Observations on the resumption of development of inhibited Ostertagia, Cooperia and Nematodirus infections in calves stabled overwinter.

Two groups of three month old, parasite-free calves grazed a permanently infected pasture for 14 days, Group A during the first two weeks of September and Group B during early November. Half of each group was killed 14 days after removal from the pasture and the remainder stabled overwinter before slaughter and parasitological examination. Marked inhibition of development occurred for Cooperia oncophora with a variable lower level of inhibition for Ostertagia ostertagi and practically none for Nematodirus helvetianus in those calves grazing late in the fall. Under the conditions of this study, inhibited Cooperia larvae resumed development in several calves soon after they were stabled while small numbers of Ostertagia resumed development regularly during the winter and spring with a considerable number of Ostertagia still present when the calves were slaughtered at the end of the stabling period. On the other hand, Nematodirus and practically all Cooperia worms were lost during the stabling period. In three of seven calves grazing late fall pastures, large Cooperia infections were either not established or failed to become patent.     

Experimentally induced Cooperia oncophora infection in calves: lymphocyte blastogenic and delayed hypersensitivity responses

Lymphocytic responses in peripheral blood and visceral lymph to Cooperia oncophora antigen and skin tests were determined in 35 Holstein male calves that were inoculated orally with single or multiple doses of C oncophora infective larvae. Several calves were vaccinated or given immune serum before larvae were inoculated. Antigen-specific in vitro blastogenesis of blood and lymph lymphocytes and delayed-type hypersensitivity reactions were observed in several inoculated, vaccinated, and/or passively immunized calves. Most calves that had delayed skin reactions also had in vitro lymphocyte responses to C oncophora antigen. The lymphocyte and skin responses were inconsistent and variable in time of onset--the earliest lymphocyte response occurring 7 days after calves were inoculated. A cellular immune response was induced by both dermal vaccination and oral inoculation; however, passive immunization by IV administration of immune serum simultaneously with inoculation did not have an apparent effect on the cellular response, as measured by the lymphocyte blastogenesis test or dermal testing. Although cellular immune responses were observed in several calves infected with C oncophora, there was no apparent relationship between the specific responses and number of nematodes establishing infection in calves after either single- or multiple-dose oral inoculations.     

The residual effect of treatment with ivermectin after experimental reinfection with nematodes in calves

The residual effect of treatment with ivermectin after experimental reinfection in calves was tested. Twenty-four calves were divided into 6 groups of 4 calves each. All calves received a primary infection of 50,000 larvae of both Ostertagia ostertagi and Cooperia oncophora and 1000 Dictyocaulus viviparus larvae. Calves of group 1 remained untreated, and all other calves were treated 21 days after primary infection (0.2 mg/kg injected subcutaneously). Calves of groups 1 and 2 were slaughtered 7 days later. Calves of groups 3-6 were reinfected with the same number of larvae 3 days, 1, 3 and 6 weeks after treatment respectively. Slaughter was 21 days after reinfection. Based on post-mortem worm counts the efficacy of ivermectin after primary infection was 99.7% for O. ostertagi, 95.1% for C. oncophora and 100% for D. viviparus. A residual effect was present for at least one week, but could not be observed 3 weeks after treatment.     

Strains of cattle parasites in the Netherlands with different propensities for inhibited development

A field study was undertaken of the possible differences in propensity for inhibited development in strains of cattle nematodes from two different locations in The Netherlands. In one location (Lelystad) the strains were thought to lack the ability for inhibited development as a result of environmental stimuli on the infective larvae, while in the other location (Utrecht) this ability was presumed to be present. The Lelystad strains were kept at the original location and were also transferred to a pasture in Utrecht, while the Utrecht strains remained in Utrecht. At both locations all permanent grazing calves showed high proportions of inhibited early fourth stage larvae of Ostertagia ostertagi, the proportion of the Utrecht strain being higher than the local and the transferred Lelystad strain. For Cooperia oncophora and Nematodirus helvetianus no clear differences between locations were observed in the permanently grazed calves. No inhibition occurred in tracer calves turned out in August in either location, but differences were seen among tracer calves turned out in October. The Lelystad strains, kept in Lelystad or transferred to Utrecht, showed no inhibition, while in the tracer calves grazed on pastures contaminated by the Utrecht strains a marked inhibition of O. ostertagi and N. helvetianus was observed. The Lelystad strains appear to lack the ability, possessed by the Utrecht strains, to inhibit their development in autumn in response to environmental stimuli.     

Persistent efficacy of doramectin and moxidectin against Cooperia oncophora infections in cattle

Duplicate studies in France and Northern Ireland were carried out to determine the persistent efficacy of topical moxidectin and doramectin against natural infections of Cooperia oncophora. In each study, groups of 15 nematode-na?ve calves were treated either with topical doramectin or moxidectin, and put out to graze on pasture contaminated with C. oncophora infective larvae. The persistent efficacy for preventing establishment of infection was assessed by the time to faecal egg excretion of C. oncophora eggs. It was found that the moxidectin treatment prevented infection for less than 10 days and the effect of doramectin lasted for 24 days.     

Experimental infections with Cooperia oncophora in calves. A study with two different larval dose levels and dosing regimens.

The effect of different larval dose level and dosing regimens on the course of Cooperia oncophora infection in calves was studied. Four groups each of 4 calves were experimentally infected either with 50,000 or 200,000 C. oncophora larvae (L3) given either as single infections or as daily trickle infections. An additional group of calves remained as uninfected controls. The animals were necropsied on week 4 after infection. Mild to moderate clinical signs of parasitic gastroenteritis developed among calves given high doses of larvae, but liveweight gains were not significantly different from those of the uninfected controls. Serum pepsinogen levels of dosed animals were within normal ranges but rose slightly, and on day 14 p.i. they differed significantly from those of the controls. On that occasion, the levels of serum pepsinogen in the trickle infected groups significantly exhibited the levels of the single infected groups. Hypoalbuminaemia was not a feature on any occasion. The various groups did not differ significantly with regard to total worm counts and adult worm counts, but the groups receiving high larval dose harboured significantly more fourth stage larvae than the group receiving low doses of larvae, both in terms of absolute counts and in terms of percentages of total worm burdens. Within the same dose level, there was a tendency of a more even distribution of worms along the small intestine when the infections was given as a single infection compared with a trickle infection. The results indicate that C. oncophora larval dose and dosing regimens may influence the pathogenic effects and to some extent the distribution of the parasite in the small intestine.     

Increased establishment of lungworms after exposure to a combined infection of Ostertagia ostertagi and Cooperia oncophora

Three-month-old calves were infected three times weekly during a 5-week period with Cooperia oncophora, Ostertagia ostertagi or a combination of these two species. For each type of infection two dose levels were applied. In addition one group of calves was kept uninfected. After removal of the primary infection by anthelmintic treatment all calves were challenged with lungworm larvae and slaughtered 5 weeks later. The groups receiving either C. oncophora or O. ostertagi as a monospecific infection did not differ from the na?ve controls. The group receiving the combination of both species differed significantly from the other groups, the establishment of the lungworms being 177%, and the faecal excretion of larvae being 325% of that of the other groups.     

Studies on ticks of veterinary importance in Nigeria. VI. Comparisons of oviposition and the hatching of eggs of Hyalomma species

Fully engorged Hyalomma spp. in Nigeria oviposited greater numbers of eggs than those partially engorged. Hyalomma impressum was a more prolific egg layer than H. rufipes, H. impeltatum and H. truncatum. The variations in the egg output as well as the recognizable peaks in the number of eggs during oviposition were described for each species. No species of Hyalomma below the engorged weight of 0.2 g oviposited; oviposition started with ticks of weight 0.3 g. Eggs produced by ticks weighing below 0.3 g did not hatch; the highest percent egg eclosion occurred with ticks of weight 0.6 g (H. rufipes) and 0.7 g (other Hyalomma species). The pre-oviposition, oviposition and eclosion periods were shortened when eggs were laid and incubated at high temperatures, although the number of oviposited eggs did not increase significantly. At the standard temperature of 24 degrees C, the longest eclosion period was seen in the eggs of H. rufipes (41 days) while those of H. truncatum, H. impressum and H. impeltatum were similar to each other (29 days). Only eggs of H. rufipes hatched at an incubation temperature of 15 degrees C. Eggs of Hyalomma species laid at the same time hatched over a 2--4 day period, except at 15 degrees C when the hatching period of H. rufipes lasted 10 days. The eclosion period was longest in the earlier ovipositions and shorter in the later ones. It is suggested that some intrauterine larval development might have started in the eggs before they were released at a later oviposition period. The percentage mortality of eggs at various temperatures showed that eggs of H. rufipes were more tolerant of low temperatures than those of H. impressum, H. truncatum and H. impeltatum, while the eggs of the latter 3 species were more tolerant of high temperatures than those of H. rufipes. The relevance of these results of the distribution and abundance of the Hyalomma species in Nigeria was discussed.     

Assessing resistance against macrocyclic lactones in gastro-intestinal nematodes in cattle using the faecal egg count reduction test and the controlled efficacy test

The main objective of the present study was to evaluate the accuracy of the faecal egg count reduction test (FECRT) to assess the resistance status of ivermectin (IVM)-resistant isolates of the cattle nematodes Ostertagia ostertagi and Cooperia oncophora, using the controlled efficacy test (worm counts) as a reference. The second objective was to investigate whether both IVM-resistant isolates showed side-resistance against moxidectin (MOX) under controlled conditions. Thirty male Holstein calves were experimentally infected with 25,000 L3 of an IVM-resistant O. ostertagi isolate and 25,000 L3 of an IVM-resistant C. oncophora isolate. Twenty-eight days later the calves were randomly divided into 2 treatment groups and 1 untreated control group. Animals in groups 1 and 2 received MOX (Cydectin(?) 1%, Pfizer) and IVM (Ivomec(?) 1%, Merial) respectively, by subcutaneous injection at a dose rate of 0.2mg/kg bodyweight. Faecal samples were collected 7 and 14days after treatment and animals were necropsied 14/15days post-treatment. Both the FECRT and the controlled efficacy test demonstrated that the O. ostertagi and C. oncophora isolates were resistant against IVM, with efficacies below 90%. The IVM-resistant O. ostertagia isolate was still susceptible to MOX treatment, as shown by over 99% reduction in egg counts and worm burden. The FECRT suggested borderline resistance against MOX in the IVM-resistant C. oncophora isolate, with egg count reductions between 97% (95% CI: 76; 100) at day 7 and 86% (95% CI: 49; 96) at day 14. However, the controlled efficacy test clearly showed MOX-resistance, with a decrease of only 31% (95% CI: -12; 57) in C. oncophora worm numbers. After MOX treatment, a significantly lower number of eggs per female C. oncophora worms was counted compared to the control group (43% reduction). Due to this reduced fecundity, the FECRT may fail to detect MOX-resistance.     

Efficacy of ivermectin administered via sustained-release bolus against gastrointestinal nematodes in cattle

Twelve calves (mean weight, 175.5 kg) were used to confirm efficacy of ivermectin delivered from a prototype sustained-release bolus against naturally acquired gastrointestinal nematodes including early fourth-stage (inhibited) larvae of Ostertagia ostertagi. The calves were allocated by restricted randomization on weight to 1 of 2 groups: controls, to which a placebo bolus was given orally, and treated calves, to which a sustained-release bolus designed to deliver 8 mg of ivermectin/day at a steady rate was given orally. After treatment, the 2 groups were housed in separate pens with concrete flooring. Twenty-eight days after treatment, all calves were euthanatized and necropsied. The ivermectin-treated calves had no larval or adult Ostertagia spp and significantly (P less than 0.01) fewer adult Trichostrongylus axei and adult Cooperia (C oncophora, C punctata and C surnabada) than control calves. Efficacy of ivermectin was greater than 99% for Cooperia spp, and 100% for other parasites. Drug-related adverse reactions were not observed.     

Development and survival of free-living stages of equine strongyles under laboratory conditions

In a series of laboratory studies the optimum conditions for the development and survival of the free-living stages of strongyle parasites occurring in horses in tropical north Queensland were determined. No differences in behaviour were noted between the strongyle species. Development to the infective stage occurred only between 10 and 35 degrees C. The rate was affected by temperature, taking 15-24 days and 3 days, respectively, at the lowest and highest temperatures for the developing stages to reach the infective third stage. Yields of infective larvae were very low outside the range 20-33 degrees C, and were highest at 28 degrees C. Survival of infective larvae was good between 20 and 33 degrees C, and large numbers were recovered after 3 months in faeces incubated at 20-28 degrees C. At 33 and 37 degrees C larval survival was affected by the moisture content of the faeces, with infective larvae surviving better in dry than in moist faeces; even a residual moisture level of 40% significantly reduced the number of larvae recovered from faeces incubated at 37 degrees C for 1 month. Moisture also affected larval development, especially at the higher temperatures of 25-39 degrees C. When faecal moisture content fell to less than or equal to 20% by 3 days, larvae which had not yet reached the infective stage were still pre-infective at 7 days, while all larvae in faeces with adequate moisture had reached the infective third stage. It was not possible to determine the critical faecal moisture level below which larval development ceased, however, 28 degrees C (range 25-33 degrees C) was found to be the optimum temperature. Larval development was very rapid and yields of infective larvae highest at this temperature.     

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.     

Inactivation of eggs and larvae of the cattle nematodes Ostertagia ostertagi and Cooperia oncophora after passage in pigs

The study investigated the effect of gastrointestinal passage in pigs on free-living stages of bovine nematodes. Two Landrace x Yorkshire pigs, A and B, were fed fresh eggs of Ostertagia ostertagi and Cooperia oncophora while two other pigs, C and D, were fed third stage larvae (L3) of the same parasites. Faeces from the pigs were collected for 48 h after ingestion. In pigs A and B, 15 and 66% of the eggs were recovered after passage, respectively. However, only 0.003 and 0.002% of the ingested eggs developed into third stage larvae (L3) after subsequent culturing. In pigs C and D, 0.01 and 0.02% of the L3 survived the passage of the gastrointestinal tract. Fresh O. ostertagi and C. oncophora eggs were cultured in parasite free porcine and bovine faeces. Only 0.05% L3 developed in porcine faeces, whereas 21% of the eggs developed into L3 in the bovine culture. Our results demonstrate an extremely poor rate of development and survival of both bovine nematode eggs and infective larvae after passage in pigs. It may imply that pigs can play an important role in reducing transmission of cattle nematodes if the two species are grazed together or alternately.     

Effect of temperature on development of the free-living stages of Ostertagia circumcincta

Hatching of the eggs of Ostertagia circumcincta was studied by recovering them from faeces and incubating them in distilled water at temperatures of 4, 16, 25 and 35 degrees C. Hatching occurred at all the temperatures. The rate of hatching increased with the rise in temperature. Development of larvae to the infective third stage (L3) was studied in faecal cultures incubated at 4, 16, 25, 30, 35, 40 and 45 degrees C. Except at 4 degrees C, L3 developed at all temperatures, the optimum temperature being 16 degrees C. The rate of development of L3 increased with the rise in temperature. This resulted in a corresponding decrease in the percentage recovery of larvae.     

Effect of time and temperature on growth of Salmonella enteritidis in experimentally inoculated eggs

Influence of time and temperature on Salmonella enteritidis multiplication in experimentally injected eggs was examined. There was an increase in the number of S. enteritidis with the increase in temperature of egg storage. There was less increase of S. enteritidis in eggs stored at 4 degrees C than in eggs held at temperatures higher than 4 degrees C (P less than 0.05). These results suggest a possible method for monitoring commercial eggs for the presence of S. enteritidis. It was concluded that the chances of recovery of S. enteritidis can be increased 10(6)-fold or more by holding the eggs at temperatures of 21 or 27 degrees C for more than 20 days and culturing their contents.     

Antigenic differences between the life cycle stages of Cooperia oncophora

The antigenic differences among the life cycle stages of Cooperia oncophora were studied by SDS-gel electrophoresis and Western blotting. Sodium dodecylsulphate polyacrylamide gel electrophoresis of the different life cycle stages of C oncophora revealed a complex protein pattern with a decreasing number of protein bands towards the adult stages. Several bands of the fourth stage larvae were in common with both the third stage and the adult nematode. Western blotting with sera from C oncophora monoinfected calves showed that the antigens of the fourth stage larvae were recognised predominantly and the presence of stage specific antigens in all stages. Strong cross reactivity was demonstrated when serum from Ostertagia ostertagi infected calves was used.     

Parasites of domestic and wild animals in South Africa. XLV. Helminths of dairy calves on dry-land Kikuyu grass pastures in the Eastern Cape Province

Successive pairs of approximately 4-month-old Friesland bull calves, raised under worm-free conditions, were exposed to helminth infection for 14 days on dry-land Kikuyu grass pastures at 28-day to monthly intervals, on a coastal farm in a non-seasonal rainfall region of the Eastern Cape Province. With the exception of one pair of calves exposed for 28 days, this procedure was repeated for 28 consecutive months from December 1982 to March 1985. The day after removal from the pastures one calf of each pair was slaughtered and processed for helminth recovery and the other 21 days later. Both members of the last four pairs of calves were killed 21 days after removal from the pastures. Sixteen nematode species were recovered from the calves, and infection with Ostertagia ostertagi was the most intense and prevalent, followed by Cooperia oncophora. The calves acquired the greatest number of nematodes from the pastures from June to October of the first year and from June to August of the second year of the survey. Few worms were recovered from the tracer calves examined from November or December to March or April in each year of the survey. The seasonal patterns of infection with Cooperia spp., Haemonchus placei, Nematodirus helvetianus, Oesophagostomum spp., O. ostertagi and Trichostrongylus axei were all similar and were negatively correlated to atmospheric temperature and evaporation. Slight to moderate arrest in the development of fourth stage larvae occurred from July to September in Cooperia spp., April to July in H. placei, and August to October in O. ostertagi and Trichostrongylus spp. during the first year of the survey. Too few worms were present in the second year to determine a seasonal pattern of arrest. Species survival during the hot and windy summer months appeared to be achieved via a combination of arrested larval development and an ageing residual population of adult worms in the host, and a small extant population of infective larvae on the pastures.     

Establishment and pathogenicity of two strains of Ostertagia ostertagi and Cooperia oncophora in calves in different locations.

An experiment was carried out simultaneously in Glasgow and in Wageningen to investigate possible differences between the local strains of Ostertagia ostertagi and Cooperia oncophora. In each location calves of the local Friesian breed were infected with 100,000 larvae of either the Glasgow or Wageningen strain of O ostertagi or C oncophora. At both locations the calves received the same diet. The Glasgow strain of O ostertagi was more pathogenic than the Wageningen strain and a larger proportion of the worm burden was found in the abomasal mucosa. The number of ova per female was greater in the Wageningen strain. For C oncophora the Wageningen strain gave rise to higher worm burdens and longer worms. Differences were also present between locations. The British Friesians had higher worm burdens of C oncophora and the worms of this species were longer in this host. Compared with the Dutch Friesians the British calves had a higher proportion of O ostertagi in the mucosa. This experiment showed how difficult it is to compare data from the literature because of differences in parasite and host strains and laboratory techniques.     

Some observations on the effects of age of calves on the phagocytosis and killing of Staphylococcus aureus by polymorphonuclear leucocytes

The effects of age on bovine polymorphonuclear (PMN) cell function were investigated by comparing the efficiencies of phagocytosis and killing of Staphylococcus aureus by peripheral blood leucocytes sequentially obtained from 15 calves between the ages of less than 1 and 84 days. One group of seven calves was kept in a controlled environmental chamber with air temperature of 5 degrees C and 58% relative humidity (RH) and another group of eight calves was kept at 16 degrees C and 58% RH. The calves were given a diet a liquid milk substitute and dry food, and were weaned abruptly from the liquid diet at 35 days of age. The in-vitro efficiencies of phagocytosis, and of killing, Staphylococcus aureus by peripheral blood leucocytes were similar for calves in air temperatures of 5 degrees C and 16 degrees C (P greater than 0.05). Peripheral blood leucocytes obtained from calves of less than 1 day of age were more efficient in phagocytosing S. aureus than those obtained when the same calves were 14-84 days of age (P less than 0.001). Peripheral blood leucocytes obtained when the calves were 42 and 56 days of age were significantly less efficient in phagocytosing and killing S. aureus than those obtained when the same calves were less than 1, 14, 28, 70 and 84 days of age (P less than 0.001).     

Interactions between lungworms and gastrointestinal worms in calves

Interactions between gastrointestinal worms (Ostertagia ostertagi, Cooperia oncophora) and lungworms (Dictyocaulus viviparus) in calves were studied by assessing the effect of primary infections with either group of worms on the development of homologous or heterologous challenge infections. Primary infections with lungworms resulted in some degree of resistance to challenge with gastrointestinal worms, but this resistance was lower than that found after homologous infection. Primary infections with gastrointestinal worms did not confer any resistance to challenge with lungworms. On the contrary, an indication was found of some enhancing effect of previous gastrointestinal worm infection on the establishment of lungworms. The highest degree of resistance against lungworm challenge was found where calves have been primarily infected with lungworms. Lungworm infections produced some elevation of serum pepsinogen levels. Gastrointestinal worms evoked a rise in circulating eosinophils, although this rise was smaller and occurred later than in lungworm-infected calves. Under the conditions of the experiment, the effect of 6000 infective lungworm larvae on weight gain was larger than the effect of 100,000 L3 of Ostertagia ostertagi and 100,000 L3 of Cooperia oncophora.