investigated in the feces of 20 thoroughbred mares in Kentucky, USA. Thirty-three species and six morphotypes belonging to 21 genera were identified. The average density of hindgut ciliates in mares was 13.5 ± 13.7 × 104 cells ml-1, whereas mean number of ciliate species per host was 14.4 ± 5.3. Bundleia nana, B. elongata, B. piriformis, Blepharoconus hemiciliatus, Holophryoides ovalis, H. macrotricha, Blepharoprosthium polytrichum, Prorodonopsis coli, Paraisotrichopsis composita, Blepharocorys microcorys, and Ochoterenaia appendiculata were the first identified species in horses inhabited in USA. Scanning electron microscopic images of Holophryoides macrotricha, Paraisotrichopsis composita, Cycloposthium dentiferum m. corrugatum, C. edentatum m. edentatum, C. edentatum m. scutigerum, Tetratoxum parvum m. parvum, T. parvum m. sulcatum, T. excavatum m. excavatum, Allantosoma intestinale, and Arcosoma brevicorniger were studied. Binary fission in Polymorphella ampulla was also studied.

A field study addressing causes of mortality in freshly dead northern elephant seals ( Mirounga angustirostris, Gill, 1866) was conducted on San Miguel Island, California, in February 2015. Necropsies were performed on 18 pups ranging in age from stillbirths to approximately 7 to 8 weeks. The primary gross diagnoses in these pups included trauma, myopathy, starvation/emaciation, infections, congenital anomalies, and perinatal mortality. However, 6 (33%) had a previously unrecognized myopathy characterized by multiple white streaks that were most obvious within the inner layer of the abdominal wall and the small innermost ventral intercostal muscles. Following histological examination, 2 more pups from San Miguel Island and 6 pups from The Marine Mammal Center (Sausalito, California) were found to have similar lesions. Histologically, the lesions within the skeletal muscles were characterized by a multifocal polyphasic, mild to severe, acute to subacute necrotizing myopathy with mineralization. Acute necrosis and degeneration characterized by pyknotic nuclei, eosinophilic cytoplasm and cytoplasmic vacuolization were found in smooth muscle myocytes within the urinary bladder and digestive system. Degeneration of myocytes was present in the tunica media of a few small- to medium-sized vessels and was characterized by a vacuolar degeneration and occasionally necrosis. This condition has been termed multifocal necrotizing myopathy. A cause of this myopathy was not identified.

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., (Copyright © 2018. Published by Elsevier B.V.)

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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

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., (Copyright © 2017 Elsevier B.V. All rights reserved.)

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., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Equine cyathostomin parasites are ubiquitous in grazing horses and have been shown to cause severe inflammatory disease in the large intestine of horses. Decades of intensive anthelmintic therapy have led to widespread anthelmintic resistance in cyathostomins across the world. In Cuba, no anthelmintic products are formulated and sold for equine usage and little is known about anthelmintic efficacy of ruminant and swine formulations used. A strongyle fecal egg count reduction test was used to assess the efficacy of a liquid formulation of ivermectin labelled for use in swine, ruminants and carnivores and a pelleted formulation of albendazole labelled for usage in ruminants. Nine farms in the province Camagüey were enrolled in the study comprising 149 horses in total. Albendazole efficacy was reduced on five farms and with the other four farms having no signs of reduced efficacy. Mean farm efficacies were ranging from 41.7% to 100% on the tested farms. Coprocultures found large strongyle larvae present on all farms, but all larvae identified post-treatment were cyathostomins. Ivermectin was found 100% efficacious on all studied farms. This study provided evidence of reduced albendazole efficacy in the study population. Further work is needed to evaluate whether these findings reflect true resistance or if they are due to pharmacokinetic or pharmacodynamic characteristics of the pelleted formulation tested here., (Published by Elsevier B.V.)

Cyathostomins (small strongyles) are ubiquitous in grazing horses and are known pathogens as cause of larval cyathostominosis. As part of their life cycle, cyathostomin larvae invade the mucosal walls of the large intestines and undergo encystment. Newly ingested third stage larvae are known to undergo arrested development and this can lead to an accumulation of encysted burdens over the course of a grazing season. It is believed that the host immune system plays a significant role in triggering this arrestment. Little is known about the development and progression of larval stages in foals that are not expected to mount a pronounced immune response to ingested cyathostomin larvae. This study evaluated counts of encysted larvae measured in 37 foals. The foals were born in 2013, 2014, and 2015 into a parasitology research herd kept without anthelmintic intervention and were humanely euthanatized between 50 and 293days of age as part of an ongoing parasite transmission study. A mucosal digestion technique was performed to enumerate encysted early third stage (EL3) and late third stage/fourth stage (LL3/L4) larvae in the cecum, ventral colon and dorsal colon. Counts were analyzed statistically to evaluate the influence of foal age, sex, and seasonality on the counts. Total counts as well as LL3/L4 counts were significantly higher during the grazing season (March-November). Three defined age groups (>100, 100-200, >200days) did not have statistically different counts. Male foals had significantly higher total counts compared to females, and this has not been reported before. The study found that 41% of the recovered larvae were EL3s, but no indication of arrested development of these was observed. This indicates that cyathostomin infection in foals progresses in a manner substantially different from mature horses., (Copyright © 2017 Elsevier B.V. All rights reserved.)

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., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Activity of fenbendazole (FBZ), oxibendazole (OBZ), piperazine (PIP) and pyrantel pamoate (PRT) alone and combinations of OBZ and PIP and of OBZ and PRT was evaluated against parascarids, strongyles and strongyloides in horse foals (n=281).This was on two farms - Farm A - mixed light-horses (n=26) and Farm B-Thoroughbreds (n=255) - in Central Kentucky in field tests in 2014 and 2015. Foals on both farms were treated one to three times each; an exception was that seven foals on Farm A were nontreated controls. Before treatment, the foals on Farm A were all weighed on scales but weights for Farm B were estimated. Evaluation of the anthelmintics was by recording the number of foals passing specific types of nematode eggs before and after treatment using qualitative and/or quantitative (EPG) methods. Results are: 1) ascarids (parascarids) - efficacy was excellent for OBZ, PIP, OBZ-PIP, and OBZ-PRT; likewise for PRT for one group (in 2014) of Farm A foals but not another group (2015) there. FBZ was inactive against these parasites. 2) strongyles - activity was lacking or incomplete for all of the compounds alone or in combination; it was the best for OBZ-PIP, OBZ-PRT and PRT alone. 3) strongyloides - data indicated no or incomplete activity; it was highest for OBZ and OBZ-PIP. The level of drug activity against parasites in Farm B foals may in some instances have been different if exact rather than estimated body weights had been obtained before treatment like for Farm A foals., (Copyright © 2016 Elsevier B.V. All rights reserved.)

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., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The diversity and ecology of cestodes from the northern fur seals, Callorhinus ursinus (NFS), were examined using newly collected material from 756 humanely harvested subadult males between 2011 and 2014. NFSs were collected from five different haul-outs on St. Paul Island, Alaska. A total of 14,660 tapeworms were collected with a prevalence of 98.5% and intensity up to 107 cestodes per host (mean intensity 19.7 ± 16.5 SD). Three species of tapeworms were found: Adenocephalus pacificus (Diphyllobothriidea) was the most prevalent (prevalence 97.4%), followed by Diplogonoporus tetrapterus (49.7%), and 5 immature specimens of Anophryocephalus cf. ochotensis (Tetrabothriidea) (0.5%). Most of the cestodes found in the NFS were immature (69.7%). However, only 0.9% of cestodes were in larval (plerocercoid) stages. The species composition, prevalence and intensity of cestodes from these NFSs were not statistically different between the five separate haul-outs. Significant increases in the intensity of NFS infections were observed during the study period.

Reasons for Performing Study: Anthelmintic treatments have been associated with local inflammatory reactions. Since each class of anthelmintic has unique mechanisms of action affecting different subpopulations of parasites, we hypothesised that they will also induce characteristic proinflammatory responses., Objectives: To determine the effect of anthelmintic class on the proinflammatory response post treatment., Study Design: Ponies naturally infected with cyathostomins and other parasites after pasture grazing were left untreated or treated with representatives of 3 different classes of anthelmintics: fenbendazole (benzimidazole); pyrantel tartrate (pyrimidine); and moxidectin (macrocyclic lactone). All were monitored for the expression of proinflammatory genes in the peripheral blood using real-time PCR., Methods: The ponies were divided into 4 treatment groups: Group 1 (n = 4) were untreated controls; Group 2 (n = 5) received 5 daily doses of fenbendazole (10 mg/kg bwt); Group 3 (n = 4) received daily treatment of pyrantel tartrate 2× (2.65 mg/kg bwt); and Group 4 (n = 5) received a single dose of moxidectin (400 μg/kg bwt). Blood samples were collected daily for 2 weeks to determine the effect of deworming on proinflammatory gene expression. Faecal egg counts were used to evaluate the efficacy of each drug., Results: While treatment with the benzimidazole significantly reduced egg counts up to 14 days post treatment, it also stimulated proinflammatory gene expression. Treatment with pyrantel salt also reduced faecal egg counts with less of a proinflammatory response. Treatment with the macrocyclic lactone was the most successful in reducing faecal egg counts and produced no signs of increased proinflammatory cytokine expression., Conclusions: This study revealed pronounced differences in the cytokine responses to anthelmintic treatment. This inflammatory reaction may play a role in the development of parasitic disease post anthelmintic treatment., (© 2014 EVJ Ltd.)

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%; overall mean 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.

The equine ascarid parasite Parascaris equorum is well known as a ubiquitous parasite infecting foals. A sibling species, Parascaris univalens, was first described over 130 years ago, but very little attention has been given to its existence and possible implications for anthelmintic resistance, clinical disease, or host age spectrum. P. univalens only possesses one germ line chromosome pair as opposed to two for P. equorum, but the two species are otherwise considered morphologically identical. For the present study, live worms obtained from the University of Kentucky parasitology horse herd were dissected and identified using karyotyping techniques. With no exception, all specimens (n = 30) were identified to be P. univalens. Further, the karyotyping technique was adapted to ascarid eggs derived from fecal samples and carried out on samples collected from 25 Thoroughbred foals from three farms in Central Kentucky. P. equorum was not identified among these, whereas P. univalens was found in 17 samples, with the remaining being inconclusive. The mitochondrial genome was sequenced, assembled, and annotated from one male worm identified as P. univalens, and comparison with available sequence reads labeled as P. equorum revealed only 0.16% nucleotide differences. However, it is unlikely that the sequences available in public databases have been unequivocally identified to species level by karyotyping. Taken together, these data suggest that P. univalens is likely the main species now observed in equines and that perhaps the designation Parascaris spp. should be used unless cytological characterization has confirmed the species.

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.

The main purpose of the research was to determine if three weekly samplings would produce consistent results for low strongyle egg shedders, generally associated with older horses. Strongyle egg counts per gram of feces (EPG) were done for 661 Thoroughbred mares on 14 farms in 2012-2013 once a week for three consecutive weeks. None of the mares had been treated with ivermectin or moxidectin for more than 90 days and with fenbendazole, oxibendazole, or pyrantel pamoate for more than 60 days. Strongyle EPG count data indicated that 38.3% of the mares were positive at first and second sampling, and 37.8%--at third sampling. There were 5.6-63.3% of horses positive per farm. Correlations between EPG count data in all three weeks are quite high (Rs = 0.84, p<0.001). Level of horse infection was found to be dependent on horse age, type of farm and individual characters of horse. This study documented that strongyle egg counts (EPGs) from individual horses did not differ significantly between weeks. Thus, a single strongyle EPG count can be interpreted as indicative of the level of egg shedding for a given horse. These data support the current trend of not deworming all adult horses on a farm, so that high shedders can be identified and treated. This saves horse owners money and reduces development of potential drug resistance of parasites because of less frequent usage of antiparasitic compounds. Information from this study helps to conclude recommendations for parasite control in horses globally., (Copyright © 2014 Elsevier B.V. All rights reserved.)

During an ongoing physiological ecology study on pups and adult female northern elephant seals (Mirounga angustirostris, Gill, 1866) on the mainland rookery at Año Nuevo State Reserve (California), an opportunity was afforded to collect fresh dead pups for parasitology and necropsy. The investigation was undertaken to delineate the causes of death of northern elephant seals recovered from Año Nuevo State Reserve. Prior to this study, there was no evidence of increased mortality or health problems on this rookery. Necropsies, histology, and ancillary diagnostic studies were conducted on 21 fresh dead preweaned pups. Ages ranged from 1 stillbirth to pups approximately 2 weeks of age. Gross lesions included varying degrees of bruising, hemorrhage, lacerations, and fractures attributed to blunt force trauma to the head, chest, and/or abdomen in 16 pups; starvation in 6 pups; bite wounds in 2 pups; generalized icterus in 2 pups; presumptive drowning in 2 pups; and 1 stillbirth. Most pups had multiple gross lesions. Following light microscopic examination, pups could be assigned into 4 general diagnostic categories: 1) trauma, 2) nutritional status, 3) infectious conditions, and 4) congenital anomalies. This investigation of preweaned pup mortality of northern elephant seals in California further refines diagnostic categories for perinatal pup mortality.

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., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Hookworms of the genus Uncinaria have been widely reported from juvenile pinnipeds, however investigations of their systematics has been limited, with only two species described, Uncinaria lucasi from northern fur seals (Callorhinus ursinus) and Uncinaria hamiltoni from South American sea lions (Otaria flavescens). Hookworms were sampled from these hosts and seven additional species including Steller sea lions (Eumetopias jubatus), California sea lions (Zalophus californianus), South American fur seals (Arctocephalus australis), Australian fur seals (Arctocephalus pusillus), New Zealand sea lions (Phocarctos hookeri), southern elephant seals (Mirounga leonina), and the Mediterranean monk seal (Monachus monachus). One hundred and thirteen individual hookworms, including an outgroup species, were sequenced for four genes representing two loci (nuclear ribosomal DNA and mitochondrial DNA). Phylogenetic analyses of these sequences recovered seven independent evolutionary lineages or species, including the described species and five undescribed species. The molecular evidence shows that U. lucasi parasitises both C. ursinus and E. jubatus, whereas U. hamiltoni parasitises O. flavescens and A. australis. The five undescribed hookworm species were each associated with single host species (Z. californianus, A. pusillus, P. hookeri, M. leonina and M. monachus). For parasites of otarids, patterns of Uncinaria host-sharing and phylogenetic relationships had a strong biogeographic component with separate clades of parasites from northern versus southern hemisphere hosts. Comparison of phylogenies for these hookworms and their hosts suggests that the association of U. lucasi with northern fur seals results from a host-switch from Steller sea lions. Morphometric data for U. lucasi shows marked host-associated size differences for both sexes, with U. lucasi individuals from E. jubatus significantly larger. This result suggests that adult growth of U. lucasi is reduced within the host species representing the more recent host-parasite association. Intraspecific host-induced size differences are inconsistent with the exclusive use of morphometrics to delimit and diagnose species of Uncinaria from pinnipeds., (Copyright © 2013 Australian Society for Parasitology Inc. All rights reserved.)

Cyathostomins can cause a severe inflammation of equine large intestine characterized by substantial ventral edema and pronounced protein loss. Anthelmintic treatment of horses can result in a localized inflammatory response in the colonic mucosa of clinically normal horses. The aim of this study was to evaluate the systemic inflammatory response of ponies naturally infected with cyathostomins to single dose representatives of three anthelmintic drug classes, namely, oxibendazole, pyrantel pamoate, and moxidectin. Thirty ponies aged between 1 and 18 years of age were allocated to one of three anthelmintic treatments groups. Anthelmintic efficacy was evaluated using the fecal egg count reduction test performed weekly between 2 and 8 weeks post-treatment. Inflammatory responses were evaluated on days 0, 1, 3, 5, and 14 after treatment using hematology, measurement of the acute phase inflammatory markers serum amyloid A, fibrinogen, haptoglobin, and iron, and real-time PCR measurement of expression of the genes for interleukins 1-β and -10, tumor necrosis factor-α, and interferon-γ. There were subtle inflammatory responses to treatment, but cytokine expression was significantly associated with the interaction term between treatment group and anthelmintic efficacy (P<0.05). Of the acute phase markers, only fibrinogen associated with treatment group. The findings suggest that systemic inflammatory responses subsequent to anthelmintic treatment of cyathostomin infection are minimal. It is possible that this response is 'buffered' by anti-inflammatory products of the parasites and/or the anti-inflammatory effects of the macrocyclic lactones., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

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.

Brucellosis, a frequent bacterial zoonosis, can produce debilitating chronic disease with involvement of multiple organs in human patients. Whereas acute brucellosis is well studied using the murine animal model, long-term complications of host-pathogen interaction remain largely elusive. Human brucellosis frequently results in persistent, chronic osteoarticular system involvement, with complications such as arthritis, spondylitis and sacroiliitis. Here, we focused on identifying infectious sites in the mouse that parallel Brucella melitensis foci observed in patients. In vivo imaging showed rapid bacterial dispersal to multiple sites of the murine axial skeleton. In agreement with these findings, immunohistochemistry revealed the presence of bacteria in bones and limbs, and in the lower spine vertebrae of the axial skeleton where they were preferentially located in the bone marrow. Surprisingly, some animals developed arthritis in paws and spine after infection, but without obvious bacteria in these sites. The identification of Brucella in the bones of mice corroborates the findings in humans that these osteoarticular sites are important niches for the persistence of Brucella in the host, but the mechanisms that mediate pathological manifestations in these sites remain unclear. Future studies addressing the immune responses within osteoarticular tissue foci could elucidate important tissue injury mediators and Brucella survival strategies.

Background: Strongyle parasites are ubiquitous in grazing horses. Strongylus vulgaris, the most pathogenic of the large strongyles, is known for its extensive migration in the mesenteric arterial system. The lifecycle of S. vulgaris is characterised by a long prepatent period where the migrating larvae are virtually undetectable as there currently is no test available for diagnosing prepatent S. vulgaris infection. Presence of S. vulgaris larvae in the arterial system causes endarteritis and thrombosis with a risk of non-strangulating intestinal infarctions. Emergence of anthelmintic resistance among cyathostomins has led to recommendations of reduced treatment intensity by targeting horses that exceed a predetermined strongyle faecal egg count threshold. One study suggests an apparent increase in prevalence of S. vulgaris on farms where reduced anthelmintic treatment intensity has been implemented. These issues highlight the need for an accurate and reliable assay for diagnosing prepatent S. vulgaris infection., Methods: Immunoscreening of a larval S. vulgaris cDNA library using hyperimmune serum raised against S. vulgaris excretory/secretory antigens was performed to identify potential diagnostic antigens. Immunoreactive clones were sequenced, one potential antigen was characterised, expressed as a recombinant protein, initially evaluated by western blot (WB) analysis, the diagnostic potential of the IgG subclasses was evaluated by ELISA, and the diagnostic accuracy evaluated using serum from 102 horses with known S. vulgaris infection status., Results: The clone expressing the potential antigen encoded a S. vulgaris SXP/RAL2 homologue. The recombinant protein, rSvSXP, was shown to be a potential diagnostic antigen by WB analysis, and a target of serum IgGa, IgG(T) and total IgG in naturally infected horses, with IgG(T) antibodies being the most reliable indicator of S. vulgaris infection in horses. Evaluation of diagnostic accuracy of the ELISA resulted in a sensitivity of 73.3%, a specificity of 81.0%, a diagnostic odds ratio of 11.69; a positive likelihood ratio (LR) of 3.85 and a negative LR was 0.33. The area under the ROC curve was 0.820., Conclusion: IgG(T) antibodies to recombinant SvSXP show potential for use as an antigen for prepatent diagnosis of migrating stages of S. vulgaris with moderate to good diagnostic accuracy.

Critical tests were performed in 2011 in four weanling horses (L-1, L-2, L-29, and L-30) treated with ivermectin paste at 200 μg/kg. They were born in 2011 and raised together on a farm (MC) in Central Kentucky. The horses had not been treated previously with an antiparasitic drug. However, ivermectin had been administered repeatedly to the horse herd for several years and strongyle eggs per gram of feces (EPGs) returned sooner posttreatment than after initial usage. Critical tests in a recent previous study in this horse herd indicated that the reason for the early return of strongyle EPGs after ivermectin treatment probably was because of lowered drug activity on immature (L(4)) small strongyles in the lumen of the large intestine. Therefore, the life cycle was shortened. The main purpose of the present study was to obtain further data on the activity of ivermectin on small strongyle immature stages, in addition to adults, in the intestinal lumen. Twelve species of small strongyles were present. Combined data for immature and adult small strongyles for the four ivermectin-treated horses demonstrated efficacy of 68 to 83 %. Removal of adults was 100 % for all four horses, and on immatures, it ranged from 0 to 16 %. Efficacy on immature small strongyles was even lower than in the previous study. This supported the earlier finding of apparent reduced time for maturation of the parasites because of incomplete elimination of immature small strongyles in the intestinal lumen of horses after ivermectin treatment. Data on five other helminth species were recorded.

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., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Strongyle parasites are ubiquitous in grazing horses, and the large strongyle Strongylus vulgaris is considered the most pathogenic helminth parasite of horses. Recent investigations have suggested an association between occurrence of this parasite and usage of selective therapy based on regular fecal egg counts. The established diagnostic method for S. vulgaris involves larval culture and subsequent morphological identification of third stage larvae under the microscope. Recently, a real-time PCR assay was developed and validated for the detection and semi-quantification of S. vulgaris eggs in equine fecal samples. The purposes of the present study were (a) to determine the presence of S. vulgaris by real-time PCR in Danish and American horses on farms using vastly different anthelmintic treatment regimens and (b) to evaluate the association between larval culture results and the PCR. A total of 991 horses representing 53 different horse farms in Denmark and Central Kentucky were studied. Fresh fecal samples were collected from all horses, and strongyle eggs retrieved for DNA extraction and subsequent real-time PCR analysis. Individual larval cultures were performed on the Danish part of the data set (663 horses on 42 farms). On the Danish farms, the S. vulgaris PCR prevalence was found to be 9.2% on farms not basing parasite control on fecal egg counts, and 14.1% on farms using selective therapy. No horses were PCR positive in the American part of the study (328 horses on 11 farms). Kappa-values indicated a moderate agreement between PCR and larval culture results, while McNemar tests revealed no statistical difference between the paired proportions. Significant associations were found between PCR cycle of threshold (Ct) value groups and larval culture counts. Results indicate that both diagnostic methods can be useful for determining the occurrence of S. vulgaris on horse farms, but that they both are affected by potential sources of error. The PCR results confirmed previous findings suggesting that S. vulgaris can reemerge under selective therapy regimens., (Copyright © 2012 Elsevier B.V. All rights reserved.)

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.

Northern elephant seals (Mirounga angustirostris Gill, 1866), inhabiting rookeries on the mainland of Año Nuevo State Reserve in central California, were investigated in 2012 for presence of hookworms (Uncinaria spp.). Material collected and examined for hookworms included: blubber (n = 15), stomach and intestines (n = 21) from dead pups; feces from the rectum of weaned pups (n = 23); sand containing apparent feces in areas of weaned pups (n = 28) and sand without apparent feces in areas of weaned pups (n = 54); milk from females (n =23) at 5 days and about 23 to 26 days postpartum; and placenta from one female. Evidence of hookworm presence was not detected in any of the samples examined. Possible reasons why hookworms were not found in northern elephant seals on the mainland of Año Nuevo State Reserve are discussed.

Prevalence of hookworms (Uncinaria lucasi Stiles, 1901) was determined in the northern fur seal (Callorhinus ursinus Linnaeus, 1758) on St. Paul Island (SPI), Alaska in July and August, 2011. Three of 61 (4.9%) dead pups harbored 1 to 13 adult hookworms each in their intestines. Parasitic larvae (L(3)) of hookworms were recovered from the blubber of 4 of 133 (3%) of subadult males (SAMs) examined. One parasitic L(3) was detected from each infected SAM. Adult U. lucasi (n = 3) were found in the intestine of 1 of 105 SAMs examined (0.95%). This is the first documented finding of adult U. lucasi in SAMs of the northern fur seals. Continued low prevalence of hookworms the last several years parallels the tremendous decline in the number of fur seals on SPI over a similar time period.

Monitoring studies of acanthocephalans in northern fur seals (Callorhinus ursinus Linnaeus, 1758) (NFSs) and a harbor seal (Phoca vitulina Linnaeus, 1758) were performed on St. Paul Island, Alaska, in July-August 2011. Gastrointestinal tracts of 105 humanely harvested NFS subadult males (SAMs) (3-4 years old) were collected during the annual Aleut subsistence harvest at four haul-out areas (HOAS): Lukanin (n = 26 NFSs), Polovina (n = 28), Gorbatch (n = 30), and Morzhovyi (n = 21). One gastrointestinal tract collected from a harbor seal (about 3-4 years old) found dead at Morzhovyi HOAS was also examined. The total prevalence of infection in NFSs with acanthocephalans was 29.52 % with variations from 7.69 % to 47.62 % between the four different HOAS. Eight acanthocephalan species of two genera-Corynosoma Lühe, 1904 (Corynosoma strumosum, Corynosoma alaskensis, Corynosoma cameroni, Corynosoma semerme, Corynosoma similis, Corynosoma validum, and Corynosoma villosum), and Bolbosoma Porta, 1908 (Bolbosoma nipponicum)-were found in the NFSs and a harbor seal. This is a new record of C. alaskensis for the NFSs. Short biological notes of the species found are presented. Differences in species composition as well as in prevalence of acanthocephalans parasitizing NFSs were observed in subpopulations from four different HOAS on St. Paul Island. The highest biodiversity of acanthocephalans and infection were found in subpopulations on Polovina and Morzhovyj HOAS, the lowest was on Lukanin HOAS. From 3.2 % (for C. validum) to 19.4 % (for C. villosum) of NFSs were infected by one acanthocephalan species; two species were found in 22.6 %; three in 9.7 %; and four in 3.2 %. Further studies of NFS parasites are necessary to follow the trends in parasitic infection rates and diversity in NFS population on the Pribilov Islands and for monitoring the influence of various ecological factors on NFS populations in Alaska.

A parasite study was done in 1,300 horse mares on 26 farms in Central Kentucky over a 5-month period in 2010 and 2011. The mares included 1,114 Thoroughbreds (TB) on 24 farms, 64 Standardbreds (SB) on 1 farm, and 122 mixed light horse types (MLH) on 1 farm. The objective of this research was to determine strongyle eggs per gram of feces (EPG) counts for evaluation by known age, number positive, and level of egg shedding by the mares. This was done to establish strongyle EPG profiles for the mares to aid in determining whether antiparasitic treatment was necessary. Eggs found were considered those of small strongyles. (A) For the Thoroughbred mares, (1) 362 (32%) were EPG-positive, and (2) the average (percentage) of EPG-positive mares by year of age was 3-5 (54%), 6-10 (36%), 11-15 (24%), 16-20 (17%), and >20 (21%); (3) EPG average counts were similar for all age categories except for the 6- to 10-year-olds, which were higher; (4) and the average (percentage) of positive mares by 100 units of EPG counts was ≤100 (50%), ≤200 (62%), ≤300 (70%), ≤400 (76%), ≤500 (80%), and >500 (20%). (B) For the Standardbred mares, 31 (48%) were EPG-positive; (2) the average (percentage) of EPG-positive mares by year of age (no >20 sampled) was lowest for the 3-5 and 16-20 categories and highest for the 6-10 and 11-15 groups; (3) EPG average counts by years of age were lowest for 3-5, 11-15, and 16-20 groups and highest for the 6-10 group; and (4) the average (percentage) of positive mares by 100 units of EPGs was 62% for the ≤100 category, 71-84% for ≤200 to ≤400 units, and the highest (97%) for the ≤500 unit. (C) For the mixed light horse type mares: (1) 94 (77%) were EPG-positive, (2) the average (percentage) of EPG-positive mares by age was lowest for the two oldest age groups, higher for the 11 to 15-year-old age group, and highest for the two youngest age groups; (3) EPG average counts by year of age were lowest for the 16-20 group, higher for the 6-10 and >20 groups, and highest for 3-5 and 11-15 groups; (4) the average (percentage) of positive mares by 100 units of EPG counts was lowest for the ≤100 category (23%), increasing about 10% progressively in ≤200 to ≤500 categories, but lower (37%) for the >500 category. (D) For all the three mare types (TB, SB, and MLH), 37% of the mares were EPG-positive, and 63% of the mares were EPG-negative; for the age (years) of positive mares, about one half belonged to the 3-5 category and a progressive decrease was seen for the 6-10, 11-15, and 16-20 groups, and 36% for the >20-year-olds; the mean strongyle EPGs highest range was seen in the 11- to 15-year-olds; the highest mean was in the 6- to 10-year-olds, and the lowest mean in the 3- to 5- and 16- to 20-year-olds. For the grouping of the strongyle EPG values by units of 100, three-fourths were in the ≤500 category, and the lowest percentage was for the >500 category. This research showed the value of strongyle EPG profiling for the mares. It was most useful for TBs where data from a large number of horses showed that over two-thirds were EPG-negative, indicating that there would be no known problem in deciding not to deworm them. While data were not as clear-cut on the SB and MLH mares, several of those which were negative and with "low" EPG values could be excluded from antiparasitic treatment.

Macrocyclic lactones (MLs) revolutionized parasite control in horses and other animals. They are unique in that they are effective against arthropods and nematodes. The first of the widely used avermectins was ivermectin. In 1983, it was marketed for use in horses as an injectable formulation but was withdrawn in 1984 after about a year and half on the market because of adverse problems. It was replaced by a paste formulation and an oral/stomach tube liquid formulation. Ivermectin is highly active on bots, ascarids, large and small strongyles, pinworms, strongyloides, stomach worms, and some other internal parasite species. Another ML, moxidectin, became available in 1997 as a gel formulation for oral administration. The parasiticidal activity of this compound is similar to ivermectin except efficacy is less on the common bot (Gastrophilus intestinalis) but high on encysted small strongyles. Recently however lower than initial activity on ascarids and small strongyles has been found for both ivermectin and moxidectin.

Early development and maturation of species of small strongyles have not been studied extensively. Most information is on the first appearance of strongyle eggs in feces of foals. However, species cannot be determined in this manner because of similarity of the morphological features of the eggs. To determine more definitive knowledge on development and species composition of natural infections of small strongyles, eight equid foals were necropsied and examined at various ages (31 to 92 days of age). The entire contents of the large intestine were examined, and all worms (6,671) recovered were identified. This was done to recover small strongyle specimens in an effort to find the stage of development of the worms for different time periods. The primary interest was to obtain data on the potential prepatent period. A total of 17 species of small strongyles were recovered. The earliest gravid female was Cylicostephanus longibursatus in a 57-day-old foal; then Cylicostephanus goldi was positive at 64 days, Cyathostomum catinatum at 68 days, Coronocyclus coronatus and Cylicostephanus calicatus at 70 days, Coronocyclus labiatus, Cylicocyclus leptostomus, Cylicocyclus nassatus, Cylicostephanus hybridus, and Cylicostephanus minutus at 74 days, and Cylicostephanus bidentatus at 92 days. While it is not known exactly when the foals began eating pasture vegetation (typically they begin to graze a short time after birth) and first ate infective free-living third-stage larvae, the results give an indication of the natural prepatent period of several species of small strongyles. Additional data are presented on the number of worms per foal, distribution of the worms in the parts (cecum, ventral colon, and dorsal colon) of the large intestine, and proportion of species found.

The activity of three anthelmintics (fenbendazole-FBZ; oxibendazole-OBZ; and pyrantel pamoate-PRT) was ascertained against the ascarid Parascaris equorum in horse foals on eight farms in Central Kentucky (2009-2010) in field tests. A total of 316 foals were treated, and 168 (53.2%) were passing ascarid eggs on the day of treatment. Evaluation of drug efficacy was determined qualitatively by comparing the number of foals passing ascarid eggs in their feces before and after treatment. The main purpose was to obtain data on current activity of these compounds against ascarids. Additionally, the objective was to compare these findings with those from earlier data on the efficacy of these three compounds on nematodes in foals in this geographical area. Efficacies (average) for the foals ranged for FBZ (10 mg/kg) from 50% to 100% (80%), for OBZ (10 mg/kg) from 75% to 100% (97%), and for PRT at 1× (6.6 mg base/kg) from 0% to 71% (2%) and at 2× (13.2 mg base/kg) 0% to 0% (0%). Although the efficacy varied among the drugs, combined data for all farms indicated a significant reduction of ascarid infections for FBZ (p < 0.0001) and OBZ (p < 0.0001) but not for PRT (p = 0.0953).

The peritoneal cavity (PNC) and intestine of northern fur seal (Callorhinus ursinus) pups and California sea lion (Zalophus californianus) pups that died in late July and early August, 2003, on San Miguel Island, California, were examined for hookworms. Prevalence and morphometric studies were done with the hookworms in addition to molecular characterization. Based on this and previous molecular studies, hookworms from fur seals are designated as Uncinaria lucasi and the species from sea lions as Uncinaria species A. Adult hookworms were found in the PNC of 35 of 57 (61.4%) fur seal pups and of 13 of 104 (12.5%) sea lion pups. The number of hookworms located in the PNC ranged from 1 to 33 (median = 3) for the infected fur seal pups and 1 to 16 (median = 2) for the infected sea lion pups. In addition to the PNC, intestines of 43 fur seal and 32 sea lion pups were examined. All of these pups were positive for adult hookworms. The worms were counted from all but one of the sea lion pups. Numbers of these parasites in the intestine varied from 3 to 2,344 (median = 931) for the fur seal pups and 39 to 2,766 (median = 643) for the sea lion pups. Sea lion pups with peritoneal infections had higher intensity infections in the intestines than did pups without peritoneal infections, lending some support for the hypothesis that peritoneal infections result from high-intensity infections of adult worms. There was no difference in intestinal infection intensities between fur seal pups with and without peritoneal infections. Female adult hookworms in the intestines of both host species were significantly larger than males, and sea lion hookworms were larger than those in fur seals. Worms in the intestine also were larger than worms found in the PNC. Gene sequencing and (RFLP) analysis of (PCR) amplified (ITS) ribosomal DNA were used to diagnose the species of 172 hookworms recovered from the PNC and intestine of 18 C. ursinus and seven Z. californianus hosts. These molecular data revealed that U. lucasi (hookworm of C. ursinus) and Uncinaria species A (of Z. californianus) infrequently mature in the intestine of the opposite host species in California rookeries. However, there is no support from molecular data for the hypothesis that cross-infection with "the wrong" Uncinaria species is a contributing factor in these cases of host peritonitis. The major significance of this research is the unusual finding of adult hookworms in the PNC of so many dead pups. No obvious explanation for this occurrence could be determined. Further research, like in the present study, should help understand and monitor the apparent ever changing role of hookworm disease in the health of northern fur seal and California sea lion pups on SMI.

The objective of this article is to review knowledge on the hookworm Uncinaria lucasi Stiles, 1901 in northern fur seals, Callorhinus ursinus Linnaeus, 1758. Emphasis is placed on research on this host-parasite system in the Pribilof Islands, AK, USA where the bulk of the studies has been performed.

Two field tests (A and B) were conducted in Thoroughbred weanlings born in 2008 and 2009 on a farm (BC) in Central Kentucky. The purpose was to determine the current efficacy of moxidectin (MOX) and ivermectin (IVM) on small strongyles by pre- and posttreatment counts of eggs per gram of feces (EPGs) in this geographical area. In previous parasite studies in horses on this farm and in horses on numerous other farms in this area, cultures of feces indicated the strongyle larvae were all small strongyles. Therefore, the strongyle eggs in the EPG counts are considered small strongyles. Designation of the location of the horses in the current study is by barn number. However, the horses were kept on pastures except when gotten up in the specific barns for treatment and collection of feces. The number of horses for study A (2008-2009) was 40 in a total of ten barns (four in each location) and that for study B (2009-2010) was 49 in a total of nine barns (four each in six locations and seven to nine each in three locations). The nine barns for the study-B horses were the same ones used for the study-A horses; the tenth barn for study A was not used for study B. The number of horses treated was 20 with MOX and 20 with IVM in study A and 24 with MOX and 25 with IVM in study B. Drugs given to horses in barns in study B were switched from those used in barns in study A. That is, horses in barns where MOX was used in study A were given IVM and vice versa for those in study B. Study-A horses in some of the barns had unexplained either negative or delayed return of EPG counts after treatment with either drug, compared to horses in other barns. No reason for inconsistencies could be determined. Study B was initiated to compare results with study-A horses, relative to their location in the same barns. Study- B horses, contrary to horses in study A, had mostly consistent results between barns. Most of the MOX- and IVM-treated horses in study B and some in study A had strongyle EPG counts returning almost twice as quickly as when these compounds were first marketed.

Three species of cyathostomes--Cylicocyclus ashworthi, Cylicostephanus bidentatus, and Cylicostephanus hybridus were identified recently in horses in Kentucky. General characteristics and distinguishing description of these species are presented. Distribution of these species and their role in the horse strongylid community are discussed. The importance of examining the entire contents of the large intestine or alternatively a high number of specimens in order to recover and identify species residing in low numbers is stressed.

Efficacy of ivermectin (IVM) and moxidectin (MOX) against small strongyles was evaluated in horses (n=363) in field tests on 14 farms in Central Kentucky between 2007 and 2009. Most of the horses were yearlings but a few were weanlings and mares. The number of horses treated with IVM was 255 and those treated with MOX was 108. Horses on six farms were allotted into two groups. One group was treated with each of the two drugs, whereas horses on the other eight farms were treated with only one of the two drugs--IVM on six farms and MOX on two farms. Strongyle eggs per gram of feces (EPGs) compared to initial use of IVM and MOX returned almost twice as quickly after treatment of horses on all of the farms. IVM has been used much more extensively in this geographical area than MOX. Reduced activity of MOX was evident even on farms with rare or no apparent previous use of MOX but with probable extensive use of IVM.

Increasing levels of anthelmintic resistance in equine nematodes have led to recommendations of more sustainable anthelmintic treatment protocols with emphasis on parasite surveillance and diagnosis, rather than prophylactic calendar-based treatments. This requires knowledge of the diagnostic test performance of techniques for counts of eggs per gram of feces (EPG) as well as methods for culturing, counting and identifying third stage (L(3)) strongyle larvae per gram of feces (LPG). For horses, such information does not exist in the published literature. The aim of this study was to examine the relationship between worm count and fecal egg count (FEC) data for strongyle and Parascaris equorum infections as well as larval culture counts for diagnosing Strongylus spp. infections. Necropsy data from 693 horses used for critical or controlled tests, including information on total worm counts, fecal egg counts (FEC) and larval culture results collected at the University of Kentucky over a period of 50 years were analyzed. Sensitivity, specificity, positive and negative predictive values, and receiver operator characteristic (ROC) curves were generated for the larval cultures and ascarid egg counts. For the strongyle egg counts, potential FEC cutoff values for treatment were evaluated statistically by comparing the total strongyle worm counts below and above chosen cutoff values. All tests had high positive predictive values (>0.95), but moderate negative predictive values (<0.70). The negative predictive values of the larval counts were negatively affected by increasing egg count levels. Strongyle FEC cutoff values up to the level of 500 EPG yielded significantly higher strongyle worm counts in the treatment group, whereas no differences were found at higher cutoffs. This supports usage of cutoffs for treatment in the 0-500 EPG range. Altogether, the present study yields unique and useful information of widely used methods for parasite surveillance and diagnosis in equine establishments., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Critical tests were performed in 2009 and 2010 in four 2-year-old horses naturally infected with internal parasites. The horses were from a herd (Farm MC) where reduced activity of ivermectin and moxidectin on small strongyles was demonstrated previously from EPG (eggs/gram of feces) data in field tests. Also, in critical tests in horses from the same herd, ivermectin was less effective on immature small strongyles in the lumen of the large intestine than when the drug was first marketed. The main interest in the present critical tests was to determine the efficacy of moxidectin (400 μg/kg) on small strongyles. This was done to try and find indications of why there has been a return of strongyle EPG counts sooner after treatment in field tests than when moxidectin was first commercially available. Removal of adult small strongyles for the four treated horses was >99% to 100%. Efficacy on immature (L(4)) small strongyles was 82%, 96%, 98%, and >99% for the individual horses. Identification of small strongyles recovered from two of the horses revealed that three genera and 11 species were present. Specimens of Cylicocyclus ashworthi are reported for the first time in horses in Kentucky although eggs of this species have been identified. Moxidectin, in the present study, was excellent on removing adult small strongyles but was less effective on immatures (L(4)) in the intestinal contents. The question as to why moxidectin efficacy on small strongyles has declined in field tests may have been answered at least to a certain extent. It seems that a significant factor is "quick development" of a few remaining immatures in the gut lumen of horses. Also, possible activity may have decreased on encysted stages in the large intestinal lining. In any event, after treatment of some horses with moxidectin, the life cycle of small strongyles is shorter now than at the onset of usage of this compound.

Deworming horses with anthelmintics that have activity against encysted small strongyle larvae (L(3) and L(4)) is a common practice in parasite control programs. The two drugs currently available for this use are moxidectin (MOX) administered in a single dose of 0.4 mg/kg and fenbendazole (FBZ) given at the larvicidal dose (10mg/kg for 5 days). Here, we report the efficacy of MOX and the larvicidal dose of FBZ for reducing counts of strongyle-type eggs per gram of feces in naturally infected horses. Fecal egg counts (FECs) of 15 yearlings were observed following deworming. On day 0, 6 of the 15 yearlings were administered a larvicidal dose of FBZ; 14 days later, all 15 yearlings received MOX at a single dose of 0.4 mg/kg. Feces were collected on day 0 for pre-treatment egg counts. Feces were collected at weekly intervals thereafter during FEC observation periods. FECs of FBZ-treated horses were compared at day 0 and 14 days post-treatment. The difference in means pre- and post-treatment with FBZ was not statistically significant (p=0.65). On days 0 and 42 of the MOX treatment observation period the mean FEC of the yearlings that had not received the FBZ treatment did not differ significantly from that of the FBZ-treated yearlings. MOX was effective in reducing fecal egg counts to 0 EPG for 21 days. At day 35 all but 2 of the yearlings had some eggs present (range=4-361 EPG) and at day 42 all but 1 yearling had eggs present (range=3-432 EPG). At day 42 the group mean FEC reduction had fallen from 100% to 67%. Results of this study do not support the use of the larvicidal dose of FBZ for small strongyle control. Larger field studies will be needed to investigate whether egg reappearance periods are shortening for MOX-treated horses., (Copyright © 2010 Elsevier B.V. All rights reserved.)

A sensitive and specific PCR hybridization assay was applied for species-specific monitoring of the small strongyle (Strongylida: Cyathostominae) populations in horses in a herd before and after treatment with the anthelmintic drug ivermectin. Fecal samples were collected pre- and post-treatment weekly from eight individual horses (four foals and four yearlings) for 6 weeks to determine counts of strongyle eggs per gram of feces (EPGs). Additionally, one foal and one yearling were nontreated controls. Also, one horse, from another herd known to be infected with Strongylus spp., was a positive control for these parasites. Genomic DNA was obtained from eggs in groups of approximately 6000-7000 eggs except for two samples containing low EPGs in which 450 eggs were used. Amplification of the intergenic spacers (IGSs) of ribosomal DNA (rDNA) of small and large strongyles followed by reverse line blot (RLB) assay were performed to identify the presence of the 12 most common equine small strongyle species and to discriminate them from Strongylus spp. Overall, 11 small strongyle species were identified in pretreatment samples. In the samples collected at 4 weeks after ivermectin treatment, eight small strongyle species were identified and four of them were predominant (Cylicocyclus nassatus, Cylicostephanus longibursatus, Cylicostephanus calicatus and Cylicostephanus minutus). At 5 and 6 weeks post-treatment, the RLB assay analysis showed almost the same composition in the small strongyle population as before treatment. Strongylus spp. were identified only in samples collected from the positive control horse for these parasites. These data confirm the ability of the PCR-RLB technique for simultaneous species-specific differentiation of equine strongyle eggs, indicating a valuable way of furthering drug-resistance studies.

In the course of a vaccine experiment on horses, microfilariae were observed in cultures of peripheral blood mononuclear cells (PBMCs) isolated from eleven of fifteen study horses. The microfilariae were clearly viable as evidenced by their vigorous movements in the cultures, thus indicating that they had survived the Ficoll gradient purification and the cryopreservation method used for retaining the PBMCs. The microfilariae were identified as Setaria equina, which is a vector-borne filarial nematode that causes a relatively benign infection of equids in which the adult worms reside in the peritoneal cavity. Although it is not possible to definitely state where the infections were acquired, the horses originated from Saskatchewan, Canada and spent a relatively short period of time in the United States prior to blood sampling. Therefore, it is likely that the infections occurred in Canada. Interestingly, assays conducted to determine levels of cytokine mRNA transcripts in the isolated PBMCs seemed to be largely unaltered by the presence of the microfilariae in the cell cultures. These findings demonstrate that a standard method used to purify and cryopreserve PBMCs from blood can result in the unintended co-isolation of worms from microfilaremic animals. Furthermore, the presence of the microfilariae did not appear to alter significantly the results of our immunologic assays, suggesting either that the nematode antigens were not recognized or that immunological tolerance may have developed in these horses. Although notable effects on the assays were not observed in this study, it seems possible that microfilarial contamination could represent a confounding variable for experiments examining cellular immunity.