Characterization of IL-10-producing neutrophils in cattle infected with Ostertagia ostertagi.

IL-10 is a master regulator of immune responses, but its cellular source and function in cattle during the initial phase of immune priming have not been well established. Despite a massive B cell response in the abomasal draining lymph nodes in Ostertagia ostertagi (OO)-infected cattle, protective immunity is slow to develop, and partial protection requires years of repeated exposure. In addressing this problem, our initial hypothesis was that B cells produce IL-10 that downregulates the host protective immune response. However, our results showed that neutrophils made up the majority of IL-10-producing cells in circulation and in secondary lymphoid tissues, particularly the spleen (80%). Conversely, IL-10-producing B cells were rare. In addition, approximately 10% to 20% of the neutrophils in the blood and spleen expressed MHC II and were IL-10 negative, suggesting that neutrophils could also participate in antigen presentation. In vitro investigation of bovine neutrophils revealed that exposure thereof to OO extract increased IL-10 and MHC II expression in these cells in a dose-dependent manner, consistent with IL-10+/MHC II+ neutrophils detected in cattle shortly after experimental OO infection. Co-culture of untreated neutrophils with anti-CD3 antibody (Ab)-stimulated CD4+ T cells led to enhanced T cell activation; also, IL-10 depletion with neutralizing Ab enhanced the stimulatory function of neutrophils. OO extract depressed neutrophil stimulation of CD4+ T cells in the presence of IL-10-neutralizing Ab, suggesting that OO utilizes both IL-10-dependent and independent mechanisms to manipulate the bovine immune response. Finally, contact and viability were required for T cell-stimulatory neutrophil function. This report, to the best of our knowledge, is the first to demonstrate that neutrophil-derived IL-10 is directly involved in T cell regulation in cattle. Our data suggest that neutrophils and neutrophil-derived IL-10 are co-opted by nematode parasites and other pathogens to attenuate host immune responses and facilitate pathogen survival.

The association between Ostertagia ostertagi antibodies in bulk tank milk samples and parameters linked to cattle reproduction and mortality.

In Western Europe, gastrointestinal nematodes are widespread in dairy cattle. This study was carried out to evaluate the relationship between optical density ratio (ODR) measured on bulk tank milk with an indirect Ostertagia ostertagi ELISA and reproduction/mortality parameters. Data were collected between 2008 and 2010 from monitoring carried out on 1643 dairy herds (Normandy, Western France). ODR values of 3 samples from each farm taken from November 2008 to 2010 were averaged and then transformed into a categorical variable. Reproductive and mortality data were obtained from 1444 herds using cow records from government databases. Statistical analysis was carried out using ordinary logistic regression (OLR). The outcome variables were the case-control status of a herd for reproductive factors, age at first calving and inter-calving intervals, and mortality ratios of various age classes. The effect of the categorical ODR variable was studied and several potential confounder herd factors were used to improve the model fit. A significant relationship was found between high Ostertagia ODR levels and a late age at the first calving (>34.5 months) (odds ratio (OR)=1.94, p<0.001). No significant relationship was observed with OLR for inter-calving intervals although bivariate analysis showed that herds with high ODR levels had longer inter-calving intervals than herds with low ODR level (first inter-calving interval in herds with low vs. high ODR levels=412 days vs. 422 days, p<0.001; other inter-calving intervals=408 days vs. 413 days, p<0.01). A high ODR level was also associated with high mortality of calves between 0 and 30 days of life (mortality ratio>6%) (OR=1.43, p<0.05) and between 91 and 365 days (ratio>3%) (OR=1.72, p<0.01). No significant relationship was observed with multivariate approach for mortalities in other classes by age, but bivariate analysis showed that herds with high ODR level had higher mortalities than herds with low ODR levels (mortality between 31 and 90 days in herds with low vs. high ODR levels=1.89% vs. 2.91%, p<0.001; mortality after 365 days=1.67% vs. 2.93%, p<0.001). In conclusion, our results confirm the usefulness of ELISA as an indicator for production losses in dairy herds. This inexpensive tool could be advantageous, used to aid farmers and veterinarians to carry out appropriate control measures.

Do the changes in the behaviours of cattle during parasitism with Ostertagia ostertagi have a potential diagnostic value?

We investigated the magnitude of temporal changes in activity, posture and feeding behaviour of cattle infected with Ostertagia ostertagi, and their reversal after treatment with an anthelmintic. Twenty-six, 3-month-old, Holstein-Friesian bulls were allocated to one of three treatment groups. Bulls in two of those (groups P and PA) received 100,000 larvae on three occasions (Days 0, 7 and 14) and the remaining animals served as controls (C). The PA group also received an anthelmintic on Day 31. Parasite eggs appeared in the faeces of P and PA bulls from Day 17; from approximately the same time blood pepsinogen levels increased and body weight (BW) gain decreased (P<0.001). The reduction in BW gain persisted until Day 45 for P animals only. There was a decrease in the number of steps taken for P and PA animals, as well as lying and standing episode frequency, by 41 and 44% respectively (P<0.001) from Day 21 onwards. The average lying and standing episode duration increased by 52 and 55% respectively (P<0.001) from the same time in P and PA compared to C bulls. In addition, meal frequency showed a tendency to decrease for P animals only (P=0.039) from Day 39, and this was the only aspect of feeding behaviour affected by parasitism. All behaviours, returned to control levels within a week of anthelmintic drenching of PA bulls, apart from the number of steps taken. Although BW gain and pepsinogen also started to recover after drenching, these had not returned to control levels by Day 45. The magnitude of the changes in activity, and standing and lying episode frequency and duration suggest that these might have a diagnostic value, especially as all can now be monitored by automated means. However, these behaviours did not show the rapid changes we expected before parasitism manifested clinically and following recovery.

Exposure to pasture borne nematodes affects individual milk yield in Swedish dairy herds.

The association between exposure to gastrointestinal nematode (GIN) parasitism, indicated by elevated individual optical density ratio (ODR) to Ostertagia ostertagi, and individual milk production was analysed in dairy cows. The multivariable analysis also accounted for other sources of variation than exposure to GIN parasitism. Intra- and inter-herd variation in parasite exposure was also evaluated. Organic and conventional herds located in south-east Sweden (13 herds per system) were visited during the housing period 2009-2010. Levels of all major pasture-borne helminths in Sweden (O. ostertagi, Fasciola hepatica and Dictyocaulus viviparus) were determined in bulk tank milk, cow milk and serum (only for O. ostertagi). Their specific antibodies were detected using three different ELISAs (Svanova Biotech) and the optical density of the sample was expressed as ODR. Positive cases of Dictyocaulus and Fasciola were seldom found. Variation in ODR in serum samples for Ostertagia was higher within herds than between herds. Correlations between ODR for Ostertagia in serum and milk parameters were significant and negative. In the multivariable analysis, two models were performed separating data from primiparous and multiparous cows. Results from mixed models showed that daily milk yield was significantly influenced by Ostertagia ODR for multiparous cows. No interactions were found between Ostertagia ODR values and herd type.

Effects of weaning and infection with Teladorsagia circumcincta on mucin carbohydrate profiles of early weaned lambs.

Differences in mucin glycosylation in milk-fed and early weaned lambs may influence susceptibility to parasitism, particularly the greater cellular content and higher sulphation of mucins in young and unweaned lambs. Weaning also reduced the percentage of Gal (p<0.05) in fundic mucin and galactosamine (GalN) (p<0.01) in duodenal mucin, but had no noticeable effect on fucosylation or sialylation. Four experimental groups of lambs were studied (n=3): (1) 3 days old; (2) 9 weeks old milk-fed; (3) 9 weeks old weaned at 3 weeks-of-age on to lucerne chaff and cereal-based pellets (4) 9 weeks old weaned and infected with 1000 Teladorsagia circumcincta L3 twice weekly for 5 weeks. Fundic and duodenal mucin monosaccharides were analysed chemically and fundic, antral and duodenal tissues were stained with lectins, periodic acid Schiff (PAS), Alcian blue/PAS (AB/PAS) and high iron diamine. Age-related maturation of mucin glycosylation was prominent in young lambs: reduced total fundic mucins and increasing fucosylation and decreasing sialylation and sulphation of all mucins, as well as changes in the types of linkages of Fuc and sialic acids. By 9 weeks-of-age, there were no longer sialylated mucins in fundic surface mucus cells, only neutral mucins, while in Brunner's glands, there was reduced sialylation and large amounts of neutral mucins. In the neonates, both fundic and duodenal tissues contained only small amounts of mucins terminating with alpha-1,2-linked Fuc, which became the principal linkage in 9 weeks old lambs. Duodenal mucins in 3 days old lambs contained both alpha-2,6- and alpha-2,3-linked sialic acids, whereas the alpha-2,3 linkage was not present in older lambs. Parasitism increased the percentage of galactose, but reduced total and neutral fundic mucins, as well as sulphation and sialylation. There was both decreased sialylation and sulphation in duodenal mucins. Although no change in fucosylation was apparent from chemical analysis, infection reduced lectin staining for alpha-1,2-linked fucose in antral and duodenal tissues and alpha-1,6- and alpha-1,3-linked fucose in the duodenum. These changes in fundic and duodenal mucins were similar to those previously seen on Day 28 p.i. after a single infection of 4-9 months old sheep with T. circumcincta larvae.

Measurement of antibodies to gastrointestinal nematodes and liver fluke in meat juice of beef cattle and associations with carcass parameters.

In an effort to develop non-invasive methods for the diagnosis of Ostertagia ostertagi and Fasciola hepatica infection in beef cattle, this study was undertaken to evaluate antibody-detection ELISAs in meat juice samples and to investigate the associations between test results and carcass parameters. Preliminary tests were carried out to determine optimal working dilutions of meat juice samples. The Pearson correlation coefficients between ELISA results (expressed as ODR) of serum and meat juice samples from 90 to 100 cows were R=0.82 and 0.75 for O. ostertagi and F. hepatica, respectively. Next, an abattoir survey in Belgian Blue suckler cows was performed, analysing meat juice samples from 726 animals in spring and 724 animals in autumn 2008, originating from a total of 480 herds. There was a large variation in the observed O. ostertagi and F. hepatica ODRs and inter-seasonal differences were observed for F. hepatica (spring>autumn), but not for O. ostertagi. The relationships between individual parasite-specific ELISA results and carcass parameters (warm carcass weight, conformation score, fat coverage) were investigated by linear or logistic mixed models with herd as a random effect, while the relationship between herd average ELISA results and herd averaged carcass weight was investigated by linear regression with mean cow age and season as covariates. An increase in individual O. ostertagi ODR over the interquartile range was associated with an increased likelihood by 1.3 of a low conformation score (score E vs. S), but no significant associations were found with other carcass parameters. However, herd mean ELISA results were negatively associated with a decrease in the herd average carcass weight (P=0.001 and P=0.09 for O. ostertagi and F. hepatica, respectively). An increase in the herd mean O. ostertagi or F. hepatica ODR over the interquartile range was associated with a decrease in herd mean carcass weight of 9.1 and 3.4 kg, respectively. The implications of these results and the value of monitoring parasitic infections by analysis of meat juice samples from the abattoir are discussed.

Abomasal parasite syndrome in North American elk (Cervus elaphus canadensis).

CASE HISTORY: An 18-month clinical course of chronic ill-thrift, weight loss and emaciation, and eventual death occurred in a group of 520 translocated elk of mixed age and sex. Translocation was carried out without regard to animal welfare or health risks associated with the translocation. Mortality was approximately 84% (436/520) despite supportive nutritional and medical treatment. PATHOLOGICAL FINDINGS: General clinical and postmortem examinations indicated only Se and Cu imbalances and nutritional inadequacy. Additional purposeful post-mortem examination and histological evaluation of tissue sections from four of the affected adult elk demonstrated elevated abomasal pH and proliferative abomasal lesions as the most significant findings, consistent with Type-II ostertagiosis; intra-lesional nematodes were seen in the abomasum of two animals. DIAGNOSIS: Fading elk syndrome, or abomasal parasite syndrome in elk. CLINICAL RELEVANCE: Abomasal parasite syndrome initiated by Type-II ostertagiosis should be considered as a differential diagnosis in cases of ill-thrift and wasting in elk or elk-red deer hybrids. Changes to the architecture and secretory function of the abomasal wall lead to apparently irreversible digestive pathophysiology and nutritional disease.

Effects of Teladorsagia (Ostertagia) circumcincta infection on lambs selected for high fleece weight.

The physiological processes leading to the expression of the resilient phenotype, which allow animals to maintain a relatively higher production level during infection, have been investigated in lambs from a closed flock selected for 40 generations for high fleece weight (HFW), but with higher FEC and worm burdens than their unselected control (C) flock run in parallel. After recovery from surgery to implant abomasal cannulae, eight parasite-naive lambs from each flock were infected intraruminally at 4.5 months-of-age with 50,000 Teladorsagia circumcincta L3. Blood, abomasal fluid and faecal samples were collected daily for measurement of serum gastrin and pepsinogen concentrations, blood eosinophils, abomasal pH and FEC. Four lambs from each flock were euthanased on Day 8 post-infection and the other four on Day 28 post-infection. At necropsy, abomasal contents and tissues were collected for worm counts, abomasal lymph nodes and fundic tissue for cytokine gene expression and fundic tissue for histopathology. Expression of resilience appeared to be age-dependent as there were no significant differences in either FEC or worm burden between lambs from the two flocks, unlike older HFW lambs in a previous study. Abomasal secretion did not differ between flocks. Histopathological changes were typical of parasitism: inflammatory cells, mainly eosinophils and lymphocytes, were numerous in nodular areas and there were fewer TGF-alpha positive parietal cells, many of which were vacuolated. By Day 28 p.i., globule leucocytes were present. Mucosal thickness was significantly greater on Day 8 than Day 28 p.i. (p=0.000) and in C than HFW lambs. There were fewer parietal cells on Day 28 than on Day 8 p.i. (p=0.003) for pooled data. Circulating eosinophil counts increased moderately in both groups, significantly less in the HFW lambs. Fewer tissue and blood eosinophils in the HFW than C group on Day 8 p.i. were consistent with cytokine gene expression patterns, particularly lower IL-5 levels. Worm count decreased by 90% by Day 28 p.i., along with declining tissue eosinophil counts and IL-13 gene expression and increasing IL-10 and IL-4 gene expression. Food intake was depressed less in the HFW lambs, suggesting that maintenance of appetite could be an important aspect of the physiological basis for resilience. Although the resilient phenotype was not apparent at the younger age, lesser effects on food intake, differences in ALN cytokine profiles and lower blood and tissue eosinophil numbers in the HFW lambs may lead to the expression of resilience when older.

Excretory/secretory products of sheep abomasal nematode parasites cause vacuolation and increased neutral red uptake by HeLa cells.

Excretory/secretory (ES) products of Ostertagia (Teladorsagia) circumcincta and Haemonchus contortus have been implicated in the inhibition of gastric acid secretion and vacuolation, and the loss of parietal cells associated with abomasal parasitism. Vacuolation of epithelial (HeLa) cells caused by adult O. circumcincta or L3 O. circumcincta or H. contortus ES products have been examined by differential interference contrast microscopy and by the neutral red uptake assay. ES products caused visible vacuolation of HeLa cells, and this effect was enhanced by 8 mM NH4Cl. Some parasite ES products caused a marked detachment of cells from the coverslip. At lower concentrations of ES products, neutral red uptake was usually increased above the control, but at higher concentrations of ES products, uptake was often decreased, probably because of cell detachment. Although generally consistent with direct observations of HeLa cell vacuolation by parasite chemicals, neutral red uptake was not a satisfactory quantitative assay.

Osteopenia and reduced serum alkaline phosphatase activity in grazing lambs naturally infected with gastrointestinal nematodes.

The effect of gastrointestinal nematode infections on bone development was investigated in growing sheep on pasture. Forty-five weaned lambs from six groups in a two-factorial design incorporating stocking rate (SR; low, medium and high) and presence or absence of infection on pasture were sampled in the late grazing season. Worm counts were performed at slaughter, and the left metacarpal bones were excised for bone assessment. Faecal egg counts and worm burdens, primarily of Ostertagia circumcincta and Trichostrongylus vitrinus, were considerably higher in the high SR infected group ("I-High") than in comparable animals at low or medium SRs, whereas uninfected groups showed negligible egg excretion. Clinical biochemistry revealed significantly reduced serum concentrations of albumin, calcium and alkaline phosphatase in infected lambs. Nematode infections were associated with significant reductions in bone mineral density (30% at high SR), measured by dual energy X-ray absorptiometry, and in bone size (9%). Histomorphometry indicated thinning of the trabecular structure and reduced bone formation in the infected groups, particularly the I-High group. Bone mineral density, bone tissue volume and structural changes were strongly associated with log-transformed worm counts. The study showed that lambs suffering from moderate to heavy degrees of naturally acquired gastrointestinal nematode infection developed marked osteopenia after weaning, i.eduring the later part of the grazing season.

A sequential study of the pathology associated with the infection of sheep with adult and larval Ostertagia circumcincta.

Disturbances in the physiology of the abomasa of sheep infected with either adult Ostertagia circumcincta given via abomasal cannulae, or larvae (L3) given intraruminally were matched by pathological changes in tissues collected by repeated mucosal biopsy. Within 2-3 days of the transplant of adult worms, abomasal pH had increased markedly in five out of six animals, and there also had been rapid increases in serum gastrin and pepsinogen concentrations in all animals. Reductions in parietal cell number were recorded as early as 1 day after the transplant of adults and were associated with the rapid accumulation of many neutrophils and eosinophils. Mucosal hyperplasia, with increased numbers of cells closer in appearance to mucous/mucous neck cells, was a relatively late development, being most pronounced in the latter part of the infection. In sheep given larvae, changes in secretory physiology were again matched by a concurrent fall in parietal cell number and by the accumulation of inflammatory cells. Changes became maximal when most worms could be expected to be present as adults, confirming the role of adults in the natural disease. Some abnormalities were detected in biopsies collected from animals maintained free of parasites and, although milder in degree, there were similarities to those observed in parasitised tissues, there being fewer parietal cells, a modest degree of mucous cell hyperplasia and inflammatory infiltrates of predominantly neutrophils. These changes were the likely result of trauma to the tissues in the immediate vicinity of the cannula, due either to the presence of the cannula itself or to the frequent collection of biopsy material from areas close to it.

Infection of sheep with adult and larval Ostertagia circumcincta: gastrin.

Gastric endocrine cell populations and serum and tissue gastrin have been examined in sheep which were infected either intraruminally by tube with 150,000 Ostertagia circumcincta larvae followed by a trickle infection of 10,000 larvae thrice weekly for 8 weeks or by the transfer of 15,000 adult worms directly into the abomasum and killed 8 days later. Depletion of both antral gastrin and somatostatin was evident in both groups: tissue gastrin concentrations were reduced by 85% in the trickle infection and both G cells (gastrin-containing) and D cells (somatostatin-containing) were pale and fewer after adult worm transfer. The concurrent depletion of antral gastrin and somatostatin supports the contention that the hypergastrinaemia in parasitised sheep is largely secondary to the increase in abomasal pH. Although there was no change in the proportions of G34 and G17 in the tissues, there was an increase in the longer form of gastrin in the circulation of the larval-infected sheep, suggesting that there may be differential secretion of G17 and G34 which may be exaggerated as the rate of secretion increases. Although the fundic mucosa was thicker following trickle infection, there was no evidence of enterochromaffin-like cell hyperplasia in either infected group. It is suggested that hyper-gastrinaemia may be beneficial to the host, as it may allow the abomasum to regain the ability to acidify its contents during continued exposure to the parasites.

Infection of sheep with adult and larval Ostertagia circumcincta: abomasal morphology.

The infection of parasite-naive sheep with approximately 15,000 adult Ostertagia circumcincta via abomasal cannulae resulted in marked changes in the structure and function of the abomasum. The functional changes, which have been characterised previously, included elevated abomasal pH and increased serum concentrations of pepsinogen and gastrin. Eight days after the transplant of adult worms, the abomasa of recipient animals were significantly heavier than those of controls (P < 0.001), the thickness of the fundic mucosa was greater (P < 0.01), there were fewer parietal cells (P < 0.01) and increases in the numbers of mitotic figures and mucus-producing cells. Mucous cell hyperplasia was also evident in the fundic mucosae of sheep receiving a trickle infection of infective, third-stage O. circumcincta larvae and was prominent within nodules associated with larval development. In non-nodular mucosa, there was hyperplasia of mucous cells and changes in the distribution of parietal cells. Decreases in the number of parietal cells at the gland base were offset by increases at a mid-gland level, probably due to chronic hypergastrinaemia, so that, overall, total parietal cell number was unaffected. Mucous cell hyperplasia and the diminution of parietal cell number are seen in a diverse range of disease states and may be mediated by host growth factors such as Transforming growth factor-alpha. Alternatively, the cellular and/or the secretory changes in response to the presence of adult worms are mediated by chemicals that are cytotoxic/inhibitory for parietal cells, and released by the parasites themselves.

Cytokine profile induced by a primary infection with Ostertagia ostertagi in cattle.

Changes that occur in the local draining lymph nodes including, changes in cell surface markers and cytokine gene expression were studied over the first 4 weeks of a primary, Ostertagia ostertagi infection of the abomasum. Cells recovered from the abomasal lymph nodes (ABLN) after infection showed a decrease in the percentage of CD3+ cells, and an increase in the percentage of IgM+ cells and cells bearing the TcR1 marker. These changes were coincident with an increase in the proportion of activated cells (II-2R). Analysis of mitogen-stimulated ABLN cells by RNase protection assay (RPA) showed a dramatic reduction in IL-2 and IFN-gamma transcription after infection. In addition, analysis of unstimulated ABLN cells by competitive RT-PCR showed a similar decrease in demonstrable levels of IL-2 mRNA, but IL-10, IL-4 and IFN-gamma mRNA levels were elevated.

Isolation and phenotypic characterization of abomasal mucosal lymphocytes in the course of a primary Ostertagia ostertagi infection in calves.

Isolation and characterization of surface marker phenotypes of abomasal intraepithelial (IEL), lamina propria (LPL) and abomasal lymph node lymphocytes (ABLN) from uninfected calves were conducted, and the dynamics of change in these populations during the course of a primary Ostertagia ostertagi infection were defined. To obtain viable IEL and LPL from the abomasal mucosa of cattle, a modified isolation method was developed. The phenotypic characterization of abomasal lymphocytes was accomplished by indirect immunofluorescence staining. In uninfected animals, numbers of T cells exceeded the number of immunoglobulin-bearing cells in IEL, LPL and ABLN. The predominant T cell type in IEL and LPL was CD8+ cells, while the CD4+ T cell predominated in ABLN. Levels of activated cells and T cell receptor-1 gamma delta T cells were higher in IEL and LPL compared to ABLN. Within 3 weeks of infection, the number of lymphocytes recovered from the abomasal lamina propira and the mass of the ABLN was dramatically increased when compared to uninfected animals. Laser flow cytometric analysis demonstrated increased levels of immunoglobulin-bearing cells, gamma delta T cells, and activated T cells in IEL, LPL and ABLN in the infected animals. The greatest changes in LPL and ABLN took place during the first days of infection, and these changes were apparent throughout the 28 days covered by the experiment.

[Ostertagiasis in bulls used for bullfights]. / Ostertagiosis en 'Toro de lidia'.

In this paper, an outbreak of ostertagiosis in fighting-bull calves is described. Of 95 calves, 12 were affected and 10 died. Grossly, the abomasal mucosa was aedematous and showed multiple whitish tiny nodules. Differential diagnoses at a macroscopic level included parasitization by E. gilruthi. However, histologically the presence of Ostertagia spp. larvae in the lumen of the gastric glands was confirmed. The inflammatory reaction observed in the abomasal mucosa, together with the clinical picture, correspond to type-II ostertagiosis, with metaplasia, hyperplasia, dilatation, and parasitic forms in the gastric glands.

Different patterns of faecal egg output following infection of Scottish blackface lambs with Ostertagia circumcincta.

Faecal nematode egg counts were monitored in 184 Scottish Blackface lambs during natural exposure to a mixed, predominantly Ostertagia circumcincta infection and 12 lambs were selected which showed consistently zero (low count group; nine lambs) or consistently positive faecal egg counts (high count group; three lambs). These lambs were then treated with anthelmintic and challenged with 50,000 infective larvae of O. circumcincta and monitored thrice-weekly for 38 weeks; they were then re-challenged with another 50,000 infective larvae of O. circumcincta and monitored for a further 8 weeks. All sheep gave positive egg counts following deliberate infection. However, there was considerable variation among sheep in the size and timing of the peak in egg production. In particular, the pattern of mean values for faecal egg counts was different in the two groups. Egg counts were lower in the later periods of the extended infection in both groups of sheep. During the first half of the extended infection, egg counts were lower in sheep from the low count group, but during the second half of the infection the pattern was reversed and egg counts were lower in sheep from the high count group. There was a weak positive correlation between egg counts following anthelmintic treatment and 28 days exposure to natural infection and egg counts 28 days after a deliberate infection. Egg counts in the later stages of the deliberate infection were strongly but negatively correlated with egg counts following natural infection. The results of these studies show that differences in egg count following natural infection can be reproduced in experimental infections and that there is substantial variation in the pattern of egg production over time in different sheep. They also suggest that naturally resistant lambs are better able to delay worm development than naturally susceptible lambs.