Negative covariance between parasite load and body condition in a population of feral horses.

In wild and domestic animals, gastrointestinal parasites can have significant impacts on host development, condition, health, reproduction and longevity. Improving our understanding of the causes and consequences of individual-level variation in parasite load is therefore of prime interest. Here we investigated the relationship between strongyle fecal egg count (FEC) and body condition in a unique, naturalized population of horses that has never been exposed to anthelmintic drugs (Sable Island, Nova Scotia, Canada). We first quantified variation in FEC and condition for 447 individuals according to intrinsic (sex, age, reproductive status, social status) and extrinsic (group size, location, local density) variables. We then quantified the repeatability of measurements obtained over a field season and tested for covariance between FEC and condition. FECs were high relative to other horse populations (mean eggs per gram ± SD = 1543·28 ± 209·94). FECs generally decreased with age, were higher in lactating vs non-lactating females, and unexpectedly lower in males in some part of the island. FECs and condition were both spatially structured, with patterns depending on age, sex and reproductive status. FECs and condition were both repeatable. Most notably, FECs and condition were negatively correlated, especially in adult females.

Variation and covariation in strongyle infection in East African shorthorn zebu calves.

Parasite burden varies widely between individuals within a population, and can covary with multiple aspects of individual phenotype. Here we investigate the sources of variation in faecal strongyle eggs counts, and its association with body weight and a suite of haematological measures, in a cohort of indigenous zebu calves in Western Kenya, using relatedness matrices reconstructed from single nucleotide polymorphism (SNP) genotypes. Strongyle egg count was heritable (h(2) = 23.9%, s.e. = 11.8%) and we also found heritability of white blood cell counts (WBC) (h(2) = 27.6%, s.e. = 10.6%). All the traits investigated showed negative phenotypic covariances with strongyle egg count throughout the first year: high worm counts were associated with low values of WBC, red blood cell count, total serum protein and absolute eosinophil count. Furthermore, calf body weight at 1 week old was a significant predictor of strongyle EPG at 16-51 weeks, with smaller calves having a higher strongyle egg count later in life. Our results indicate a genetic basis to strongyle EPG in this population, and also reveal consistently strong negative associations between strongyle infection and other important aspects of the multivariate phenotype.

Association between large strongyle genera in larval cultures--using rare-event poisson regression.

Decades of intensive anthelmintic treatment has caused equine large strongyles to become quite rare, while the cyathostomins have developed resistance to several drug classes. The larval culture has been associated with low to moderate negative predictive values for detecting Strongylus vulgaris infection. It is unknown whether detection of other large strongyle species can be statistically associated with presence of S. vulgaris. This remains a statistical challenge because of the rare occurrence of large strongyle species. This study used a modified Poisson regression to analyse a dataset for associations between S. vulgaris infection and simultaneous occurrence of Strongylus edentatus and Triodontophorus spp. In 663 horses on 42 Danish farms, the individual prevalences of S. vulgaris, S. edentatus and Triodontophorus spp. were 12%, 3% and 12%, respectively. Both S. edentatus and Triodontophorus spp. were significantly associated with S. vulgaris infection with relative risks above 1. Further, S. edentatus was associated with use of selective therapy on the farms, as well as negatively associated with anthelmintic treatment carried out within 6 months prior to the study. The findings illustrate that occurrence of S. vulgaris in larval cultures can be interpreted as indicative of other large strongyles being likely to be present.

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.

Strongyle egg reappearance period after moxidectin treatment and its relationship with management factors in UK equine populations.

Parasitic nematodes, particularly cyathostomins, are ubiquitous in grazing horses world-wide. Considerable burdens of cyathostomin larvae can encyst in the large intestinal wall. The most recommended treatment against these pathogenic stages is moxidectin. Information is required on how effective moxidectin is against cyathostomin populations in different regions. The objectives here were to determine the efficacy of moxidectin treatment and estimate the strongyle egg reappearance period (ERP) after treatment in several equine populations, to confirm the type of strongyle nematodes present and to identify other (i.e. management) factors associated with shortened ERP. Eight yards were recruited and moxidectin in combination with praziquantel administered to all horses (n=261). Faecal egg count (FEC) analysis was performed at weeks 0, 2, 6, 10 and 12 after treatment to determine efficacy and ERP. The ERP was estimated using two previously published methods. Morphological identification of cultured third stage larvae from the sample population was compared to a Strongylus vulgaris-specific end-point PCR to examine the presence of S. vulgaris in samples before and after treatment. Strongyle egg shedding patterns were also compared to worm management practices at each site. At 2 weeks post-treatment, moxidectin was highly effective (faecal egg count reduction range, 99.9-100%). The strongyle ERP ranged from 6 weeks to >12 weeks depending on the calculation method applied. Only cyathostomin larvae were detected by morphological identification. The results from the coprocultures and PCR showed that S. vulgaris was absent before and after treatment. Analysis revealed that regular faecal removal from pasture was associated with lower average FEC and lower prevalence of egg shedding.

Production and characterization of monospecific adult worm infections of Strongylus vulgaris and Strongylus edentatus in ponies.

Since 1978, 20 surgical implantations of either Strongylus vulgaris or Strongylus edentatus have been performed in our laboratory for the purpose of obtaining single species cultures of these parasites. Following surgical implantation peak EPG values of 13-327 (S. vulgaris) and 363-1284 (S. edentatus) generally occurred during the first 3 weeks post-implantation. Duration of infections was as long as 5 years. Successful outcome of such surgeries appears to be related to the total number of parasites used (> or = 38) and the ratio of female to male worms implanted (1:1 or 2:1).

Observations on the epidemiology and control of Strongylus vulgaris infections.

The epidemiology and control of helminth infections in the horse were studied in four small grazing experiments between 1981 and 1984 at the University of Utrecht. At autopsy in November or December negligible Strongylus vulgaris burdens were found in the cranial mesenteric artery of four groups of ponies, which had been treated with an anthelmintic in July and subsequently transferred to a clean pasture. Considerable arterial S. vulgaris burdens were seen in three groups of ponies which were treated with an anthelmintic in July without a move to clean pasture, and in another group of ponies in 1984, which was set stocked on a pasture used for horses in 1983 and which was treated with an anthelmintic (albendazole) 2 days before turnout in April and subsequently in May, June and July. A tracer pony, grazed with this group between the middle of September and the middle of November, harboured an even higher burden of arterial S. vulgaris larvae. The arterial S. vulgaris in the latter group could not be the result of contamination of the pasture with S. vulgaris eggs before July, as in the three other groups with considerable arterial S. vulgaris burdens. Pasture larval counts showed that S. vulgaris larvae do not only overwinter, but are able to survive in considerable numbers until autumn, longer than most other gastrointestinal nematodes. There were some indications that translation of infective larvae, which overwintered on pasture in some free living stage, occurred between May and July.

Seasonal translation of equine strongyle infective larvae to herbage in tropical Australia.

Longevity in faeces, migration to and survival on herbage of mixed strongyle infective larvae (approximately 70% cyathostomes: 30% large strongyles) from experimentally deposited horse faeces was studied in the dry tropical region of North Queensland for up to 2 years. Larvae were recovered from faeces deposited during hot dry weather for a maximum of 12 weeks, up to 32 weeks in cool conditions, but less than 8 weeks in hot wet summer. Translation to herbage was mainly limited to the hot wet season (December-March), except when unseasonal winter rainfall of 40-50 mm per month in July and August allowed some additional migration. Survival on pasture was estimated at 2-4 weeks in the summer wet season and 8-12 weeks in the autumn-winter dry season (April-August). Hot dry spring weather (pre-wet season) was the most unfavourable for larval development, migration and survival. Peak counts of up to 60,000 larvae kg-1 dry herbage were recorded. The seasonal nature of pasture contamination allowed the development of rational anthelmintic control programs based on larval ecology.

Dose titration of moxidectin oral gel against migrating Strongylus vulgaris and Parascaris equorum larvae in pony foals.

Moxidectin was tested for efficacy in ponies against experimental infections of 56 day Strongylus vulgaris larvae and 11 day Parascaris equorum larvae. Three dosages of moxidectin were tested: 300 micrograms per kg live body weight, 400 micrograms per kg, and 500 micrograms per kg, and the vehicle served as control. Ponies were first infected with 600 S. vulgaris third-stage larvae (L3) on Experiment Day 0 and then with 3000 embryonated P. equorum eggs on Day 45. Moxidectin treatments were administered on Day 56 and necropsy examinations were performed on Day 91. Strongylus vulgaris fourth-stage (L4) and fifth-stage (L5) larvae were recovered at necropsy from the control ponies, in dissections of the cranial mesenteric artery and its branches (L4 and L5), and recovered from nodules in the wall of the cecum and ventral colon (L5). Parascaris equorum larvae were recovered from the small intestine of control ponies. Moxidectin was highly efficacious against S. vulgaris L4 and L5 at all three doses tested (99.6-100%), and appeared to be equally efficacious against P. equorum larvae (100%); however, control ponies had low levels of P. equorum infections compared to previous experimental infections performed using identical methods. This suggests that the prior S. vulgaris infection on Day 0 may have influenced the subsequent experimental P. equorum infection on Day 45 and contributed to the lower recovery.

Role of lipids in the transmission of the infective stage (L3) of Strongylus vulgaris (Nematoda: Strongylida).

Infective larvae (L3) of Strongylus vulgaris have limited energy stores for host finding and for infection. For transmission to occur, the larvae must have sufficient energy to (a) migrate onto grass, where they are ingested by their equine host (host finding), and (b) penetrate into the host gut. This study is designed to test the hypothesis that L3 larvae of S. vulgaris partition their energy stores between locomotory activity (used in host finding) and infection activity (penetration). Chronic locomotory activity was stimulated by incubating S. vulgaris L3 larvae at a constant temperature (38 C). After 8 days of treatment, locomotory activity ceased (exhaustion). Exhausted L3 larvae had significantly decreased total lipid when compared to controls (P < 0.05), but there was no decrease in levels of protein of carbohydrate. Lipids of S. vulgaris L3 larvae are comprised of 9 fatty acids, some of which are depleted in exhausted worms (14:0, 14:1, 16:0, 16:1, 18:1, 18:2), whereas others (18:0, 20:4, 24:0) remain unchanged. These data suggest that specific fatty acids provide the energy source for locomotory activity in S. vulgaris. Exhausted L3 larvae were also less able to penetrate host cecal tissue in in vitro penetration assays when compared to controls (P < 0.05), suggesting that the depletion of individual fatty acids during locomotory activity also reduced infectivity. These data do not support the hypothesis that S. vulgaris L3 larvae partition their energy stores between host-finding and infection activities. A comparison of lipid storage profiles in the L3 larvae of 4 nematode species with similar transmission strategies (S. vulgaris, Strongylus edentatus, Strongylus equinus, and Haemonchus contortus) revealed similarities in the fatty acid composition of these species. These data suggest a relationship between transmission patterns and energy storage strategies in the L3 larvae of nematode parasites of vertebrates.

Impact of mixed strongyle infections in foals after one month on pasture.

Twelve Standardbred foals (age 3-6 months), with little previous exposure to parasites, were allocated to 2 groups and put onto pasture with low (Group L) or high (Group H) levels of larval contamination of large strongyles and cyathostomes. After 4 weeks grazing in September, the foals were housed indoors until necropsy 15 weeks later. Foals in Group H became clinically more affected than those of Group L in that they showed loss of vigour, weight gain depression, intermittent soft faeces and inappetence. One foal of Group H had persistent diarrhoea and was subjected to euthanasia 12 weeks after housing. Signs of colic were not observed. Faecal egg counts were significantly higher in Group H than in Group L (P<0.05). At necropsy, the mean number of S. vulgaris and cyathostomes was 20 and 18,000, respectively, in Group L, and 167 and 25,000 in Group H. Routine blood chemistry did not specifically reveal presence of S.vulgaris in pre-patency. A transient neutrophilia and eosinophilia, most prominent in Group H, was seen 2-8 weeks after start of exposure and anaemia was observed later in Group H. Serum albumin and albumin/globulin ratio were reduced, particularly in Group H, and a marked hyperbetaglobulinaemia was observed at 16-20 weeks in Group H. In conclusion, heavy infections with strongyles including S. vulgaris may become established in weaned foals after a brief period on pasture. Infections may be expressed clinically as debilitation, inappetence and intermittent diarrhoea without colic, and the need for control is imperative.

Transrectal ultrasonography of the cranial mesenteric artery of the horse.

Transrectal ultrasonography was performed on the cranial mesenteric artery and its major branches in 23 conscious adult horses. Ultrasonographically, 25 arterial segments were classified as either normal or abnormal. These ultrasonographic classifications were later compared with the gross and histologic evaluations of each artery following necropsy of each horse. In this study, transrectal ultrasonography as a diagnostic test for verminous arteritis had a 90% sensitivity for detecting normal arteries and an 86% specificity for detecting abnormal arteries, suggesting that ultrasonography may be useful in the antemortem diagnosis of verminous arteritis.

In vitro ultrasonographic appearance of the normal and verminous equine aorta, cranial mesenteric artery, and its branches.

Ninety-one equine aortic and cranial mesenteric arterial segments were evaluated ultrasonographically in a water bath. On the basis of pathologic evidence of verminous arteritis, arterial segments were classified into 4 categories, and the ultrasonographic characteristics of each group were evaluated. Normal arteries (class 1) were ultrasonographically characterized by a smooth luminal surface layer and uniform wall thickness and echogenicity. Arteries with only histopathologic evidence of verminous arteritis (class 2) were ultrasonographically characterized by a smooth luminal surface layer, uniform hyperechoic luminal layer. Arteries with both gross and histopathologic evidence of verminous arteritis (class 3) were characterized ultrasonographically by an irregular luminal surface layer, varying wall thickness, varying wall echogenicity, and the presence of a hyperechoic luminal layer. The ultrasonographic characteristics of arteries with luminal thrombosis (class 4) were an irregular luminal surface, varying wall thickness, and nonuniform echogenicity.

Occurrence of Strongylidae (Nematoda: Strongyloidea) in Polish horses "tarpans" from Popielne Reserve.

The study was performed to evaluate intensity of strongylid infection in stabled tarpans using the method of collecting worms after anthelmintic treatment and in wild tarpans from Popielne reserve on the basis of faecal examination. After treatment with ivermectin 12 cyathostomes and one large strongyle species were recovered from the faeces of two tarpans. The most abundant cyathostome species were C. catinatum, C. pateratum, C. longibursatum, C. nassatum and C. coronatus. A higher intensity of infections with small strongyles (cyathostomes) was found in stabled group than in the group from the reserve. Large strongyles were more prevalent in wild tarpans, with Strongylus vulgaris as most common species (66.7%), but the intensity of infection was low. The present results are compared with earlier studies of horses in Poland and other countries. The results confirm the stability of cyathostomes in different breed of horses over the world.

Strongylus vulgaris (Looss, 1900) in horses in Italy: is it still a problem?

A post-mortem survey was carried out on 46 Sardinian horses to evaluate the presence of Strongylus vulgaris and associated pathology. Horses were from local farms and had been treated with broad-spectrum anthelmintics at least 3 times a year. Examination of the cranial mesenteric arterial system (CMAS) showed parasite-induced lesions in all horses. S. vulgaris larvae were found in 39% of examined arteries, while their detection rate in coprocultures was 4%. Histology, carried out on 26 horses, showed mainly chronic and chronic-active lesions. Histometry showed a significant increase in thickness of the arterial wall, in particular of the intima tunic and adventitia tunic of the ileocolic artery and its colic branch. MCV, MCHC and alpha2, beta and gamma globulins were increased in horses with S. vulgaris larvae in the arteries, while the albumin/globulin ratio was decreased. Horses that were positive on faecal examination showed decreased values for RBC, PCV and the albumin/globulin ratio. Although several studies have shown a dramatic decrease of S. vulgaris infection worldwide, our data show that this parasite continues to exert its pathogenic role, even when its detection rate is quite low within the strongyle population infecting horses.

Ultrastructural aspects of feeding and secretion-excretion by the equine parasite Strongylus vulgaris.

Light, scanning, and transmission electron microscopy were employed to provide further data on the putative origins of the immunogenic secretory-excretory product (ESP) of Strongylus vulgaris (Looss 1900). The sharply delineated but superficial attachment to the equine caecum by the mouth leaves behind an oval area devoid of epithelial cells. Attachment does not extend deeply enough to reach the muscularis mucosa layer of the equine intestine. The progressive digestion of the ingested plug of tissue (epithelial cells, blood cells and mucous) was visualized. The coelomocytes, floating cells and membranous structures located in the pseudocoelom and intimately associated with the digestive, excretory and reproductive systems, and with the somatic muscles are described. The secretory-excretory system comprises two, ventrally-located, secretory-excretory glands connected to tubular elements. These glands synthesize granules of various sizes and densities that are delineated.

Plasma thromboxane B2 levels in horses experimentally infected with Strongylus vulgaris.

Plasma thromboxane B2 (TXB2) the stable inactive metabolite of thromboxane A2 (TXA2), was measured daily by specific radioimmunoassay in three groups of animals before and after experimental infection with Strongylus vulgaris. Infection of four 'parasite naive' foals produced a typical acute syndrome with intermittent but statistically insignificant rises in TXB2 levels. Interpretation of results was complicated by the presence of a non-septic peritonitis associated with implantation of the foals with electrodes for recording myoelectrical activity. In two foals of similar age, with some natural exposure to S. vulgaris, there was little or no clinical response to infection and increases in TXB2 were absent. Baseline levels were also much lower, indicating that the peritonitis may have affected the results obtained in the first group of foals. Severe mesenteric arteritis was confirmed at necropsy in all six foals. A third group of yearling horses, all with natural exposure to the parasite, were generally resistant to infection. One animal developed arteritis with clinical signs of diarrhoea and mild colic, and also showed intermittent increases in TXB2. The mean plasma TXB2 level after infection was significantly higher than in the control period, although absolute levels were lower than those recorded in the 'parasite naive' foals. Other animals in this group had low TXB2 levels and minimal arteritis was found at necropsy. These results indicate that although infection appears to have an effect on plasma TXB2, the changes are inconsistent and not reliable indicators of the presence of verminous arteritis. The results also confirm the difficulty in establishing infection and the variability of the response in animals with previous exposure.

The effect of Strongylus vulgaris larvae on equine intestinal myoelectrical activity.

The myoelectrical activity of the ileum, caecum and large colon was monitored from Ag-AgCl bipolar recording electrodes in four conscious 'parasite-naive' weanling foals. All foals were inoculated with 1000 infective 3rd-stage Strongylus vulgaris larvae and alterations to the myoelectrical activity observed. The frequencies of caecal and colonic spike bursts increased significantly in all post infection periods coinciding with assumed larval penetration into the intestinal mucosa and migration through the vasculature. Peaks in caecal and colonic activity occurred at Days 1 to 5 post infection. In the caecum, peaks occurred again at Days 15 and 31 post infection, preceding similar rises in colonic spike burst frequency at Days 19 and 35. Longer term changes indicated a return towards pre-infection levels of activity suggesting smooth muscle adaptation to decreased blood flow. The analysis of caecal and colonic spike burst propagation indicated that the increases in burst frequency were not attributable to an increase in the propagation of spike bursts in any particular direction, but rather to proportional increases in all directions of activity. There was a slight decrease in the simple ileal spike burst frequency immediately post-infection. None of the experimental animals exhibited signs of abdominal pain during the trial, and there was no evidence of bowel infarction at post mortem examination despite the presence of severe parasite-induced arterial lesions. The results suggest that increased caecal and colonic motility is an important host response in susceptible foals exposed to S. vulgaris larvae.