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.     

Ostertagia ostertagi infections in angora goats

Angora goats grazing on a paddock contaminated with Ostertagia ostertagi and O. circumcincta were found to have infections consisting of equal numbers of both species. Subsequently, goats were infected with O. ostertagi larvae of bovine origin. The infection became patent by day 28 and egg counts reached a plateau about 7 weeks after infection. Worm numbers did not decline during the 12 weeks of the experiment. It was concluded that Angora goats are suitable hosts for O. ostertagi.     

Experimental infections of Ostertagia ostertagi in goats

In order to determine the usefulness of the goat as a model host for Ostertagia ostertagi, a series of experiments was conducted in which young goats and calves were experimentally infected with L3 of calf-source and goat-source isolates. The goat-source isolate was derived from a continuous passage of the bovine parasite in goats. Patent infections resulted in 73 out of 86 inoculated goats (85%). The largest number of patent infections was observed when inoculation consisted of a single dose of goat-source larvae. Percentage establishment of infection was generally low in goats inoculated with either larval source. Time taken to achieve patency in goats was frequently within the range normal for cattle infections, but was often extended (21-67 days). With the exception of the generally higher level of establishment of goat- or calf-source isolates in calves and the low frequency of the vulval flap in adult female worms established in goats, little difference was observed in percentage establishment or worm population characteristics of the two isolates in goats.     

Control of Ostertagia ostertagi infections in Australia

The control of Ostertagia ostertagi infections in Australia is aimed specifically at young cattle in their first and second year of grazing after weaning. Mature breeding stock are not routinely treated. The recommended strategy is preventive, using an integrated approach of timed anthelmintic treatments in relation to the epidemiology of ostertagiasis in different environments. Best results are obtained when treatments, given at weaning and 6 months later, are combined with a move to 'safe' pastures which have not been grazed by cattle for the previous 6 months. Good results are also obtained in the winter rainfall regions when set-stocked cattle are treated before and during autumn to prevent contamination of pastures at this time. More complicated grazing management, involving the spelling of pastures during autumn and winter, combined with anthelmintic treatments, is needed in some summer rainfall regions where weather conditions are especially favourable for the development and survival of the free-living stages of O. ostertagi.     

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.     

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.     

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.     

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.     

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.     

The efficiency of levamisole, thiabendazole and fenbendazole against naturally acquired infections of Ostertagia ostertagi in cattle

In two experiments, conducted in cattle with naturally acquired infections of Ostertagia ostertagi, comparative assessments were made of the anthelmintic efficiency of levamisole, thiabendazole and fenbendazole, each at 1.0, 1.5 and 2.0 times the recommended dose rate. Variable efficiencies of 81 and 49 per cent for levamisole, 86 and 56 per cent for thiabendazole were obtained against adult O ostertagi. Neither drug showed substantial activity against early fourth stage larvae. Efficiency of fenbendazole against adult O ostertagi was consistently high; 85 and 89 per cent in the two experiments respectively. In the first experiment in which cattle were slaughtered two to three days after treatment, only 22 per cent of inhibited early fourth stage larvae were removed whereas in the second experiment when slaughter took place 10--11 days after treatment, this efficiency was 89 per cent. There was no increased effect of increased dose rates on treatments with thiabendazole or fenbendazole. The activity of levamisole against adult worms and inhibited larvae was increased at twice the recommended dose rate.     

Pathophysiological and parasitological studies on Ostertagia ostertagi infections in calves

Friesian calves given a low level infection of the abomasal parasite Ostertagia ostertagi over a six week period displayed a mild diarrhoea with high faecal egg counts and elevated plasma pepsinogen values. At necropsy on day 23 abomasal lesions characteristic of ostertagiasis were widespread. At 42 and 84 days oedema and congestion were also prominent. Total worm burdens on days 23 and 42 were similar but a marked decrease had occurred by day 84. Feed digestibility and nitrogen economy were not markedly affected but radioisotopic measurements demonstrated an increase in albumin disappearance and catabolic rates, and plasma faecal clearance during the course of the infection. Prior administration of a morantel sustained release bolus to a group of similarly infected calves reduced the total worm burdens to less than 50 per cent of those recorded in the infected calves on days 23 and 42 and this fell to 3 per cent on day 84. Abomasal damage and the adverse pathophysiological changes associated with infection were prevented in this group.     

Pathogenesis of simulated natural infections with Ostertagia ostertagi in calves

The possibility of a mucosal hypersensitivity reaction and its relationship to the pathogenesis of simulated natural infections with Ostertagia ostertagi were studied in calves. Four groups of 4 calves each were used. One group was used as noninfected control; a 2nd group was given increasing doses of infective larvae; a 3rd group was given increasing doses of larvae and these were removed by succeeding treatment with an anthelmintic; and a 4th group was given an initial dose of larvae which was then eliminated with an anthelmintic. All calves given larvae became sensitized, as shown by an intradermal skin test. The continuously infected calves had significantly (P less than 0.05) higher fecal egg counts, eosinophil counts, plasma pepsinogen values, and worm burdens and significantly (P less than 0.05) lower lymphocyte counts than did the other groups of calves. These animals also had the most extensive mucosal pathologic changes. The group given intermittent larval challenge exposures followed by an anthelmintic showed decreased lymphocyte values, but these were not significant. Plasma pepsinogen values of this group increased between every challenge exposure and treatment, a 3-day period. This indicated that a mucosal hypersensitivity reaction had occurred in these calves at these times, because they were shown to have been sensitized, and challenge-exposure infections were not present for sufficient time to have produced direct pathologic effects. It therefore seems that a part of the pathologic changes in O ostertagi infections may be the result of the continuous challenge exposure experienced by the animals through a constant intake of larvae from pasture and the intestinal reaction to this challenge exposure.     

Applying a kinetic method to an indirect ELISA measuring Ostertagia ostertagi antibodies in milk

Indirect enzyme-linked immunosorbent assays (ELISAs) are frequently run as endpoint ELISAs (e-ELISAs). However, kinetic ELISAs (k-ELISAs) have certain advantages over e-ELISAs. The objective of this study was to understand the relationship between e-ELISA and k-ELISA results. Specifically, to determine whether it was possible to run both k-ELISA and e-ELISA on the same plate and establish an appropriate time interval for k-ELISA measurements. A normalization method for k-ELISA slopes (slope ratio) is proposed. Using an indirect e-ELISA test measuring antibodies against Ostertagia ostertagi in milk from dairy cattle, we found that running a k-ELISA had no effect on optical density ratio results of an e-ELISA on the same plate, and that agreement was very strong at 10, 15, and 28 min, allowing for a reduction in the total processing time for ELISA tests.     

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.     

Development of immunity to Ostertagia ostertagi (Trichostrongylidae: Nematoda) in pastured young cattle.

This experiment comprised 3 groups of calves, (+P2), (-P2) and (-P1), which all started their first grazing season as parasite-free calves. The (+P2)- and (-P2)-group grazed 2 seasons. In the first season the (-P2)-group of calves was grazing a pasture with no detectable trichostrongyles and treated with anthelmintics every second week. The untreated (+P2)-group grazed an Ostertagia ostertagi contaminated pasture. During the second grazing season these 2 original groups grazed together with a new group of first-year grazing calves (-P1) on paddocks infected with O. ostertagi. Parasitological analyses showed that (+P2)-group had negligible egg excretions in the second year in comparison with (-P2) and (-P1). This indicated, that the egg output may be regulated through acquired immunity. The difference in egg excretions was not reflected in the serum pepsinogen levels, which were only slightly elevated for all groups in the second year. Post mortem examination at the end of the experiment showed that only the (-P1)-group harboured relatively high numbers of worms in the abomasa at that time. Antibodies of 3 immunoglobulin classes were investigated: IgA, IgG1 and IgG2. The IgA and IgG1 responses correlated with the presence of developing and adult worms in the abomasa and they remained elevated in the (+P2)-group throughout the experiment, perhaps indicating an involvement of these antibodies in a protective immune response. In the (-P2)-group the IgA and IgG1 showed fast and sharp rises during the second season that most likely were age-related and as such a result of maturation of the immune system. The role of IgG2 is unclear as the IgG2 response was weak in all groups of calves and difficult to relate to the parasitological data.     

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.     

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)     

Assessment of the repeatability of a milk Ostertagia ostertagi ELISA and effects of sample preparation

An indirect Ostertagia ostertagi ELISA on milk is a promising diagnostic tool in bovine parasitology. Interpretation of the test results requires a good knowledge of the test characteristics. In this study, border effects, the repeatability of the ELISA and the effect of different factors such as storage, skimming and freeze–thaw cycles of the milk samples were investigated. The border effects trial showed that significant border effects can occur. The repeatability trial was conducted over 3 days. An alternative graphical technique to assess the repeatability over a large number of ELISA plates measured over different days was developed. From these graphs, it was obvious that the ODR values obtained on the third day were deviating from the values on the first and second day. On the third day, also abnormal control values were observed. When the control values were normal, 94% of the variability was explained by the milk sample and 6% by assay variability. The expected 95% range of the difference of 2 ODR readings of the same sample on the same plate and the same sample on different plates was −0.14 to 0.14 and −0.16 to 0.16. No extra variability was observed when samples were tested on a different day, however these results are based on the measurement of 2 days. Storage for 2–4 days at 4 °C, using whole milk instead of skimmed milk and up to 2 extra freeze–thaw cycles of the milk samples did not significantly affect the test results.