Development of free-living stages of equine strongyles in faeces on pasture in a tropical environment

The development of the free-living stages and yields of infective third stage strongyle larvae in faeces from a horse with a mixed natural infection deposited on pasture plots were studied over a 2-year period in a coastal area in tropical north Queensland. Two sets of faecal masses (one exposed to, and the other protected from the action of a natural population of dung beetles) were deposited monthly and after 7 days faecal samples were taken for larval recovery and counts. Hatching and development of the free-living stages occurred in faeces on pasture throughout the year. Development was rapid as infective stages were reached within a week of faecal deposition in all months. Yields of infective larvae were affected by the season and the action of dung beetles on the faecal masses. Highest yields were obtained from both beetle-exposed and protected faeces during winter (June to August) and lowest yields were in spring (September to November). High temperatures in spring and summer resulted in low yields of larvae, however, the dry conditions in spring made this season the most unfavourable period. In autumn and winter the temperatures were never low enough to stop or markedly slow down the rate of development, and allowed the development of large numbers of infective larvae. Dung beetle activity was observed throughout the year, and exposed faeces were usually completely dispersed within 24 h of deposition. This resulted in lower yields of infective larvae from these than from protected faeces. Though larval yields were lower, the actual numbers were still substantial so as to cast doubt on the usefulness of these beetles as biological control agents for equine strongylosis in the dry tropics.     

Climatic influences on development and survival of free-living stages of equine strongyles: implications for worm control strategies and managing anthelmintic resistance

Development of resistance to anthelmintic drugs by horse strongyles constitutes a growing threat to equine health because it is unknown when new drug classes can be expected on the market. Consequently, parasite control strategies should attempt to maintain drug efficacy for as long as possible. The proportion of a parasite population that is not exposed to anthelmintic treatment is described as being "in refugia" and although many factors affect the rate at which resistance develops, levels of refugia are considered the most important as these parasites are not selected by treatment and so provide a pool of sensitive genes in the population. Accordingly, treatment should be avoided when pasture refugia are small because such treatments will place significant selection pressure for resistance on worm populations. Given this new paradigm for parasite control, it has become important to identify seasons and circumstances wherein refugia are diminished. Free-living stages of equine strongyles are highly dependent on climatic influences, and this review summarises studies of strongyle development and survival under laboratory and field conditions in Northern (cool) temperate, Southern (warm) temperate and subtropical/tropical climates. In Northern temperate climates, refugia are smallest during the winter. In contrast, refugia are lowest during the summer in warm temperate and subtropical/tropical climates. Although adverse seasonal changes clearly have significant effects on the ability of free living stages of strongyle nematode parasites to survive and develop, available data suggest that climatic influences cannot effectively "clean" pastures from one grazing season to the next.     

Development of Hyalomma lusitanicum under laboratory conditions

At the constant temperature of 25 degrees C and relative humidity (RH) of 84%, the average pre-oviposition period of Hyalomma lusitanicum was 47 days, the oviposition lasted an average of 26 days and the total egg production was an average of 6320 per female. At 16 degrees C the females did not lay eggs at all, but those which survived for 1 year and were transferred thereafter to 25 degrees C and 84% RH laid viable eggs. At 35 degrees C, the oviposition was identical at all levels of RH tested (25, 62 and 93%). At 25 degrees C, the pre-oviposition period was shorter at 93% RH, and the number of eggs laid was fewer at 25% RH. The eggs hatched in 32-40 days, the hatching percentage being lower in batches of eggs laid at the beginning and at the end of the oviposition period. The larval and nymphal moultings were not influenced by the type of host. As the temperature increased, the pre-moult period became shorter. The engorged larvae were more sensitive to the low RH than the engorged nymphs, whose moulting percentage was always greater than 72 in all regimes. Low temperature and high humidity had a favourable effect on the survival of unfed nymphs. The female-to-male ratio was 1:2. Hyalomma lusitanicum always behaved as a 3-host tick. The adults did not engorge on rabbits. The female ticks engorged on calves weighed an average 543 mg. Ticks maintained at 25 degrees C and 84% RH and engorged on calves completed the life cycle in 138-196 days, which does not include the period of chitinization of about 30 days. More than half of this period was spent in egg laying and hatching.     

Development and survival of free-living stages of gastrointestinal nematodes of sheep and goats on pasture in the Nigerian derived savanna

Four grass plots were sequentially contaminated with goat faeces containing known numbers of unembryonated eggs of predominantly Haemonchus contortus and Trichostrongylus spp. between October 1982 and April 1983. Four other plots were similarly contaminated with sheep faeces between February and May 1987. An additional plot was repeatedly contaminated with sheep faeces from February to April 1987. Populations of free-living stages in faeces and of infective larvae (L₃) in the herbage were subsequently monitored until the end of April and June of 1983 and 1987 respectively. During February and May 1987 two control cultures of sheep faeces were incubated in the laboratory at 25°C–30°C and at a constant temperature of 50°C and the free-living development was also monitored. L₃ developed very readily in the faeces cultured at 25°C–30°C and in those spread on a grass plot in October, at the end of the wet season, but developed less on the plot contaminated in May at the start of the wet season. Worm eggs in faeces deposited on plots during the hot dry season (December to April) or incubated at 50°C died and disintegrated after 24–48 h exposure to the high environmental temperatures. The results indicate that it is unlikely that gastrointestinal nematodes of sheep and goats can develop or survive on open pasture during the dry season in the Nigerian derived savanna zone.     

Phenothiazine in the origin of benzimidazole resistance in population-B equine strongyles

Clinical trials on a central Kentucky Thoroughbred Farm (B) during 1960-1961 involved nine test treatments of mares, yearlings and weanlings (n = 124), and compared efficacies of phenothiazine (PTZ), piperazine (PPZ), mixtures of PTZ + PPZ and thiabendazole (TBZ) for (1) percent reduction and (2) complete clearance or graded reduction of strongyle egg per gram (epg) counts. Three grades of PTZ (N.F. green, micronized purified and 2-3 microns purified) at the dose rate of 55 mg kg-1 were ineffective in six tests; reduction of strongyle epg count were low grade (26%) and complete clearance was found in only 1 of 32 horses. Piperazine alone at 88 mg base kg-1 was efficacious in two tests; epg counts were effectively reduced (98%), but complete clearance was found in only 2 of 10 horses. Several mixtures of PTZ + PPZ with various dosages of PTZ (27.5-55 mg kg-1) plus PPZ base (88 mg kg-1) were quite effective in six tests, as epg counts were reduced by 96% and complete clearance occurred in 25 of 38 horses. Four trials with TBZ at dose rates between 13 and 44 mg kg-1 in 1961 reduced epg counts by 98%, but cleared only 15 of 36 horses. These data indicated: (1) a large segment of the Farm B small strongyle population was resistant to PTZ at the outset of these observations in 1960; (2) TBZ-resistant individuals were also present in this population when TBZ was first used in 1961. During the ensuing 4-year study (1962-1965) on Farm B mares (n = 15 per year) and yearlings (n = 15-17 per year), biweekly epg and larvae per gram (lpg) counts were carried out to compare the efficacy of three treatment programs: (1) monthly doses of TBZ at 26 mg kg-1; (2) monthly doses of TBZ at 13 mg kg-1; (3) bimonthly doses of a mixture of PTZ + PPZ and carbon disulfide (CS2) at dose rates of 27.5 + 88 and 78 mg kg-1, respectively, or a mixture of PTZ + PPZ + trichlorfon (TCF) at dose rates of 27.5 + 88 + 40 mg kg-1, respectively. Emergence of a TBZ-resistant segment of the small strongyle infections was evaluated by three criteria: (1) annual mean strongyle epg counts; (2) efficacy of individual treatments for epg reductions (%); (3) complete clearance or graded reductions of strongyle epg counts.4+ PTZ-resistant small strongyles. Initial use of TBZ in 1961 produced effective epg reductions, but complete clearance data revealed the presence of TBZ-resistant small strongyles at the outset...     

Triterpenoid saponins affect the function of P-glycoprotein and reduce the survival of the free-living stages of Heligmosomoides bakeri

We studied the effect of triterpenoid saponins on the development of free-living stages of Heligmosomoides bakeri, a parasitic nematode of the mouse intestine. We evaluated the effectiveness of oleane-type glucuronides (GlcUAOA) isolated from Calendula officinalis and Beta vulgaris. The rhodamine 123 retention assay was used to detect dysfunctions of the major membrane transporter for xenobiotics, P-glycoprotein (Pgp). Both C. officinals and B. vulgaris GlcUAOA affect the development of the free living stages and function of Pgp in H. bakeri. The GlcUAOA inhibits egg hatching and moulting of larvae and also changes their morphology. These saponin fractions reversed the toxic effect of thiabendazole on the nematode; the function of Pgp was also inhibited.     

Effect of infection by Babesia spp. on the development and survival of free-living stages of Boophilus annulatus

Oviposition, egg hatching and survival of newly-hatched larvae of Boophilus annulatus were studied in relation to infection by Babesia species and different temperature regimens. Infection of female ticks by Babesia bigemina or B. bovis had no effect on the time elapsed between engorgement and oviposition. The duration of oviposition was shorter in infected females incubated at 25 degrees C or 35 degrees C and infected females laid fewer eggs than the controls. No larvae hatched at 16 degrees C. B. bigemina-infected eggs hatched more quickly than uninfected eggs at 35 degrees C. The hatching percentage of B. bigemina-infected eggs was reduced by 50% at an incubation temperature of 25 degrees C and by 75% at 35 degrees C. At 16 degrees C there was no difference in the duration of survival of infected and non-infected larvae but at 25 degrees C and 35 degrees C the mean survival period of infected larvae was significantly lower than those of controls.     

Prevalence,abundance and site distribution of equine small strongyles in Normandy,France

Forty-two horses from Normandy (France) were examined post-mortem for small strongyle infections from October to March. In the positive horses, total worm numbers ranged from 234 to 90,247 (mean 11,297). Encysted larvae represented the major part of the total cyathostome burdens with a high percentage (83%) being early third stage larvae. They were mostly recovered from the caecum (48%) and ventral colon (40%) and were less present in the dorsal colon (12%). Adult cyathostomes were mainly located in the ventral colon (64%) and less frequently in the dorsal colon (27%) and caecum (9%). Twenty species of Cyathostominae were identified. The 10 most prevalent species (in sequence of prevalence) were Cyathostomum coronatum, Cylicocyclus nassatus, Cylicocyclus insigne, Cyathostomum catinatum, Cylicostephanus goldi, Poteriostomum imparidentatum, Cyathostomum labiatum, Cylicocyclus ultrajectinus, Cylicostephanus calicatus and Cylicostephanus minutus which comprised 84% of the total adult population. Twelve species showed a site preference in the ventral colon, five in the dorsal colon and only one in the caecum while two species were collected in nearly equal numbers from the ventral and dorsal colon. The number of species per horse ranged from 1 to 12 with a median of 5. Infections with singletons occurred in 12.5% of the positive horses while multiple infections were encountered in 87.5%. A positive correlation was found between the intensity of cyathostome infection and its diversity, measured either by the number of occurring species or Shanon indexes.     

Benzimidazole resistance of equine strongyles: critical tests of several classes of compounds against population B strongyles from 1977 to 1981

From 1977 to 1981, critical tests were conducted on 10 horses naturally infected with population B strongyles. Drugs tested were: oxibendazole (OBZ), 10 mg/kg of body weight (n = 1); albendazole, 10 mg/kg (n = 1); a mixture of thiabendazole (TBZ), 44 mg/kg and trichlorfon, 40 mg/kg (n = 1); a mixture of TBZ at 44 mg/kg with piperazine (PPZ) at 55 mg of base/kg (n = 1); febantel (FBT), 6 mg/kg (n = 3), 12 mg/kg (n = 1), or 24 mg/kg (n = 1); and pyrantel (PRT) pamoate, 6.6 mg of base/kg (n = 1). Large strongyles, Strongylus vulgaris (9 horses) and S edentatus (5 horses), were effectively removed (100%) by each compound and mixture. Five species of small strongyles (Cyathostomum catinatum, Cyathostomum coronatum, Cylicocyclus nassatus, Cylicostephanus goldi, and Cylicostephanus longibursatus), previously singled out in this population as resistant to 5 benzimidazoles (TBZ, mebendazole, cambendazole, fenbendazole, and oxfendazole), but not OBZ, were efficaciously removed by OBZ, albendazole, the mixture of TBZ plus PPZ, FBT (24 mg/kg), or PRT. These 5 small strongyle species were resistant to FBT (6 mg/kg) in 3 foals and Cylicostephanus minutus was also resistant in 2 of 3 foals to FBT (6 mg/kg). Doubling the dose of FBT to 12 mg/kg increased the average removal of the 5 basic species plus Cylicostephanus minutus to 88%. The mixture of TBZ and trichlorfon was relatively ineffective against 3 species (C catinatum, C coronatum, and Cylicostephanus longibursatus), but unexpectedly efficacious (86% to 99%) for 2 species (Cylicocyclus nassatus and Cylicostephanus goldi).(ABSTRACT TRUNCATED AT 250 WORDS)     

The elimination of equine strongyles and hematological and pathological consequences following larvicidal doses of thiabendazole

Twelve horses wer divided into three groups and given various doses of a mixed species strongyle inoculum, representing light, moderate, and heavy infctions. Three weeks after the larval inoculations, three animals from each group were given larvicidal doses of thiabendazole (TBZ) (440 mg kg^?1 on two consecutive days); one animal from each group served as a non-medicated control. Treatment was repeated three weeks later. One treated animal from each group was designated for long-term study; others were necropsied to study adult and larval parasite loads.Six of the twelve animals with strongylosis developed moderate eosinophilia.TBZ given at 440 mg kg^? on two consecutive days caused depression, lethargy, and anorexia which lasted for five days. Eosinopenia, lymphopenia, and neutrophilia occured in treated animals, and lasted for three days. During the course of TBZ treatment, one horse died from what appeared to be a mis-dosing or an anaphylactic reaction.At necropsy, active thrombi of the anterior mesenteric artery were seen in parasitized animals, but not in those treated with TBZ. Five out of seven medicated horses were completely free of adult and larval strongyle parasites. One had a few STrongylus edentatus larvae and another had small strongyles. No Strongylus vulgaris larvae or adults were recovered from any horse treated with TBZ.     

Factors affecting the survival and migration of the free-living stages of gastrointestinal nematode parasites of cattle in central queensland

Faecal pats containing parasitic nematode eggs were deposited monthly on worm-free pasture, from mid-1975 to early in 1979, near Rockhampton in central Queensland. Pasture samples were collected monthly from beside these pats and the number of infective larvae on the samples was counted.

Cooperia spp. were the most numerous larvae on pasture all year round and Haemonchus placei were commonly present in low numbers. Small numbers of Oesophagostonum radiatum larvae were found, mostly during summer.

Dung beetle activity and rainfall influenced larval populations on pasture, but temperature did not. Beetles were not active in winter, and pats deposited in spring, summer and autumn when beetles were active yielded only 42, 44 and 26%, respectively, as many larvae per 1000 eggs deposited as winter pats. Pats in which beetle activity was minimal (feeding only), moderate and intense (complete destruction), yielded 43, 10 and 6%, respectively, as many larvae per 1000 eggs as intact pats.

Larval densities on pasture were highest after the first saturating rains during the spring-summer period and most of these larvae migrated from unattacked pats deposited in winter. Beetle numbers and activity increased with the summer rains and so few larvae were available to migrate onto pasture during late summer and autumn when the highest falls of rain were recorded. The regression of larval recovery on rainfall was positive and statistically significant when data collected soon after these very heavy rainfall periods were omitted from the analysis.

In 1977, drought-breaking rains increased the normal larval density on pasture 10-fold because larvae in pats deposited in the last 4 months of the drought migrated onto pasture immediately after the rains.

This work suggests that in summer rainfall areas where dung beetles are active, helminth control may be achieved by reducing the worm egg output from cattle during the winter.