Persistence of dead Ostertagia ostertagi in the abomasal mucosa following anthelmintic treatment

Anthelmintic trails, conducted with albendazole, fenbendazole and ivermectin for efficacy against gastrointestinal nematodes, principally inhibited early fourth larval stages of Ostertagia ostertagi in naturally infected cattle. Cattle wee slaughtered seven to 20 days after treatment. O ostertagi was the predominant abomasal nematode recovered with occasional small numbers of Haemonchus species and Trichostrongylus axei. Control calves uniformly had very large O ostertagi infections, primarily early fourth stage larvae. Viable surviving worms and variable numbers of dead and degenerate worms were recovered in abomasal contents and washings. These O ostertagi larvae and adults were characterised by adherent debris or proteinaceous material, degenerated cuticles and distortion of internal structures. This study demonstrated the necessity for proper timing of slaughter for anthelmintic trial evaluation to allow clearance of dead nematodes, specifically O ostertagi larvae which are sequestered in the abomasal glandular tissue. Nematode collection within seven to 12 days after treatment will include dead and degenerate larval nematodes. The peripheral coating of larvae was suggestive of the Splendore-Hoeppli effect which has been associated with immunological responsiveness. The antigenic stimulus for this material and the lymphocyte and eosinophil infiltration was suspected to be early fourth stage O ostertagi larvae within the mucosa but was not identified definitively.     

Epidemiology of abomasal nematodes of dairy cattle in Hokkaido, northern Japan

The prevalence and intensity of infection with abomasal nematodiasis was studied in dairy cattle of Hokkaido, northern Japan, for successive two years. During the period of March in 1985 to September in 1987, a total number of 393 abomasa of Holstein-Friesian cows was examined for nematode parasites. Nematodes were detected from 75% of the cows. The prevalence of nematode species detected was Ostertagia ostertagi 250 (63.6%), Mecistocirrus digitatus 181 (46.1%), Trichostrongylus axei 85 (21.6%) and Haemonchus sp. 1 (0.3%). The prevalence and population composition of each growth stage varied seasonally in O. ostertagi and M. digitatus. The large percentage of arrested larvae, early L4 O. ostertagi and immature L5 M. digitatus, detected during the mid-winter and the increasing percentage of matured adult populations of both species in early spring revealed the occurrence of the autumn associated arrested development (hypobiosis) phenomenon in bovine abomasum nematodes of Japan.     

Efficacy of febantel against abomasal nematodes and lungworms in cattle

The efficacy of febantel at a dosage of 5 mg/kg (45.5% paste formulation) against inhibited early 4th-stage larvae (EL4) of Ostertagia ostertagi, other nematodes of the abomasum, and Dictyocaulus viviparus was investigated in 4- to 6-month-old Holstein calves that grazed on pasture heavily contaminated with parasites from February 24 to April 1, 1986 (36 days). In Louisiana, this is the first month of a 3-month period in which increasing numbers of inhibition-prone O ostertagi larvae are acquired, and infection risk with D viviparus may remain high. Three of 4 calves that died of lungworm infection during the pasture-exposure period were necropsied. Large numbers of abomasal nematodes, including inhibited O ostertagi larvae, and large numbers of D viviparus were recovered. Twenty-five calves were randomly allotted by equal distribution of body weight to 2 groups and treated on April 4: placebo-treated calves (n = 13) and febantel-treated calves (n = 12). Equal numbers of treated and control calves were killed at 6 and 7 days, respectively, after treatment. Mean numbers of O ostertagi in control cattle were: adults, 4,931; developing 4th-stage larvae (DL4), 1,119; and inhibited EL4, 3,410. Ostertagia lyrata, Trichostrongylus axei, Haemonchus sp, and D viviparus were well distributed in nearly all control calves. Percentage reduction of O ostertagi in treated calves, when compared with controls, was: adults, 83.6%; DL4, 57.8%; and inhibited EL4, 34.8%. Percentage reductions of other species were: O lyrata, 92.6%; T axei adults, 99.3% and 4th-stage larvae (L4), 100%; Haemonchus sp adults, 66.7%, and L4, 64%; D viviparus adults 90.6%, and immature forms, 97.1%.(ABSTRACT TRUNCATED AT 250 WORDS)     

Exsheathment of Ostertagia ostertagi infective larvae following exposure to bovine rumen contents derived from low and high roughage diets

The objective of this study was to characterize the exsheathment kinetics of Ostertagia ostertagi infective larvae (L3) following in vivo exposure to bovine rumen contents derived from low and high roughage diets. O. ostertagi L3 were placed in disposable dialysis bags and incubated for various time points between 0 and 360 min in the rumen of a fistulated steer maintained on a 71% grain diet or a 100% grass diet. The maximum percentage of exsheathed L3 was observed 120 min post-exposure to grass-derived rumen contents, while maximum exsheathment for L3 exposed to grain-derived rumen contents did not occur until 360 min. This work provides the first report of the in vivo exsheathment kinetics for O. ostertagi in its bovine host. Results of this study also support earlier reports that rumen pH may affect the exsheathment efficiency of abomasal trichostrongylids.     

Immune modulation by Ostertagia ostertagi and the effects of diet

IgG1 antibody responses to Ostertagia ostertagi third stage larvae (L3) and the third party antigen, keyhole limpet haemocyanin (KLH), and faecal egg counts were determined in calves infected with a single dose of O. ostertagi and in uninfected, pair-fed calves. The infected and uninfected calves were given diets either high (H) or low (L) in protein and energy. The diets were within the normal range of husbandry practice in the UK. IgG1 antibody responses to L3 antigen were significantly greater from 6 weeks post-infection in infected calves given the L diet than in infected calves given the H diet (P less than 0.05). The effects of diet and infection on anti-KLH IgG1 responses were independent of each other. IgG1 responses to KLH were decreased by infection and by the L diet compared with the H diet.     

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.     

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 levamisole against Ostertagia ostertagi in Louisiana cattle during maturation of inhibited larvae (September) and during minimal inhibition (December/January).

Levamisole (LEV) was tested in four experiments to compare efficacy values against Ostertagia ostertagi when larval maturation was occurring (September), following inhibition and also when populations were expected to be largely adult (winter). A primary objective was to determine the importance of developing fourth-stage larvae (DL4) and inhibited, early fourth-stage larvae (EL4) in replacing adult worms lost through treatment and the effect of this on reduced efficacy against adult worms. Young crossbred beef calves ranging in weight from 150 to 230 kg were used in the first (September 1981), second (September 1983) and third experiments (January 1987). Jersey calves of 110 kg average weight were used in the fourth experiment (December 1988). Calves were randomized to groups according to weight and group sizes ranged from three to five calves. All parasite infections were naturally acquired, but a mixture of nematode third-stage larvae (L3) (22,500 per calf), including 20% Ostertagia ostertagi, was inoculated into Jersey calves of Experiment 4 following a 2 week exposure to natural infection. All LEV treatments were by subcutaneous injection at dosages of 6 and 8 mg kg-1. Treatment with ivermectin was used only in Experiment 3 as an efficacy reference. All calves were killed at 8-10 days after treatment. The efficacy of LEV against all developmental stages of Ostertagia ostertagi was consistently low in all experiments and a dose-dependent response was not evident. Large numbers of all Ostertagia ostertagi developmental stages were present in non-treated calves in both September experiments. Percent reduction of adults, DL4 and EL4 at the 6 mg kg-1 and 8 mg kg-1 dosages, respectively, were adults, 51.7 and 23.6 (1981), 8.7 and 51.3 (1983); DL4 40.3 and 13.2 (1981), 37.9 and 33.1 (1983); EL4, 19.6 and 0 (1981), 59.6 and 42.9 (1983). Smaller numbers of Ostertagia ostertagi were present in winter experiments and adult worms greatly outnumbered larval stages. Percent reductions of adults, DL4 and EL4, respectively, were (1987) LEV 6 mg kg-1, 40.2, 0 and 0; ivermectin 200 micrograms kg-1, 98.7, 97.7 and 100.0; (1988) LEV 6 mg kg-1, 62.4, 100.0 and 100.0; LEV 8 mg kg-1, 49.1 65.0 and 74.1. Too few larval stages were present in the latter experiment for valid efficacy values.(ABSTRACT TRUNCATED AT 400 WORDS)     

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.     

Efficacy of moxidectin against gastrointestinal nematodes of cattle.

Three groups of 11 naturally infected crossbred beef calves were injected subcutaneously with moxidectin 1 per cent injectable at 0.2 or 0.3 mg moxidectin/kg bodyweight or with the unmedicated vehicle. Nematode infections had been acquired during grazing from December to April. Based on the faecal egg counts and total worm counts of the control calves at necropsy (11 to 13 days after treatment) most of the calves had heavy parasitic burdens. Ostertagia ostertagi was predominant and the mean numbers of adults, developing fourth stage larvae (L4) and inhibited early L4 were 45,906, 10,061 and 68,918, respectively. Haemonchus placei and Trichostrongylus axei were also present in the abomasa. Three species of Cooperia, Oesophagostomum radiatum L4 and T colubriformis adults were found in the intestinal tract. Both dosages of moxidectin were equally effective (P < 0.05) against all the abomasal nematodes (99.9 to 100 per cent) and the intestinal tract nematodes (99.4 to 100 per cent). No adverse reactions to the moxidectin treatment were observed. Abomasal pathology characteristic of heavy O ostertagi infection was observed in the control calves, but not in the treated calves.     

Efficacy of ivermectin delivered from a sustained-release bolus against inhibited early fourth-stage larvae of Ostertagia ostertagi and other nematodes in cattle.

The anthelmintic efficacy of ivermectin (IVM) delivered from a sustained-release (SR) bolus was evaluated against natural infections with gastrointestinal tract nematodes in 12 crossbred beef heifers in spring. The 12 calves were randomly allotted to 2 groups of 6 calves each. Group-1 calves were treated with an SR bolus designed to deliver 8 mg of ivermectin/d. Group-2 calves were nontreated controls. Cattle groups were kept in separate concrete-floored pens (grass hay nutrition) and slaughter was performed at 35 days after treatment. Fecal egg counts for group-1 calves remained zero after treatment, except for detection of less than 1 egg/g of feces in 1 calf at the time of slaughter; counts in nontreated calves increased. Mean and range of Ostertagia ostertagi inhibited larvae in nontreated calves were 27,093 and 10,622 to 56,368, respectively. Efficacy of the IVM SR bolus was 100% against O ostertagi developing fourth-stage larvae (L4) and inhibited early L4, Haemonchus placei adults, Cooperia punctata and C spatulata adult males, Cooperia spp adult females, Cooperia spp L4, Trichostrongylus colubriformis adults, Bunostomum phlebotomum adults, and Oesophagostomum radiatum adults. Efficacy for O ostertagi and T axei adults was 99.9%. Numbers of nontreated calves infected with C pectinata adult males and Oes radiatum L4 were too low to evaluate efficacy. Calves treated with the IVM bolus gained 10.2 kg, whereas nontreated calves lost 1.8 kg. Abomasal lesions were clearly greater in nontreated calves on the basis of index comparisons of abomasal weight and total live weight and gross pathologic features.     

Efficacy of abamectin against natural infections of gastrointestinal nematodes and lungworm of cattle with special emphasis on inhibited, early fourth stage larvae of Ostertagia ostertagi.

The anthelmintic efficacy of abamectin (avermectin B1) was evaluated against gastrointestinal nematodes, including Ostertagia ostertagi inhibited larvae and lungworm, in yearling crossbred beef heifers during late spring. The calves were grazed on contaminated pasture for 10 weeks and then held under conditions free of nematode infection for 3 weeks prior to allotment and treatment on 5 June. Thirteen calves were randomly assigned to two groups of six by restricted randomization on body weights; the extra lightest calf was assigned to the non-treated control group. Group 1 calves were treated with abamectin at 200 micrograms kg-1 body weight by s.c. injection and Group 2 calves were not treated; all were killed at 14 days after treatment. Ostertagia ostertagi was present in all controls; arithmetic mean numbers of adults, developing fourth stage larvae (L4) and inhibited EL4 were 7683, 605 and 36,102, respectively. Other nematode genera present in controls in sufficient numbers for the experiment were Haemonchus placei adults, Trichostrongylus axei adults, Cooperia spp. adults, Oesophagostomum radiatum adults, Bunostomum phlebotomum adults, Dictyocaulus viviparus adults and E5 (immature adults). Abamectin was highly effective (consistently greater than 99% efficacy and P less than 0.05) in removing all nematodes present in treated calves as represented in non-treated controls, including the primary target of Ostertagia ostertagi inhibited EL4. The lowest efficacy was 93.8%, against D. viviparus E5.     

Celiac trunk cannulation for obtaining abomasal lymph from cattle

Cannulation of the celiac trunk was surgically performed in 26 Holstein steers. The procedure was successful in 23 (88.5%) of the steers. Twenty-two of the steers were infected either naturally or experimentally with the abomasal nematode, Ostertagia ostertagi and/or other gastrointestinal parasites. The remaining 4 steers were not infected. Lymph obtained after surgery was used in various immunologic and biochemical assays. Daily lymph flow rate and total and differential WBC counts were determined after surgery in 4 of the infected and 3 of the noninfected steers. Steers were euthanatized for tissue specimen collection 7 days after surgery. At the time of euthanasia, lymph was still flowing from the cannula of 13 (56.5%) of the steers in which surgery was successful. This surgical procedure represents a valuable technique for studying at the local level, immunologic and physiologic responses of cattle to infection with O ostertagi.     

Effects of Ostertagia ostertagi and omeprazole treatment on feed intake and gastrin-related responses in the calf

Infection with the bovine abomasal nematode, Ostertagia ostertagi, results in a loss of acid-secreting parietal cells and an increase in gastric pH. The effects of an experimental infection with Ostertagia and/or daily treatment with omeprazole (OMP) at 2mgkg(-1) bodyweight for four consecutive days (experiment days 24-27, inclusive) on voluntary feed intake, blood and tissue gastrin concentrations, abomasal G-cell numbers, gastric pH, and blood cholecystokinin (CCK) and pepsinogen concentrations were investigated in the calf. Ostertagia-infected calves demonstrated a significant drop in feed intake between days 24 and 27 post-infection (38%; P<0.001) and in G-cell numbers (42%; P<0.05) and significant increases in abomasal pH (P<0.001), fundic mucosal weight (99%; P<0.01), and blood gastrin (P<0.05) and pepsinogen (P<0.0001). OMP treatment of worm-free animals resulted in a significant drop in intake between days 24 and 27 (30%; P<0.001) and in G-cell numbers (17%; P<0.05) and significant increases in abomasal pH (P<0.01) and blood gastrin (P<0.001). OMP treatment of Ostertagia-infected animals with an existing hypergastrinaemia had no effect on feed intake, abomasal pH, blood gastrin or pepsinogen or abomasal G-cell numbers. Blood CCK concentrations were also unaffected by either Ostertagia infection or OMP treatment. These data suggest that: (a) the depression in feed intake associated with OMP in worm-free calves was not due to a side effect of drug treatment; (b) inappetance in Ostertagia-infected animals is closely associated with the parasite-induced hypergastrinaemia; and (c) the elevation in abomasal pH was a major factor responsible for the elevated blood gastrin concentrations seen in parasitised and OMP-treated animals.     

Gastrointestinal nematode immunization trials in cattle

Three experiments were conducted to immunize calves against Ostertagia ostertagi, Trichostrongylus axei, and T colubriformis. Calves were given intraperitoneal injections with in vitro-grown parasitic larvae of the three species and IV and intraperitoneal injections of exoantigens obtained from culture media used to grow O ostertagi. None of the treatments provided immunity to subsequent oral challenge exposure with normal infective larvae.     

Stimulated pepsinogen secretion from dispersed abomasal glands exposed to Ostertagia species secretion

Carbachol and secretions from Ostertagia species parasites significantly (P less than 0.001) stimulated isolated preparations of dispersed gastric glands from bovine and ovine abomasal mucosa to secrete pepsinogen. Atropine reduced the response to both secretagogues. Live adult and larval stages of Ostertagia ostertagi and O circumcincta and homogenates of these parasites did not significantly (P greater than 0.05) increase pepsinogen production from bovine or ovine gland preparations.     

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

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

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.     

Factors influencing rain splash dispersal of infective larvae of Ostertagia ostertagi (Trichostrongylidae) from cow pats to the surroundings

Laboratory experiments were conducted to investigate rain splash dispersal of infective Ostertagia ostertagi larvae (L3) from cow pats to the surroundings during simulated rainfall. Simulated rainfall on dry cow pats resulted in dispersal of only very few L3. When cow pats were watered prior to the experiments, many infective larvae were stimulated to migrate up onto the surface of the pats, where they might be hit by water drops. When pre-watered pats were hit by falling drops, a considerable proportion of L3 was translocated. More than 90% of the translocated larvae were transported passively by splash droplets and only a minor part of the larvae migrated actively in water films or were transported passively by water run-off to the soil surrounding the pats. A reduction of the fall height of the simulated raindrops resulted in a significant reduction of the number of translocated L3. When cow pats were hit by drops of diameters between 2 and 5 mm, it was found that small raindrops were just as effective as large drops in splash dispersal of L3, provided the amount of rain was the same; but individual small drops were less effective in this respect than individual large drops. When splash droplets could travel freely, most of the splash-dispersed L3 were found within a horizontal distance of 90 cm from the pats; this applied for all sizes of drops. Most splash droplets carrying L3 were travelling within a vertical distance of 30 cm from the ground.     

Identification of Ostertagia ostertagi specific cells in bovine abomasal lymph nodes

To investigate the contribution of different bovine cell subpopulations in the development of in vitro induced responses by Ostertagia ostertagi third larval antigen extract (L3), bovine abomasal lymph node cell suspensions were depleted of specific cell populations. The depleted cell suspensions were subsequently assayed for their proliferative responses to O. ostertagi L3 antigen extract. Proliferative responses to O. ostertagi L3 antigen extract were restricted to a CD2+ CD4- CD8- cell population and MHC II+ cells different from B-cells were of major importance. Depletion of CD4, CD8, CD4CD8, IgM or CD21 positive cells did not decrease proliferation to L3 antigen extract. Depletion of gammadelta T-cells, which also comprise a subpopulation of CD2+ CD4- CD8- cells, reduced proliferation to L3 antigen extract only in one animal. The results suggest that either gammadelta T-cells could be involved in the proliferation or that another as yet unidentified population is important for proliferation. The precise role of these populations during infection with O. ostertagi and the mechanism by which these cells may influence the host immune response are important issues that remain to be elucidated.