Suspected resistance of Ostertagia ostertagi in cattle to levamisole

Observations on a beef cattle farm in Flanders led to the suspicion of resistance to levamisole in a strain of Ostertagia ostertagi. After treating a group of six animals with levamisole (5 mg kg-1 L.W., i.m.) the reduction in the number of trichostrongylid eggs per gram of faeces varied between 0 and 66.6%, whereas a similar group treated with fenbendazole (7.5 mg kg-1 L.W., p.o.) showed a reduction in worm burdens of 100%. Coproculture showed that the remaining eggs in the first treatment group were all Ostertagia sp. The suspected field strain was compared with a reference strain of O. ostertagi by means of the in vitro larval paralysis test. This test showed LC95 values of 9.12 micrograms ml-1 and of 99.04 micrograms ml-1 for the reference and the field strain respectively, which indicates a resistance factor for the latter of 10.9. These results were not unequivocally confirmed by the post mortem findings on a tracer calf necropsied 4 days after treatment with levamisole.     

Serum pepsinogen levels and Ostertagia ostertagi populations in clinically normal adult beef cattle

Serum pepsinogen levels and Ostertagia ostertugi populations in clinically normal grass-fed bullocks were investigated in three groups of 10 prime cattle aged between 2.5 and 2.75 years slaughtered in late summer (February), early autumn (March) and late autumn (May) respectively. Apart from occasional foci of mucosal hyperplasia abomasa were grossly normal. Serum pepsinogen levels ranged between 0.2 and 2.5 i.u./l with group means of 1.4,l.S and 1.3 i.u./l. O. ostertagi counts ranged between 0 and5,194 with group means of 734,630 and 701 worms. The composition of the worm populations varied with a higher proportion of adults recovered in February and very few worms from most cattle in March, suggesting the termination of a parasite generation. An increase in numbers of early fourth-stage larvae in May indicated exposure to a new generation. These changes were not reflected in the pepsinogen levels.

The findings are discussed in relation to the adequacy of the pepsinogen assay as a diagnostic aid in field infections, animal age, and correlations between pepsinogen levels and parasite populations.     

Fenbendazole against inhibited early-fourth-stage Ostertagia ostertagi in cattle

Abstract

Extract

Sir — I am grateful for the opportunity to reply to the criticisms raised by Mr Vincent ⁽⁵⁾ Vincent, K. 1977. (Correspondence). N.Z. vet.J., 25: 226–226. [Taylor &; Francis Online] , [Google Scholar]. It seems to me that the bulk of his argument is concerned with the statistical treatment of the data that I presented ⁽³⁾ Elliott, D. C. 1977. The effect of fenbendazole in removing inhibited early-fourth-stage Ostertagia ostertagi from yearling cattle. N.Z. vet. J., 25: 145–147. [Taylor &; Francis Online] , [Google Scholar]. However, he has apparently misunderstood the purpose of statistical analysis of results, and also seems to have missed the point of my paper. For example, he states that my results “are contradictory to all previous trials (sic) results”. My interpretation. arising from the method of analysis chosen, is that under the conditions of my trials, any apparent effect of fenbendazole could be due to chance.     

Serum pepsinogen levels and Ostertagia ostertagi populations in clinically normal adult beef cattle

Serum pepsinogen levels and Ostertagia ostertagi populations in clinically normal grass-fed bullocks were investigated in three groups of 10 prime cattle aged between 2.5 and 2.75 years slaughtered in late summer (February), early autumn (March) and late autumn (May) respectively. Apart from occasional foci of mucosal hyperplasia abomasa were grossly normal. Serum pepsinogen levels ranged between 0.2 and 2.5 i.u./l with group means of 1.4, l.5 and 1.3 i.u./l. O. ostertagi counts ranged between 0 and 5,194 with group means of 734,630 and 701 worms. The composition of the worm populations varied with a higher proportion of adults recovered in February and very few worms from most cattle in March, suggesting the termination of a parasite generation. An increase in numbers of early fourth-stage larvae in May indicated exposure to a new generation. These changes were not reflected in the pepsinogen levels. The findings are discussed in relation to the adequacy of the pepsinogen assay as a diagnostic aid in field infections, animal age, and correlations between pepsinogen levels and parasite populations.     

Attempts to produce protection against Ostertagia ostertagi in cattle.

Various single or multiple doses of Ostertagia ostertagi were administered to young calves, and the production of protection phenomena elicited by single challenge inoculations ranging from 50,000 to 300,000 larvae or multiple challenge inoculations totaling 98,000 and 300,000 larvae was investigated. With some regimens, the vaccinations apparently resulted in protection against challenge exposure, as reflected by 36 to 56% fewer worms becoming established in challenge-exposed vaccinated calves than in challenge-exposed nonvaccinated, control calves. Other protection phenomena were elicited by some vaccinated calves of significantly more female worms lacking the distinctive vulval flap of O ostertagi and harboring significantly fewer eggs per female. Challenge exposure with a pathogenetic dose of 300,000 larvae produced the same degree of retarded weight gain in vaccinated as in nonvaccinated calves, and at necropsy, visceral lesions and pathologic alterations were equally severe in both groups of calves.     

Synergistic influence of Ostertagia ostertagi and Trichostrongylus axei on Ostertagia ostertagi larval inhibition and abomasal lesions in cattle

Parasite-free 4-month-old calves were inoculated with Ostertagia ostertagi and/or Trichostrongylus axei followed 6 weeks later by increasing doses of O ostertagi for 8 weeks. Clinical signs of parasitism, fecal egg counts, and plasma pepsinogen concentrations were monitored, and gross lesions and parasite burdens were determined postmortem. Clinical signs of parasitism were not observed and weight gains were not affected in experimentally infected calves. In calves infected with O ostertagi, mean plasma pepsinogen concentrations were greater than for control calves and were diagnostically significant 4 weeks after inoculation and during the last 4 weeks of serial inoculations with O ostertagi. In calves that were given O ostertagi and T axei, abomasal pH was significantly increased, and abomasal lesions were more pronounced than in control calves or in calves inoculated with only O ostertagi or T axei. Abomasal lymph nodes were enlarged in all parasitized calves; other lymph nodes in the calves inoculated with both O ostertagi and T axei were usually smaller than in calves inoculated with only O ostertagi or T axei. Numbers of O ostertagi-inhibited larvae were small in all inoculated calves, but the percentage inhibition was significantly greater in calves inoculated with both O ostertagi and T axei. The percentage inhibition was 3.53% for the O ostertagi-inoculated calves and 7.07% for calves inoculated with both O ostertagi and T axei. These percentages indicated a synergistic effect of concurrent abomasal parasitism, whereas a synergistic effect on T axei worm burden was not observed. The low percentage of larval inhibition indicated that factors other than host resistance are involved in naturally occurring pretype II ostertagiosis.     

Immunity to Ostertagia ostertagi

Control of ostertagiasis is a major economic problem in the temperate areas of the world. An immunological control scheme for cattle is an important aspect of integrated control and its achievement is dependent on our understanding of how immunity is acquired. Also, host mechanisms regulated by Ostertagia need to be understood in order to address immunological questions. Little is known about acquired immunity to Ostertagia infection in cattle. The degree of immunity is incomplete and its development slow. Evidence is accumulating for impairment of antibody and cellular immune responses by Ostertagia which is responsible for the survival of the parasite in the young host. Other experimental work suggests that local factors, such as eosinophil accumulation, mucosal mast cell kinetics and other cellular and humoral immune responses, may play a role in pathogenesis and immunity. At the present time these areas are also totally unexplored in the study of Ostertagia infection.     

Further studies on the response to transplanted adult Ostertagia ostertagi in calves

Calves infected by surgical transplantation of adult Ostertagia ostertagi had raised levels of plasma pepsinogen and those in which the largest number of worms established also had elevated plasma gastrin concentrations. Despite the elevated plasma pepsinogen values, the abomasal pH of the animals did not change significantly, and there was no significant difference in the percentage establishment of adult parasites in calves previously infected with O ostertagi third stage larvae and those which had been maintained parasite-naive before transplant.     

Study on the inductive factors of hypobiosis of Ostertagia ostertagi in cattle

Two experiments were carried out to determine the causes producing the Ostertagia ostertagi hypobiosis phenomenon in cattle. In the first experiment, the effect of time on third-stage larvae in the environment was studied during a 2-year period. Three experimental paddocks contaminated with O. ostertagi eggs at different times of the year were used, and the levels of hypobiosis were recorded by using 'indicator' and 'tracer' calves. The results suggest that time as such is not a hypobiosis-inductive factor. The second experiment was conducted under laboratory conditions, where the effects of temperature and light on infective larvae were studied. Infective larvae were subjected to different conditions of temperature and light during 6 weeks, and then inoculated to parasite-naive calves, which were slaughtered after 4 weeks. Percentages of hypobiotic larvae in these calves varied from 3.5 to 94.8%, depending on the different storage conditions the larvae underwent before inoculation. Results suggest that increasing temperature and increasing time of light exposure simulating spring conditions would be the factors which act upon third-stage larvae inducing them to a later hypobiotic stage in the host.     

Plasma pepsinogen levels in some experimental infections of Ostertagia ostertagi in cattle

Two groups of calves were infected with larvae of Ostertagia ostertagi to establish large numbers of adults and arrested larvae. In one group symptoms of ostertagiasis were seen and there was a loss of three months growth; in the other, in which adult worms were removed by a single anthelmintic treatment, there was only a transient reduction in live-weight gain. Plasma pepsinogen levels were however the same in the two groups and followed the same course. Even after 25 weeks, when calves had been growing normally for up to three months, plasma pepsinogen values were still around 5 iu per litre, well above the level generally regarded as diagnostic of ostertagiasis. The relevance of these findings to the use of the test in the diagnosis of ostertagiasis is discussed. The literature is reviewed.     

Efficacy of oxfendazole against inhibited Ostertagia ostertagi in naturally infected cattle.

Oxfendazole was administered to pregnant cows at 2.5 and 5 mg/kg body weight to determine the anthelmintic efficacy against naturally acquired larvae which became inhibited at the early 4th stage. The experimental design included three groups of orally-treated cows, that is, 10 placebo treated control cows, 11 cows treated with 2.5 mg/kg of oxfendazole and 10 cows treated with 5.0 mg/kg of oxfendazole. Oxfendazole at 2.5 mg/kg body weight was 82 and 94% effective against EL-4 and adult O. ostertagi, respectively. At 5 mg/kg, Oxfendazole was 95 and 99% effective against EL-4 And adult O. ostertagi, respectively. The results suggested the use of a field dosage level of 5 mg/kg body weight oxfendazole where inhibited larvae may be encountered.     

Attempts to immunize cattle against Ostertagia ostertagi infections employing 'normal' and 'chilled' (hypobiosis-prone) third stage larvae.

Immunity to Ostertagia ostertagi infections in calves develops slowly and only becomes manifest towards the end of a grazing season in which they have been exposed to the parasite. In an attempt to hasten the onset of immune reactions, three immunization protocols were set up. Twenty four heifers were allocated into four groups. Beginning in January, animals in two of the groups were inoculated with four 1-monthly increasing dosages of either 'normal' or 'chilled' (hypobiosis-prone) larvae, those in the third group received a single large infection with 'chilled' larvae and those in the fourth group served as non-infected controls. All animals were turned out on a common pasture in late April. Development of immunity was evaluated through determinations of faecal egg counts, live weight gains, serum pepsinogen levels and specific serum antibody responses of three isotypes (IgG1, IgG2 and IgA). Significantly reduced egg excretions in the immunized groups were apparent early in the season, indicating that the immunizations had, in this respect, been efficacious. The 'chilled' and 'normal' larvae seemed equally efficient given as multiple and single infections. A single large dosage of 'chilled' larvae seemed to have adverse effects. Only moderate antibody responses were elicited probably because of low challenge infection level on pasture. Considerable variation in responses existed between and within the four groups, for which reason conclusions regarding correlations between antibody isotype responses and immune effects on parasites could not be made.     

Efficacy of fenbendazole against inhibited larvae of Ostertagia ostertagi in yearling cattle

Efficacy of fenbendazole, at doses of 7.5 and 10.0 mg/kg of body weight, against inhibited early 4th-stage larvae of Ostertagia ostertagi and other nematodes of the abomasum and intestinal tract, was investigated in naturally infected yearling heifers in late May 1982. In Louisiana, this is near the end of the period (March to May) in which maximal numbers of inhibition-prone larvae are acquired. The mean numbers of O ostertagi in 10 untreated control cattle were: adults, 4,880; developing 4th-stage larvae, 12,546; and inhibited early 4th-stage larvae, 167,931. At the 7.5 mg/kg dose level (10% liquid suspension) in 10 cattle, percentage reduction of O ostertagi in comparison with controls was: adults, 95.7%; developing 4th stages, 91.1%; and inhibited 4th stage, 55.0%. Percentage reductions of other genera were as follows: abomasum--Trichostrongylus axei, 99.6%; Haemonchus sp, 95.1%; intestinal tract--Cooperia spp, 97.8%; Trichostrongylus colubriformis, 100.0%; and Oesophagostomum radiatum 4th stage and adults, 100.0%. At the 10.0 mg/kg dose (10% liquid suspension) in 11 cattle, the percentage reduction of O ostertagi in comparison with controls was: adults, 98.6%; developing 4th stages, 92.9%; and inhibited 4th stage, 80.0%. Percentage reductions of other genera were: abomasum--T axei, 99.9%; Haemonchus sp, 98.8%; intestinal tract--Cooperia spp, 99.3%; T colubriformis, 100.0%; and Oes radiatum 4th stage and adults, 100.0%. Variability of efficacy against inhibited larvae was observed, particularly at the 7.5 mg/kg dose; at this dose, 7 of the 10 heifers in the group yielded in excess of 54,000 surviving larvae.(ABSTRACT TRUNCATED AT 250 WORDS)     

Response to transplanted adult Ostertagia ostertagi in calves

Plasma pepsinogen levels became elevated in groups of recipient calves immediately after transplant with adult Ostertagia ostertagi. These rises occurred in both previously parasite-naive calves and in calves which had experienced prior infection terminated with an anthelmintic either seven or 21 days before transplant. From the results it appears that adult O ostertagi play a significant role in the elevated plasma pepsinogen levels associated with bovine ostertagiasis.     

Blood gastrin and pepsinogen responses to subclinical infection with Ostertagia ostertagi in adult dairy cattle

Blood gastrin and pepsinogen responses to a single infection with 100,000 Ostertagia ostertagi infective larvae in lactating dairy cows were investigated. None of the infected cows showed signs of clinical ostertagiasis, nor was there any difference in live weight gain, milk yield or faecal egg count between groups. Pepsinogen levels of the infected group were significantly elevated between days 3 and 24 after infection (peak 1041 mU tyrosine; day 14). In contrast, there was no significant difference in blood gastrin levels between infected and control animals suggesting that few adult worms had become established in the former group. These data are compared with the increases in both gastrin and pepsinogen levels recorded in susceptible calves exposed to the same level, pattern and strain of ostertagia infection in a previous experiment. It is suggested that gastrin assay may be of value in adult cattle for indicating when elevated pepsinogen levels are merely associated with a rise in larval intake and not with the establishment of large adult worm burdens.     

Development of a copro-antigen capture ELISA for detecting Ostertagia ostertagi infections in cattle

The present study reports on the development of a copro-antigen capture ELISA for detecting Ostertagia ostertagi infections in cattle. The ELISA was based on polyclonal rabbit antibodies, which recognize O. ostertagi excretory/secretory antigens (ES). ES antigens are released by the metabolic active stages of the parasite in the abomasum, and passed in the faeces of the host. The detection limit of pure ES material was 30 ng ml(-1) in sample buffer and 125 ng ml(-1) in faecal extract. The test was evaluated using a follow up from six artificially infected calves. Elevated levels of Ostertagia coproantigens could be measured from 21 days after infection, indicating that only the presence of adult parasites can be detected. To evaluate the capacity of the assay to measure levels of infection, three groups of cattle were tested: 38 artificially infected calves, 17 naturally infected first grazing season calves and 16 naturally infected adult dairy cows. Optical densities were significantly correlated to the worm burdens of the animals and the ELISA had an overall sensitivity of 91% and a specificity of 45%. The test gave negative readings for faeces of animals carrying patent mono-infections with Cooperia oncophora.     

Improved methods for isolating DNA from Ostertagia ostertagi eggs in cattle feces

A multiplex PCR assay for differentiating strongyle eggs from cattle has recently been described; however, the egg disruption and DNA extraction procedures, though effective, are inadequate for large studies or clinical application. The purpose of this research was to evaluate methods for disrupting trichostrongyle eggs, then assess commercial kits for extracting egg DNA using Ostertagia ostertagi as a model species. Egg disruption procedures tested included probe sonication, bath sonication, bead beating, boiling, microwaving, proteinase K/SDS digestion, freezing, and various combinations of the above with the incorporation of sodium dodecyl sulfate. These procedures were evaluated in conjunction with four commercial DNA extraction kits: DNA Stool mini kit and DNeasy Plant kit (Qiagen), Fast DNA kit (QBiogene), and the MAP extraction kit (Tetracore). Results showed that egg disruption was best accomplished with the bead beater and ceramic beads, resulting in 100% disruption within 1min. When DNA extraction was preceded by the isolation of eggs from feces, all procedures except the Fast DNA kit produced PCR-ready DNA from at least two eggs. The DNeasy Plant kit allowed consistent detection of DNA released from one egg. Due to the morphological similarities among trichostrongyle eggs in ruminants, strongyle eggs in equids, and hookworm eggs, the methods described herein may have broad application to other nematodes.     

The removal of inhibited early fourth stage Ostertagia ostertagi from yearling cattle by MK 933, an ivermectin formulation

Abstract

An ivermectin? ? Stated to be 22,23-dihydroavermectin B₁. Supplied by Merck Sharp and Dohme (N.Z.) Ltd., as test material. formulation, MK 933, when administered subcutaneously at the rate of 200 ?g/kg to steers aged 12-14 months, gave a significant (P<0.01) removal of adult, inhibited early fourth stage and later fourth stage Ostertagia (predominantly O. ostertagi) spp.

The lowest, median and highest post-treatment counts of early and later fourth stage worms combined were: controls 33,000, 143,500, 233,000; animals treated with MK 933; <500, 2,500,12,000. For adult worms, the lowest, median and highest counts in the controls were 23,000, 43,000, 68,000, whereas in treated steers no count exceeded 500. The treatment was also highly effective against Trichostrongylus axei.