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PubMed1992

Systemic immunization of sheep with surface antigens from Haemonchus contortus larvae.

Sheep immunized systemically with a surface extract from third-stage H. contortus larvae showed high serum antibody reactivity against surface antigens and whole, viable larvae. After a first infection, no significant difference was found between the mean egg counts of the vaccinates and controls although most vaccinated sheep seemed to show an increased susceptibility to infection. The local abomasal response was stimulated by giving both vaccinated and control sheep a large, abbreviated infection cured after 11 days by drenching. Thereafter, a second challenge infection was given. This immunization regime resulted in seven of the nine vaccinated sheep showing clear protection against the second challenge infection.

Animals↗
PubMed1993

An outbreak of haemonchosis associated with anthelmintic resistance in sheep.

An outbreak of haemonchosis associated with anthelmintic resistance was recorded in a flock of 150 crossbred lambs. Closantel and ivermectin were 100% effective against Haemonchus contortus while fenbendazole and morantel reduced faecal egg counts by 87 and 29%, respectively. Levamisole showed an efficacy of 95%. At double the recommended dose (10 mg kg-1), fenbendazole had an efficacy of 99% against H. contortus infection. This appears to be the first report of an outbreak of haemonchosis associated with fenbendazole and morantel resistance in a strain of H. contortus in sheep.

Animals↗
PubMed1993

Kinetics of expulsion of Haemonchus contortus from sheep and jirds after treatment with closantel.

Experiments were conducted in sheep after intramuscular treatment with closantel and in jirds after oral treatment with closantel to determine when expulsion of established H. contortus commences. Expulsion starts at about 8 h in sheep and coincides with the onset of reduced motility in worms recovered from the abomasum. In jirds, expulsion starts by 2 h after treatment. Experiments also conducted in jirds showed that infective larvae are first killed by circulating closantel 3 days after infection, when blood feeding starts, and that by 8 days 80% of larvae are lost.

Animals↗
PubMed1993

The B cell dependence of Haemonchus contortus antigen-induced lymphocyte proliferation.

The proliferative responses of ovine lymphocytes to soluble antigens of third-stage larvae of Haemonchus contortus (HcAg) were examined in vitro. Lymphocytes in mononuclear cell cultures responded consistently to HcAg even if the lambs were naive to H. contortus. Lymphocytes from sheep that were immune to reinfection with H. contortus also consistently responded to HcAg with a response significantly greater than that recorded from naive lambs. When the responses of T cell or B cell enriched fractions were examined, neither enriched cell population responded to HcAg, even though the T cell populations were shown to contain monocytes by histochemistry. Studies using the accessory cell dependent T cell mitogen Con A demonstrated that such T cell enriched populations had normal Con A-induced responses compared to unfractionated mononuclear cells. In addition, neither the T cell nor B cell fractions responded to HcAg in the presence of autologous mononuclear cells which contained monocytes and dendritic cells. Responses to HcAg in the T cell enriched population were restored, however, by the presence of mononuclear cells that were cultured with HcAg for 48 hours and then irradiated before the addition of the T cells. These results were seen with cells from both H. contortus naive lambs and immune sheep. This provides evidence that B cells and not dendritic cells or monocytes are acting as antigen presenting cells in Haemonchus antigen-induced lymphocyte responses in both naive lambs and immune sheep.

Animals↗
PubMed1993

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↗
PubMed1993

The potential value of integral membrane proteins in the vaccination of lambs against Haemonchus contortus.

An extract of adult Haemonchus contortus enriched in the parasite's intestinal microvillar membrane protein H11 and other integral membrane proteins but free of the protein contortin was evaluated as a potential vaccine in two breeds of sheep. The worm burdens of Clun Forest sheep injected with the extract and challenged with 25,000 infective larvae were reduced 89% by weight compared to the average for the controls. The worm burdens of Dorset sheep (challenged with 10,000 infective larvae) were reduced 72%. In both breeds the reduction in the number of female worms, 92 and 71.8%, respectively, was greater than the reduction in the males (86.5 and 46%). Parasite egg output, determined only for the Dorsets, was reduced 92% protection correlated with serum antibody titre. Most of the antibodies were directed against H11.

Animals↗
PubMed1993

Purification and evaluation of the integral membrane protein H11 as a protective antigen against Haemonchus contortus.

A detergent extract of adult Haemonchus contortus enriched in the integral membrane protein H11, previously shown to give protective immunity against the parasite, was fractionated by lectin and ion-exchange chromatography. The fractions were evaluated for their ability to immunize Clun Forest and Dorset Horn sheep against experimental haemonchosis. Most of the protective activity was associated with H11. Used in an approximately 95% pure form it gave a mean reduction in parasite egg output of 94.6% and reduced male and female worm numbers by 86.5 and 93.5%, respectively. Level of protection correlated with serum antibody titre to H11.

Animals↗
PubMed1995

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↗
PubMed1995

Relationship of blood loss to worm numbers, biomass and egg production in Haemonchus infected sheep.

Chromium51 labelled erythrocytes were used to measure blood loss in sheep infected with either H. contortus, H. placei or the F1 of their reciprocal cross hybrids (H. contortus males x H. placei females = CXP, H. placei males x H. contortus females = PXC). CXP consisted of sterile males and semi-sterile females and sheep infected with this hybrid had nil egg counts; while PXC consisted of semi-sterile males and females and their hosts had some eggs in their faeces. Immediately after their blood loss was estimated the sheep were killed and their worms counted. One hundred male and 100 female worms from each sheep were dried at 60 degrees C overnight and then weighed to give an estimate of the biomass in each infection. Total daily egg production was calculated from egg counts made on 3 random 2 g samples from the 24 h faecal collection. The relationship of the log blood loss to log worm number, to log biomass of infection in each sheep and to log total egg production in the parental species was linear. Biomass was no better an estimator of blood loss than was worm number. There was no improvement in the estimation of blood loss by regressing the male and female worms separately. In all of the analyses, there was no improvement by regressing H. contortus and H. placei or their hybrids separately. It appeared that egg production was not a major factor in the requirement for blood by Haemonchus. The size of individual female worms was negatively correlated with the number of worms in the infection.(ABSTRACT TRUNCATED AT 250 WORDS)

Anemia↗
PubMed1995

The epidemiology of helminth infections of growing sheep in Argentina's western pampas.

Seasonal population trends of helminth parasites of growing lambs were investigated over 4 years. Successive worm-free lambs were grazed together with untreated lambs for 20-30 days and then slaughtered for helminth counts 2 weeks after their removal from pasture. Likewise untreated lambs from the same flock were slaughtered from 2 to 15 months of age for worm counts in the same way as tracer lambs. The predominant parasites were Haemonchus, Nematodirus and Trichostrongylus. Haemonchus contortus was found to be of major importance and the flock acquired massive worm burdens from summer to mid-autumn. Minimum burdens were seen from winter to early spring and maximum L4 stages were found from mid-autumn to early winter. Nematodirus burdens increased from December with a peak in late summer and then decreased to low values. Maximum larval availability was in autumn to early winter. Trichostrongylus (mainly T. colubriformis) populations increased in autumn and peaked in June-July, while the highest larval availability was in autumn. The minor genera recovered were Ostertagia, Cooperia, Trichuris, Oesophagostomum and Moniezia. Dictyocaulus, Chabertia and Teladorsagia were noted occasionally and liver flukes were not detected. No important and pathogenic numbers of L4 stages were seen and all predominant species were able to survive over summer or winter in pasture.

Animals↗
PubMed1995

Protection against multiply drug-resistant and geographically distant strains of Haemonchus contortus by vaccination with H11, a gut membrane-derived protective antigen.

H11 is a "hidden antigen" derived from adult Haemonchus contortus which is capable of conferring a high degree of protection against subsequent challenge. To be commercially useful, any vaccine based on this antigen must protect against field strains and drug-resistant worms. The results show that vaccination with H11 affords high levels of protection against the multiply drug-resistant (MDR) Lawes strain of Haemonchus contortus. Moreover, there was no difference in protection whether the H11 was extracted from drug-resistant or susceptible worms, indicating that multiple drug-resistance does not significantly alter the immunogenicity of H11. Ideally, a commercial vaccine based on this antigen should also be capable of protecting against strains from geographically separate areas, so that it is not necessary to manufacture "region-specific" vaccines. Vaccination with H11 purified from either Australian or U.K. Haemonchus contortus was effective in protecting against subsequent challenge with Australian larvae, indicating that one type of H11 should have world-wide application.

Animals↗
PubMed1995

Interaction between Ostertagia circumcincta and Haemonchus contortus infection in young lambs.

Twenty-one-week-old, worm-free, pen-reared lambs were infected with either 6000 O. circumcincta L3 per week, or 3000 H. contortus L3 per week, or both (9000 L3 per week). Egg counts were monitored throughout the experiment, and worm burdens and larval establishment rates of both worm species were estimated after 4, 7, 10 and 13 weeks of infection. After 10-13 weeks of infection with H. contortus only, establishment of O. circumcincta was lower than in previously uninfected controls, demonstrating that a high level of immunity to H. contortus affords some cross-protection against O. circumcincta. Total H. contortus worm burdens and egg counts (about 2000 worms and 3000 e.p.g., respectively) in sheep infected with both worm species were less than half those observed in sheep infected with H. contortus alone (about 5000 worms and 10,000 e.p.g., respectively). Cross-protection between the two species was observed, but was probably less important than the reduction in H. contortus establishment that was caused by O. circumcincta disrupting abomasal physiology.

Animals↗
PubMed1995

Analysis of proteins related to conditioning for arrested development and differentiation in Haemonchus contortus by two-dimensional gel electrophoresis.

The abundance of the majority of proteins of infectious third-stage larvae (L3) of Haemonchus contortus, conditioned for arrested development, remained unaltered. Only seven proteins showed quantitative differences as observed by two-dimensional gel electrophoresis. These differences were also observed in a laboratory strain which has lost the ability for arrested development. The abundance of two of these proteins increased dramatically during conditioning of larvae for 5-10 weeks. This coincided with the highest percentage of inhibited larvae in experimental infections. Moreover, the abundance of these proteins decreased again after prolonged conditioning (22 weeks). The abundance of the other 5 proteins was not correlated to the percentage of inhibition. We therefore conclude that these proteins are involved in the aging process of larvae. The changes in protein between free-living (L3) and parasitic stages (L4) were large and seem to reflect the large environmental changes experienced by the larvae when entering a mammalian host. Early fourth- (EL4) and late fourth- (LL4) stage larvae differed in 9 proteins. One protein was stage-specific for EL4. These results imply that only minor alterations do occur in these stages notwithstanding the large morphological differences between these larvae.

Animals↗
PubMed1995

Variation in immune responsiveness of sheep to the antigens of intestinal nematodes and blowfly larvae.

The total and IgG1 antibody responses to the intestinal nematode parasites Haemonchus contortus and Trichostrongylus colubriformis were measured in the serum of 160 lambs, 4 months of age. These antibodies had developed as the result of natural exposure to the parasites on pasture. Three sires were examined and strong sire effects on half-sib progeny were found. Plotting of ELISA antibody results in two dimensions revealed clustering of responses within sire groups. Bimodal antibody distributions were also observed within sire groups and the whole population for T. colubriformis. A bimodal distribution of antibodies to H. contortus was found for one sire group but not for the whole population. The injection of blowfly larvae (Lucilia cuprina) extract into 42/160 lambs at a later age (12 months) was followed by increased antibodies to L. cuprina and an apparent increase in antibodies to T. colubriformis. A bimodal distribution for antibodies to L. cuprina was found in one sire group and in the whole population. These bimodal distributions of antibodies to L. cuprina did not coincide with the distribution of antibodies to T. colubriformis or H. contortus, measured on the same serum samples. It was concluded that high and low responder sire groups could be differentiated in lamb populations for all three parasites. These effects persisted during lamb maturation and appeared to be genetic effects. Finally, cross-reacting antibodies between L. cuprina and T. colubriformis appear to be stimulated by injection of L. cuprina antigens.

Animals↗
PubMed1995

Characterisation of an avermectin resistant strain of Australian Haemonchus contortus.

A strain of Haemonchus contortus (CAVR) isolated in Australia was found to be resistant to ivermectin (IVM) with 0.4 mg kg-1 of the anthelmintic failing to significantly reduce worm burdens. Resistance to IVM was sex-influenced in the CAVR strain with adult males showing a greater sensitivity to IVM. Cross resistance to moxidectin was evident with approximately 15% of the population surviving a dose of 0.1 mg kg-1. The free-living stages of the CAVR isolate had a reduced sensitivity to avermectin (AVM) inhibition of development and motility. Similar structure-activity patterns and resistance factors were obtained for a series of related AVMs as inhibitors of larval development and L3 motility in CAVR and White River II, an IVM-resistant H. contortus isolate from South Africa. Further, both isolates were found to be 3 times more sensitive to paraherquamide than a susceptible H. contortus isolate. This suggest that the same resistance mechanism is operating in both isolates. The CAVR strain is susceptible to the benzimidazoles, levamisole and closantel.

Animals↗
PubMed1995

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↗
PubMed1995

Vaccination of sheep against haemonchosis with H11, a gut membrane-derived protective antigen from the adult parasite: prevention of the periparturient rise and colostral transfer of protective immunity.

Pregnant ewes were immunised with a fraction highly enriched in the membrane glycoprotein antigen H11, isolated from the intestinal brush border of adult Haemonchus contortus. Immunity induced by immunisation was able to abolish almost completely (98-99%) the worm egg output from pregnant ewes challenged with ca. 10,000 infective larvae of H. contortus during the last trimester. Furthermore, lambs born and reared on vaccinated ewes had substantial antibody levels to H11 derived from maternal transfer. This antibody conferred moderate protection against a bolus challenge of ca. 3000 infective larvae of H. contortus in 5-week-old lambs.

Animals↗
PubMed1994

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↗