Enhancement of resistance of lambs to Haemonchus contortus by previous infection with Ostertagia circumcincta

Two experiments were performed to determine whether previous infection of young lambs with Ostertagia circumcincta increased their resistance to the development of Haemonchus contortus and to explore alterations in the microenvironment of the abomasum related to the interaction. The parasitic infections were monitored with periodic fecal egg counts and by recovery and enumeration of parasites at necropsy of the lambs. Alteration of the microenvironment was examined by measurement of serum gastrin concentrations at critical times during both experiments. The results indicated that the numbers of H contortus were significantly (P less than 0.05) less in lambs which had O circumcincta infections before their exposure to H contortus. There was less reduction in lambs from which O circumcincta was removed (anthelmintic therapy) before their exposure to H contortus. Fecal egg counts also were significantly (P less than 0.05) less in the former group of lambs (exposed to both parasites) than those in lambs exposed only to H contortus. Serum gastrin values were significantly (P less than 0.05) increased in lambs infected with O circumcincta and indicated that the interaction observed could be due, in part, to alteration of abomasal pH.     

Activity of ivermectin against natural infections by abomasal nematodes in lambs in controlled tests: evaluation of equine and bovine injectable formulations administered intraorally

The efficacy of 2 injectable formulations of ivermectin, administered intraorally at the dosage of 200 micrograms/kg of body weight, was evaluated against naturally occurring infections by abomasal nematodes in lambs in 2 controlled tests. One test (A) included 17 lambs treated with the equine formulation and 16 nontreated lambs. For the other test (B), 14 lambs were treated with the bovine formulation, and 12 were nontreated. In controlled test A, only mature nematodes were recovered, and removals were 98% to 100% for Haemonchus contortus, Ostertagia circumcincta male, O trifurcata male, Ostertagia spp female, and Trichostrongylus axei. For controlled test B, removals were 99% to 100% for Haemonchus spp (immature), H contortus (mature), Ostertagia spp (immature), O circumcincta male (mature), O trifurcata male (mature), Ostertagia spp female (mature), Trichostrongylus spp (immature), and T axei (mature). Toxicosis was not evident in treated lambs.     

Decrease in establishment of Haemonchus contortus caused by inoculation of a Taenia hydatigena larvae vesicular concentrate

The effect of Taenia hydatigena larvae vesicular concentrate (ThLVC) on the establishment of an experimental infection by Haemonchus contortus was evaluated. The lambs that received ThLVC showed a greater (P<0.05) average of blood eosinophils (BE) than the lambs that did not receive ThLVC. Lambs that were only infected with H. contortus larvae showed a fecal egg count (FEC) and an adult phase (AP) number greater (P<0.05) than lambs that received ThLVC prior to infection. No effect was observed in size and prolificacy of AP after the administration of ThLVC. The infection with H. contortus caused an increase (P<0.05) in CD4+ lymphocytes in abomasal lymph node (ALN) and the combination of ThLVC plus the infection with H. contortus caused an increase (P<0.05) in CD4+ lymphocytes in the abomasal wall (AW). In addition, a positive correlation between gamma-delta lymphocytes of ALN (r=0.73, P<0.05) with the presence of AP in the abomasum was observed. The quantity of plasma cells in ALN and AW was not affected by the administration of ThLVC nor related to the resistance observed. The results shown in this work leave no doubt that ThLVC administration prior to inoculation produces eosinophilia and partially protects against the establishment of H. contortus. However, this protection is not only attributable to the role of eosinophils, since ThLVC can function stimulating other immune response cells, such as T lymphocytes, both contributing to prevent the presence of worms.     

Effect of cimetidine on abomasal pH and Haemonchus and Ostertagia species in sheep

Cimetidine, a histamine H2-receptor blocking agent which reduces gastric acid flow in simple stomach animals was administered directly into the abomasum of sheep at 100 mg kg-1 twice daily for five consecutive days and again on the eighth day. The drug caused a significant increase in the pH values of the abomasal contents and was highly effective in reducing the adult population of Haemonchus contortus (100 per cent) and Ostertagia circumcincta (70 per cent) in the abomasum of sheep. The possibility of using cimetidine as a method of treating abomasal nematode parasitism is discussed.     

Effect of experimental Ostertagia circumcincta infection on the performance of grazing lambs

Groups of 10 three-and-a-half-month-old Suffolkcross lambs were infected daily, five times a week, for 12 weeks with either zero, 500, 1500, 3000 or 5000 larvae of Ostertagia circumcincta and grazed together on a series of 'clean' paddocks. All the lambs were moved at seven to 10 day intervals to a fresh paddock to prevent autoinfection and were killed after 14 weeks. Another group was killed at the outset as a control for carcase analysis. Mean O circumcincta populations were 34 (grazing control) and 1224, 473, 1092 and 4331 for the groups receiving 500, 1500, 3000 and 5000 larvae per day respectively. The percentage of fourth stage larvae increased with increasing larval intake. Extensive abomasal damage with mucosal hypertrophy and depletion of parietal cells occurred in the lambs receiving 3000 and 5000 O circumcincta larvae per day. Intakes of 1500 larvae per day and above depressed growth rate by 24 to 37 per cent, reduced the deposition of protein in the carcase by 34 to 52 per cent and lowered calcium/phosphorus deposition by 23 to 48 per cent. Plasma pepsinogen concentrations were consistently elevated in lambs receiving 3000 or 5000 larvae per day and may be a useful aid to recognising abomasal damage sufficient to affect productivity.     

Reindeer as hosts for nematode parasites of sheep and cattle

The reindeer husbandry range of Scandinavia overlaps with sheep, goat, and cattle pastures. The aim of this study was to determine whether reindeer are suitable hosts for ovine or bovine nematode parasites, and thus may spread these parasites into the reindeer husbandry regions. To render worm-free, twelve 4-month-old male reindeer calves, six lambs, and six bovine calves were given ivermectin at 200 microg/kg body weight. Five weeks post-treatment, six reindeer calves were each artificially dosed with 10,000 third-stage larvae (L3) of gastrointestinal nematodes derived from sheep, and an additional six reindeer with L3 derived from cattle. Lambs and bovine calves received the same dose of ovine and bovine larvae as reindeer, from the same larval source, respectively. Faecal samples collected on five occasions after the larval dosing revealed that by the fourth week, all reindeer calves, lambs, and bovine calves were infected. Animals were slaughtered on days 40 (reindeer) or 47 (lambs and bovine calves) after the larval dosing. Reindeer calves were most susceptible to L3 derived from sheep. The overall mean intensity of Haemochus contortus, Trichostrongylus axei, and Teladorsagia circumcincta, did not differ between reindeer and sheep; however, early fourth-stage larvae of H. contortus were more abundant in reindeer (p = 0.002). The establishment of bovine-derived Ostertagia ostertagi was similar in reindeer (62%) and bovine calves (57%), but larval inhibition was much higher in reindeer (91%, p < 0.001) than in cattle (31%). Very poor establishment of bovine derived Cooperia oncophora was recorded in reindeer calves (2%) compared with bovine calves (59%). These results show that young reindeer are susceptible hosts to the important gastrointestinal parasites of sheep (T. circumcincta, H. contortus) and cattle (O. ostertagi), as well as being a suitable host for T. axei.     

Vagal and splanchnic afferent nerves are not essential for anorexia associated with abomasal parasitism in sheep

Heavy burdens of the abomasal nematode, Ostertagia (Telodorsagia) circumcincta, in growing lambs result in a reduction in liveweight gain due largely to a drop in voluntary feed intake. The present study investigated: (1) the role of subdiaphragmatic vagal and non-vagal visceral afferent nerves in mediating a reduction in voluntary feed intake, using subdiaphragmatic vagal deafferentation (vagotomy) either alone or in combination with coeliac-superior mesenteric ganglionectomy (vagotomy and sympathectomy); and (2) the association between appetite, abomasal pH, selected blood values (amidated gastrin (G-17-amide), glycine-extended gastrin (G-17-Gly), pepsinogen and leptin) and worm burden, in sheep experimentally infected with 100,000 O. circumcincta infective larvae per os. Neither vagotomy alone nor vagotomy and sympathectomy in combination adversely affected the establishment or course of development of the parasite burden, when compared with a control group subject to sham surgery. Furthermore, neither surgical procedure prevented the drop in appetite seen 5-10 days post-infection, although combined vagotomy and sympathectomy did reduce voluntary feed intake prior to the start of the study. Ostertagia infection resulted in a significant increase in abomasal pH in all three groups, which was accompanied by an increase in blood G-17-amide and in G-17-Gly, the latter reported for the first time in parasitized ruminants. There were no significant differences in blood leptin, also reported for the first time in parasitized sheep, either between groups or in comparison with pre-infection levels, though weak negative correlations were established between blood leptin and appetite from day 5 to the end of the study in all three groups and a positive correlation with blood G-17-amide in the control group over the same period. These data suggest that neither intact subdiaphragmatic vagal afferent nerves or coeliac-superior mesenteric ganglion fibres, nor changes in circulating gastrin and leptin concentrations play a major role in mediating the hypophagic effects of O. circumcincta in parasitized sheep.     

Reduced Ostertagia circumcincta burdens in milk-fed lambs

AIMS: To compare the susceptibility to parasitism by Ostertagia circumcincta of lambs fed entirely with bovine milk or weaned on to solid feed at 3 weeks of age. In addition, the effect of a single daily feed of milk on worm burdens was assessed. METHODS: Eight lambs were assigned to each of the 3 diets: milk (M), milk plus solid feed (cereal-based pellets and lucerne chaff) (MS), or solid feed only (S). Those to be fed solid feed were converted from complete milk feeding to the designated diet during their third week of life. From 3 weeks of age, all lambs were infected with 1000 O. circumcincta larvae twice weekly for 6 weeks; 4 lambs from each diet group were given normal sheathed L3 and another 4 were infected with exsheathed larvae. Faecal egg counts (FEC) and serum gastrin and pepsinogen concentrations were monitored from Day 17 after first infection, and worm burdens and abomasal pH and morphology were determined at necropsy. RESULTS: Total worm burdens and FEC were significantly lower in the M than MS and S groups, whereas there was no significant difference between those receiving sheathed and exsheathed larvae. The milk-fed lambs had a smaller reticulo-rumen and omasum and a more acidic abomasal pH. Serum gastrin and pepsinogen were increased in all groups, irrespective of diet or type of larvae used for infection. CONCLUSIONS: The cause of the lower worm establishment in lambs fed only milk was probably not failure to exsheath in the immature gastro-intestinal tract, as there were similar worm burdens in lambs whether sheathed or exsheathed larvae were administered. The lower pH of the abomasal contents of the preruminant lambs may have been a factor, as the parasites have previously been shown to die more rapidly in vitro at low pH. Alternatively, the milk itself had adverse effects on the parasites, but was ineffective when combined with solid feed. There was no benefit from feeding a milk plus solid diet over a solid diet.     

Cellular profiles in the abomasal mucosa and lymph node during primary infection with Haemonchus contortus in sheep

Cellular changes in the abomasal tissue and draining abomasal lymph nodes were examined after primary infection of lambs with Haemonchus contortus for 3, 5 or 27-36 days.Infection with H. contortus larvae resulted in a rapid and selective increase in the percentage of CD4(+) T-cells in the abomasal lymph node at 3 days post-infection (PI). By 5 days PI, the lymph node weight had increased two-fold; however, the percentage of lymphocyte populations in the abomasal lymph node resembled that seen in uninfected sheep. Lymph node weights remained at increased levels in the adult nematode infected sheep and down-regulation of B-cell surface markers (sIg and MHC Class II) was apparent in this group. Significant increases in the percentage of CD4(+) T-cells co-expressing MHC Class II, but not CD25, were observed in the larval infected groups except in adult nematode infected sheep. Increased numbers of eosinophils, CD4(+), gamma delta(+) T-cells and B-cells were found in the abomasal tissue by 5 days PI, but no further increases in these cell populations were observed in the adult nematode infected group. In contrast, the level of both lamina propria and intraepithelial mast cells observed in the abomasal mucosa was highest in the sheep carrying an adult nematode burden. These findings indicate that sheep are able to generate an early immune response to infection with H. contortus larvae, characterised by the activation of CD4 T-cells and B-cells in the draining lymph nodes and recruitment of eosinophils, CD4(+) and gamma delta-TCR,WC1(+) T-cells and B-cells in larval infected tissues. However, these changes do not seem to be maintained during infection with the adult parasite where increases in mast cell numbers dominate the local response, indicating that different parasite stages may induce distinct and possibly counteractive immune responses.     

Stocking rate effects on the worm burdens of Angora goats and Merino sheep

Worm egg counts of yearling Angora or Merino wethers grazing in mixed flocks at either 4/ha or 8/ha were observed over a 5 month period. Based on faecal egg counts infection levels in the sheep increased for the first 3 months and then decreased as the animals developed resistance. Egg counts in the goats did not decrease during the experiment. Post mortem worm counts indicated that there was a significant host species--stocking rate interaction on worm burdens of Haemonchus contortus adults and Ostertagia circumcincta adults and larvae. Goats had more H. contortus and O. circumcincta adults than sheep at the 8/ha stocking rate. At 4/ha goats had fewer O. circumcincta fourth stage larvae than sheep. There was no stocking rate effect on the numbers of Trichostrongylus axei or T. colubriformis. Sheep had more T. axei and fewer T. colubriformis than goats.     

Effect of copper oxide wire particle treatment on establishment of major gastrointestinal nematodes in lambs

Eighty-four 10-week-old lambs were used to investigate the effect of copper oxide wire particle treatment on the establishment of major gastrointestinal nematodes. They were maintained on pasture previously treated to minimise larval contamination. Five grams of uniform sized copper oxide wire particles were given orally five days before infection with either 20,000 Trichostrongylus colubriformis larvae, 20,000 Ostertagia circumcincta larvae or 3000 Haemonchus contortus larvae given as three doses at three-day intervals. The animals were slaughtered 21 or 22 days after the last infective dose. Parasite burdens in the lambs treated with copper oxide wire particles were reduced by 96 per cent in the case of H contortus and by 56 per cent in the case of O circumcincta compared to burdens in controls. There was no significant effect of copper particles on the establishment of T colubriformis.     

Significance of serum pepsinogen and abomasal pH levels in a field infection of O circumcincta in lambs.

Serum pepsinogen estimations from serially bled lambs grazing on pasture from spring to autumn showed correlations with the availability of Ostertagia larvae on pasture, with faecal egg counts of O circumcincta, and with Ostertagia worm counts in similar lambs slaughtered fort-nightly from the same pasture. In the slaughtered lambs correlations were recorded between worm count, serum pepsinogen level and abomasal pH. The value of serum pepsinogen estimations as a diagnostic test is discussed with reference to these findings.     

How widespread is resistance to invermectin among gastrointestinal nematodes in sheep in The Netherlands?

In Autumn 2009, a faecal egg count reduction test (FERCT) was carried out on three sheep farms. Groups of 8-11 lambs were treated with ivermectin or moxidectin, with a 14-day interval between treatment and sampling. Ivermectin resistance was present on all three farms. Treatment with ivermectin resulted in a reduction in faecal egg numbers of 94.6%, 63%, and 59%. On two farms, 14 days after treatment pooled faecal samples yielded predominantly larvae of Hamonchus contortus (100% and 98%, respectively). On the third farm, H. contortus and (probably) Teladorsagia circumcincta were resistant to ivermectin (64% and 36% of the larvae, respectively). Treatment with moxidectin resulted in a 100% reduction in egg output in sheep on all three farms. More sensitive culture techniques failed to detect any larvae in samples taken from two farms, but a few Ostertagia-type larvae, probably of T. circumcincta, were detected in samples from the third farm. It can be concluded that gastrointestinal nematodes in sheep from these three farms were resistant to ivermectin, whereas resistance to moxidectin was not detected.     

Abomasal secretion in sheep receiving adult Ostertagia circumcincta that are prevented from contact with the mucosa

AIMS: Both adult and larval Ostertagia circumcincta infections raise abomasal pH and serum gastrin and pepsinogen concentrations, either because of physical effects of the worms or from their chemical secretions. The study was designed to examine whether adult worms require contact with the gastric mucosa to effect changes in gastric secretion. METHODS: Abomasal pH and serum gastrin and pepsinogen concentrations were measured in four groups of sheep: Group A (n = 4): abomasal contents containing about 18,000 adult O. circumcincta were obtained from donor sheep, concentrated and infused into 5 microm mesh porous bags attached to surgically implanted abomasal cannulae. A second worm transplantation was carried out 1 week later to assess worm survival after 16 hours. Group B (n = 4): about 9000 adult worms, recovered after migration out of abomasal contents set in agar, were placed in small 5 microm mesh bags which were inserted through indwelling abomasal cannulae and left for 3 days. Group C (n = 2): about 3000 adult worms from the population recovered from agar were infused through abomasal cannulae which allowed. free movement in the abomasum. Group D (n = 3) was left uninfected. RESULTS: Worms transplanted directly into the abomasum (Group C) caused rapid and marked effects on abomasal secretion. Adult O. circumcincta died within 16 hours of transfer into the abomasum when they were restrained within porous bags. Nevertheless, in sheep receiving 18,000 worms, abomasal pH increased soon after new feed was presented on Days 1, 2 and 4 after worm transfer; serum gastrin was elevated in three sheep from 113 hours and serum pepsinogen increased in one animal. Sheep receiving 9000 worms showed similar trends but the results were equivocal. CONCLUSIONS: Adult O. circumcincta prevented from physical contact with the gastric mucosa by restraint in porous bags are able to raise abomasal pH. This study implicates parasite excretory-secretory products in mediating changes in gastric secretion caused by adult abomasal worms.     

Abomasal contents of parasitised sheep contain an inhibitor of gastrin secretion in vitro

Serum gastrin concentrations are typically elevated in parasitised sheep; however, in some animals serum gastrin concentrations may fall abruptly despite a very high abomasal pH. Although proliferating abomasal bacteria in culture generate a potent inhibitor of in vitro gastrin secretion, this inhibitor has not been detected in abomasal contents of unparasitised sheep. In sheep parasitised by O. circumcincta, all abomasal fluid samples of pH 5 and above were inhibitory to gastrin release in vitro. Inhibitory activity and abomasal pH were correlated in two separate experiments; the model best fitting the data being sigmoidal in each case, with zero activity at pH 3.6 and 4.6, respectively. There was no clear evidence that the presence of a gastrin inhibitor in the abomasal contents reduced the serum gastrin concentration in parasitised sheep. Serum gastrin was correlated with abomasal pH (log(10) serum gastrin concentrations conformed to log-linear sigmoidal models).     

Pepsinogen types in worm-free sheep and in sheep infected with Ostertagia circumcincta and Haemonchus contortus

Pepsinogen isolated from the plasma of Dorset wethers with abomasal burdens of Ostertagia circumcincta was found to have the same proportions of three zymogen subtypes as that isolated from the plasma of Merino wethers with abomasal burdens of Haemonchus contortus. It appears that the underlying pathophysiological response which causes increased plasma pepsinogen concentrations during abomasal parasitism may be the same irrespective of the breed of sheep or species of parasite.     

The epidemiology of abomasal nematodes of sheep in Sweden, with particular reference to over-winter survival strategies

In May 2002, studies on the seasonal patterns of nematode infection of sheep were undertaken on four commercial sheep farms in southern Sweden, which had previously reported problems with nematode parasitism, especially due to Haemonchus contortus. One farm was used for intensive investigation. This entailed the establishment of two replicate groups of sheep, each consisting of 20 ewes and their lambs, on adjacent pasture paddocks. The seasonal patterns of nematode infection were followed by regular (approximately monthly) sampling of both ewes and lambs for nematode faecal egg counts and larval differentiation, and the sequential use of replicate groups of tracer lambs. H. contortus and Teladorsagia circumcincta were the most abundant nematode species, with the former most prevalent in the post-parturient faecal egg counts of ewes. Tracer worm counts showed almost 100% arrested development in the early fourth larval stage for H. contortus as early as mid-summer and the numbers of parasites progressively increased during the season. T. circumcincta also showed high levels of arrested development, but not as early, or as absolute, as for H. contortus. Tracers allocated to the paddocks at the time of turn-out following winter in May 2003, showed virtually a total absence of H. contortus in contrast to exceedingly high infections with T. circumcincta. Results of the three additional monitoring flocks supported these findings. It can be concluded that under Swedish sheep farming conditions, H. contortus has evolved to survive the long, cold winters entirely within the host as the arrested larval stage, relying on the lambing ewe to complete its life cycle. The peri-parturient relaxation of resistance in the ewes triggers the resumption of development to the adult egg-laying parasites to result in pasture contamination and the completion of just one parasite generation/year. In contrast, T. circumcincta can survive well over-winter, both on pasture and within the host.     

Effect of Ostertagia ostertagi on lamb performance and cross resistance to O circumcincta

Fifteen worm-free lambs (two-and-a-half to three months or four to four-and-a-half months old) were infected with 3500 or 4000 Ostertagia ostertagi larvae on five days each week for six weeks, and their performance compared to that of controls. Eleven lambs were killed after eight weeks and four were challenged with O circumcincta to determine whether any cross resistance had developed. A feature of the O ostertagi infection was the considerable variation in response. Overall liveweight gain was lowered by 24 per cent in the two-and-a-half to three-month-old infected lambs, mainly due to reductions of 27 to 40 per cent in four of the seven lambs. There was no consistent effect in the older lambs. The worm populations consisted mainly of early fourth stage larvae and developing worms, but a small percentage reached sexual maturity and these produced a low faecal egg count (1 to 63 eggs per gram). Numerous intraluminal refractive crystals were present in the gut of both adult worms and developing stages, possibly reflecting degenerative changes. Hypertrophy of the abomasal mucosa with patchy loss of differentiation was a feature of the infection, and in four lambs serum pepsinogen concentrations were elevated. Exposure to O ostertagi did confer some protection against challenge with O circumcincta in that worm counts were reduced to about 60 per cent of those in controls, although no increase was observed in the numbers of arrested larvae. The successful passage of O ostertagi through young lambs could be important in mixed or alternate grazing systems by providing a reservoir of infection for the alternate host.     

Efficacy of the nematode-trapping fungus Duddingtonia flagrans against three species of gastro-intestinal nematodes in laboratory faecal cultures from sheep and goats

The ability of the nematode-killing fungus Duddingtonia flagrans to reduce number of infective larvae of three species of gastro-intestinal parasitic nematodes developing in dung was investigated in both goats and sheep. Groups of lambs and kids (12-20 weeks old) were given mono-specific infections of Haemonchus contortus, Ostertagia (Teladorsagia) circumcincta or Trichostrongylus colubriformis. Following patency of the infections (t1) faecal samples were collected for determination of faecal nematode egg count (FEC) and culture of parasite larvae. Groups of animals were then dosed on 2 consecutive days with one of the two dose rates of the fungus (250,000 or 500,000 spores/kg liveweight). One (t2) and 5 (t3) days after the second dose of fungus samples were again collected for FEC and culture. The number of larvae recovered from the faecal cultures at t1 and t3 were used as controls to assess the efficacy of the experimental treatment at t2. Average efficacy was 78% with group means ranging from 40 to 93%. Dose rate of fungus appeared to influence efficacy against O. circumcincta but not against H. contortus or T. colubriformis. Overall, there were no differences in the efficacy of the fungus against any of the parasite species or in either host animal. The results of this trial indicate the potential use of this fungus as a broad spectrum anti-parasite agent for use in both goats and sheep.