Label-free resistive-pulse cytometry.

Numerous methods have recently been developed to characterize cells for size, shape, and specific cell-surface markers. Most of these methods rely upon exogenous labeling of the cells and are better suited for large cell populations (>10,000). Here, we review a label-free method of characterizing and screening cells based on the Coulter-counter technique of particle sizing: an individual cell transiting a microchannel (or "pore") causes a downward pulse in the measured DC current across that "pore". Pulse magnitude corresponds to the cell size, pulse width to the transit time needed for the cell to pass through the pore, and pulse shape to how the cell traverses across the pore (i.e., rolling or tumbling). When the pore is functionalized with an antibody that is specific to a surface-epitope of interest, label-free screening of a specific marker is possible, as transient binding between the two results in longer time duration than when the pore is unfunctionalized or functionalized with a nonspecific antibody. While this method cannot currently compete with traditional technology in terms of throughput, there are a number of applications for which this technology is better suited than current commercial cytometry systems. Applications include the rapid and nondestructive analysis of small cell populations (<100), which is not possible with current technology, and a platform for providing true point-of-care clinical diagnostics, due to the simplicity of the device, low manufacturing costs, and ease of use.

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.

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.

One season of pasture exposure fails to induce a protective resistance to cyathostomes but increases numbers of hypobiotic third-stage larvae.

The development of acquired resistance to cyathostome challenge after 1 season's exposure to a cyathostome-contaminated pasture was investigated using 17 parasite-naive ponies, which were 2-3 yr of age. These were divided into 3 groups: 1 to graze a cyathostome-contaminated pasture for 4 mo (exposed ponies), 1 to graze a "clean" pasture not previously grazed by parasitized animals (nonexposed ponies), and 1 group to remain in the barn under helminth-free conditions (parasite-free ponies). After pasture exposure all ponies were housed in stalls in the barn dewormed with ivermectin (200 micrograms/kg) and oxibendazole (100 mg/kg), a treatment that eliminated most cyathostomes encysted in the mucosa as well as all luminal parasites, on the basis of necropsies of 5 animals, after 17 days. Remaining ponies were challenged with 100,000 cyathostome-infective third-stage larvae (L3) per os 3 wk after anthelmintic treatment. Necropsies were performed 7 wk after the challenge. Total cyathostome burdens (luminal plus encysted stages) were not significantly different among any of the groups. However, a significantly higher percentage of hypobiotic early L3 (EL3) and a lower percentage of adults were found in exposed ponies. This observation supports the hypothesis that resistance acquired through exposure promotes cyathostome hypobiosis. This increase in EL3 in exposed ponies was associated with a significant increase in weight of cecum and ventral colon biopsies.

Re-evaluation of ivermectin efficacy against equine gastrointestinal parasites.

Two trials were conducted to confirm the efficacy of ivermectin paste against endoparasites of horses. In these trials, 20 ponies were treated with ivermectin oral paste at 200 mcg x kg body weight once on Day 0, and 20 ponies served as unmedicated controls. The animals carried naturally acquired parasite infections as confirmed by pretrial fecal examination. The animals were necropsied for worm recovery on Days 14, 15 or 16. Parasites recovered were identified to species. Horses treated with ivermectin had significantly (P<0.05) fewer (>99.0% reduction) adult small strongyles (Coronocyclus spp including C. coronatus, C. labiatus, C. labratus; Cyathostomum spp including C. catinatum, C. pateratum; Cylicocyclus spp including C. ashworthi, C. elongatus, C. insigne, C. leptostomum, C. nassatus, C. radiatus; Cylicodontophorus bicoronatus; Cylicostephanus spp including C. asymetricus, C. bidentatus, C. calicatus, C. goldi, C. longibursatus, C. minutus; Gyalocephalus capitatus; Parapoteriostomum spp including P. euproctus, P. mettami; Petrovinema poculatum; Poteriostomum spp including P. imparidentatum, P. ratzii) and adult large strongyles (Strongylus edentatus, S. vulgaris; Triodontophorus spp including T. brevicauda, T. serratus; Craterostomum acuticaudatum) than the controls. Ivermectin was also highly effective (94% to >99%, P<0.05-0.01) against Gasterophilus intestinalis larvae, Habronema spp., Oxyuris equi, Parascaris equorum. The data from these two trials confirm that ivermectin paste administered to horses orally at 200mcg x kg(-1) continues to be highly effective for treatment and control of a broad range of small and large strongyle species as well as other species of gastrointestinal parasites.

Seasonal transmission of gastrointestinal parasites of equids in southern Louisiana.

Parasite-naive pony foals were used as sentinels to monitor transmission of gastrointestinal parasites of equids in Louisiana during 4 seasons of the year. Two annual periods were studied, 1988-1989 and 1992-1993. Two or 3 foals each season were turned out to graze a contaminated pasture along with resident parasitized mares and their foals. After a grazing period of 8 wk, sentinel ponies were held in a parasite-free box stall for a period of 6 wk to allow parasites to develop, thus enhancing the evaluation of hypobiotic stages. Following this holding period, necropsies were performed for complete parasite recoveries. Data show that transmission of large and small strongyles occurs during all seasons in southern Louisiana, with highest levels of transmission occurring in the winter and only minimal transmission taking place in the summer. Numbers of mucosal cyathostomes, as well as total cyathostome numbers, were highest in the winter, and luminal cyathostome numbers were highest in the spring. Transmission of Anoplocephala perfoliata and Parascaris equorum occurred during all seasons of the year, although numbers of P. equorum were reduced in spring 1989 and 1993. Gasterophilus intestinalis instars were recovered from fall and winter sentinels only. Oxyuris equi L4 were found all seasons 1 yr, but only during the fall and winter of the final year.

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).

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.

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.

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.

Transglutaminase activity in equine strongyles and its potential role in growth and development.

Transglutaminases (E.C. 2.3.3.13) are a family of Ca(2+)-dependent enzymes that stabilize protein structure by catalyzing the formation of isopeptide bonds. A novel form of transglutaminase has been identified and characterized that seem to play an important role in growth, development, and molting in adult and larval stages of filarial nematodes. The aim of this study was to identify the ubiquitous nature of this enzyme in other nematodes and to measure its significance to larval growth, molting, and development. For this purpose, equine Strongylus spp. were used. Activity of this enzyme was identified in extracts of larvae and adults of Strongylus vulgaris, S. edentatus, Parascaris equorum and Cylicocyclus insigne. The significance of transglutaminase in the early growth and development of Strongylus vulgaris, S. edentatus and S. equinus was tested by adding specific inhibitors, monodansylcadaverine (MDC) or cystamine (CS), to in vitro cultures of third (L3) and fourth stage larvae (L4). The viability, molting and growth of these nematode species were affected by both inhibitors. Cystamine promoted abnormal development of Strongylus edentatus L3, resulting in an aberrant expansion of the anterior end. Addition of these inhibitors to cultures of L4 also reduced growth of the three species. The results indicated that transglutaminase is present in a wide array of nematode parasites and may be important in growth and development of their larval stages.

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.

Efficacy of moxidectin 2% oral gel against second- and third-instar Gasterophilus intestinalis De Geer.

In a trial designed to evaluate the efficacy of the recommended dosage of moxidectin 2% oral gel against the gastric stages of Gasterophilus spp., 14 ponies were selected from a herd on the basis of the inclusion criterion of the presence of Gasterophilus spp. eggs attached to their hair coats. After random allocation, the ponies were treated with 1 of 2 treatments, moxidectin 2% equine gel in a single dose at the commercial dosage of 400 microg moxidectin/kg body weight or placebo gel. The animals were necropsied 14 days posttreatment. Efficacies against second- and third-instar Gasterophilus intestinalis De Geer were 100% and 99.5%, respectively.

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.

EpsR modulates production of extracellular polysaccharides in the bacterial wilt pathogen Ralstonia (Pseudomonas) solanacearum.

Ralstonia solanacearum is the causal agent of bacterial wilt of many agriculturally important crops. Exopolysaccharide synthesized by products of the epsI operon is the major virulence factor for R. solanacearum. Expression of epsI has been demonstrated to be under the control of several proteins, including several two-component regulators. Overexpression of EpsR was found previously to reduce the amount of synthesis specifically from the epsI promoter. Here we present data that a single chromosomal copy of epsR activates the epsI promoter, suggesting that EpsR is a concentration-dependent effector of epsI gene expression. Furthermore, the ability of EpsR to modulate epsI expression is dependent on the phosphorylation state of EpsR. Gel mobility shift assays suggest that EpsR can specifically bind the epsI promoter and that this binding requires a phosphorylated form of EpsR.

Sequences 5' of the conserved tRNA-like promoter modulate the initiation of minus-strand synthesis by the brome mosaic virus RNA-dependent RNA polymerase.

Each of the brome mosaic virus (BMV) genomic RNAs contains a conserved tRNA-like structure that is sufficient to direct minus-strand RNA synthesis in vitro. The tRNA-like promoters, tB1 and tB3, direct approximately equal amounts of synthesis in vitro. However, 5' sequences were found to affect the amount of minus-strand synthesis, suggesting that sequences beyond the tRNA-like structure are important in moderating minus-strand synthesis. Consistent with this, sequences upstream the tRNA-like structure are able to partially suppress mutations at or near the initiation site. This activity is observed in the 5' sequences of both BMV and CMV (cucumber mosaic virus) templates. However, a chimeric RNA containing the CMV tRNA-like promoter fused to the 5' sequences of BMV was not able to suppress mutations at the initiation site, suggesting that homologous 5' and 3' sequences are required to affect initiation. The ability to suppress mutations at the initiation site was correlated with a slight increase in the ability of the BMV RNA-dependent RNA polymerase to interact with the RNA.

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.

Identification and characterization of a pyrantel pamoate resistant cyathostome population.

Three fecal egg count reduction assays (FECR) and one critical trial were performed to determine the efficacy of pyrantel pamoate (PP) at 6.6 mg base kg-1 on a well managed stud farm in Louisiana where a loss of efficacy was suspected. Efficacy of PP based on FECR varied from 25% in mares to 83% in yearlings. Second treatments with PP 2 weeks following an initial treatment failed to reduce eggs per gram (EPG). A critical trial was performed to determine the cyathostome species resistant to PP. Three strongyle-naive ponies which acquired infections on the farm were used for this purpose. Following treatment with PP at the recommended dose, 11 species of cyathostomes remained in the intestine of the tracer ponies. Reduced efficacies (62%-88%) were noted for seven species. Resistance to oxibendazole (OBZ), which was > 90% effective on this farm in 1982, was also evaluated by FECR and found to exist. The results of one experiment indicate that dual resistance of parasites to PP and OBZ also exists.

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.