Effect of silver nanoparticles (AgNP's) from Duddingtonia flagrans on cyathostomins larvae (subfamily: cyathostominae).

The in vitro effect of silver nanoparticles of the Duddingtonia flagrans filtrate enriched with chitin was evaluated on infective larvae of cyathostomins (L3). After biosynthesis, an assay was carried out with two experimental groups in microtubes, for a period of 24 h: G1 (AgNP's-D. flagrans (43.4 µg/mL) + 120 L3) and G2 (distilled water + 120 L3). At the end of this period, AgNP's-D. flagrans (G1) demonstrated an effect on L3 with a 43% reduction (p < 0.01) in relation to G2. Thus, the authors suggest new designs with AgNP's-D. flagrans for the control of cyathostomins.

Entomopathogenic nematodes increase predation success by inducing cadaver volatiles that attract healthy herbivores.

Herbivore natural enemies protect plants by regulating herbivore populations. Whether they can alter the behavior of their prey to increase predation success is unknown. We investigate if and how infection by the entomopathogenic nematode Heterorhabditis bacteriophora changes the behavior of healthy larvae of the western corn rootworm (Diabrotica virgifera), a major pest of maize. We found that nematode-infected rootworm cadavers are attractive to rootworm larvae, and that this behavior increases nematode reproductive success. Nematode-infected rootworms release distinct volatile bouquets, including the unusual volatile butylated hydroxytoluene (BHT). BHT alone attracts rootworms, and increases nematode reproductive success. A screen of different nematode and herbivore species shows that attraction of healthy hosts to nematode-infected cadavers is widespread and likely involves species-specific volatile cues. This study reveals a new facet of the biology of herbivore natural enemies that boosts their predation success by increasing the probability of host encounters.

A model for the dynamics of the parasitic stages of equine cyathostomins.

A model was developed to reproduce the dynamics of the parasitic stages of equine cyathostomins. Based on a detailed review of published literature, a deterministic simulation model was constructed using the escalator boxcar-train approach, which allows for fully-overlapping cohorts of worms and approximately normally distributed variations in age/size classes. Key biological features include a declining establishment of ingested infective stage larvae as horses age. Development rates are constant for all the parasitic stages except the encysted early third stage larvae, for which development rates are variable to reflect the sometimes extended arrestment of this stage. For these, development is slowed in the presence of adult worms in the intestinal lumen, and when ingestion of infective larvae on herbage is high or extended. In the absence of anthelmintic treatments, the life span of adult worms is approximately 12 months, and the presence of an established adult worm burden largely blocks the transition of luminal fourth stage larvae to the adult stage, resulting in mortality of the larvae. This inhibition is removed by effective anthelmintic treatment allowing the rapid replacement of adult worms from the pool of mucosal stages. Within the model, the rate and seasonality at which infective stage larvae are ingested strongly influences the dynamics of the pre-adult stages. While the adult worm burden remains relatively stable within a year, due to the negative feedback they have on developing stages, the numbers and proportions of larval stages relative to the total worm burden increase with the numbers of infective larvae ingested. Further, within the model, the seasonal rise and fall of encysted stages is largely driven by the seasonal pattern of infective larvae on pasture. Because of this, the model reproduces the contrasting seasonal patterns of mucosal larvae, typical of temperate and tropical environments, using only the appropriate seasonality of larvae on pasture. Thus, the model reproduces output typical of different climatic regions and suggests that observed patterns of arrested development may simply reflect the numbers and seasonality of free-living stages on pasture as determined by different management practices and weather patterns.

Identification of third stage larval types of cyathostomins of equids: An improved perspective.

Cyathostomins comprise around 50 parasite species of equids, offering a great challenge regarding their individual identification. The objective of our work was to improve identification of infective third stage larvae (L3) with a morphological key supplemented with detailed scientific illustrations based on our research and available literature. The highlighted features were; the number, arrangement, and shape of intestinal cells (IC), general features and the total body length of the eight different Cyathostomin sensu latum types (Type A, B, C, D, E, F, G, H), Gyalocephalus capitatus, and Posteriostomum spp. Due to variability, we were unable to define final body length measurements to differentiate L3 of cyathostomins. However, IC characteristics displayed a higher difference between L3 types than total body length. Through the number and arrangement of IC, 14 species were classified within three larval types. The classification of L3 into distinct larval types sensu latum gives us the advantage of reducing the number of probable species presented in equine faecal samples using a low-cost technique when monitoring the parasite fauna present in individual horses or on the farm level. The present improved identification key shall increase the diagnostic capabilities of classical equine parasitology techniques, using general L3 morphology thereby pragmatically improving regional and transnational epidemiological and biodiversity studies. The present key may also assist in defining the cyathostomin community in cyathostominosis clinical cases and within drug resistant populations across different management systems and geographical locations.

The behaviour of the nematode, Steinernema feltiae (Nematoda: Steinernematidae) in sand contaminated with the industrial pollutant chromium VI.

This study set out to determine the suitability of the nematode Steinernema feltiae as a bioindicator for heavy metal pollution, specifically chromium VI. Nematodes were introduced into sand contaminated with concentrations of Cr VI+, in a range between 10 and 100 ppm, in increments of 10. Reproductive potential, development times and infectivity vs exposure times to Cr VI were employed as endpoints. It was observed that infective juveniles (IJ) from this nematode can survive and successfully infect host insects in the presence of Cr VI for as much as 13 days, and that the nematode increases its reproductive potential at concentrations up to 100 ppm Cr VI+. Conversely, development times (time in days taken for progeny to emerge after larval host death) and IJ infectivity rates were observed to reduce with increasing concentrations of Cr VI. The ability of this nematode to survive in the presence of high concentrations of Cr VI, and its ability to increase progeny numbers at the early stages of Cr VI exposure may provide a survival advantage for this nematode at contaminated sites. It may also demonstrate potential for development as a model species for toxicological assessment in in-situ field sampling.

Anthelmintic therapy of equine cyathostomin nematodes - larvicidal efficacy, egg reappearance period, and drug resistance.

Cyathostomins are ubiquitous in grazing horses across the world, and anthelmintic resistance has been reported with increasing levels over past decades. The aims of the present study were (i) to investigate the efficacy against encysted larval stages of moxidectin (0.4 mg/kg) and fenbendazole (10 mg/kg daily for five consecutive days) and compare these regimens at 2 and 5 weeks post-treatment, (ii) to investigate individual cyathostomin species associated with shortened egg reappearance periods, and (iii) to document species exhibiting decreased susceptibility to the evaluated compounds. Thirty-six ponies were allocated to treatment groups with half euthanatized 2 weeks post-treatment, and the remainder necropsied after 5 weeks. Luminal and mucosal worm counts were conducted and strongyle egg counts were determined at weekly intervals. At 2 weeks, mean reductions of early L3s were 50.4% and 73.8% for fenbendazole and moxidectin, respectively. At 5 weeks, the respective efficacies were 51.3% and 71.8%. Two week efficacies against late L3s and L4s (LL3s/L4s) were 70.8% and 74.6% for fenbendazole and moxidectin, respectively, whereas very low numbers were found in all three groups at 5 weeks. None of the mucosal counts were significantly different between treatment groups. Fenbendazole and moxidectin reduced luminal worm counts by 93.2% and 98.3% at 2 weeks following administration, with moxidectin group adult counts being significantly lower than the other two groups (P < 0.0001). Both treatment groups had increased counts 3 weeks later (P = 0.0415). A moxidectin ERP of 4 weeks was associated with surviving luminal L4s, and adult species contributing to this were Cyathostomum catinatum, Cylicostephanus longibursatus, Cylicocyclus ashworthi and Cylicocyclus nassatus. This study documented (i) larvicidal efficacy of fenbendazole much lower than historical standards, (ii) survival of luminal immatures (L4) following moxidectin administration, and (iii) new information about cyathostomin species associated with these phenomena.

Anthelminthic activity of methanol extracts of Diospyros anisandra and Petiveria alliacea on cyathostomin (Nematoda: Cyathostominae) larval development and egg hatching.

Methanol extracts of plant structures are promising alternatives to traditional pharmaceutical anthelminthic treatments. An in vitro evaluation was done of how methanol extracts of Diospyros anisandra bark and leaves, and Petiveria alliacea stems and leaves, collected during the rainy and dry seasons, effected cyathostomin larval development and egg hatching. Seven concentrations (600, 300, 150, 75, 37.5, 18.7 and 9.3µg/ml) were tested using the egg hatch assay. An ANOVA was applied to identify differences between the concentrations and the controls. Fifty percent lethal concentration (LC50) and the 95% confidence interval were calculated with a probit analysis. At and above 37.5µg/ml, the D. anisandra bark extracts from both seasons exhibited ≥95% egg hatch inhibition (EHI), while the D. anisandra leaf extracts had >90% EHI at and above 75µg/ml. For P. alliacea, the extracts from leaves and stems from either season exhibited >97% EHI at and above 300µg/ml, although similar efficacy was also observed at lower concentrations with the rainy season stems (75µg/ml) and leaves (150µg/ml). Values for LC50 were lowest for the rainy season D. anisandra bark (10.2µg/ml) and leaf extracts (18.4µg/ml), followed by the rainy season P. alliacea stems extract (28.2µg/ml). In the D. anisandra extracts, EHI was largely due to its ovicidal activity (≥96% beginning at 37.5µg/ml), whereas in the P. alliacea extracts it was due to L1 larval hatch failure (≥90% beginning at 75µg/ml). Overall, the rainy season D. anisandra bark extracts had a strong in vitro anthelminthic effect against cyathostomins by inhibiting larval development, and the rainy season P. alliacea stem extracts had a strong effect by preventing egg hatching. Both are possible control alternatives for these nematodes.

Life history adjustments to intestinal inflammation in a gut nematode.

Many parasitic nematodes establish chronic infections. This implies a finely tuned interaction with the host immune response in order to avoid infection clearance. Although a number of immune interference mechanisms have been described in nematodes, how parasites adapt to the immune environment provided by their hosts remains largely unexplored. Here, we used the gastrointestinal nematode Heligmosomoides polygyrus to investigate the plasticity of life history traits and immunomodulatory mechanisms in response to intestinal inflammation. We adopted an experimental model of induced colitis and exposed worms to intestinal inflammation at two different developmental stages (larvae and adults). We found that H. polygyrus responded to intestinal inflammation by up-regulating the expression of a candidate gene involved in the interference with the host immune response. Worms infecting mice with colitis also had better infectivity (earlier adult emergence in the intestinal lumen and higher survival) compared with worms infecting control hosts, suggesting that H. polygyrus adjusted its life history schedule in response to intestinal inflammation.

Encysted cyathostomin larvae in foals - progression of stages and the effect of seasonality.

Cyathostomins (small strongyles) are ubiquitous in grazing horses and are known pathogens as cause of larval cyathostominosis. As part of their life cycle, cyathostomin larvae invade the mucosal walls of the large intestines and undergo encystment. Newly ingested third stage larvae are known to undergo arrested development and this can lead to an accumulation of encysted burdens over the course of a grazing season. It is believed that the host immune system plays a significant role in triggering this arrestment. Little is known about the development and progression of larval stages in foals that are not expected to mount a pronounced immune response to ingested cyathostomin larvae. This study evaluated counts of encysted larvae measured in 37 foals. The foals were born in 2013, 2014, and 2015 into a parasitology research herd kept without anthelmintic intervention and were humanely euthanatized between 50 and 293days of age as part of an ongoing parasite transmission study. A mucosal digestion technique was performed to enumerate encysted early third stage (EL3) and late third stage/fourth stage (LL3/L4) larvae in the cecum, ventral colon and dorsal colon. Counts were analyzed statistically to evaluate the influence of foal age, sex, and seasonality on the counts. Total counts as well as LL3/L4 counts were significantly higher during the grazing season (March-November). Three defined age groups (>100, 100-200, >200days) did not have statistically different counts. Male foals had significantly higher total counts compared to females, and this has not been reported before. The study found that 41% of the recovered larvae were EL3s, but no indication of arrested development of these was observed. This indicates that cyathostomin infection in foals progresses in a manner substantially different from mature horses.

Equine Cyathostominae can develop to infective third-stage larvae on straw bedding.

BACKGROUND: Domesticated grazing animals including horses and donkeys are frequently housed using deep litter bedding systems, where it is commonly presumed that there is no risk of infection from the nematodes that are associated with grazing at pasture. We use two different approaches to test whether equids could become infected with cyathostomines from the ingestion of deep litter straw bedding. METHODS: Two herbage plot studies were performed in horticultural incubators set up to simulate three straw bedding scenarios and one grass turf positive control. Faeces were placed on 16 plots, and larval recoveries performed on samples of straw/grass substrate over 2- to 3-week periods. Within each incubator, a thermostat was set to maintain an environmental temperature of approximately 10 °C to 20 °C. To provide further validation, 24 samples of straw bedding were collected over an 8-week period from six barns in which a large number of donkeys were housed in a deep litter straw bedding system. These samples were collected from the superficial bedding at 16 sites along a "W" route through each barn. RESULTS: No infective larvae were recovered from any of the plots containing dry straw. However, infective cyathostomine larvae were first detected on day 8 from plots containing moist straw. In the straw bedding study, cyathostomine larvae were detected in 18 of the 24 samples. Additionally, in the two barns which were sampled serially, the level of larval infectivity generally increased from week to week, except when the straw bedding was removed and replaced. CONCLUSIONS: We have demonstrated that equine cyathostomines can develop to infective larvae on moist straw bedding. It is therefore possible for a horse or donkey bedded in deep litter straw to become infected by ingesting the contaminated straw. This has implications for parasite control in stabled equids and potentially in housed ruminants, and further investigation is required in order to establish the relative infective pressure from pasture versus straw bedding.

Arrested development of experimental Cyathostominae infections in ponies in Czech republic.

Nine ponies, aged 12 months at the time of infection, were randomly allocated to one of three groups. Each pony was infected with 30,000 infective cyathostome larvae stored 4 weeks under the natural environmental conditions of the Czech Republic. Horses of Group A were infected with larvae conditioned from August 16 to September 12 (the first phase of the experiment). Horses of Group B were inoculated with larvae conditioned from October 10 to November 4 (the second phase of the experiment). In the third phase, horses from Group C received larvae stored under natural conditions (from 8.11 to 18.11.2011). The larvae were then kept under laboratory conditions (5 °C, the light reduction according to the nature condition). In the first and second phase of the experiment, 98.57% and 95.48% of adult nematodes were found, respectively. However, only 29.07% of adults were found in the third phase of experiment, and the proportion of arrested larvae (EL3) was 67.73%. The share of these larvae in the cecum during last phase of the experiment comprised 92.59% of all cyathostomes.

Further indication of lowered activity of ivermectin on immature small strongyles in the intestinal lumen of horses on a farm in Central Kentucky.

Critical tests were performed in 2011 in four weanling horses (L-1, L-2, L-29, and L-30) treated with ivermectin paste at 200 µg/kg. They were born in 2011 and raised together on a farm (MC) in Central Kentucky. The horses had not been treated previously with an antiparasitic drug. However, ivermectin had been administered repeatedly to the horse herd for several years and strongyle eggs per gram of feces (EPGs) returned sooner posttreatment than after initial usage. Critical tests in a recent previous study in this horse herd indicated that the reason for the early return of strongyle EPGs after ivermectin treatment probably was because of lowered drug activity on immature (L(4)) small strongyles in the lumen of the large intestine. Therefore, the life cycle was shortened. The main purpose of the present study was to obtain further data on the activity of ivermectin on small strongyle immature stages, in addition to adults, in the intestinal lumen. Twelve species of small strongyles were present. Combined data for immature and adult small strongyles for the four ivermectin-treated horses demonstrated efficacy of 68 to 83 %. Removal of adults was 100 % for all four horses, and on immatures, it ranged from 0 to 16 %. Efficacy on immature small strongyles was even lower than in the previous study. This supported the earlier finding of apparent reduced time for maturation of the parasites because of incomplete elimination of immature small strongyles in the intestinal lumen of horses after ivermectin treatment. Data on five other helminth species were recorded.

Contamination of the environment by strongylid (Nematoda: Strongylidae) infective larvae at horse farms of various types in Ukraine.

Analysis of the influence of horse-keeping conditions by contamination of the environment (pastures, paddocks, and stalls) by the strongylid infective larvae (L(3)) was carried out at various types of horse farms, hippodromes, and riding clubs in Ukraine. A total of 1,237 horses from three types of horse-keeping conditions were examined. Epidemiological studies of stall and grazing area (pasture and paddocks) contamination by L(3) were performed at hippodrome (stalled horse-keeping) and horse farms with stall/paddock-keeping and stall/pasture-keeping conditions. Grass and stall litter samples were examined by the Baermann procedure. It was found that horses of stall-keeping conditions had the lowest level of strongylid infection (prevalence 46.4-77.8%, average infection 25.6-92.9 eggs per gram of feces (EPG)) and lowest proportion of large strongyle L(3) in coprocultures (1.6-11.3%). Horses of stall/pasture-keeping conditions were the most infected (prevalence 95.1-100%, average infection 198.2-453.7 EPG), and the proportion of large strongyle L(3) was 17.3-24.7%. Strongyle L(3) were found in litter of all parts of individual stalls; areas at the stall center, "toilet", and entrance were the most contaminated. The highest L(3) number in stall litter was registered in summer. Contamination of permanent pasture grass by L(3) was notably lower than grass in paddocks (86.3-161.4 L(3)/kg compared with 305.9-409.1 L(3)/kg). The highest level of pasture grass contamination was observed in the middle of summer (July)--up 970.7 L(3)/kg. The results obtained confirmed importance of environmental contamination in epidemiology of horse strongylidosis at various types of horse-keeping conditions.

Isolation of potentially useful antigens from cyathostomin third-stage larvae by using a fast protein liquid chromatography one-step method.

Three major protein complexes (51, 29, and 15 kDa, named P1 to P3, respectively) were resolved by gel filtration of the excretory/secretory antigens collected from a mixture of horse cyathostomin third-stage larvae (L3s). The potential application for the detection of infected horses was assessed with an enzyme-linked immunosorbent assay (ELISA) by the comparison of the serological and copromicroscopical results. The value of the area under the receiver operating characteristic (ROC) curve was higher than 0.9 when the three peaks were used. Elevated values (>90%) for the sensitivity, specificity, and the positive-likelihood ratio were also observed for all the antigen complexes. A significant increment in the IgG antibody levels 4 weeks prior to the observation of eggs in the feces of weanlings naturally infected was recorded. Our results indicate that the evaluation of chemotherapy is possible by using immunoenzymatic probes and fast protein liquid chromatography (FPLC)-purified antigens. Data collected in the present investigation indicate that FPLC isolation offers a very helpful one-step method for collecting antigens with diagnostic potential to be employed in immunoenzymatic probes.

Morphology and diagnosis of the fourth-stage larva of Coronocyclus labratus (Looss, 1900) (Nematoda: Strongyloidea) parasitising equids.

The fourth larval stage (L4) of Coronocyclus labratus (Looss, 1900) Hartwich, 1986, one of the common species of the Cyathostominae found in equids, is identified and described. The larvae found were identified as C. labratus by finding moulting forms possessing characters of both larval and adult stages. The larvae are similar to those of Cylicocyclus leptostomum (Kotlán, 1920) Foster, 1936, Cyathostomum catinatum Looss, 1900 and Cylicostephanus goldi (Boulenger, 1917) Lichtenfels, 1975. The buccal capsule (BC) wall of the L4 of Cylicocyclus leptostomum is thinner than that of Coronocyclus labratus, and the ring of the oesophageal funnel is comparatively less well developed. In C. labratus the dorsal tooth protrudes only slightly into the buccal cavity, and this larva possesses a characteristically elongate, pyriform oesophagus. In Cyathostomum catinatum, the BC walls are thicker than in Coronocyclus labratus, and the dorsal tooth is more bluntly pointed. Sublateral teeth, if present, are bluntly pointed in the former species, but usually absent or indistinct in C. labratus. Larvae of Cylicostephanus goldi differ from those of Coronocyclus labratus in the larger size of the BC.

Selective anthelmintic therapy of horses in the Federal states of Bavaria (Germany) and Salzburg (Austria): an investigation into strongyle egg shedding consistency.

For 9 consecutive months (March-November 2008), faecal samples were collected monthly from 129 horses residing within 40 km of Salzburg, Austria. Samples were analysed quantitatively using a modified McMaster egg counting technique. Whenever a faecal egg count (FEC) result exceeded 250 eggs per gram (EPG), the horse was treated with pyrantel, ivermectin or moxidectin. In 52 of 129 horses (40.3%), no strongyle eggs were ever detected over the course of 9 months. In 39 horses (30.2%), strongyle eggs were detected in at least 1 sample, but the egg count never exceeded 250 EPG. The remaining 38 (29.5%) horses were treated at least once in response to a FEC that exceeded 250 EPG. As a result of this selective anthelmintic scheme, the total number of anthelmintic treatments was reduced to 54% of the number of treatments administered to the same horses in the previous year. Both the maximum and mean FEC dropped significantly after initiation of the study. A statistically significant, negative correlation was demonstrated between the maximum and mean FEC of a horse and its age. Pasture hygiene appeared to reduce FECs, but the effect was not statistically significant. The magnitude of the initial FEC was significantly correlated with the maximum FECs in the subsequent 8 months (p<0.01). The same relationship was observed for the maximum FEC of the first 2 samples. Furthermore, horses which required several anthelmintic treatments had a higher initial FEC and a greater maximum FEC in the first 2 samples than horses which received only one or no treatment. These results suggest that selective anthelmintic treatment accomplished a reduced pasture contamination with strongyle eggs, while simultaneously decreasing the number of anthelmintic treatments. Sustained implementation of a selective treatment strategy has the potential to reduce selection pressure for anthelmintic resistance. These results reported herein will assist equine practitioners in designing and monitoring sustainable anthelmintic treatment programs.

Biological control of horse cyathostomin (Nematoda: Cyathostominae) using the nematophagous fungus Duddingtonia flagrans in tropical southeastern Brazil.

The viability of a fungal formulation using the nematode-trapping fungus Duddingtonia flagrans was assessed for the biological control of horse cyathostomin. Two groups (fungus-treated and control without fungus treatment), consisting of eight crossbred mares (3-18 years of age) were fed on Cynodon sp. pasture naturally infected with equine cyathostome larvae. Each animal of the treated group received oral doses of sodium alginate mycelial pellets (1g/(10 kg live weight week)), during 6 months. Significant reduction (p<0.01) in the number of eggs per gram of feces and coprocultures was found for animals of the fungus-treated group compared with the control group. There was difference (p<0.01) of 78.5% reduction in herbage samples collected up to (0-20 cm) between the fungus-treated group and the control group, during the experimental period (May-October). Difference of 82.5% (p<0.01) was found between the fungus-treated group and the control group in the sampling distance (20-40 cm) from fecal pats. During the last 3 months of the experimental period (August, September and October), fungus-treated mares had significant weight gain (p<0.01) compared with the control group, an increment of 38 kg. The treatment with sodium alginate pellets containing the nematode-trapping fungus D. flagrans reduced cyathostomin in tropical southeastern Brazil and could be an effective tool for biological control of this parasitic nematode in horses.

In vitro development of cyathostomin larvae from the third stage larvae to the fourth stage: morphologic characterization, effects of refrigeration, and species-specific patterns.

A mixed population of equine cyathostomin (Nematoda, Strongyloidea) infective third stage larvae (L3) was cultured in vitro using a cell-free medium. Some L3 were cultured immediately after Baermann collection from fecal cultures, while others were kept in water at 4 degrees C for 7 days before initiating the in vitro cultures. Cultures were examined daily for viability. At days 2, 7, 14 and 21 larvae were collected for identification of developmental stage and morphological changes, using both light and scanning electron microscopy. Larvae were classified as early L3 (EL3), developing L3 (DL3), late L3 (LL3) and fourth stage larvae (L4) on the basis of morphological features. Viability remained high throughout the entire study period in cultures of both non-refrigerated (84.7%) and refrigerated (77.4%) larvae. However, viability of the non-refrigerated was significantly greater from 7 through 21 days of culture. Significant differences were also observed in the percentage of DL3 between the non-refrigerated and refrigerated larval cultures by day 7. The highest percentage of DL3 larvae (22.5%) was reached at the end of study in those larvae that were not previously refrigerated. The data suggests that prior refrigeration decreases viability and slows L3 development. At day 21 LL3 larvae were only a small percentage of the DL3: 6.9 and 5% in non-refrigerated and refrigerated cultures, respectively. Few of these larvae freed themselves from the L3 cuticle and moulted to L4 stage. Characteristics of individual species in vitro developmental patterns were determined by the molecular identification of individual larvae in pools of larvae randomly collected at days 0 and 21. Seven species (Coronocyclus coronatus, Cylicostephanus goldi, Cylicostephanus longibursatus, Cyathostomum catinatum, Cylicocyclus nassatus, Cylicocyclus ashworthi, Petrovinema poculatum) were identified in the day 0 pool. The greatest tendency to develop in vitro was shown by the genus Cylicostephanus with the species C. goldi and C. longibursatus that developed to the LL3-L4 stages. C. nassatus, C. ashworthi and C. coronatus did not progress in their development beyond the EL3 stage, while no apparent signs of development were registered for C. catinatum.

Comparison of two fecal egg recovery techniques and larval culture for cyathostomins in horses.

OBJECTIVE: To compare the McMaster and centrifugal flotation techniques and larval culture for recovery of cyathostomin (small strongyle) eggs from the feces of horses. SAMPLE POPULATION: Fecal samples from 101 horses. PROCEDURES: In experiment I, homogenized fresh feces from a single horse were randomly subsampled by use of each technique for 10 replicates. In experiment II, samples from 43 horses that had no anthelmintic treatment were analyzed by use of McMaster, centrifugal flotation, and larval culture techniques. In experiment III, 57 horses were treated with an anthelmintic by owners, and fecal samples were analyzed as for experiment II. RESULTS: In experiment I, use of the McMaster technique recovered 72% of the eggs obtained by use of centrifugal flotation from paired subsamples. In experiment II, use of the McMaster technique recovered 81% of the eggs obtained by use of centrifugal flotation. Only cyathostomins resulted from individual larval cultures. In experiment III, 24 samples had negative results for all 3 tests, 18 samples had positive results only with larval cultures, and 15 samples had positive results of centrifugal flotation (only 5 of which had positive results via the McMaster technique). CONCLUSIONS AND CLINICAL RELEVANCE: Centrifugal flotation consistently was superior to the McMaster technique, especially at low fecal egg numbers. The combination of centrifugal flotation and larval culture may provide the best accuracy for evaluation of anthelmintic efficacy.

A survey of seasonal patterns in strongyle faecal worm egg counts of working equids of the central midlands and lowlands, Ethiopia.

A study was conducted for two consecutive years (1998-1999) to determine the seasonal patterns of strongyle infection in working donkeys of Ethiopia. For the purpose 2385 donkeys from midland and lowland areas were examined for the presence of parasitic ova. A hundred percent prevalence of strongyle infection with similar seasonal pattern of strongyle faecal worm egg output was obtained in all study areas. However, seasonal variations in the number of strongyle faecal worm egg output were observed in all areas. The highest mean faecal worm egg outputs were recorded during the main rainy season (June to October) in both years in all areas. Although an increase in the mean strongyle faecal egg output was obtained in the short rainy season (March-April) followed by a drop in the short dry season (May), there was no statistically significant difference between the short rainy season and long dry season (Nov-Feb) (P > 0.05). A statistically significant difference however, was obtained between the main rainy season and short rainy season, and between the main rainy season and dry season (P < 0.05). Based on the results obtained it is suggested that the most economical and effective control of strongyles can be achieved by strategic deworming programme during the hot dry pre-main rainy season (May), when the herbage coverage is scarce and helminthologically 'sterile', and the arrested development of the parasites is suppose to be terminating. This could insure the greatest proportion of the existing worm population to be exposed to anthelmintic and also reduces pasture contamination and further infection in the subsequent wet season.