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.     

Efficacy of levamisole pour-on compared with levamisole subcutaneous injection against Dictyocaulus viviparus infection in calves.

The efficacy of levamisole pour-on against Dictyocaulus viviparus was compared to that of subcutaneous levamisole injection. Eighteen calves were raised individually and artifically infected with D. viviparus larvae. Faecal samples were collected 27 and 28 days later and larvae per gram (l.p.g.) determined. The animals were then divided into three comparable groups. Group 1 animals remained untreated as controls. Group 2 animals received levamisole 10% w/v subcutaneous injection at a dose of 5 mg kg-1 and Group 3 received levamisole pour-on 20% w/v at a rate of 10 mg kg-1 applied transdermally. Results of l.p.g. measurements from faecal samples taken 7 and 8 days post-treatment indicated a dramatic reduction in the worm burden of animals in both treatment groups. Necropsies at 14 days post-treatment revealed few adult worms in these groups, indicating a 99 and 98% kill rate for pouron and subcutaneous injection, respectively.     

Dictyocaulus species: cross infection between cattle and red deer

Aim: To discover whether cross infection between red deer (Cervus elaphus) and cattle is possible with either a bovine isolate of the cattle lungworm, Dictyocaulus viviparus, or with a cervine isolate of the lungworm, Dictyocaulus eckerti which is thought to be maintained primarily in deer. Method: Twelve cattle and 12 red deer were reared parasite-free from birth. At 3-4 months of age, half of each species (n=6) were experimentally infected with D. viviparus and the other half with D. eckerti. The course of infection was monitored for 34 days, after which the animals were slaughtered and the lungs removed to assess levels of infection. Results: Faecal larval counts demonstrated that patent Dictyocaulus infections occurred in all groups. At necropsy, adult worms were found in the lungs in all groups except the cattle that were infected with D. eckerti. The largest numbers of adult worms were found in the red deer infected with D. eckerti. Conclusion: It was demonstrated that both cattle and red deer could be infected with either D. viviparus or D. eckerti. However, D. eckerti larvae that originated from deer established more successfully in deer and D. viviparus larvae that originated from cattle established more successfully in cattle.     

Comparison by experimental infections in cattle of a Dictyocaulus species occurring naturally in red deer and a dictyocaulus of bovine origin

Dictyocaulus species larvae were obtained from young red deer which had become infected on pastures considered to be carrying the Dictyocaulus species indigenous to the red deer of Scotland. These larvae were cultured to third stage and transmitted to five bovine calves. Five other bovine calves were infected with third stage Dictyocaulus viviparus larvae of bovine origin. Microscopic appearances of both groups of larvae were indistinguishable and their lengths were similar. Results indicated that the Dictyocaulus species derived from deer induced milder though similar clinical and pathological responses in cattle than did the D viviparus derived from cattle. It was concluded that there are strains of different pathogenicity within the species D viviparus, that the deer derived Dictyocaulus species was a strain of D viviparus, and that the hazards to animal health associated with infection by D viviparus in farming systems where red deer and cattle may graze alternately are likely to be acceptable.     

Arrested development of bovine Ostertagia spp. and Cooperia spp. in New Zealand. Effect of temperature and duration of storage on infective larvae

Infective larvae of Ostertagia spp. and Cooperia spp. derived from naturally infected dairy calves were subjected to periods of storage of up to 16 weeks at 4 degrees C or 15 degrees C to determine if this treatment would influence their propensity for arrested development in previously worm-free calves. Results showed no significant increase in the propensity of Ostertagia spp. for arrested development in response to the treatments, but a small increase in the case of Cooperia spp.     

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.     

Survival of Aujeszky's disease virus in infected pig slurry

It has been demonstrated that after experimental infection of pig slurry from the space under the slatted floor (infection dose of 10(6)PFU per ml), the Aujeszky's disease virus (ADV) survived for 72 hours at the temperature of 15 degrees C and at pH 6.5, but was inactivated after 96 hours. When technologically treated pig slurry from the storage tanks was saturated with water and infected with ADV at the dose of 10(5)PFU per ml, the virus survived for 23 days when kept at 15 degrees C and 4 degrees C and at pH 6.8, but was inactivated under the same conditions after 30 days. When the infective ADV dose in the technologically treated pig slurry in the storage tanks was reduced to 10(4)PFU per ml, the virus survived 16 days at +4 degrees C and pH 7.0 and 8.0 but was inactivated within 23 days after infection.     

Effect of condensed tannins extracted from four forages on the viability of the larvae of deer lungworms and gastrointestinal nematodes

The inhibitory activity of condensed tannins extracted from four forage legume plants were evaluated by using a larval migration inhibition assay. The first (L1) and third (L3) stages of deer lungworm (Dictyocaulus viviparus), and the third stage (L3) of deer gastrointestinal nematodes were incubated with tannins extracted from Lotus pedunculatus, Lotus corniculatus, sulla (Hedysarum coronarium) and sainfoin (Onobrychus viciifolia). The tannins extracted from all the forages had inhibitory activity as measured by their ability to paralyse the larvae and inhibit them from passing through sieves. At the highest concentration used (1200 microg/ml) the tannins extracted from sainfoin had the highest activity against ensheathed L1 lungworm larvae (58 per cent), followed by L. pedunculatus (45 per cent), sulla (42 per cent) and L. comiculatus (35 per cent) when the larvae were incubated at 37 degrees C. The same trend, but with lower activities, was observed when the larvae were incubated at 22 degrees C. Anthelmintic activity against L3 lungworm larvae was evaluated by measuring the death rate of ensheathed L3 larvae after incubation with condensed tannins for two, 24 and 48 hours at room temperature (22 degrees C). The death rate was significantly higher (P<0.001) after 48 hours incubation than after two hours or 24 hours, and significantly higher (P<0.001) after 24 hours than after two hours incubation. Condensed tannins from sainfoin had the highest inhibitory activity followed by L. pedunculatus, sulla and L. comiculatus. The tannins from sainfoin also had the highest activity against L3 larvae of gastrointestinal nematodes, followed by L. pedunculatus, sulla and L. comiculatus. Exsheathed larvae of gastrointestinal nematodes were significantly more susceptible to the action of the tannins than ensheathed larvae.     

Survival of Clostridium difficile and its toxins in equine feces: implications for diagnostic test selection and interpretation.

Although Clostridium difficile is recognized as a cause of enterocolitis in horses and humans, there has been little work published regarding the lability of C. difficile and its toxins in feces. A significant decrease in recovery of C. difficile from inoculated equine fecal samples occurred during storage. Recovery after storage in air at 4 degrees C decreased from 76% (37/49) after 24 hours to 67% (33/49) at 48 hours and 29% (14/ 49) after 72 hours. In contrast to aerobic storage, 25 of 26 samples stored anaerobically at 4 degrees C yielded growth of C. difficile for 30 days, whereas the organism was only detected for 2.5 +/- 2.52 days (x +/- SD) in paired samples stored aerobically. The use of an anaerobic transport medium was effective in maintaining viability of C. difficile. These findings indicate that poor aerotolerance is the reason for the rapid decrease in culture yield. In contrast to C. difficile organisms stored aerobically at 4 degrees C, C. difficile toxins were considerably more stable and could be detected by enzyme-linked immunosorbent assay in both broth and inoculated fecal samples for at least 30 days. The poor survival of C. difficile but the stability of its toxins when feces are stored aerobically must be considered when submitting samples for diagnosis of C. difficile-associated enterocolitis in horses and when interpreting laboratory results.     

Induction of protective immunity to Dictyocaulus viviparus in calves while under treatment with endectocides

Three groups of five parasite-naive calves were used. The treatments were: (a) Group 1 calves were weighed on Day 0 and injected with doramectin at 200 microg/kg. From Day 1 to 19 they were dosed orally with 2000 infective larvae of Dictyocaulus viviparus. On Day 28 they were again injected with doramectin, and infected with D. viviparus larvae from Days 33 to 41. They were then left untreated until Day 81 when they were infected with 20 infective larvae of D. viviparus per kg body weight. They were killed on Day 110 and lungworms were counted; (b) Group 2 calves were immunised with oral lungworm vaccine on Days 0 and 28, and infected and slaughtered as Group 1 on Days 81 and 110, respectively; (c) Group 3 calves acted as infection controls. Blood samples were taken at Days 0, 21, 49, 77 and 110 for antibody tests to D. viviparus. At autopsy there were no significant differences between the number of lungworms from Groups 1 and 2 (Means 17.4 and 31.3, respectively); Group 1 had significantly less value than Group 3 (Mean 228) (p < 0.05). Increased antibody titres to the larval sheath of the infective larvae were observed from Groups 1 and 2, showing that the larvae in Group 1 had penetrated the intestine before being killed by the circulating anthelmintic. This experiment shows that if calves are exposed to infective larvae while under systemic endectocide cover, an immune reaction is stimulated.     

Tolerance to low temperatures of Toxocara cati larvae in chicken muscle tissue

Infectivity of Toxocara cati larvae in muscle tissue of chickens after storage at 4°C and -25°C was assessed in a mouse bioassay to provide information on the risk of meat-borne toxocarosis. Muscle tissue samples of 30-day old T. cati infections were stored at 4°C for 14 and 28 days and at -25°C for 12, 24 and 48h, whereafter, larvae were released by digestion. For each experimental group, the released larvae were inoculated in six mice. After 15 days, mice were euthanized and larval burden was assessed by digestion. In the control group (no storage of the infected chicken meat), 47.9% of the inoculated larvae established in mice, whereas storage of meat at 4°C for 14 days or 28 days reduced the recovery to 24.1% or 3.3%, respectively. Muscle larvae exposed to -25°C for 12, 24 or 48h did not establish in the mice. The observation that larvae retain infective after refrigeration at exposure in 4°C for 28 days, emphasize the zoonotic potential of poultry meat as a causative agent of human toxocarosis.     

Biology of cestodes of domestic ducks and wild water birds

Life cycles of 16 cestode species of the families Hymenolepididae and Diploposthidae were studied in 52 ponds of the State Fishery in Bohemia and Moravia. The parasites were recovered from domestic ducks kept on the ponds and from wild birds, mostly of the genera Anas and Aythya, occurring in these water biotopes. Cestode larvae were found in 9 species of Copepoda (9669 out of 1 600 200 examined specimens, i. e. 0.6%) and 3 species of Ostracoda (500 out of 272 300 examined specimens, i. e. 0.18%). Five species of water snails serving as reservoir hosts harboured cysticercoids of 5 cestode species (680 out of 10 212 examined specimens, i. e. 6%). The studies of cestode life cycle under natural conditions were supplemented with 680 successful experiments with intermediate hosts and 179 successful experiments with definitive hosts. The development of larvae was related to the temperature and lasted approximately 13-20 days at 18-25 degrees C. The highest intensity of infection occurred incrustaceans under experimental conditions, a lower intensity was found in crustaceans from duck farms and the lowest in crustaceans from other parts of ponds (e. g., in the case of Fimbriaria fasciolaris in Copepoda, 37, 22 and 10 cysticercoids). The intermediate hosts infected with cestode larvae can live for 3-6 weeks (Copepoda) and 4-7 weeks (Ostracoda) and after invagination of cysticercoids for 20-25 days, on the average. The cysticercoids survive their hosts for 14-18 h at 4-6 degrees C, for 10-11 h at 12-14 degrees C, for 6-7 h at 18-20 degrees C and for 3-4 h at 24-26 degrees C. If they are swallowed by water snails at that time (dead crustaceans are a component of their food), they survive in their digestive tract even for two years (the longest period demonstrated experimentally) and after this time they are able to develop into an adult cestode in the definitive host. The infectivity of cysticercoids increases with their age. It is the lowest immediately after invagination (10-15%), during the stay of cysticercoids in crustaceans it gradually increases (30-60% on days 10-20 after invagination) and it is the highest in cysticercoids from snails (45-55% after 20-50 days in snails, 60-80% after 50-100 days in snails and 70-90% after more than 100 days in snails).     

Trapping efficacy of Duddingtonia flagrans against Haemonchus contortus at temperatures existing at lambing in Australia

The aim of this study was to determine the trapping efficacy of Duddingtonia flagrans against Haemonchus contortus at the temperature ranges experienced around lambing in the major sheep producing regions of Australia. Faeces were collected from Merino wethers, maintained in an animal house and which had received either D. flagrans chlamydospores for a 6-day period (DF) or not (NIL). Faeces were incubated at one of four daily temperature regimens which were composed of hourly steps to provide 6-19 degrees C, 9-25 degrees C, 14-34 degrees C and 14-39 degrees C to mimic normal diurnal air temperature variation. Enumeration of the number of preinfective and infective larvae that had migrated from or remained in faecal pellets was used to calculate percentage recovery and trapping efficacy of D. flagrans. Recovery of H. contortus larvae of both stages was significantly lower in DF faeces but the magnitude of the effect was considerably greater for infective larvae. Mean recovery of infective larvae from NIL and DF faeces was 10.6 and 0.4%, respectively, indicating a mean trapping efficacy of 96.4%. The lowest trapping efficacy (80.7%) was observed at 6-19 degrees C but total recovery of infective larvae, from DF faeces, was greatest at the two highest temperature regimens, although still less than 0.9%. The results of this study indicate that typical Australian lambing temperatures should not be a barrier to the use of D. flagrans as an effective biocontrol of H. contortus in Australia.     

A survey on Dictyocaulus viviparus antibodies in bulk milk of dairy herds in Northern Germany

Parasitic bronchitis caused by the bovine lungworm, Dictyocaulus viviparus, occurs worldwide in temperate areas. The parasite is found predominantly in calves and heifers, but dairy cattle can suffer from lungworms when they become infected for the first time or if they have lost immunity due to lack of exposure to lungworm larvae during the grazing season. The present study was performed to determine the D. viviparus bulk milk antibody prevalence in dairy herds in the East Frisian region of northwestern Germany, Lower Saxony, by analysing bulk milk samples collected in January (860 samples), September (866 samples) and November (860 samples) 2008, thereby representing 906 dairy farms. These samples were tested for antibodies against D. viviparus by a milk ELISA. This test detects patent infections only since it is based on recombinant major sperm protein as antigen. While in January 12.8% of dairy farms were positive for D. viviparus antibodies, the bulk milk samples collected in September and November revealed 6.9% and 6.6% positive dairy herds. From the 906 dairy farms included in the study, 191 (21.1%) tested positive at least once for antibodies against lungworm. From 810 dairy farms from which bulk milk samples were obtained during all three samplings, 146 (18.0%) farms were positive at one sampling date, 27 (3.3%) at two, and 4 (0.5%) on all three sampling dates. The majority of the farms represented in the study belonged to four districts of East Frisia, which showed no significant difference in the proportion of positive dairy farms.     

The effects of Dictyocaulus viviparus infection on energy metabolism of calves

To determine the effects of lungworm infection on energy metabolism and rate of weight gain, five 3-mo-old male Friesian calves were infected orally twice each week with 640, third-stage larvae of Dictyocaulus viviparus (D.v.) over an 8-wk period. Infected calves were matched with uninfected controls on the basis of similar rates of feed consumption and weight gain during the acclimation period before infection. Infected calves were fed 2 kg of concentrates daily (88% DM), about 8.5 Mcal/d. Controls each received approximately 250 g less, about 7.5 Mcal/d. Similar amounts of hay (5.6 to 5.8 Mcal/d) were provided to all calves. Clinical, serum chemical, hematological and parasitological criteria, weight gain and utilization of energy were monitored on a weekly basis. Serum chemical and hematological analyses and clinical examinations of infected animals revealed signs typical of lungworm infection. Fecal and sputum sample examinations for infected calves were positive for D.v. larvae and ova, respectively. Control animals gained approximately 80 g.animal-1.d-1 more than infected calves. Lungworm infection had no significant effect on digestibility of energy or protein. Metabolizability of energy ingested was somewhat higher in the infected calves due to a higher dietary concentrate to roughage ratio. Utilization of metabolizable energy and protein tended to be less efficient for infected animals. Results showed that D.v.-infected calves need more feed for gain than do uninfected animals. This extra requirement is due to an increased maintenance requirement and probably to a reduced protein retention from digested protein.     

Effects of Storing Pig Ovaries at 4 or 20°C for Different Periods of Time on the Morphology and Viability of Pre-Antral Follicles

The purpose of this study was to evaluate the effect of cooling ovarian tissue on pig pre-antral follicles. Ovaries were maintained in saline solution (0.9%) at 4 or 20 degrees C for 6, 12 or 18 h. After storage, pre-antral follicles were morphologically evaluated. While primordial follicles were not affected by the storage, the percentage of morphologically normal growing follicles was significantly reduced in ovarian tissue stored at 20 degrees C for 12 or 18 h. To test the viability of stored follicles, growing follicles isolated from ovaries stored at 4 degrees C for 18 h and at 20 degrees C for 6 h were cultured for 3 days. Follicles stored in either condition presented the same growth pattern in vitro as fresh follicles. We conclude that storage of pig ovaries at 4 degrees C for up to 18 h or at 20 degrees C for up to 6 h does not affect the morphology of growing follicles or their ability to grow in vitro.     

Factors affecting recovery of Dictyocaulus viviparus third stage larvae from herbage and growth of Pilobolus on dung pats

Significantly greater Dictyocaulus viviparus stage three larval recoveries from herbage samples adjoining first stage larval infected dung pats with Pilobolus, and the effects of biotic and mechanical factors on dung pat integrity supported previous findings. Several meteorological factors including sunshine hours, relative humidity, rain total and wind speed showed significant correlations with growth of Pilobolus on dung surfaces. The influence of monitored variables on the recovery of third stage larvae from herbage and the growth of Pilobolus was modified by height of the sward surrounding the dung pat. Multiple regression analysis showed that 60 per cent of variation in D viviparus third stage larvae recovery from herbage was accounted for by known variables.     

Immunodiagnosis of experimental Parelaphostrongylus tenuis infection in elk

Elk infected with the meningeal worm, Parelaphostrongylus tenuis (Protostrongylidae), do not consistently excrete larvae in feces, making the current method of diagnosing live animals using the Baermann fecal technique unreliable. Serological diagnosis could prove more useful in diagnosing field-infected animals but depends on the identification and availability of good quality antigen. To mimic field infections, 2 elk were inoculated with 6 infective L3 larvae of P. tenuis, and another 2 with 20 L3 larvae. Fecal samples were examined for nematode larvae using the Baermann technique and serum samples taken were tested for anti-P. tenuis antibody with ELISAs by using the excretory-secretory (ES) products of L3, and sonicated adult worms as antigens. One animal passed first-stage larvae in its feces 202 days postinoculation, but passed none thereafter. The remaining 3 inoculated animals did not pass larvae. In contrast to parasite detection, antibodies against larval ES products were detected in all animals starting from 14 to 28 days postinoculation and persisted until the termination of the experiment on day 243 in 2 animals that harbored adult worms. Antibodies against somatic antigens of the adult worm were not detected until day 56 but also persisted until the end of the experiment in the animals with adult worms. In 2 elk that had no adult worms at necropsy, anti-ES antibodies were detected transiently in both, while anti-adult worm antibodies were present transiently in one. These findings confirm the superiority of P. tenuis larval ES products over somatic adult worm antigens as serodiagnostic antigens, as previously observed in studies of infected white-tailed deer, and extend the application of the newly developed ELISA test in diagnosing and monitoring cervids experimentally infected with P. tenuis.     

Treatment of inhibited Dictyocaulus viviparus in cattle with ivermectin

Fifteen calves, each infected with approximately 3000 third stage larvae, were used to compare the tendencies of two strains of Dictyocaulus viviparus to inhibit at the fifth larval stage and to evaluate the efficacy of ivermectin. There were notable differences between the strains. While greater than 99% of worms developing from infection with an Alpine strain remained inhibited 42 days after infection, only 0-26% of those recovered following infection with a U.K. laboratory strain were arrested. Neither adult nor immature D. viviparus were present in the lungs of animals treated with ivermectin subcutaneously at 200 micrograms/kg body weight 28 days after infection with the Alpine larvae.     

Verminous Pneumonia in Adult Dairy Cows in Southern Ontario due to Dictyocaulus viviparus

An outbreak of verminous pneumonia due to Dictyocaulus viviparus in a herd of mature, lactating, dairy cows in southern Ontario is described. The fact that the herd had been closed to new additions for ten years and had never experienced clinical disease due to D. viviparus in the past makes the occurrence of this herd problem difficult to explain. Correlation of fecal Baermann analysis for D. viviparus larvae with the progress of anthelmintic treatment is discussed. It is suggested that certain climatological variations in combination with unique, immunological aspects of D. viviparus infection may have contributed to the development of clinical disease in this herd.