Helicobacter equorum is highly prevalent in foals

Faecal samples of sixty-six 3-day- to 6-month-old foals were screened for Helicobacter equorum DNA by means of a PCR amplifying a 1074 bp fragment of the 23S rRNA gene with primers specific for this enterohepatic Helicobacter species. H. equorum DNA was demonstrated in faeces from 28.6% of the less than 1-month-old foals, while 67.8% of foals from 1 to 6 months of age tested positive. In a previous study, H. equorum was demonstrated in faeces of 0.8-7.9% of adult horses. These results indicate that the prevalence of H. equorum in horses differs with the age of the investigated horse population. The organism seems highly prevalent in foals between 1 and 6 months of age but the possible association with intestinal disease in this age group needs further investigation.     

The development of immunity to Parascaris equorum infection in the foal

Following infection with 8000 Parascaris equorum eggs in two- to four-week-old foals reared under worm-free conditions a high percentage of the infective dose completed its tissue migration and returned to the small intestine. Patent infections were establisehd between 81 and 104 days after infection and high faecal egg counts were recorded. A group of six- to 12-month-old foals, which had been either reared under worm-free conditions or exposed to natural ascarid and strongyle infections on pasture, received a similar infection of 8000 P equorum eggs. Compared with the younger foals there was a marked reduction in the number of larvae reaching the small intestine. Also, patent infections were established less frequently (50 per cent of cases) and, when present, the faecal egg counts remained low.     

Prevalence of Helicobacter equorum in faecal samples from horses and humans

Recently, a new enterohepatic Helicobacter species, H. equorum, was isolated from faecal samples of two clinically healthy horses. At the onset of this study, nothing was known about the prevalence of this organism in horses, nor was there any information available on the possible zoonotic character of this agent. This study aimed to determine the prevalence of H. equorum in faecal samples from equine and human origin. Therefore, faecal samples of 120 healthy privately owned horses, 227 healthy riding-school horses and 239 hospitalised horses were screened for H. equorum-DNA by means of a PCR amplifying a 1074-bp fragment of the 23S rRNA gene with primers specific for H. equorum. The vast majority of the hospitalised horses were under treatment with an antimicrobial agent at the moment of sampling, while the other horses had not been treated with an antimicrobial agent in the 14 days preceding the sampling. Stool samples of 531 humans suffering from gastro-intestinal disease and 100 clinically healthy humans were likewise examined. H. equorum-DNA was demonstrated in faeces from 0.8% of the privately owned horses, 3.1% of the riding-school horses and 7.9% of the hospitalised horses. The prevalence of H. equorum was significantly higher in hospitalised than in healthy, privately owned horses (P=0.02). H. equorum-DNA was not detected in human samples. These results indicate that the prevalence of H. equorum in horses may be influenced by the health status of the investigated horse population and/or by antimicrobial treatment. We may additionally assume that this micro-organism does not commonly infect humans.     

Anthelmintic efficacy on Parascaris equorum in foals on Swedish studs

Background

In the last few years stud farms have experienced increasing problems with Parascaris equorum infections in foals despite intensive deworming programs. This has led to the question as to whether the anthelmintic drugs used against this parasite are failing. This study aimed to investigate the efficacy of ivermectin, fenbendazole and pyrantel on the faecal output of ascarid eggs of foals.

Methods

A Faecal Egg Count Reduction Test (FECRT) was performed on nine large studs in Sweden. Anthelmintic drugs were given orally and faecal samples were examined for ascarid eggs on the day of deworming and 14 days later. Faecal Egg Count Reductions (FECRs) were calculated on arithmetic means of transformed individual FECRs and on arithmetic means of individual FECRs.

Results

Seventy-nine (48%) out of a total of 165 foals sampled were positive for P. equorum eggs before deworming and 66 of these met the criteria for being used in the efficacy assessment. It was shown that there was no, or very low activity of ivermectin on the output of ascarid eggs in the majority of the foals, whereas for fenbendazole and pyrantel it was >90%.

Conclusion

Ivermectin resistance was shown in 5 out of 6 farms. Therefore, ivermectin should not be the drug of choice in the control of P. equorum infections in foals. According to the results of this study, fenbendazole or pyrantel are still effective and should be used against this parasite.     

Determination of anthelmintic efficacy against equine cyathostomins and Parascaris equorum in France

This article reports the results of a faecal egg count reduction test on 4 farms in France, as an integrated part of the routine deworming strategy against horse cyathostomins and Parascaris equorum. Treatment with fenbendazole (FBZ) or ivermectin (IVM) was evaluated in yearlings on Farms 1 and 2 and treatment with pyrantel embonate (PYR) was tested on Farms 3 and 4. Calculation of the arithmetic mean faecal egg count reduction and the 95% confidence intervals (95% CI) around the mean was performed using bootstrap analysis. For equine cyathostomins, resistance to FBZ was found with an arithmetic mean reduction of 48.8% (95% CI: 1.9–69.3%). On Farms 1 and 2, horses with reduced efficacy were identified. PYR was found to be effective against cyathostomins, with an arithmetic mean reduction of 95.3% (95% CI: 84.6–99.8%), as well as IVM (100%). For P. equorum, both FBZ and PYR were effective (100% reduction). The efficacy of IVM, however, was low (45.5%; 95% CI: 0–96.3%). These results confirm that FBZ resistance in equine cyathostomins is present in France and that anthelmintic resistance to IVM is present in P. equorum. This study underlines the necessity to evaluate the efficacy of horse deworming strategies on a regular basis under field conditions.     

Acute in vivo interactions of Helicobacter equorum with its equine host

REASONS FOR PERFORMING STUDY: A novel urease-negative Helicobacter species has been isolated from faecal samples of clinically healthy horses, but no information is available about the main sites of colonisation in the equine gastrointestinal tract nor is the pathogenic potential of this microorganism known. An experimental infection in horses was therefore carried out. METHODS: Four horses were infected with H. equorum strain CCUG 52199T and subjected to euthanasia at 10 (n = 2) and 30 days (n = 2) post inoculation. A fifth animal was inoculated with phosphate buffered saline and used as control. Gastrointestinal samples were examined histologically and bacteriologically. These samples, as well as faecal material collected at regular intervals, were also subjected to PCR analysis. RESULTS: All horses remained clinically healthy and no specific macroscopic lesions were identified, nor were there any microscopic changes. H. equorum-DNA was detected in the faeces during the whole experiment in all infected animals but not in the negative control. Sites of colonisation were caecum, colon and rectum. CONCLUSIONS: H. equorum is able to colonise the equine lower bowel and is excreted in faeces without apparent pathology. No association between the presence of the organism and gastrointestinal disease was demonstrated.     

Evaluation of ivermectin paste in the treatment of ponies for Parascaris equorum infections

Twenty ponies less than 18 months of age and infected with Parascaris equorum were treated with either 0.2 mg of ivermectin/kg of body weight (n = 10) or a placebo (n = 10; controls). Five control and 5 ivermectin-treated ponies were euthanatized 14 and 35 days after treatment, respectively. At necropsy, the small intestinal contents, lungs, and liver were examined for larvae and/or adult P equorum. Significantly (P less than 0.02) higher mean total numbers of P equorum were found in the small intestinal contents of the controls on day 14 (51) and on day 35 (21) than in the ivermectin-treated ponies on days 14 (0) and 35 (3). The efficacy of ivermectin in removing adult and intestinal larvae of P equorum at 14 days after treatment was 100%. The efficacies of ivermectin in removing adults and intestinal larvae of P equorum at 35 days after treatment were 100% and 76.9%, respectively. Gross examination of liver and lung tissues revealed damage as a result of P equorum infections in all ponies. The Baermann technique used on liver and lung tissues did not yield any P equorum larvae. Adverse reactions attributable to treatment were not observed.     

Efficacy of ivermectin in the treatment of induced Parascaris equorum infection in pony foals

Eighteen pony foals inoculated with 1,500 +/- 109 infective Parascaris equorum eggs were given 0.02 ml of ivermectin vehicle (liquid)/kg of body weight, PO, (control); 0.2 mg of ivermectin paste/kg, PO; or 0.2 mg ivermectin liquid/kg, PO, on postinoculation day (PID) 28. Foals were euthanatized on PID 42, and the small intestinal contents were examined for P equorum larvae. The mean number of fourth-stage P equorum larvae in foals treated with ivermectin paste and liquid were 3.5 and 6, respectively. Significantly (P less than 0.01) higher mean numbers of larvae (1,250) were detected in foals treated with ivermectin vehicle. Larvae recovered from foals treated with ivermectin vehicle were of significantly (P less than 0.002) longer mean length than those from foals treated with ivermectin paste or liquid. Gross examination of lungs and liver revealed similar pathologic changes from the migration of P equorum in all foals. Adverse reaction to treatment was not observed.     

Controlled trials of fenbendazole and febantel in ponies with parascaris equorum infections

The activity of fenbendazole and febantel was evaluated in 12 pony foals whichwere inoculated with 2600 P. equorum eggs. The foals were not maintained free of parasites before or after inoculation. Once patent for P. equorum, the foals were randomly assigned to groups and treated one time intraorally with either 0.5 ml corn syrup/kg of body weight (controls; n=4), 10 mg fenbendazole/kg (n=4), or 6 mg febantel/kg (n=4). Foals were necropsied and examined for parasites 10 days after treatment. Fenbendazole and febantel were highly effective against adult and immature P. equorum. Grosslesions attributed to P. equorum were evident in all foals. P. equorum were not foundin any of the fenbendazole-or febantel-treated foals. The mean number of adult and immature P. equorum found in the controls was 66.8 (15–166) and 65.0 (21–147), respectively. Strongyle infections were insufficient for efficacy evaluations to be done. Neither anthelmintic was effective against mature Drasehia megastoma, mature Habronema majus, immature H. muscae, Gasterophilus intestinalis or late 4th-stage larvae of Strongylus vulgaris. Adverse side effects due to treatment were not observed.     

Effects of low and high temperatures on viability of Parascaris equorum eggs suspended in water

Microcentrifuge tubes containing 5000 eggs of Parascaris equorum suspended in water were frozen at -5, -10, -15, -20, and -80 degrees C for 1-168 h and then thawed at a room temperature. Other samples of P. equorum eggs suspended in water were inserted into wells in the heated metal block of a thermal DNA cycler. Block temperatures were set at 5 degrees C incremental temperatures from 40 to 100 degrees C. At each temperature setting microcentrifuge tubes containing P. equorum eggs were removed 1 and 5 min later. Both, frozen and heated egg suspensions as well as untreated control suspensions were then incubated to test of viability based on the development of infective larvae inside viable eggs. We found out that eggs of P. equorum in water can retain viability and infectivity after freezing and that eggs survive longer at higher freezing temperatures. Our results also indicated that when water containing P. equorum eggs reached temperatures of 60 degrees C or higher within 1 min, the viability of eggs was lost.     

Identification of foals infected with Parascaris equorum apparently resistant to ivermectin

During September 2002, routine fecal examinations performed on 16 Thoroughbred foals residing on a farm outside Toronto, Ontario, Canada, revealed low to moderate numbers of Parascaris equorum eggs in feces from 9 of the 16. All foals were then treated with ivermectin at a dose of 220 to 280 microg/kg (100 to 127 microg/lb), p.o., and fecal egg counts were repeated 12 days later. Fecal P. equorum egg counts increased between the first and second fecal examination in 7 foals, were unchanged in 1, and decreased in 5. Fecal samples were collected 13 days after treatment from 21 additional foals that had been treated with ivermectin at the same dose, and P. equorum eggs were detected in 12 of the 21. For all 37 foals, high P. equorum egg counts (> or = 100 eggs/g of feces) 12 to 13 days after ivermectin treatment were significantly more likely in foals that had been regularly treated with ivermectin since birth and permanently resided on the farm, compared with foals that had been treated with other anthelmintics or had an unknown deworming history. Collectively, these data suggested that P. equorum in these foals was resistant to ivermectin administered at the recommended dose.     

The distribution ofParascaris equorum eggs in the soil profile of bare paddocks in some Norwegian studs

Fifteen bare paddocks, which consisted of soil only and were located on 12 different studs, were examined for their content ofParascaris equorum eggs in the upper 15 cm of the soil profile. The paddocks were classified into three different groups according to the type of soil: clayey soil (A), morainic soil (B) and gravel or gravel-like sand (C). Soil profiles were collected down to a depth of 15 cm and were divided into three layers: 0–5 cm (D1), 5–10 cm (D2) and 10–15 cm (D3). The eggs in each layer were counted and identified as infective or noninfective eggs. The paddocks in group C, which had good drainage conditions, had significantly lower numbers of eggs in the whole profile and in D1 and D2 than the paddocks in groups A or B. Furthermore, there was a significantly higher proportion of the total egg count present in D3 in the group C paddocks. This may have been due to a higher degree of passive transportation of eggs down the profile in the gravel or gravel-like sand. Even though there was a significantly higher proportion of infective eggs in the soil from the group C paddocks, the lower total numbers of eggs resulted in a lower total number of infective eggs in those paddocks. The study showed that the soil type was an important factor in determining the content ofP. equorum eggs in the upper layer of the soil profile in bare paddocks and consequently for the potential infestation of horses withP. equorum.     

The effects of windrow composting on the viability of Parascaris equorum eggs

Parascaris equorum generally infects horses less than 18 months old and its pathological effects can be severe. Infection occurs when larvated eggs, present in pastures, paddocks, stalls, and on feeding and watering equipment are ingested. The purpose of this study was to examine the effects of windrow composting on the viability of P. equorum eggs at a cooperating central Kentucky horse farm. Three grams of feces containing 2216 P. equorum eggs per gram were sealed in filter bag sentinel chambers. Chambers were exposed to 1 of 3 treatments: constant exposure or intermittent exposure to the interior of the windrow; controls were stored at 4 °C. At day 0, all chambers in the experimental treatments were placed in the center of 10 locations of the windrow. On subsequent days when the windrow was turned, chambers in the constant exposure treatment were returned to the interior of the windrow and chambers in the intermittent exposure treatment were alternated between resting on top of, or inside, the windrow. Chambers from each treatment and control chambers were removed at days 2, 4, 6, 8, 10, 12, 14, and 18; and incubated for 21 days at room temperature (24 °C). After incubation, eggs were recovered from the chambers using double centrifugation flotation. Eggs were evaluated microscopically, staged according to development and classified as viable or nonviable based on whether embryonation to the larval stage had occurred. Results were reported as the mean percent viable eggs for each treatment and time point. A mixed linear model with repeated measures was used to evaluate the influence of experimental day and treatment on the percent viability of P. equorum eggs. Chambers treated with constant exposure contained 10.73% (SD = 0.29) viable eggs on day 2 and declined to an average of 0.00% by day 8. Chambers exposed to the intermittent treatment contained 16.08% (SD = 0.26) viable eggs on day 2 and decreased to 0.00% by day 6. Control chambers for days 2, 4, 6, 8, 10, 12, 14, and 18 all had viabilities above 79.00%. A significant fixed effect of experimental day (p < 0.0001) and compost treatment (p < 0.0001) was observed. There was no significant interaction between experimental day and compost treatment (p > 0.7459). The results of this study demonstrate that windrow composting was effective at rendering P. equorum eggs nonviable when it was tested under the conditions at a working horse farm.     

Evaluation of ivermectin for larvicidal effect in experimentally induced Parascaris equorum infections in pony foals

A controlled test was carried out on 15 pony foals inoculated with 1,500 +/- 108.8 infective Parascaris equorum eggs. The foals were assigned to 3 treatment groups. Treatments given on postinoculation day 11 included 0.2 mg of ivermectin/kg of body weight, formulated as paste (n = 5), or liquid (n = 5), or no treatment (controls; n = 5). The foals were euthanatized on postinoculation day 25, and examined for larvae in the small intestine, lungs, and liver. Larvae were not found in foals treated with ivermectin liquid or paste, whereas significantly (P less than 0.05) higher mean numbers (960.9; range, 379 to 1,736) of 4th-stage larvae were found in the controls. Histologic and gross examination of lungs and liver revealed pathologic changes attributable to P equorum migration that were similar in all foals. Adverse reactions to treatment were not observed.