A multicenter evaluation of the effectiveness of Quest Gel (2% moxidectin) against parasites infecting equids.

Controlled trials with a common protocol were conducted in Idaho, Illinois and Tennessee to evaluate anthelmintic effectiveness of Quest Gel (QG; 2% moxidectin) against lumenal parasites in horses. Candidate horses were required to have naturally acquired nematode infections, as confirmed by presence of strongylid eggs in feces. At each site, 24 equids were blocked on the basis of pretreatment strongyle fecal egg counts (EPG) and randomly assigned to treatments within blocks. Within each block of two animals, one received QG on Day 0 at a dosage of 0.4 mg moxidectin/kg b.w. and one was an untreated control. Body weights measured the day before treatment served as the basis for calculating treatment doses. Horses assigned to treatment with QG received the prescribed dose administered orally with the commercially packaged Sure Dial syringe. Horses were necropsied 12-14 days after treatment, and lumenal parasites and digesta were harvested separately from each of five organs, including the stomach, small intestine, cecum, ventral colon and dorsal colon. Parasites from stomachs and small intestines were identified to genus, species and stage. Micro- (i.e., < 1.5 cm) and macroparasites (i.e., > 1.5 cm) in aliquots from the cecum, ventral colon and dorsal colon were examined in aliquots of approximately 200 parasites until at least 600 parasites had been identified to genus, species and stage or until all parasites in the 5% aliquot were examined, whichever occurred first. Data were combined across sites and analyzed by mixed model analysis of variance to assess the fixed effect of treatment and random effects of site and block within site. Because QG does not contain a cestocide, efficacy of QG against tapeworms was not significant (P > 0.05). Based on geometric means, however, efficacy of QG was greater than 90% (P < 0.05) against 38 species and developmental stages of cyathostomes, strongyles, bots, larval pinworms and ascarids encountered in at least 6 of 36 control horses in the combined data set. None of the horses treated with moxidectin exhibited evidence of adverse effects. Study results demonstrate QG, administered to horses with naturally acquired endoparasite infections at a dosage of 0.4 mg moxidectin/kg b.w., was highly effective against a broad range of equine parasitic infections.

A minimal RNA promoter for minus-strand RNA synthesis by the brome mosaic virus polymerase complex.

The approximately 150 nt tRNA-like structure present at the 3' end of each of the brome mosaic virus (BMV) genomic RNAs is sufficient to direct minus-strand RNA synthesis. RNAs containing mutations in the tRNA-like structure that decrease minus-strand synthesis were tested for their ability to interact with RdRp (RNA-dependent RNA polymerase) using a template competition assay. Mutations that are predicted to disrupt the pseudoknot and stem B1 do not affect the ability of the tRNA-like structure to interact with RdRp. Similarly, the +1 and +2 nucleotides are not required for stable template-RdRp interaction. Mutations in the bulge and hairpin loops of stem C decreased the ability of the tRNA-like structure to interact with RdRp. Furthermore, in the absence of the rest of the BMV tRNA, stem C is able to interact with RdRp. The addition of an accessible initiation sequence containing ACCA3' to stem C created an RNA capable of directing RNA synthesis. Synthesis from this minimal minus-strand template is dependent on sequences in the hairpin and bulged loops.

Thermoluminescence in hydroxyapatite.

A thermoluminescent (TL) glow peak has been shown to exist at 80 degrees C in synthetic hydroxyapatite, in amorphous calcium phosphate, and in deproteinated rat bone mineral. This TL glow peak is excited by a thermal pretreatment at elevated temperature, followed by exposure to ultraviolet (UV) light. When measured on a cylic TL-readout UV-exposure basis after an original thermal pretreatment, the intensity of this 85 degrees C glow peak displays a logarithmic decay curve characteristic of a transfer thermoluminescence process: A deep trap is activated during the annealing process, and the subsequent UV exposure causes charge transfer to the 85 degrees C glow peak, as is verified using electron spin resonance. Exposure to the atmosphere appears to decrease the TL intensity levels, presumably due to water adsorption; vacuum drying and additional thermal pretreatment reverse this effect on the TL intensities. TL measurements are applied to the comparison of bone mineral samples of varying age. Increases in overall TL intensity are demonstrated between a four-week old normal rat group and eight- and fourteen-week-old normal rat groupings. The TL intensity is also increased by metabolic disorder which causes changes in bone mineral chemistry and crystallography.

Generation and partial characterization of an eosinophil chemotactic cytokine produced by sensitized equine mononuclear cells stimulated with Strongylus vulgaris antigen.

Supernatants generated by stimulation of peripheral blood mononuclear cells (PBMC) from Strongylus vulgaris sensitized or immunized ponies were assayed in vitro for eosinophil chemotactic activity (ECA) using the filter system in blind well chambers. The supernatants from these cultures were chemotactic for eosinophils, but not for neutrophils. Supernates from cultures of unsensitized PBMC stimulated with S. vulgaris antigen were not chemotactic for eosinophils. ECA was first detected in culture supernatants after 1.5 h of incubation and was dependent on both antigen and PBMC concentrations, but independent of serum concentrations. Both female and male S. vulgaris worm antigens stimulated ECA production from sensitized PBMC. ECA was not induced by in vitro stimulation of sensitized S. vulgaris PBMC by female Strongylus edentatus worm antigen. Partial characterization of the eosinophil chemotactic cytokine showed it to be nondialyzable, greater than 8000 molecular weight (MW), and sensitive to heating (56 and 95 degrees C), trypsin, and sodium metaperiodate treatments, suggesting that the cytokine is a protein containing some essential carbohydrate moieties. The cytokine described in this paper could partially contribute to the in vivo blood and tissue eosinophilia in experimental S. vulgaris infection.

In vivo activation of equine eosinophils and neutrophils by experimental Strongylus vulgaris infections.

Eosinophils and neutrophils from ponies with Strongylus vulgaris-induced eosinophilia (eosinophilic ponies; activated eosinophils and neutrophils) were assayed in vitro for chemotactic and chemokinetic responses to zymosan-activated serum (ZAS) using the filter system in Boyden chambers, for Fc and complement (C) receptors using the EA and EAC-rosette assays, respectively, and for phagocytic and bactericidal activities using opsonized Escherichia coli and the acridine orange method. The responses of activated eosinophils and neutrophils in the above assays were compared with those of eosinophils and neutrophils from S. vulgaris-naive ponies without eosinophilia (noneosinophilic ponies; nonactivated eosinophils and neutrophils). Differences in cell density following centrifugation in a continuous Percoll gradient were used to further characterize the heterogeneity of activated eosinophils and neutrophils. Activated and nonactivated eosinophils demonstrated similar chemotactic responses to ZAS while activated and nonactivated neutrophils demonstrated similar chemokinetic responses to ZAS. A higher percentage of activated eosinophils and neutrophils expressed Fc and C receptors compared with nonactivated cells (P less than 0.05). Generally, higher percentages of eosinophils and neutrophils expressed C than Fc receptors. However, the percentage of neutrophils with both receptors was higher than that of eosinophils. Phagocytosis and killing of E. coli by either type of eosinophil were not consistently observed. Both activated and nonactivated neutrophils phagocytized E. coli and significant differences between the two cell types were not observed. The bacterial activity, however, of activated neutrophils was significantly greater than that obtained using nonactivated neutrophils (P less than 0.05). Activated eosinophils and neutrophils were both separated into two distinct fractions based on differences in cell densities. A higher percentage of band 2 eosinophils (density of 1.106) expressed C receptors than did band 1 eosinophils (density of 1.049) (P less than 0.05). A higher percentage of band 1 neutrophils (density of 1.072) expressed both Fc and C receptors and these neutrophils were more phagocytic and bactericidal than were band 2 neutrophils (density of 1.082) (P less than 0.05). These data suggest that equine eosinophils and neutrophils are activated by chronic S. vulgaris infections.

Immunity in equine cyathostome infections.

Defining the characteristics of immunity and immune responses to equine cyathostome infections is clearly important to advancing our understanding of the development of these nematodes within the host, the clinical conditions attributed to them, and in developing more rational and novel strategies for their control. Nonetheless, little is currently known on this topic. Current data based on field observations, worm burdens and fecal egg counts suggest that horses acquire a resistance to cyathostome infection with age. This response is slow to develop and incomplete in that most horses regardless of age harbor significant populations of these nematodes. More convincing evidence has been obtained from experimental infections which indicate that mature horses previously exposed to infection are resistant to re-infection and this resistance is directed at all stages of the parasite life cycle. Further, some immunity against the developing stages within the mucosa appears to require less exposure and occurs in younger animals. Some non-specific events which induce expulsion of all species of lumenal dwelling nematodes also appear to take place post-infection with L3. Antibodies have been detected in limited studies against somatic extracts of adult worms. Not surprisingly, titers of these antibodies do not correlate resistance to re-infection. Serendipitous observations have, however, associated a greater expression of the gene for IL-4 with the spontaneous expulsion of lumenal parasites. The development of a usable model is required to further advance our knowledge in this area.

Identification of thiabendazole-resistant cyathostome species in Louisiana.

A critical trial was performed with five ponies 6-9 months of age and raised on a horse farm with demonstrated benzimidazole-resistant cyathostomes. Eleven species of cyathostomes were recovered, seven of which had resistance to thiabendazole. Degrees of resistance varied among ponies and from species to species. Resistant species were Cyathostomum coronatum, Cyathostomum catinatum, Cylicostephanus minutus, Cylicostephanus calicatus, Cylicostephanus longibursatus, Cylicostephanus goldi and Cylicocylus nassatus. This is the first study identifying resistant cyathostome species in the Gulf Coast region of the USA and, although no new resistant species were found, results differed somewhat from those of other authors in that none of the cyathostome populations that have been studied contain the same complement of resistant species.

Equine cyathostome populations: accuracy of species composition estimations.

Historically, surveys of equine parasites either are not quantitative in regard to prevalence and intensities of cyathostome species, or if quantitative, are estimates based on the identification of a very small sample of the population. Commonly 100-200 worms are identified. In the current study cyathostomes from 10 ponies were counted and identified to species in subsets of approximately 200 worms each from 5% aliquots of the large intestine contents until all worms in the aliquot were examined. A mean of 10.9+/-4.3 species were identified by examining 200 cyathostomes from each animal. This number increased to 25.2+/-2.6 species when the 5% aliquots were totally examined, indicating that prevalence rates from species with low intensities are probably much greater than previous survey data indicate. A statistical model was used to determine how many worms need to be identified to give a 95% confidence level that all species present are identified.

Efficacy of the nematophagous fungus Duddingtonia flagrans in reducing equine cyathostome larvae on pasture in south Louisiana.

The effectiveness of Duddingtonia flagrans in reducing the free living third stage larvae (L(3)) of equine cyathostomes on pasture when fed to horses has been demonstrated in cold temperate climates. The objective of this experiment was to assess the efficacy of D. flagrans against equine cyathostomes in the subtropical environment of southern Louisiana. Fecal pats were prepared by mixing feces obtained from a parasite-free horse fed D. flagrans at a dose of approximately 2 x 10(6) spores kg(-1), with feces containing cyathostome eggs from a parasitized horse. Control pats contained feces from a parasite-free horse mixed with feces containing cyathostome eggs. The fecal pats were placed on pasture in six replicates at 4-week intervals from March 1997 until January 1998. Comparison of recoveries of L(3) from non-treated control pats in the field with non-treated coprocultures maintained in the laboratory indicated that L(3) survival on pasture was reduced during the months of May, June, July, August and September. The efficacy of the fungus was determined by L(3) recovery from grass surrounding the fecal pats of treated and control groups. D. flagrans significantly reduced L(3) during the months of April, May, and October 1997 to January 1998 (range 66-99% reduction, p=0.0001), and for the year as a whole (p=0.0001).

An experimental evaluation of methods used to enumerate mucosal cyathostome larvae in ponies.

With the increased interest in equine cyathostomes it has become apparent that some evaluations of methods currently used to count the various larval stages which occur in the mucosa would be beneficial. Experiments were conducted to investigate the effects of fixation and storage of mucosal tissues at -20 C on the accuracy of counting these larvae. The accuracy of counting developing larvae within the mucosa by transmural illumination (TMI) and by artificial digestion (DIG) of the mucosa was also compared. The data indicate that fixation of digested mucosa in PBS-buffered 5% or 10% formalin did not effect the enumeration of either early hypobiotic L3 or larger developing L3 or L4. Although not optimal, counting these larvae by either TMI of DIG after freezing did not significantly differ from counts made on fresh tissues. Significant differences were also not seen between counts of developing larvae made by TMI or DIG. Because DIG must be used to count EL3 and small developing L3, it is possible that TMI is not necessary in heavily infected equids.

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).

Dose titration of moxidectin oral gel against gastrointestinal parasites of ponies.

Moxidectin was tested as an oral gel formulation during a controlled test performed to evaluate dosages against equine gastrointestinal parasites. Four groups of ten ponies were used. Ponies ranged from 1 to 20 years of age and were naturally infected in southern Louisiana or Mississippi. Fecal exams and fecal cultures were performed on all ponies to determine the strongyle egg counts and the percent distributions of large and small strongyles. Following these determinations, ponies were allocated to replicates of four ponies to provide an even distribution of strongyle infection, age, weight and gender. Members of each replicate were then randomly assigned to one of four treatment groups. The doses tested were 300, 400 and 500 micrograms kg-1 body weight. The oral gel vehicle alone served as control. Treatments were administered behind the tongue and the ponies were observed continuously for 4 h for any adverse reactions; thereafter, ponies were observed at least twice daily. Necropsy examinations were performed 14 days post-treatment for the recovery and identification of any parasites present. Moxidectin, at all doses tested, was 100% efficacious against adults of Strongylus vulgaris, Strongylus edentatus, Triodontophorus spp. and 22 species of small strongyles. Moxidectin was also 100% efficacious against larvae of Strongylus edentatus and Oxyuris equi, greater than 94% efficacious against Strongylus vulgaris larvae and Oxyuris equi adults at 14 days post-treatment. Moxidectin proved highly efficacious against luminal small strongyle larvae (> 99.9% against L4 and > 92% against L3) and moxidectin demonstrated some efficacy against encysted small strongyle larvae as well.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of ivermectin at an elevated dose against encysted equine cyathostome larvae.

The efficacy of a high dose of ivermectin (1.0 mg per kg Eqvalan liquid drench) on encysted cyathostomes was tested in a controlled study using 12 adult ponies with naturally acquired cyathostome infections. Six treated ponies and six non-treated controls were held in separate stalls for a period of 5 weeks. Cyathostome burdens, which included lumenal larvae, adults and encysted larvae, were determined at necropsy. The viability of encysted larvae, based on morphologic integrity, was assessed by observation of mural transillumination and by the histologic appearance of 12 larvae per pony. Efficacy against adult cyathostomes was 99.9%. Lumenal cyathostome larval numbers were reduced by 87%. Numbers of encysted cyathostome larvae, identified by transillumination of the large intestine, were reduced by 35%. However, this reduction was not statistically significant (P > 0.05) and differences in viability of encysted larvae were not observed. The data strongly indicated that ivermectin has little demonstrable effect on encysted equine cyathostomes.

Immune responses of pony foals during repeated infections of Strongylus vulgaris and regular ivermectin treatments.

Ten helminth-free pony foals divided into three groups were used in this study. Eight foals were each experimentally infected per os with 50 Strongylus vulgaris infective larvae weekly for 4 weeks, at which time one foal died of acute verminous arteritis. The remaining seven foals subsequently received 50 S. vulgaris infective larvae every 2 weeks for an additional 20 weeks. Four of the infected foals remained untreated (Group 1) and three of the infected foals were given ivermectin at 8, 16 and 24 weeks post initial infection (Group 2). Two foals served as controls (Group 3). Foals in Group 1 developed eosinophilia, which was sustained throughout the course of infection. A mild eosinophilia also developed in Group 2 foals; however, the eosinophil numbers were markedly reduced for 3 weeks after each ivermectin treatment. Only foals in Group 1 developed significant (P less than 0.05) hyperproteinemia, hyperbetaglobulinemia and a reversal of the albumin/globulin (A/G) ratio 4 weeks after initial infection. Significant (P less than 0.05) IgG anti-S. vulgaris ELISA titers developed in foals in Groups 1 and 2 3 weeks after infection and were sustained for the duration of the experiment. Western blot analysis of soluble somatic antigens of S. vulgaris adult female and male worms probed with sera from foals in Groups 1 and 2 revealed only subtle differences between these animals. The blastogenic reactivity of peripheral blood mononuclear cells (PBMC) to phytohemagglutinin and concanavalin A was not significantly different between groups. Peripheral blood mononuclear cells from foals in Groups 1 and 2 developed significant (P less than 0.05) blastogenic reactivity to S. vulgaris soluble adult somatic antigen when examined at 25 weeks after infection. Mesenteric lymph node cells from foals in Group 2, although not statistically significant, were more reactive to antigen than were the mesenteric lymph node cells from foals in Group 1 when examined at 27 weeks after infection. These results suggest that significant alterations in the immune response of ponies to S. vulgaris does not occur after intravascular killing of larvae by ivermectin treatments.

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.

Comparison of moxidectin oral gel and ivermectin oral paste against a spectrum of internal parasites of ponies with special attention to encysted cyathostome larvae.

Two dosages of moxidectin oral gel were evaluated and compared to a therapeutic dose of ivermectin oral paste in the control of a spectrum of gastrointestinal parasites of ponies naturally infected in southern Louisiana or Mississippi. Thirty-two mixed-breed ponies ranging in age from one to 21 years were used in this controlled test. Eight weeks prior to the experiment, ponies grazing on contaminated pasture were moved to a paddock and fed a pelleted ration, thus reducing or eliminating the potential for additional infection and ensuring the existence of a population of encysted larvae. Ponies were then allocated to replicates of four animals based on values of fecal strongyle egg counts and percent strongyle larvae composition determined from Baermann sedimentations of fecal cultures. Members of replicates were allocated to one of four treatment groups: moxidectin oral gel administered at 300 micrograms kg-1 body weight, moxidectin oral gel at 400 micrograms kg-1, the oral gel vehicle as negative control, and ivermectin oral paste at 200 micrograms kg-1. Prior to treatment, ponies were confined in pairs to covered concrete runs by treatment group. Two weeks following treatment, necropsy examinations of all animals were performed. Parasites were recovered from the lumen of the stomach, the intestinal tract, the cranial mesenteric artery and its major branches, the peritoneal body wall and from pepsin digests of mucosal scrapings taken from the cecum and large colon. Encysted cyathostome larval burdens were also compared using mural transillumination of segments of the large colon for visualization of the encysted forms. Control ponies were not uniformly infected with the spectrum of parasites; however, moxidectin, at either dosage, compared favorably with ivermectin in the control of the adults of Strongylus vulgaris, Strongylus edentatus, Triodontophorus spp., Oesophagodontus robustus, Trichostrongylus axei, Oxyuris equi, Parascaris equorum, Habronema muscae, as well as both the adult and larval Cyathostominae recovered from the lumen. Moxidectin also appears as efficacious as ivermectin against migrating large strongyle larvae at the two weeks post-treatment evaluation. Moxidectin demonstrated a trend towards greater efficacy against encysted cyathostome larvae than a therapeutic dosage of ivermectin, but this difference was not statistically significant. Moxidectin was less effective than ivermectin against Gasterophilus intestinalis and was equally ineffective as ivermectin against Anoplocephala perfoliata.

Antibody responses of ponies to initial and challenge infections of Strongylus vulgaris.

An indirect fluorescent antibody assay (IFA) was developed using Strongylus vulgaris third stage larvae (L3) as antigens. Observations using the IFA indicate that a species-specific antibody response to S. vulgaris L3 develops in S. vulgaris-infected ponies and that some surface L3 antigens are shared by adult worms. Sequential antibody levels against S. vulgaris were measured in strongyle-naive and in immune ponies following initial and challenge infections using the IFA and an indirect hemagglutination assay (IHA). Antibody levels measured by IFA increased faster following initial infections than did levels measured by IHA. Antibody levels appear to increase following challenge infections of immune ponies when measured with the IFA, but not with the IHA. Significant differences in antibody titers were not seen between ponies which developed colic following challenge infections and those that did not develop colic. Antibodies were not detectable in ponies unexposed to larval migrations, but which received surgical implantation of S. vulgaris adults into the cecum.

Seasonal development and survival of equine cyathostome larvae on pasture in south Louisiana.

Cyathostome development and survival on pasture in subtropical climates of the US have yet to be completely defined and available data on seasonal transmission are minimal. In an attempt to study this phenomenon, a group of pony mares and their foals was maintained on a naturally contaminated pasture in southern Louisiana. Fecal egg counts (FEC) and numbers of infective third stage larvae (L3) kg(-1) dry herbage were recorded biweekly during two time periods, from January 1986 through December 1988, and September 1996 through October 1997. A FEC rise occurred during the late summer-early autumn which preceded the peak of L3 on pasture during the winter season. The numbers of cyathostome L3 were reduced during the hottest months of the year due mainly to daily minimum temperatures above 18 degrees C, and in winter during short freezing spells when daily minimum temperatures dropped below 0 degrees C. Tilling of the pasture reduced the number of cyathostome L3 during the early winter months but this is an efficacious measure only if horses are given an effective anthelmintic treatment prior to being returned to pasture. The data collected suggest that parasite reduction in southern Louisiana is possible using a treatment program with treatment beginning at the end of September and continuing through the end of March.

Foals raised on pasture with or without daily pyrantel tartrate feed additive: comparison of parasite burdens and host responses following experimental challenge with large and small strongyle larvae.

Three groups of foals were raised under different management programs in this study: Group 1 (n = 6) and Group 2 (n = 6) were raised with their dams on pasture; Group 3 foals (n = 5) were raised under parasite-free conditions. Mares and foals of Group 1 received daily pyrantel tartrate (PT) treatment with their pelleted feed ration, whereas mares and foals of Groups 2 and 3 received only the pelleted ration. Pasture-reared foals were weaned and moved to a heavily contaminated pasture for 5 weeks. Group 1 foals continued to receive daily PT treatment whereas Group 2 foals received only the pelleted feed ration. Following this period, all foals were moved into box stalls. Half of each group was challenged with 10(3) Strongylus vulgaris infective third-stage larvae (L3), 5 x 10(3) Strongylus edentatus L3 and 10(5) mixed cyathostome L3; the remaining half served as unchallenged controls. Necropsy examinations were performed 6-week post-challenge for evaluation of parasite burdens and lesions. Daily PT treatment of Group 1 reduced the patent cyathostome infections of both mares and foals and was effective in reducing pasture burdens of infective larvae. Daily treatment of Group 1 foals during weaning continued to suppress EPG levels; however, it did not prevent large strongyle infections during the weaning period. Group 1 foals were more sensitive to challenge than Group 2 foals, which did not exhibit any post-challenge disturbances. Group 1 foals were equally susceptible to challenge as parasite-free foals.

Experimental cyathostome challenge of ponies maintained with or without benefit of daily pyrantel tartrate feed additive: comparison of parasite burdens, immunity and colonic pathology.

Eighteen mixed-breed, naturally infected ponies ranging in age from 1 to 16 yr and four cyathostome-naïve ponies reared and maintained under parasite-free conditions ranging in age from 1 to 4 yr were used in this study. Naturally-infected ponies were treated with 1 dose of ivermectin (IVM) at 200 micrograms kg-1, followed by a 5-day regimen of oxibendazole (OBZ) at 20 mg kg-1 to remove existing cyathostome burdens; cyathostome-naïve control ponies were treated with IVM alone. The naturally infected ponies were matched on age and gender, then randomly assigned to one of three treatment groups of six animals per group; the four cyathostome-naïve ponies constituted a fourth group. Following OBZ treatment, Group 1 ponies were treated with pyrantel tartrate (PT) in their pelleted ration; the remaining ponies received only the pelleted ration. Beginning on experiment Day 3, a daily challenge infection of 10(4) mixed cyathostome larvae was administered orally to ponies of Group 1, Group 2 and the cyathostome-naïve controls. Group 3 ponies served as unchallenged controls to determine residual parasite burdens following IVM/OBZ treatment. Necropsy examinations were performed on three Group 3 ponies on Day 1; the remainder of the necropsy examinations began on Day 41. Cyathostome burdens were evaluated by recovery of larvae and adults from the luminal contents, by digestions of the intestinal mucosa, and by mural transillumination of full-thickness intestinal sections. Differences in postchallenge clinical responses were also compared. Necropsy examinations included comparisons of grossly visible inflammation of the large bowel, weights of biopsy specimens from each region, and histologic evaluations of these biopsies. Parasite recoveries at necropsy indicated a strong protective effect derived from daily PT treatment. Mean weights of intestinal biopsies corresponded with worm burdens, but histological evaluation did not reveal architectural or cellular changes to account for the increase in weight; therefore, edema was suspected. A strong age-related resistance to challenge infection was apparent in both the PT-treated and control groups by virtue of the lower mean worm burdens found in older ponies compared to younger ponies of the same treatment group; however, daily PT treatment of older ponies reduced the variability of their worm burdens to a uniformly low level. Comparisons of luminal and mucosal parasite burdens of age stratified nontreated controls further suggest that the age related resistance, which is acquired, targets increasing numbers of parasite stages as this resistance matures. Further, there is no evidence for an immune mediated acquisition of hypobiotic L3.