Multifocal Necrotizing Myopathy in Northern Elephant Seal ( Mirounga angustirostris) Pups, San Miguel Island, California.

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.

Encysted cyathostomin larvae in foals - progression of stages and the effect of seasonality.

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.

Efficacy of two extra-label anthelmintic formulations against equine strongyles in Cuba.

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.

Evaluation of activity of fenbendazole, oxibendazole, piperazine , and pyrantel pamoate alone and combinations against ascarids, strongyles, and strongyloides in horse foals in field tests on two farms in Central Kentucky in 2014 and 2015.

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.

The cestode community in northern fur seals (Callorhinus ursinus) on St. Paul Island, Alaska.

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.

Parascaris univalens--a victim of large-scale misidentification?

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.

Profiles of strongyle EPG values for Thoroughbred mares on 14 farms in Kentucky (2012-2013).

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.

Causes of death in preweaned northern elephant seal pups (Mirounga angustirostris, Gill, 1866), Año Nuevo State Reserve, California, 2012.

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.

Molecular systematics of pinniped hookworms (Nematoda: Uncinaria): species delimitation, host associations and host-induced morphometric variation.

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.

Characterization of the inflammatory response to anthelmintic treatment of ponies with cyathostominosis.

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-ß 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.

SvSXP: a Strongylus vulgaris antigen with potential for prepatent diagnosis.

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.

Update on the prevalence of the hookworm, Uncinaria lucasi, in northern fur seals (Callorhinus ursinus) on St. Paul Island, Alaska, 2011.

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.

Review of research on hookworms (Uncinaria lucasi Stiles, 1901) in northern fur seals (Callorhinus ursinus Linnaeus, 1758).

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.

Observations on development of natural infection and species composition of small strongyles in young equids in Kentucky.

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.

Further evaluation in field tests of the activity of three anthelmintics (fenbendazole, oxibendazole, and pyrantel pamoate) against the ascarid Parascaris equorum in horse foals on eight farms in Central Kentucky (2009-2010).

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).

Investigations of peritoneal and intestinal infections of adult hookworms (Uncinaria spp.) in northern fur seal (Callorhinus ursinus) and California sea lion (Zalophus californianus) pups on San Miguel Island, California (2003).

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.

Reduced activity of moxidectin and ivermectin on small strongyles in young horses on a farm (BC) in Central Kentucky in two field tests with notes on variable counts of eggs per gram of feces (EPGs).

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.

Field tests demonstrating reduced activity of ivermectin and moxidectin against small strongyles in horses on 14 farms in Central Kentucky in 2007-2009.

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.

Three recently recognized species of cyathostomes (Nematoda: Strongylidae) in equids in Kentucky.

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.

Critical tests evaluating efficacy of moxidectin against small strongyles in horses from a herd for which reduced activity had been found in field tests in Central Kentucky.

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.