Predacious activity of the nematode-trapping fungus Duddingtonia flagrans against cyathostome larvae in faeces after passage through the gastrointestinal tract of horses

This study was undertaken to examine the potential of the nematode-trapping microfungus Duddingtonia flagrans to survive passage through the gastrointestinal tract of horses and subsequently to destroy free-living stages of cyathostomes in faecal cultures. Three different oral dose levels were tested, two horses being used for each level. Faeces were collected twice daily and the number of parasite eggs per gram of faeces were determined. The numbers of infective third stage larvae which developed in faecal cultures were determined after the cultures had been incubated for 2 weeks at 24°C. Results showed a positive relationship between dose level and reduction in the number of infective larvae. Fungi were recovered in faeces at times which corresponded to high larval reduction.     

Recovery of different stages of Dictyocaulus viviparus from cattle lungs by a combination of a perfusion and a Baermann technique

The lungs of 71 calves used in two grazing studies were examined for lungworms with a perfusion method combined with a modified Baermann method. The numbers of worms recovered from the lungs were 11,630 in total, 7495 adults, 872 juvenile fifth stage larvae (L5) and 3263 inhibited L5. The percentages found after perfusion were 78.9 per cent in total, 91.8 per cent adults, 78.6 per cent juvenile L5 and 49.6 per cent inhibited L5. The perfusion method seemed adequate and rapid for recovering the adult and juvenile stages but not the inhibited stage. For estimating the numbers of inhibited larvae a combination of perfusion with the Baermann technique is necessary.     

Recovery of Mycobacterium avium subsp. paratuberculosis from nematode larvae cultured from the faeces of sheep with Johne's disease

A study was conducted to determine whether trichostrongylid nematode larvae become contaminated with Mycobacterium avium subsp. paratuberculosis when they develop in the faeces of sheep with Johne's disease. Nematode larvae were hatched from ova in the faecal samples of affected sheep. Larval sheaths were removed and these as well as exsheathed larvae were subjected to radiometric culture for M. paratuberculosis. The organism was recovered from washing water used to prepare the larvae, third stage larvae and larval sheaths, but not from exsheathed larvae. The recovery of M. paratuberculosis from larvae was associated with the severity of the histological lesions in affected sheep and with the results of culture of the organism from intestinal tissues and faeces. Nematode parasites of sheep might be able to act as mechanical vectors for M. paratuberculosis as the organism associates with infective third stage larvae when these develop in the faeces of sheep with Johne's disease.     

Recovery of Sarcocystis gigantea sporocysts from cat faeces

A method for the mass recovery of S. gigantea sporocysts from cat faeces involving homogenisation and centrifugation in water, passage through 250- and 53-microns sieves and floatation in 1.2 SG NaCl solution, is described. An examination of the various processes involved in this procedure showed that the greatest yields were obtained when a proportion of faeces to floatation medium of 1:20 and centrifugation at 6000 X g for at least 5 min was used. Ninety-six per cent of the sporocysts recovered were obtained from the first centrifugation in aqueous NaCl solution (SG 1.2). Although neither sieving nor additional washing of homogenised samples prior to floatation significantly affected sporocyst recovery, both reduced the amount of debris present. A considerable reduction in the amount of debris resulted from feeding infected cats on tinned fish rather than tinned meat. The addition of CCl4 to the NaCl solution also improved sporocyst purity but with a marked reduction in the numbers recovered.     

Recovery of Yersinia pseudotuberculosis from the faeces of healthy cattle

Attempts were made to recover and serologically identify Yersinia pseudotuberculosis from the faeces of groups of nine to twenty clinically healthy cattle eight to thirteen months old on each of 50 farms in the northern part of New Zealand. Yersinia pseudotuberculosis was recovered from 134 (26.3%) of 509 faeces samples from cattle on 42 (84%) farms and from nine of ten samples on two of these farms. Serotypes I, II, and III were identified, of which serotype III was by far the most common and accounted for 125 (93.2%) of the 134 isolates. Because of the common occurrence and widespread distribution of Y. pseudotuberculosis it is suggested that the clinical significance of isolation of this micro-organism from faeces samples or intestinal contents should be treated with caution.     

Observations on the population dynamics of five cyathostome nematode species of horses in northern USA

Monthly variations in the magnitude of adult and larval cyathostome burdens were observed in 55 horses necropsied over a 15-month period in the northern USA. Peak numbers of adult cyathostomes occurred in late winter (March) and late summer (September). Larval cyathostomes demonstrated peak numbers from February to April and again in October, beginning one month earlier than the spring adult peak and one month after the autumn adult peak, respectively. The reproductive status of individual female Cyathostomum catinatum, Cyath coronatum, Cylicocyclus nassatus, Cylicostephanus goldi and Cylicostephanus longibursatus was classified as immature, gravid or spent. Seasonal changes in these classifications were monitored as a marker for the age structure of these populations. Each reproductive category of female small strongyle was dominant during only one period per year and these periods were similarly distributed for all five species examined. Immature cyathostomes were most common from late winter to spring (March to May); gravid worms were predominant beginning in spring (April/May) and continuing into autumn (October to December). Spent females prevailed from autumn through winter (October to March/April).     

Comparison of two techniques for quantitation of encysted cyathostome larvae in the horse

Haustral portions of intestine of 6 horses were isolated by excising the taeniae coli from the cecum and the ventral colon. Uniform 5-cm X 5-cm sections were cut from the haustra and were illuminated from the serosal side with a strong light source (mural transillumination). Cyathostome larvae encysted in the mucosa and submucosa were observed at 15 X magnification and counted. Two separate counts of the larvae in 80 replicates of tissue by the mural transillumination technique (MTT) revealed no significant (P less than 0.05) difference between sample means. Larvae in tissue sections were counted in situ by MTT, and the mucosal scrapings of the tissue sections were digested in pepsin and HCl to determine larval yields for comparison with the MTT counts. Numbers of larvae recovered by pepsin and HCl digestion for 3 and 6 hours were significantly (P less than 0.01) lower than were numbers originally determined by MTT. Larvae recovered by tissue digestion for 3 or 6 hours were examined individually and given objective scores for morphologic damage. Distribution of scores was time-dependent; increased damage to larvae was associated with a longer time of digestion. Individual 4th-stage cyathostome larvae were dissected from cysts in the large intestinal wall and were incubated in water, 0.9% saline solution, 1.1% HCl, or pepsin (7,000 U of activity/ml). Significantly fewer (P less than 0.05) larvae were recovered from all solutions after 3 and 6 hours. The proportion of dissected larvae that were given high scores after exposure to pepsin was significantly (P less than 0.01) greater than were those held in HCl, saline solution, or water for both periods.     

Dispersal of Dictyocaulus viviparus larvae from bovine faeces in Ireland

An involvement of Pilobolus species fungus in the dispersal of Dictyocaulus viviparus third stage larvae from dung to surrounding herbage under Irish conditions was investigated. The presence of Pilobolus kleinii on artificial dung pats containing first stage larvae of D viviparus was associated with a 19-fold increase (P less than 0.05) in numbers of third stage larvae recovered from the surrounding herbage. A subjective examination of natural dung pats showed that the presence of Pilobolus species was significantly correlated with hours of bright sunshine (r = -0.5, P less than 0.01), total rainfall (r = 0.41, P less than 0.05) and the height of herbage surrounding the pats (r = 0.31, P less than 0.001). A multiple regression analysis showed that meteorological parameters and the height of surrounding herbage accounted for 38 per cent of the variation in growth of Pilobolus species on dung pats. The incidence of extensive damage to natural dung pats within five days of deposition, caused by biotic factors, another possible cause of D viviparus third stage larvae dispersal, varied from 0 to 92 per cent of the pats depending on their degree of dryness.     

Spread of equine lungworm (Dictyocaulus arnfieldi) larvae from faeces by Pilobolus fungi

Between 10 and 25% of the Dictyocaulus arnfieldi larvae excreted in faeces from a naturally infected donkey were harvested as infective stages from faecal cultures by means of Pilobolus fungi. The faeces were collected between 24 and 56 hours after drenching the donor animal with Pilobolus spores and kept at 16 +/- 2 degrees C. Most larvae were collected between the 5th and the 8th day of culturing during which period fructification and sporangium discharge also peaked. The sporangia and the adhering larvae were collected in Petri dishes inserted between the faecal mass and a light source. All recovered larvae were viable. A mean larval length of 368 microns (range 312-440 microns) and width of 14.6 microns (range 12-20 microns) was recorded for the infective stage. The method was found suitable for the recovery of infective stages for experimental purposes. The authors suggest that the Pilobolus mechanism play an important part in the spread of equine lungworm infection under field conditions similar to the situation in bovine lungworm (Dictyocaulus viviparus) infection.     

A modified procedure for the extraction of infective nematode larvae from bovine faeces

A convenient and space-saving technique for extracting nematode larvae from bovine faeces is described. The numbers of larvae recovered using this technique are similar to those obtained by conventional methods.     

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.     

Emergence from inhibited development of cyathostome larvae in ponies following failure to remove them by repeated treatments with benzimidazole compounds

The effect of three albendazole treatments at 5-week intervals, beginning at turnout in April, on cyathostome infections in Shetland ponies was compared with the effect of sequential treatments with albendazole, oxfendazole and oxibendazole. The results showed a substantial reduction in faecal egg output after the first albendazole treatment. Since faecal egg counts remained very low, no estimation of the effect of the second treatment was possible. The third treatment with albendazole and oxibendazole was followed by an increase in faecal egg counts to values of greater than 100 eggs g-1 within 4 weeks. A final albendazole treatment in December, 1 week before necropsy, failed to reduce faecal egg counts. These results suggest resistance to albendazole and oxibendazole in the cyathostome populations of the ponies. The increase in faecal egg counts after the third anthelmintic treatment in July occurred, although overwintered pasture infectivity was very low. The most likely explanation for this increase is resumption of the development of worms which overwintered as inhibited larvae in the host.     

Comparative efficiency of moxidectin gel or ivermectin paste for cyathostome control in young horses

Thirty-six young horses were allocated to three similar groups. Horses in Group 1 were treated with moxidectin gel on Days 0, 90, and 180, Group 2 horses received ivermectin paste on Days 0, 60, 120, and 180, and horses in Group 3 were untreated controls. All horses were maintained on a common pasture for the first 180 days. Immediately after the final scheduled deworming, each group was moved to a separate, clean pasture where it remained until Day 360. At monthly intervals, fecal egg counts, body weights, body condition scores, and pasture larval counts were measured. The cumulative costs of both deworming regimens were calculated. Young horses treated three times at 90-day intervals with moxidectin gel had significantly lower monthly fecal egg counts than untreated controls from Days 30 through 300. Horses given ivermectin paste four times at 60-day intervals had significantly lower egg counts than controls 30 days after each treatment and 60 days after the third dose. Average daily gains of treated horses were significantly greater than controls from Days 120 through 360 (moxidectin) and from Days 210 through 360 (ivermectin). Quarterly moxidectin treatments reduced egg counts more effectively and cost less than ivermectin given bimonthly.     

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.