Gastrin and pepsinogen changes during an Ostertagia ostertagi challenge infection in calves

Daily changes in serum gastrin and pepsinogen concentration have been studied during two types of infection with Ostertagia ostertagi in calves. In a first experiment two calves were trickle infected (10 times 10,000 L3 Ostertagia) and two animals received a single infection of 100,000 L3 Ostertagia. Gastrin and pepsinogen changes are discussed in relation to adult wormburdens. The second experiment involved 8 calves and was designed to investigate pepsinogen and gastrin changes following a challenge infection in previously sensitized calves. The high dosed group was infected with 5,000 L3 O. ostertagi during 30 days, the low dosed group received 500 L3 O. ostertagi and group 3 served as uninfected control. At day 41 post infection all animals were treated with oxfendazole and on day 61 challenged with 100,000 L3 O. ostertagi. Only in the high dosed group a distinct pepsinogen and gastrin reaction was noticed. Both parameters dropped to almost preinfection levels after treatment. Two days post challenge a moderate rise (+/- 1,000 mU tyr) of the pepsinogen concentration was observed in the previously infected animals and gastrin showed a temporary slight increase in several animals 8 to 10 days post challenge. The effect of treatment and challenge infection is discussed in relation to gastrin and pepsinogen changes and immunity.     

Identification and isolation of a specific antigen with diagnostic potential from Dictyocaulus viviparus.

The purpose of the investigation was to isolate and identify a specific antigen of Dictyocaulus viviparus that can be used to diagnose lungworm infections in cattle. Somatic, excretion and secretion antigens of adult D. viviparus and somatic antigens of L3 larvae were examined in an indirect enzyme-linked immunosorbent assay (ELISA) to determine whether they cross-reacted with sera collected from calves with mono-infections of Fasciola hepatica. Ostertagia ostertagi, Ascaris suum, or Cooperia oncophora. Serum samples containing antibodies directed against F. hepatica, A. suum, and O. ostertagi cross-reacted with somatic antigens of adult D. viviparus; these sera cross-reacted less with excretion and secretion antigens. When somatic antigens of adult D. viviparus were analysed in a Western blot, a 17-kDa protein that did not react with the heterologous sera was detected. This protein was isolated by ultrafiltration and anion chromatography. Sera collected from calves infected with D. viviparus was tested in indirect ELISAs with either somatic antigens of adult D. viviparus or with a low molecular antigen fraction of this preparation containing the 17-kDa protein. The extinction values that were measured in both assays correlated well. We conclude that the 17-kDa protein isolated from somatic antigens of adult D. viviparus may be useful in developing an improved immunoassay to diagnose lungworm infections in cattle.     

Larval migration inhibition activity in abomasal mucus and serum from calves infected with Ostertagia ostertagi

The present study investigated whether abomasal mucus from calves naturally infected with gastrointestinal nematodes possessed larval migration inhibition (LMI) activity in vitro, and whether LMI activity was greater in mucus from previously immunised animals, compared to primary infected and uninfected calves. LMI activity was also assessed in serum from calves during both natural and artificial Ostertagia ostertagi infections, in an attempt to monitor the development of acquired immunity. Both abomasal mucus and serum exhibited larval paralysing activity. Although the LMI capacity of the abomasal mucus was very variable, the highest paralysing activity was consistently observed in mucus from previously immunised calves. LMI activity in serum increased significantly during both artificial and natural Ostertagia infections. After a challenge infection, sera from immunised animals showed a significantly higher LMI capacity, compared to previously uninfected calves. Moreover, serum LMI activity was significantly negatively correlated with Ostertagia worm counts after the challenge infection. The present results suggest that LMI activity in serum and/or abomasal mucus reflects a protective immune response against O. ostertagi in the abomasal mucosa.     

EFFICACY OF ANTHELMINTICS AGAINST CATTLE NEMATODES

The efficacies of fenbendazole and thiabendazole against artifically acquired cattle nematodes and the efficacies of fenbendazole, prabendazole and levamisole against naturally acquired cattle nematodes were studied in western Victoria. Fenbendazole significantly (P less than 0.05) reduced numbers of late and early fourth stage larvae (greater than 96%), artificially acquired Cooperla spp (100%) and naturally acquired adult Ostertagia ostertagi (98%) and Trichostrongylus axei (90%). Thiabendazole significantly reduced numbers of artificially acquired late fourth stage larvae (83%) and parbendazole significantly reduced numbers of adult O. ostertagi (72%) in one group of cattle. Fenbendazole was the most effective anthelmintic.     

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.     

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.     

Analysis by ELISA and Western blotting of antibody reactivities in cattle infected with Mycobacterium paratuberculosis after absorption of serum with M phlei

Preabsorption of cattle serum with Mycobacterium phlei was of value in eliminating falsely positive reactions in an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against M paratuberculosis. Specific antibody titres from 16 animals naturally infected with M paratuberculosis were unaffected by absorption. Analysis by Western blotting indicated that a different set of antigens of M paratuberculosis were recognised by serum from falsely positive reactors compared with that from animals with established infection. After experimental infection the time required for seroconversion in the ELISA in nine calves lay between 10 and 28 months, although one animal had not seroconverted after 30 months when the experiment ended. All animals shed M paratuberculosis in their faeces before seroconversion.     

Protection studies with a globin-enriched protein fraction of Ostertagia ostertagi

The protective capacity of an adult stage Ostertagia ostertagi globin antigen was tested in four vaccination experiments in cattle. In a preliminary experiment, calves were vaccinated three times intraperitoneally with 250 microg globin in Freund's adjuvant and challenged with a trickled infection of 25,000 infective larvae. In three subsequent field studies, calves were vaccinated twice or three times intramuscularly with 80-100 microg globin in Quil A and challenged with a natural gastrointestinal nematode infection on pasture. Higher globin-specific antibody levels were detected in the vaccinated calves than in the control animals in all vaccine trials. In the preliminary experiment, geometric mean cumulative egg counts in the globin group were reduced by 52% and total worm burdens were reduced by 28%, compared to the controls. In the first field trial cumulative faecal egg counts were reduced by 63% in the vaccinated calves. However, the reduction in faecal egg output in these two experiments was not statistically significant and no reduction in faecal egg counts was observed in the vaccinated animals in the two last field trials. In conclusion, vaccination of calves with O. ostertagi globin resulted in highly variable protection levels after challenge infection.     

Serodiagnosis of bovine cysticercosis by detecting live Taenia saginata cysts using a monoclonal antibody-based antigen-ELISA

An ante mortem antigen-ELISA-based diagnosis of Taenia saginata cysticercosis was studied in artificially (n = 24) and naturally (n = 25) infected cattle with the objective of further validating the assay as a field diagnostic test. Based on total dissection as the definitive method of validity, the assay minimally detected 14 live cysticerci in artificially infected calves and 2 in naturally infected steers. In natural infections, the minimum number of live cysticerci consistently detected by Ag-ELISA was 5 while in artificial infections it was above 14. However, other animals with 12 and 17 live cysticerci in artificially infected calves, and 1 and 2 live cysticerci in naturally infected steers, escaped detection for unknown reasons. Animals harbouring dead cysticerci gave negative reactions in the assay as was the case in non-infected experimental control calves. There was a statistically significant positive linear correlation between Ag-ELISA optical density values and burdens of live cysticerci as obtained by total dissection of both artificially infected calves (r = 0.798, n = 24; P < 0.05) and naturally infected steers (r = 0.631, n = 25; P < 0.05). These results clearly show the potential effectiveness of ante mortem monoclonal antibody-based antigen detection ELISA in the diagnosis of bovine cysticercosis in cattle. Its value lies in the diagnosis of infection in cattle as a screening test in a herd, rather than as a diagnostic test at the individual level, due to false positive and negative reactions. In a herd of heavily infected cattle, the assay may, however, provide for individual diagnosis. Nevertheless, more work is recommended to increase its sensitivity so as to be able to diagnose light infections consistently in the field.     

Pharmacodynamics, pharmacokinetics and faecal persistence of morantel in cattle and goats

Morantel could not be detected (<0.05 pg/ml)in the plasma of cattle or goats followingthe oral administration of morantel tartrate at a dose rate of 10 mg/kg
bodyweight. No morantel was detected in the milk of lactating goats except in
one animal where a concentration of 0.092 pg/ml was detected at 8 h after drug
administration. Morantel was highly effective against Cooperia oncophora infec-
tions in calves treated 6, 9 or 18 days after infection; however, was highly
effective against Ostertagia ostertagi only when treated 18 days after infection.
Morantel did not affect the fecundity of adult 0.ostertagi surviving treatment 18
days after infection which had similar average numbers of eggs in their uteri
(range 13.4 f 0.73-16.8 L 0.98) as did parasites from control animals (range
12.0 k 0.70-13.6 2 0.66). Morantel could be detected at a concentration of 96 k
4.5 pgig (dry weight) in the faeces of a calf 24 h after treatment with I0 mgikg
bodyweight of morantel tartrate. The concentration of morantel in replicate
samples of this faeces exposed to natural atmosphere, but not to soil or soil
organisms, declined slowly over the following 322 days. At day 322 after the start
of the experiment 8.8 pg/g of morantel could be measured in the remaining
faecal material. Throughout the faecal degradation study the concentration of
morantel in the crusts of the replicate sample pats was lower than the
concentration in the core samples.     

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.     

High concentration of serum gastrin immunoreactivity and abomasal mucosal hyperplasia in calves infected with Ostertagia ostertagi and/or Trichostrongylus axei

Parasite-free, 4-month-old-calves were inoculated with Ostertagia ostertagi and/or Trichostrongylus axei, followed 6 weeks later by inoculation with increasing doses of O ostertagi for 8 weeks in the 2 groups (n = 4) of calves that had been given O ostertagi. Gastrin immunoreactivity concentration in serum was measured before and after infection and was correlated with changes in mucosal thickness. Gastrin immunoreactivity concentration in preinoculation control sera ranged from 95.2 to 287.1 pg/ml, and increased values were measured in all parasitized calves after 15 weeks. Significantly (P less than 0.05) increased serum gastrin immunoreactivity concentration compared with the preinfection value, was found in calves infected with O ostertagi or T axei, and highly significant (P less than 0.01) values were observed in calves infected with both parasites. Abomasal mucosal hyperplasia was observed in all parasitized calves; increased mucosal thickness and mucosal cross-sectional area were most prominent in calves infected with O ostertagi and T axei.     

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.     

The activity of albendazole against adult and larval gastrointestinal nematodes in naturally infected calves in the Netherlands

Summary The activity of albendazole against gastrointestinal helminths in naturally infected calves in the Netherlands was tested. The calves were in their fist grazing season and kept in two groups of ten. One of these groups was grazed alternately with sheep. Five out of each group were drenched with albendazole (7.5 mg/kg) on the day they were housed (November 1). Before and 2, 14, and 28 days after treatment individual faecal samples were taken from all calves and larval cultures were made. Ten calves, six treated and four untreated, were killed for post mortem studies 14 days after treatment The remaining calves were slaughtered 14 days later. The drug was highly effective in reducing the egg output, measured as the number of larvae cultured per gram of faeces. Compared with the untreated calves, the reduction was more than 99% two days after treatment, 100% at 14 days, and 99% after 28 days. It was shown that egg output 28 days after treatment came from worms which had developed from arrested larvae of Ostertagia ostertagi that had survived treatment. Post mortem results showed an efficacy of 100% against adult O. ostertagi, of almost 100% against Trichostrongylus axei, and 100% against adult and larval Cooperia oncophora. Twenty-eight days after treatment, the reduction of arrested early fourth stages of O. ostertagi was 85% in comparison with the untreated calves. Apparently less effect was found against Trichuris ovis at the given dose rate.     

Efficacy of a topical formulation of eprinomectin against endoparasites of cattle in New Zealand

Two controlled studies involving 24 cattle were conducted in New Zealand to determine the efficacy of a topical, non-flammable formulation of eprinomectin against induced and naturally acquired nematode infections. In Trial 1, nematode infections were induced on Day -5 with third-stage larvae of Cooperia spp., Haemonchus contortus, Ostertagia ostertagi and Trichostrongvlus colubriformis so that the nematodes would be at the fourth larval stage when the cattle were treated. In Trial 2, cattle had naturally acquired nematode infections as determined by faecal nematode egg counts and larval cultures. The cattle were allocated on Day 0 (Trial 1) or Day 6 (Trial 2) on a stratified random basis according to bodyweight to one of two treatments: untreated control or eprinomectin (0.5% w/v) applied topically at 1 ml/10 kg bodyweight. Necropsies were undertaken on Days 14 and 15 and total nematode counts were done. In Trial 1, cattle treated with eprinomectin had significantly (p < 0.05) fewer Cooperia spp. and O. ostertagi than the controls. Larvae of H. contortus and T. colubriformis did not establish. In Trial 2, cattle treated with eprinomectin had significantly (p < 0.05) fewer of the following parasites than the controls: Haemonchus spp. (adult), Cooperia surnabada (adult), C. oncophora (adult), Cooperia spp. (L,), Ostertagia lyrata (adult), O. ostertagi (adult), Oesophagostomum spp. (adult), T. avei (adult and L1) and Trichuris spp. (adult). Reductions of 100% were observed for Capilfaria spp. (adult), D. viviparus (adult and L,), and Nematodirus helvetianus (adult), but these were not statistically significant (p > 0.05) because four or fewer control animals were infected with these parasites. In Trial 2, efficacies of greater than 99% were observed against all species for which moderate to high burdens occurred in the untreated controls. These findings indicate that eprinomectin in a topical formulation is a highly effective nematocide in cattle.     

Activity of fenbendazole against inhibited early fourth-stage larvae of Ostertagia ostertagi

The efficacy of fenbendazole (Panacur, Hoechst-Roussel) against inhibited early fourth-stage larvae of Ostertagia ostertagi and other nematodes of the abomasum and intestinal tract was investigated in naturally infected, yearling cattle in April 1978. The time when peak levels of inhibited larvae occurred was determined by epizootiologic study which began in November 1977. All animals were removed from pasture and maintained free from further helminth infection until slaughter (19 to 21 days). The fenbendazole liquid suspension was administered as an oral drench at dose level of 10 mg/kg to 10 animals and then at dose level of 15 mg/kg to an additional 10 animals at 10 days after removal from pasture. Eleven animals were maintained as untreated controls. In cattle given the dose of 10 mg/kg, the following reductions were observed: O ostertagi adults--100%, developing stages--80%, and inhibited larvae--97%; other worm genera in the abomasum and nematodes of the intestinal tract--100%. In the cattle given the larger dose, the following reductions were observed: O ostertagi adults--100%, developing stages--98%, and inhibited larvae--99%; other worm genera in the abomasum and nematodes of the intestinal tract--100%.     

Further tests of activity of levamisole on Ostertagia ostertagi in dairy calves with notes on overwinter survival of gastrointestinal helminths on pasture

Two controlled tests were conducted in 1981 and 1982 in dairy calves on the University of Kentucky research farm to determine activity of the bolus formulation of levamisole given at the dose rate of 8 mg/kg against naturally occurring infections of Ostertagia ostertagi. Removal efficacies of mature O ostertagi were 98% in the 1981 test (3 treated and 3 nontreated calves) and 94% in the 1982 test (7 treated and 8 nontreated calves). Against immature Ostertagia spp, removal efficacies were 100% and 65% for the 1st and 2nd tests, respectively. The calves were grazed on the same pasture as dairy calves in previous controlled tests in 1979 and 1980 where activity of levamisole against mature O ostertagi (data recently published) was much less than in the present tests. It does not appear that the poor performance in the early tests can be attributed to the drug resistance phenomenon. Data on overwinter survival (119 days) of free-living stages of gastrointestinal parasites on pasture were derived from the nontreated calves in the 1982 controlled test. The calves, raised helminth-free, were placed on the pasture on Apr 5, 1982. Helminths recovered at necropsy of the calves, besides O ostertagi, included Trichostrongylus axei, Nematodirus helvetianus, Nematodirus spp, Cooperia oncophora, Trichuris spp, and Moniezia spp. The lung-worm, Dictyocaulus viviparus, previously found in cattle on the farm, was not found in these calves.     

Use of diagnostic markers to monitor fasciolosis and gastrointestinal nematodes on an organic dairy farm

A 12-month study was conducted to assess and monitor gastrointestinal tract nematodes and liver fluke in cohorts of cattle on a Scottish organic dairy farm. Various diagnostic markers for helminth parasites of cattle from different age groups were assessed monthly from April 2007 to March 2008. First season grazing stock were subjected to significant challenge from Ostertagia ostertagi nematodes as reflected in serum pepsinogen concentrations, which rose markedly in the second half of the grazing season. In addition, plasma albumin concentrations decreased and faecal egg counts (FEC) increased moderately, indicating exposure to both O ostertagi and probably Cooperia oncophora. Second season grazing animals had a peak FEC early in the grazing period, suggestive of a potential carry-over of Ostertagia species infection ('Type 2') during housing. All classes of cattle showed evidence of fluke (Fasciola hepatica) infection. Adult cow exposure to O ostertagi and fluke was estimated via the use of ELISA testing to detect antibodies to O ostertagi and F hepatica and the high levels detected suggested a significant exposure response. Despite low stocking densities and sympathetic grazing management, there was a significant challenge to all grazing stock from gastrointestinal nematodes and liver fluke.     

Humoral immune response of water buffalo monitored with three different antigens of Toxocara vitulorum

Humoral immune response of water buffalo naturally infected with Toxocara vitulorum was monitored using three different antigens of this parasite in serum and colostrum of buffalo cows and calves. Soluble extract (Ex) and excretory/secretory (ES) larval antigens and perienteric fluid antigen (Pe) of adult T. vitulorum were used to measure the antibody levels by an indirect ELISA. Serum of 7-12 buffalo cows for the first 365 days and colostrum of the same number of buffalo cows for the first 60 days of parturition, and serum of 8-10 buffalo calves for the first 365 days after birth were assayed. The ELISA detected antibodies against all three T. vitulorum antigens in the colostrum and serum of 100% of buffalo cows and calves examined. The highest antibody levels against Ex, ES and Pe antigens were detected in the buffalo cow sera during the perinatal period and were maintained at high levels through 300 days after parturition. On the other hand, colostrum antibody concentrations of all three antigens were highest on the first day post-parturition, but decreased sharply during the first 15 days. Concomitantly to the monitoring of immune response, the parasitic status of the calves was also evaluated. In calves, antibodies passively acquired were at the highest concentrations 24 h after birth and remained at high levels until 45 days coincidentally with the peak of T. vitulorum infection. The rejection of the worms by the calves occurred simultaneously with the decline of antibody levels, which reached their lowest levels between 76 and 150 days. Thereafter, probably because of the presence of adults/larvae stimulation, the calves acquired active immunity and the antibodies started to increase slightly in the serum and plateaued between the days 211 and 365. All three antigens were detected by the serum antibodies of buffalo calves; however, the concentration of anti-Pe antibody was higher than anti-EX and anti-ES, particularly after 90 days of age. By conclusion, the buffalo cows develop immunity and keep high levels of antibodies against T. vitulorum-Ex, ES and Pe antigens and these antibodies are transferred to their calves through the colostrum. This passively acquired immunity does not protect the calves against the acquisition of the infection, but these antibodies, passively or actively acquired, may have an important role during worm rejection by the calves and prevention of intestinal reinfection.     

Evaluation of diagnostic tests for Johne's disease in young cattle

OBJECTIVE: To investigate the development of immune responses in calves experimentally and naturally infected with Mycobacterium paratuberculosis and to evaluate the potential for diagnostic tests to detect infected calves. DESIGN: Sequential testing of four treatment groups of calves over a 2 year period. PROCEDURE: Twenty-nine calves were allocated to four groups. Group D calves were orally dosed with M paratuberculosis, group N calves naturally exposed to M paratuberculosis, group V calves vaccinated for M paratuberculosis, and group C were control calves (not infected or vaccinated). Blood and faecal specimens were collected from each calf at monthly intervals to 18 months of age and then every 2 months until they were slaughtered between the ages of 21 and 29 months. Specimens were tested using absorbed EIA, IFN-gamma EIA and faecal culture. The infection status of the calves was confirmed by extensive histopathological examination and tissue culture. RESULTS: M paratuberculosis infection was confirmed in 10 calves, comprising six of eight orally dosed calves, three of five naturally exposed calves and one of nine vaccinated calves. The six artificially infected calves and one naturally infected calf were detected shedding M paratuberculosis in their faeces. Results with positive absorbed EIA were obtained from one artificially infected calf, one naturally infected calf and three vaccinated calves. All calves including controls had positive results on at least one occasion using the IFN-gamma EIA. In addition, seven calves had positive bovine tuberculosis results using the IFN-gamma EIA, even though bovine tuberculosis has been eradicated from Australia. CONCLUSION: Detection of M paratuberculosis infection in young cattle continues to be difficult using current tests.