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.     

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.     

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.     

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.     

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.     

Experimentally induced ostertagiosis in rabbits inoculated with Ostertagia ostertagi of bovine origin

Weanling specific-pathogen-free rabbits were orally inoculated with 50,000 Ostertagia ostertagi 3rd-stage infective larvae cultured from bovine feces and were killed 42 days after inoculation. Two preliminary trials were done, using conventionally reared weanling rabbits inoculated with 2,500, 5,000, and 50,000 O ostertagi 3rd-stage larvae and dexamathasone in 1 of the trials; the rabbits were killed 14 and 28 days after inoculation. Fecal egg counts were monitored, and worm burdens were determined. The pH of the gastric contents was measured at necropsy. Recoveries of the parasite from the gastric mucosa comprised early 4th-stage larvae and larvae that had developed beyond the early 4th stage. The maximum worm burden established at 42 days after inoculation was 1,668 immature nematodes or 3.34% of the inoculum. Gross and microscopic gastric lesions were present. Gross nodular mucosal elevations, which tended to be confluent in the cardiac region, were observed. Microscopic changes were principally mucosal hyperplasia, focal lymphocyte accumulation with moderate glandular dilation, and slight variable eosinophil and plasma cell accumulation. The pathologic changes had characteristics similar to those in cattle infected with O ostertagi and other conditions characterized by gastric mucosal hyperplasia.     

Forms of bovine pepsinogen in plasma from cattle with three different 'syndromes' of Ostertagia ostertagi infection

Calves infected orally with third stage larvae of Ostertagia ostertagi or infected with adult O ostertagi by direct transplantation into the abomasum had raised plasma pepsinogen activity, as did four-year-old dairy cattle challenged with O ostertagi third stage larvae on five occasions. Using fast protein liquid chromatography two forms of pepsinogen; pepsinogen 1 (PG1) and pepsinogen 2 (PG2) were identified in each of the parasitic infection regimes.     

Activity against Ostertagia ostertagi of low doses of oxfendazole continuously released from intraruminal capsules in cattle.

Slow release from intraruminal capsules of 0.48 mg/kg oxfendazole per day for 5.5 days gave percentage efficiencies of 99 +/- 0.6, 86.6 +/- 6.3 and 93.1 +/- 4.5 for the removal of adult worms, developing 4th stages and inhibited larvae of Ostertagia ostertagi respectively. These efficiencies were not significantly different from those obtained from a single oral dose of 2.5 mg/kg. Peak plasma levels of oxfendazole were similar for the 2 types of administration but high levels were maintained longer in cattle given capsules.     

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)     

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.     

Immunosuppression of lymphocyte blastogenesis in cattle infected with Ostertagia ostertagi and/or Trichostrongylus axei

Twenty 4-month-old calves were infected with O ostertagi and/or T axei and the responses to phytolectins were evaluated. Whole blood cultures were incubated with phytohemagglutinin (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM). The blastogenic response was determined by tritiated thymidine uptake with results presented as counts per minute (cpm), stimulation indices (SI) and a mononuclear cell responsive index determined by dividing the phytomitogen induced cpm by the absolute mononuclear cell number per ul. The control group results were adjusted to 100 percent and changes in the percentage difference by the parasitized calves was determined. There was a decline in lymphocyte responsiveness to PHA beginning at the time of infection. Significant depression of responses to PHA was observed in all parasitized calves 8 weeks after infection although clinical signs of parasitism did not occur. Lymphocyte responses to PW, were not different in infected calves from the control, although the O ostertagi group had significantly higher PWM mean upon than the calves infected with T. axei. A slight depression in response to Con A was also observed at 8 weeks after infection followed by a significant increase after 10 weeks. The immunosuppression appeared to be a feature of gastrointestinal parasitism and related to infections with O. ostertagi and/or T. axei.     

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.     

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.     

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.     

Infection with Ostertagia ostertagi in goats and calves

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. Percent 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 percent establishment or worm population characteristics of the two isolates in goats as based on source of larval inoculum, inoculation course, and age of host at inoculation. Prolonged passage of infection in goats did not result in stabilized isolate more adapted to the goat or less adapted to calves. Fecal egg counts were generally minimal or negative in goats during the first 30 days of infection, but were often increased and not substantially lower than counts in calf infections after 60 or 90 days. Low level egg counts in goats were observed to persist for up to 17 months. During the spring of 2 years, goat kids grazed on a cattle pasture acquired O. ostertagi infections which included adult worms, but a larger number of early L4. The latter were presumed to be inhibited in development just as such inhibition occurs in cattle during spring.     

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.     

Concurrent daily infection of calves with Fasciola hepatica and Ostertagia ostertagi.

Three groups of calves were infected daily with either 1500 Ostertagia ostertagi larvae, 20 Fasciola hepatica metacercariae, or 1500 O ostertagi plus 20 F hepatica metacercariae. Weekly measurements were taken of calf weight, faecal egg output, plasma concentrations of albumin, plasma activities of sorbitol dehydrogenase, gamma glutamyl transpeptidase and pepsinogen and standard haematological indices. Calves were killed either 10 or 21 weeks after daily infections began. F hepatica infection had little influence on the size and structure of the O ostertagi worm population or vice versa. Mean worm burdens found at 20 weeks in those animals infected with both F hepatica and O ostertagi were 293 flukes and 20,641 nematodes. While this level of infection is similar to that seen in the disease complex in the field, there was no evidence of clinical disease or any difference in weight gain between the groups in this experiment. Factors other than additive worm burdens are obviously important for the expression of disease under field conditions.     

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.