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Infection of immunosuppressed mice with the abomasal nematode parasite of ruminants, Haemonchus contortus.

Infective larvae of Haemonchus contortus established in mice either immunosuppressed with the corticosteroid, dexamethasone, or the cytotoxic drug, cyclophosphamide, or treated with the histamine H2 inhibitor, cimetidine. Infections persisted for as long as the immunosuppressive treatment (7 days) and growth of larvae was similar to that seen in sheep. Virtually no larvae survived in untreated mice. Accordingly, it would appear that adaptive immunity is an important barrier against primary infection by H. contortus in mice and is a determinant of host-range for this parasite. Antibody raised in either sheep or mice against soluble extracts of adult H. contortus precipitated with different but overlapping sets of worm antigens. This suggests that the unique antigens recognized by the mouse compared with the sheep are crucial for the rapid protective responses which prevent primary infection.

Abomasum↗

Selection of Merino sheep for increased and decreased resistance to Haemonchus contortus: peri-parturient effects on faecal egg counts.

Peri-parturient worm egg counts were compared in 395 Merino ewes bred for either increased (IRH) or decreased (DRH) resistance to Haemonchus contortus, or from an unselected control flock (CH). Following a 10 month period with no anthelmintic treatment, a rise in egg counts began about 4 weeks before parturition and continued into lactation. At all stages, egg counts were significantly lower in IRH ewes, even those failing to conceive. Before lambing, IRH ewes averaged 263 epg, compared with 1113 epg in CH and 1618 epg in DRH ewes. After 2 weeks of lambing, lactating ewes averaged 1050, 2173 and 3708 epg, respectively, in the three lines. At the end of lambing, egg counts had increased to 1645, 3959 and 4124 epg, respectively. Counts appeared to have peaked in IRH ewes but not in DRH ewes. Ewes with twins had higher counts than those with singles, and ewes suckling lambs had higher counts than those losing lambs. No significant variation was associated with age of ewe (2-7 years). Although there were several nematode species present, the results suggest that, in comparison with DRH ewes, IRH ewes were particularly effective in reducing pasture contamination with H. contortus, both before and during the peri-parturient period.

Animals↗

Susceptibility of two crossbreeds of sheep to Haemonchus contortus.

Studies were conducted on 20 crossbred lambs of 4-5 months old. Ten lambs were Nali x Corriedale cross (Hisardale) and the other 10 were Nali x Lohi cross (Munjal). Seven lambs each of both crossbreeds were infected with 10,000 infective larvae of Haemonchus contortus. Three lambs of each crossbreed served as uninfected controls. Clinicopathological examinations were done before and after infection and all the lambs were necropsied 28 days post-infection. The body weight gain, haemoglobin and packed cell volume were significantly lower in Hisardale lambs and the peripheral eosinophil count was significantly higher in Munjal lambs. Significant hypoalbuminaemia and hypoglobulinaemia, a sharp increase in albumin: globulin ratio; significantly higher faecal egg counts; adult worm counts; abomasal pH, volume, congestion and oedema were observed in Hisardale lambs as compared to Munjal lambs. Clinical signs of haemonchosis were more severe in Hisardale lambs and two of them died. Munjal lambs therefore seem less susceptible than Hisardale lambs to H. contortus infection.

Animals↗

Effects of weaning, supplementation and gender on acquired immunity to Haemonchus contortus in lambs.

This study was designed to investigate whether delaying the age of weaning, or feeding a protein-rich supplement alters the rate at which lambs develop immunity to Haemonchus contortus and whether there is any interaction between nutrition, stress of weaning, and gender and the development of this immunity. Ninety-six Merino lambs were allocated to one of four treatment groups: supplemented-unweaned; supplemented-weaned; unsupplemented-unweaned; unsupplemented-weaned. There were approximately similar numbers of male and female lambs in each group. Supplemented lambs received 80 g/head/day of a protein-rich pellet from 16 to 23 weeks of age. Over the same period the lambs were drenched with 300 H. contortus larvae twice per week. Faecal worm egg counts were determined every week, and PCV and liveweight every 2 weeks for each lamb. Neither weaning nor sex had any effect on PCV (P > 0.05) but from day 50 after the start of infection, the decline in PCV was more pronounced in unsupplemented than in supplemented lambs. Faecal worm egg counts were higher (P < 0.001) in unsupplemented-weaned than in supplemented-weaned lambs and in females when compared with castrates (P < 0.03). There was a weaning x supplementation x age interaction (P < 0.03) with unsupplemented-weaned lambs developing a higher faecal egg count than supplemented-weaned lambs. There was also a significant (P < 0.02) sex x weaning x age interaction with weaned-female lambs having a higher faecal egg count than weaned-castrate lambs; egg counts increased more rapidly in unweaned-female lambs than in weaned-female lambs.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗

The sensitization of mucosal mast cells during infections with Trichostrongylus colubriformis or Haemonchus contortus in sheep.

Responses of isolated mucosal mast cells (MMC) during infections with either Trichostrongylus colubriformis or Haemonchus contortus were examined by measuring the release of sheep mast cell protease (SMCP) in a degranulation assay. MMC from sheep immune to T. colubriformis released maximal amounts of SMCP and histamine within 0.5h of incubation with larval antigen whereas maximum secretion of leukotrienes occurred 3h after addition of antigen. It was only after 8 weeks of a primary T. colubriformis infection, that MMC released significantly elevated levels of SMCP (23%); this occurred when the worm burden was being rejected. In contrast, the SMCP release from MMC of immune sheep was significantly higher at 40%, and occurred within 1-4 days after challenge (DAC). The SMCP release peaked at 6-8 DAC at 51%, and declined after 56 DAC to < 25%. MMC isolated from the duodenum and mid-small intestine of immune sheep released 2-3 times higher proportion of SMCP than did cells recovered from the terminal ileum. Mast cell numbers were similar in the 3 regions but the quantity of globule leucocytes (GL) was 2.5 times higher in the duodenum. During infections with H. contortus in the abomasum, MMC isolated from the small intestine released greater levels of SMCP when incubated with larval antigens than did abomasal MMC. There was no increased release during the first 12 weeks of a primary infection although the SMCP release (23%) from immune MMC at 7-10 DAC was significantly enhanced. Once again the release from MMC isolated from the three intestinal regions of sheep immune to H. contortus was lowest in the terminal ileum.(ABSTRACT TRUNCATED AT 250 WORDS)

Abomasum↗

The heritability and specificity of responsiveness to infection with Haemonchus contortus in sheep.

High responder (HR) and low responder (LR) lines of sheep of the Hungarian Merino breed were selected by using faecal egg counts (epg) as parameter of responsiveness after double artificial Haemonchus contortus test infections. A total of 81 lambs born to assortative matings of HR rams with HR ewes and LR rams with LR ewes, as well as 28 control lambs from unselected parent sheep, were studied to estimate the heritability of responsiveness. A relatively high level of heritability (h2 = 0.49 +/- 0.17) was observed. A comparative assessment of responder classification carried out by two subsequent and distinct H. contortus and Trichostrongylus colubriformis infections revealed a remarkable degree of agreement: identical results in 69.7% and marginal differences in 23.2% of 99 animals. These results suggest that by selecting sheep for high responsiveness to a certain species of nematodes, one can also achieve a substantial improvement of resistance against a wide range of other nematode species.

Animals↗

Developmentally regulated expression of a Haemonchus contortus surface antigen.

All life cycle stages of Haemonchus contortus were screened with antibody-secreting cell (ASC) probes generated from immune sheep 5 days after a challenge infection. An antigen with an approximate molecular weight of 70-90 kDa was found to be predominantly expressed in the 3rd-stage larvae, while a molecule with an approximate molecular weight of 28-31 kDa was recognised by the ASC-probes in the earlier developmental stages. Antisera generated against the purified 70-90 kDa molecule and antibodies eluted from the 70-90 kDa region in the 3rd-stage larvae were found to react with the 28-31 kDa region in the 2nd-stage larvae. These results suggest that the 70-90 kDa antigen, previously identified as a major 3rd-stage surface antigen, is synthesised as a lower molecular weight molecule in the earlier larval stages and assembled into a higher molecular weight complex through non-reducible covalent bonds.

Age Factors↗

The pathogenesis of experimental Haemonchus longistipes infection in goats.

Goats are highly susceptible to Haemonchus longistipes and could therefore serve as an inexpensive model to study camel haemonchosis. The course of the disease in goats is similar to that in camels and to H. contortus infection in sheep and goats. Unlike the age-dependency of camel haemonchosis, however, the severity of H. longistipes infection in goats is dose-dependent and varies from mild to hyperacute.

Animals↗

Antibody-containing cells in the abomasal mucosa of sheep with genetic resistance to Haemonchus contortus.

The frequency distribution of parasite-specific antibody-containing cells (ACC) and total immunoglobulin-containing cells (ICC) in the abomasal mucosa was examined in genetically resistant and random-bred sheep, following infection with Haemonchus contortus, to determine whether the two genotypes differed in the development of a local immune response. Immunofluorescence staining was used to detect ACC and ICC of IgA, IgG1, IgG2 and IgM classes. No ACC were found in the abomasum of any of the sheep before infection. ACC were first detected in the abomasal mucosa of both the resistant and susceptible sheep seven days after infection, reached a peak level on day 21 and then declined. At all observation times, the majority of ACC in both genotypes were of the IgA isotype, followed by IgG1 and IgM. The IgG2-ACC response was negligible compared with the IgA and IgG1 response. The comparison of genotypes showed that the resistant sheep had significantly more IgA-ACC on days 14, 21, 28 and 35, and significantly more IgG1-ACC than random-bred sheep on days 14, 21 and 28. The numbers of IgG2- and IgM-ACC did not differ between the genotypes. The abomasal mucosa of the resistant sheep was also found to have significantly more ICC than the mucosa of random-bred sheep; IgA-ICC predominated at all the observation times followed by IgG1 and IgM. These results suggest that IgA and IgG1 antibodies, produced locally in response to infection, may play a role in mediating genetic resistance to haemonchosis in sheep.

Abomasum↗

Development of a vaccine against haemonchus contortus.

Haemonchus contortus is an economically important nematode parasite of sheep and the occurrence worldwide of strains resistant to anthelmintic chemicals has emphasized the need to develop a vaccine against it. Here, Ed Munn describes the approach to this problem adopted in his laboratory. The principle developed should be applicable to other gastrointestinal parasites.

Journal Article↗

The abomasal histology of worm-free sheep given primary and challenge infections of Haemonchus contortus.

The histological changes in the abomasal mucosa of pairs of worm-free ewes and lambs were compared at various stages after infection and reinfection with 350 Haemonchus contortus L3 per kg bodyweight. In terms of worm burdens the ewes were relatively resistant to reinfection compared with the lambs and the histological changes in the abomasum at most stages after infection were more marked in the adult animals. These included increases in the numbers of mast cells, globule leucocytes, eosinophils and IgA plasma cells in the mucosa.

Abomasum↗

Parasites as stressors: plasma cortisol responses of goats infected with the stomach worm Haemonchus contortus to exogenous corticotropin (ACTH).

Ten male, juvenile pigmy goats of similar age and weight were allocated randomly to two groups. Goats in one group were each inoculated with 20,000 infective larvae of Haemonchus contortus. The other group served as uninfected controls. Goats were housed together, and precautions were taken to avoid the creation of differential, between group, stressogenic circumstances. Body weights, nematode egg production, hematocrits, and clinical signs were monitored over a 61-day period following inoculation of larvae. On Days 59 and 61, adrenal response tests (ART) were conducted by measuring the levels of plasma cortisol before and 2 h after administration of porcine ACTH at the rate of 0.35 I.U. kg-1 body weight on Day 59 and 2.2 I.U. kg-1 on Day 61. Although the infections did not reduce body weights, they were 'heavy' on the basis of egg production, and led to significant reductions in packed erythrocyte volumes. There was no significant difference between the groups of goats in the responses to ART, indicating that the infections did not produce sufficient stress to reduce the ability of the adrenal cortex to respond to exogenous ACTH.

Adrenocorticotropic Hormone↗

Experimental Haemonchus contortus infection in three breeds of sheep.

Nine- to 10-month-old Florida Native, St. Croix and Dorset/Rambouillet sheep were infected with Haemonchus contortus. The primary infection was terminated by anthelmintic treatment 9 weeks later and animals were then reinfected with H. contortus. Dorset/Rambouillet sheep showed higher fecal egg counts and decreased packed cell volumes and plasma protein levels compared with the other two breeds during the primary infection. However, no breed differences were found in total worm burdens in either primary or secondary infection. Differences between breeds were also not detected in lymphocyte responsiveness to parasite antigen, H. contortus specific mucosal antibody levels, numbers of abomasal mast cells and globule leukocytes or abomasal histamine levels.

Animals↗

Changes in liveweight gain and blood constituents in experimental infection of goats with a goat-derived compared with a sheep-derived strain of Haemonchus contortus.

Two groups of goats were experimentally infected with two different strains of Haemonchus contortus and compared with the controls. Group A animals were infected with a goat-derived strain (GDS) while Group B animals were infected with a sheep-derived strain (SDS). Changes in the liveweights and some blood constitutents between the two infected groups were compared. The pathogenic effects of the GDS larvae were generally more serious when compared with those of the SDS larvae.

Animals↗

Pathophysiology of Haemonchus placei infection in calves.

This study was conducted to investigate the pathophysiology of Haemonchus placei infection in Friesian calves. Seven calves were divided into two groups, three uninfected calves (control group) and four infected animals. The latter group were infected orally with 500 H. placei larvae kg-1 body weight. Five weeks after infection they were all housed in metabolic crates and injected with 125I-bovine albumin. 51Cr-red cells and 59Fe-transferrin, to study albumin metabolism, erythrokinetics and ferrokinetics. The results showed that there was a significant reduction in the mean haematocrit values and reduced weight gains in the infected calves compared with the controls. There was also a change in the distribution of albumin from the extravascular to the intravascular pool and a significant increase in the plasma and blood volumes of infected calves although the blood and albumin loss via the gastrointestinal tract recorded in this study was similar in both groups.

Age Factors↗

Establishment of camel isolates of Haemonchus longistipes and Trichostrongylus colubriformis in goats.

Two main nematode parasites, Haemonchus longistipes and Trichostrongylus colubriformis, have been reported to cause morbidity and mortality in camels. Although goats are usually reared with camels in the combined animal husbandry system prevailing in Northern India, these parasites have not been reported in goats. Therefore, it was planned to conduct an experimental cross-transmission study of H. longistipes and T. colubriformis isolated from camels and transferred to young goats. Twelve healthy kids (5-6 months) were divided into three groups of four each. Groups I and II were infected orally with third stage infective larvae of H. longistipes and T. colubriformis cultured in the laboratory from faecal isolates from camels, at the rate of 50,000 and 100,000 larvae, respectively; group III comprised uninfected healthy controls. Daily clinical and faecal examinations were made. Body weights and blood haematological indices were measured at weekly intervals. The infection became established in both infected groups, and the animals started passing ova 17 days postinfection. Egg counts varied from 275.00 +/- 248.74 to 6150 +/- 2830.63 eggs g-1. Animals were killed 28 days postinfection. On postmortem examination mature as well as immature worms were recovered from the abomasum and intestine with typical postmortem changes seen in both the groups. Clinical manifestations were weakness and loss of body weight but no diarrhoea. There was a marked fall in haemoglobin and packed cell volume, indicating anaemia in both infected groups.

Abomasum↗

Comparison of resistance of four genotypes of rams to experimental infection with Haemonchus contortus.

Fifty-five rams aged about 18-24 months weighing 30-42 kg were used in this study. Ten rams of each of four genotypes, S (Sumatra), H1 (50% Sumatra-50% Virgin Island), B1 (50% Sumatra-50% Barbados Blackbelly), E1 (50% Sumatra-50% Java Fat-tail), were infected orally with a newly isolated strain of Haemonchus contortus. Each animal received 2000 infective larvae 3 times week-1 for 3 weeks, with a total of 18,000 larvae. Fifteen rams belonging to Sumatra and its crosses with Virgin Island were used as uninfected controls. Peak egg counts were observed on Day 35 for genotype B1 and on Day 42 for genotype H1 and S. In genotype E1 a slow but consistent increase in EPG continued until Day 49 when the experiments terminated. Overall faecal egg counts at all sampling dates were not statistically different between genotypes (P > 0.05). There was a large variation in the EPG of individual rams within a genotype. The overall average EPG (geometric means) of individual rams within a genotype ranged from 3 to 1028 for B1, 4 to 261 for E1, 7 to 3119 for H1 and 9 to 506 for S. The analysis of packed cell volume (PCV) for four genotypes and controls from all sampling times showed significant differences (P < 0.05). The overall mean PCV was highest in S (31.1) and lowest in H1 (28.4). The ranking of four genotypes for PCV was S > E1 > B1 > H1. The decrease of PCV during the course of infection was highly significant for all genotypes of infected rams (B1, E1, H1, S) (P < 0.01). PCV of the control group did not exhibit much change during the course of the experimental period. Weight gain of infected rams was lower than those of uninfected controls (P < 0.5) but there was no significant difference between the four genotypes of infected rams. Individual variation within genotype in susceptibility to infections was generally more important than between genotype differences. Two major conclusions of the present study are: (1) The imported breeds with higher body weight, namely the Barbados Blackbelly and Virgin Island, may be used in cross breeding to increase the body size of local Sumatra sheep. (2) Based on the faecal egg counts it is possible to identify the animals for use in selective breeding programmes for higher resistance to H. contortus.

Analysis of Variance↗

Evaluation of the phenotypic performance of a Red Maasai and Dorper double backcross resource population: indoor trickle challenge with Haemonchus contortus.

Six F(1) Dorper (D) x Red Maasai (R) rams were mated to both D and R ewes to produce backcross lambs. These six double backcross resource families are being analysed to identify quantitative trait loci that may be controlling resistance to gastro-intestinal (GI) nematode parasites, mainly Haemonchus contortus. After assessing the phenotypic performance of the lambs following exposure to natural infections, the surviving lambs were drenched and moved indoors for an artificial challenge with H. contortus along with straightbred D and R lambs. A total of 1317 lambs were included in the analyses consisting of 523 3/4D, 580 3/4R, 87 D and 127 R. The D lambs were significantly and consistently heavier than R lambs and 3/4D were significantly heavier than the 3/4R lambs. The difference between the backcrosses was about half of that between the straightbreds. Resistance was assessed in terms of faecal egg counts (FEC) and total worm counts (TWC) at necropsy while packed cell volume (PCV) was used to assess resilience to weekly oral doses of 2500 infective larvae of H. contortus. No significant breed differences were observed for log transformed FEC (LFEC). A significant breed difference in PCV was recorded. The backcrosses had the higher values and while no differences were observed between the straightbreds, 3/4D had significantly higher PCV than the 3/4R. Despite the absence of breed differences in FEC the R and the 3/4R had significantly fewer worms than the D and the 3/4D. The D had significantly longer worms than the R and the 3/4D had significantly longer worms than the 3/4R. Worms recovered from D had more eggs than those recovered from R. Similarly worms from 3/4D contained more eggs than those from 3/4R. Thus, on a breed basis the breed with more worms had longer worms. In contrast, when, in a small part of the experiment two doses of larvae were used to check for any breed by dose interactions, worms from the low dose (and hence fewer worms) animals were longer. We postulate that in fast growing hosts like the D, worms also have a better potential for growth and reproduction than in hosts that have less potential for growth. The overall correlation coefficient between PCV and LFEC was -0.67 while that between LFEC and LTWC was 0.72. From these results it is clear that the R do not respond to the artificial challenge the way they do to natural infection suggesting that phenotyping of R and R cross lambs for purposes of selecting those that are resistant or susceptible is best done under natural challenge.

Animals↗