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Humoral and cellular responses following local immunization with a surface antigen of the gastrointestinal parasite Haemonchus contortus.

A purified, larval specific antigen of the abomasal parasite Haemonchus contortus was used to immunize sheep. In an attempt to induce a local immune response in the abomasum, the antigen was injected twice into the abomasal wall after one peripheral immunization. Serum antibody responses were boosted by each intra-abomasal immunization but not by the challenge infection given 3.5 weeks after the last immunization. Examination of the specific antibody secreting cells (ASC) recirculating in the peripheral blood indicated that there was an increase in blood ASC 5 days after local stimulation. This increase was maintained only after immunization and not after infection, probably reflecting the different responses induced when antigen is presented by injection in an adjuvant or by the parasite during infection. High proliferative T cell responses in the abomasal lymph nodes were only observed in one of the five sheep immunized with antigen; this was also the only sheep in this group to maintain an adult parasite burden at postmortem corresponding with the lowest antibody response. Peak faecal egg counts after infection were reduced by 54% in the immunized group compared to control sheep. Egg counts in the control sheep were, however, variable and dropped quickly, probably as a consequence of the inflammatory response induced by the injection of aluminium hydroxide into the abomasal wall.

Abomasum↗

Alveolar mastocytosis and eosinophilia in lambs with naturally acquired nematode infections of Protostrongylus rufescens and Haemonchus contortus.

Specific-pathogen-free Dorset and St. Croix lambs were placed on pasture contaminated with Haemonchus contortus third stage larvae and slugs carrying third stage larvae of Protostrongylus rufescens for an entire grazing season to evaluate breed differences in acquired resistance to these nematodes. Lambs were evaluated for clinical signs, clinical pathology and histopathologic lesions associated with these infections. Both breeds acquired natural infections with H. contortus and lungworm when allowed to graze contaminated pastures for 5 months during the summer and fall in central Maryland. Dorset sheep maintained heavy abomasal worm burdens of H. contortus throughout the grazing period when compared to St. Croix breed sheep. Seven of 12 Dorset sheep and three of 12 St. Croix sheep on pasture acquired heavy lungworm infections after at least 15 weeks of exposure, as evidenced by shedding of first stage larvae in feces and numerous subpleural lung lesions containing adult P. rufescens found at necropsy. All lungworm infected animals had mild respiratory and gastrointestinal signs, and two of five Dorset sheep with both infections had chronic anemia. All lungworm and H. contortus infected Dorset sheep had decreased numbers of circulating white blood cells. There was mastocytosis in the lungs of lungworm infected Dorset and St. Croix sheep when compared to age- and breed-matched control sheep prevented from acquiring both lungworm and trichostrongyle infections. No difference was noted in the number of mast cells in the abomasum, duodenum and skin of infected and non-infected Dorset sheep. A morphologic range of mast cell forms was observed in the lungs of infected sheep including transitional cells and globular leukocytes. The number of eosinophils was significantly greater in the lungs but not in the abomasum of infected sheep. Despite the pronounced cellular infiltrates surrounding the adult lungworms, they were viable on recovery and appeared undamaged when examined histologically.

Abomasum↗

Observations on the fumarate reductase system in Haemonchus contortus and their relevance to anthelmintic resistance and to strain variations of energy metabolism.

Fumarate reductase activity in a thiabendazole-resistant strain of Haemonchus contortus was found to be significantly lower than that from a susceptible strain. However, the fumarate reductase activity in a mebendazole-resistant strain did not differ from that in the susceptible strain, even though it was cross-resistant to thiabendazole. Published reports of fumarate reductase activity in strains of H. contortus susceptible or resistant to benzimidazoles were reassessed. A second, unrelated Australian thiabendazole-resistant strain also proved to have significantly diminished fumarate reductase activity, whereas two American strains, one resistant to thiabendazole and one to cambendazole, possess fumarate reductase activities indistinguishable from corresponding susceptible strains. It therefore appears that the phenomenon of benzimidazole resistance cannot be generally correlated with diminished fumarate reductase activity, although in the specific case of the Australian thiabendazole-resistant strains it may be a contributory factor.

Animals↗

Self-curve and immunity following infection and reinfection in ovine haemonchosis.

Egg expulsion and post mortem examination were used to compare self-cure and resistance to reinfection with Haemonchus contortus in female Merino sheep, AA and BB hemoglobin types. Animals were infected by 500 or 5000 larvae of H. contortus once, or several times, before being challenged by 500 (low dose) or 500 (high dose) larvae. In AA sheep infected once by 500 larvae self-cure was detected by a challenge of either 500 or 5000 larvae while in multi-infected sheep only a challenge of 5000 L, induced the self-cure. In BB sheep a mild self-cure was detected only by a challenge of 5000 larvae. In AA sheep receiving several infections, challenge by 500 larvae was not effective in producing resistance while 5000 L3 was effective in producing a state of resistance for up to 35 days post challenge. At post mortem examination, a strong state of resistance was evidenced in multi-infected animals by the low number of L4 and adults present in the abomasum up to the 35th day after challenge. In BB sheep no state of resistance was demonstrated by challenge with either 500 or 5000 L3, even in multi-infected animals.

Abomasum↗

The effects of transfer factor, parasitic antigen and precipitated immunoglobulin treatments in lambs infected with Haemonchus contortus.

Significant reductions in adult worm burdens in lambs less than six months old infected with 5000 third stage Haemonchus contortus larvae were achieved following the combination of a single transfer factor treatment with multiple parenteral treatments of adult parasitic antigen and precipitated immunoglobulin. Specific cellular responses in the abomasa associated with individual treatments were discernible in all the experimental groups and quantitative differences in these responses reflected the varying treatment levels and treatment combinations. The reactions associated with transfer factor were similar to immediate type hypersensitivity responses and their role as possible effector mechanisms in correcting partial immune incompetency in lambs is discussed.

Abomasum↗

Epidemiology of gastrointestinal parasites of Awassi sheep in Iraq.

Studies on the epidemiology of gastrointestinal nematodes of sheep, conducted at two different farms in the Baghdad area, in 1979, revealed two peaks in worm egg counts in ewes, one in spring and the other in autumn; the latter coincided with the lambing season. Similarly in lambs, two peaks of infection were apparent, but the spring peak was not so well defined. During the hot and dry summer months of Iraq, helminthic infection was at a low level. Faecal culture for larvae and their identification indicated the predominance of Haemonchus contortus infection in the ewes and that of Ostertagia and Trichostrongylus spp. in the lambs examined, Trichostrongylus infection predominating during the hot, dry period of the year. The self-cure phenomenon as well as preparturient rise in egg counts, as reported in other countries, was also observed during this study.

Animals↗

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↗

Wool production of sheep chronically infected with Haemonchus contortus.

A chronic infection was established in a group of twenty 18-month-old non-reproductive Merino ewes by oral administration of 3000 infective H. contortus larvae twice weekly for 12 weeks. Their live-weights and wool production were compared with those of 20 uninfected ewes grazing the same pasture. In the infected sheep, faecal egg counts increased over a period of 9 weeks to reach a mean of 5000 eggs per gram, accompanied by small, but significant effects on packed-cell volume and live-weight. The effects of infection on wool growth were also small, and not statistically significant, although there was evidence of a faster seasonal decline in wool growth in infected sheep. It was concluded that mortality induced by acute infection is the most important economic effect of this parasite.

Animals↗

Some factors influencing the immunisation of sheep with irradiated Haemonchus contortus larvae.

Over the last 6 years a number of experiments in which sheep were successfully immunised with 2 doses of 10 000 Haemonchus contortus larvae, irradiated at 60 krad in a gamma-source, have been reported by the authors. In this paper, the failure of such a regimen of immunisation, using the same strain of larvae, is reported together with an investigation of the possible causes. Technical errors were eliminated as a cause, on the grounds that the failure occurred in 2 separate laboratories using different 60Co sources, as was increased radiosensitivity of the continuously passaged strain of H. contortus as measured by the percentage development of irradiated larvae in 6 naive lambs. From experiments utilising 18 lambs and using both irradiated and normal larvae, it was postulated that the strain had lost its original degree of immunogenicity. Subsequently, in 2 experiments involving 27 lambs, it was shown that irradiation at 40 krad was completely successful in restoring immunogenicity and producing a degree of protection against challenge similar to that previously reported with 60 krad. It was concluded that, with the current strain of H. contortus, 60 krad is now too high a level of irradiation of larvae to produce a consistently high degree of protection against challenge.

Animals↗

Experimental infection of sheep by the camel stomach worm, Haemonchus longistipes.

An attempt has been made to infect sheep by Haemonchus longistipes with the objective of developing a relatively cheap and manageable model of camel haemonchosis. A large proportion of inoculated animals (28/34) developed patent infections; the prepatent period was short, lasting for 3 to 4 weeks in the majority of infected animals. Worm burdens were low and so was the total egg output. Most of the recovered worms were reduced in size.

Animals↗

Serum and abomasal antibody response of sheep to infections with Haemonchus contortus.

Serum and abomasal IgA, IgG and IgM antibody response against adult worm, L3 and egg antigens of Haemonchus contortus was monitored by the ELISA technique after one or two infections with this nematode. Following the first infection, antibody levels in serum did not change materially. After administration of a challenge dose of infective larvae, antibodies of the three immunoglobulin classes in infected animals rose slightly, but this rise appeared later than the fall in the faecal egg counts. In contrast, in abomasal mucosa, IgA anti-larval antibody levels, which did not increase materially after the primary infection, rose rapidly after a transient inhibition when sheep were challenged. A close temporal relationship was observed between the rise in local anti-worm IgA antibodies and the self-cure reaction, but antibody levels fell rapidly after worm diminution. The local antibody response was thus considered to be related to immunity of sheep to H. contortus.

Abomasum↗

The pathogenesis of experimental Haemonchus longistipes infection in camels.

The pathogenesis and clinical signs of Haemonchus longistipes infection were studied in four experimentally infected camels two of which were adults and the other two were young. In the former animals, an acute infection developed, characterized by mucoid diarrhoea, anorexia, anaemia, loss of body weight, oedema of the lower parts of the limbs, general malaise and death at 8-10 weeks post-infection. In the two younger camels, a less dramatic disease was encountered with less severe symptoms and no oedema, but also terminating fatally at 19-20 weeks post-infection. Parasitological, haematological and biochemical parameters were determined during the course of the infection and were mostly comparable with those usually encountered in haemonchosis of other animals.

Animals↗

The seasonal prevalence of inhibited development of Haemonchus contortus in sheep in Senegal.

Throughout a period of 13 months, the abomasa of 1024 sheep, living in a sahelian region, were examined for adult Haemonchus contortus and their inhibited L4 larvae. Of these 796 (78%) were found harbouring H. contortus, but with low worm burdens (geometric mean 37). Significant numbers of inhibited larvae appeared early in the dry season (November) and they represented 49% of the total in December. A similar proportion of inhibited larvae were present until April and after that their number declined abruptly. It was concluded that Haemonchus sp. survives the dry season as hypobiotic larvae as well as adults.

Abomasum↗

Haemonchus contortus and other trichostrongylid infections in parturient, lactating and dry ewes.

From autumn (April) to spring (November), groups of pregnant and dry Merino ewes grazed pasture contaminated with infective larvae of Haemonchus contortus, Ostertagia circumcincta and Trichostrongylus colubriformis. A periparturient rise in faecal egg counts occurred in the pregnant ewes, following the maturation in early spring of arrested fourth-stage larvae of H. contortus and failure of pregnant ewes to expel the resulting adult worms. Peak egg counts were seen in September, just before lambing. In dry ewes, egg counts were slightly elevated at the time of the rise in the pregnant ewes, but fell to very low levels as adult worms developing from previously arrested larvae were expelled. Lactating ewes acquired greater burdens of O. circumcincta and T. colubriformis than did dry ewes, but were equally refractory to new infections with H. contortus. It is therefore suggested that the impairment of immunity to helminth infection seen in reproductive ewes may be more specific than was previously envisaged.

Animals↗

The effect of dietary protein on the pathogenesis of acute ovine haemonchosis.

Two experiments were conducted to examine the influence of dietary protein on parasite establishment and pathogenesis in Finn Dorset/Dorset Horn lambs infected with Haemonchus contortus. The lambs were introduced to high (169 g) or low (88 g crude protein (CP) kg-1 dry matter (DM)) protein diets at 3 months of age and infected 1 month later with 350 larvae kg-1 body weight (BW). Blood and faecal samples were collected for analysis and body weights recorded weekly. In the first experiment some of the infected lambs were killed 6 weeks after infection and the remainder 5 weeks later. In the second experiment all the infected lambs were killed 4 weeks after infection. The results showed that lambs on a low protein diet were less able to withstand the pathogenic effects of infection with 350 H. contortus larvae kg-1 BW than lambs given the higher protein diet. Thus mortality was greater in the low protein group and adverse clinical signs, such as inappetence, weight loss and oedema were observed more frequently. This group also had a more severe anaemia, hypoproteinaemia and hypoalbuminaemia than the high protein group. In contrast, faecal egg counts, total daily faecal egg output and worm burdens were similar in all groups of infected lambs, indicating that the diets did not influence parasite establishment.

Acute Disease↗

The effect of dietary protein on the pathophysiology of acute ovine haemonchosis.

An experiment was conducted to examine the effect of dietary protein on the pathophysiology of acute haemonchosis in Finn Dorset/Dorset Horn lambs. Erythrokinetic and metabolic studies using 51Cr-red cells, 59Fe-transferrin and 125I-albumin and digestive efficiency and nitrogen balance measurements were conducted between 19 and 39 days after infection. Identical radioisotopic and nutritional studies were conducted later on pair-fed control lambs. The results demonstrated that lambs on a low protein diet (88 g crude protein (CP) kg-1 dry matter (DM)) were less able to withstand the pathophysiological consequences of infection with 350 Haemonchus contortus larvae kg-1 body weight than lambs given a high protein (170 g CP kg-1 DM) diet, despite similar worm burdens. Erythrokinetic studies indicated that abomasal blood loss was similar in both groups of infected lambs, and circulating red cell volumes were reduced and plasma volumes were expanded to a similar degree in both groups. The majority of infected lambs in both dietary groups responded equally well to the gastric haemorrhage by increasing the rate of red cell production and their absorption of dietary iron was greater than in the control lambs. Albumin catabolism was increased in both groups of infected lambs, particularly in those given the low protein diet. Within each dietary group there were no differences in the digestibilities of the various proximate fractions of the experimental diets and no differences in the urinary or faecal nitrogen losses between infected lambs and their pair-fed controls. However, loss of appetite was a major feature of the disease, particularly in the lambs given the low protein diet and this feature appeared to be crucial in determining the ability of the infected sheep to withstand the pathophysiological effects of the parasite.

Acute Disease↗

The effect of triflumuron (SIR8514) on the free-living stages of sheep nematodes.

Studies both in vitro and in vivo showed that the insect growth regulator, triflumuron, exhibited potent larvacidal effects against the free-living stages of Trichostrongylus colubriformis. Effects were not as marked on the closely related nematodes, Haemonchus contortus and Ostertagia circumcincta. Nevertheless, these findings suggest that growth regulators may be used to develop novel methods of nematode control, and thus offer alternatives or adjuncts to conventional anthelmintic therapy and at the same time reduce the selection for anthelmintic resistance.

Animals↗

Partial protection of lambs against Haemonchus contortus by vaccination with a fractionated preparation of the parasite.

Florida Native lambs, less than 6 months of age, were successfully vaccinated against Haemonchus contortus with a high mol. wt fraction (greater than 30,000 daltons) derived from a somatic extract of H. contortus larvae (SEL) and excretions and secretions (ES) of larvae isolated during in vitro development from the infective 3rd to 4th stage. A 59% reduction in adult worm numbers was obtained in vaccinates compared to naive lambs following challenge. The protection in vaccinated lambs was similar to that seen in lambs exposed to a primary infection of H. contortus larvae which had been cleared with anthelmintic prior to the challenge infection. The unfractionated SEL/ES preparation and a low mol. wt fraction gave no significant protection against challenge infection.

Animals↗