The effect of multiple transfers of immune serum on maturing Schistosoma bovis infections in calves.

To investigate the role of humoral factors in immunity, serum from cattle with naturally acquired immunity to Schistosoma bovis was injected intraperitoneally into calves that had been infected 4 weeks earlier with 10,000 S. bovis cercariae. Serum was injected weekly until 12 weeks post-infection to a total of 4,500 ml per calf and controls received normal serum or saline. No significant difference in worm or in faecal or tissue egg counts were seen in the three groups of recipients in spite of the observation that the serum donors had proved highly resistant to experimental challenge. In a second experiment, pre-infection or 4-, 8- or 12-week post-infection serum from donors given a single experimental infection with 10,000 S. bovis cercariae was injected intraperitoneally into groups of calves that had been infected 4 weeks earlier with 20,000 S. bovis cercariae. Injections were given weekly up to week 10 post-infection to a total of 2000-3500 ml serum per calf. In calves injected with immune serum there was a reduction in faecal and tissue egg counts and in the numbers of worms recovered as compared with the controls. In recipients of 8- and 12-week serum the reductions in faecal and tissue egg counts were higher than those in worm recovery, suggesting that 8- and 12-week post-infection sera contained factors capable of causing, in addition to worm death, suppression of worm fecundity. This provides further evidence of the importance of fecundity suppression in immunity to schistosomiasis.

Suppression of Schistosoma bovis egg production in cattle by vaccination with either glutathione S-transferase or keyhole limpet haemocyanin.

Two of the antigens which have shown vaccine potential in animal experiments against Schistosoma mansoni are glutathione-S-transferase (GST) and GP38, protective epitopes of which are shared with keyhole limpet haemocyanin (KLH). We therefore tested S. bovis GST and KLH for vaccine efficacy against S. bovis in the natural Zebu cattle host. In a preliminary experiment three vaccinations with a total of 1.39 mg of native GSTs of S. bovis induced specific antibody at the time of challenge as detected by Western blotting and ELISA and mean faecal egg counts between weeks 6-10 post-challenge were reduced by 56.4 to 82.5% compared to non-vaccinated controls. Mean adult worm recoveries and tissue egg densities in large intestine and liver samples were also reduced in the vaccinated group, but these differences were not statistically significant. In a subsequent experiment one group of calves was vaccinated with a similar schedule to that used above; a second group of calves was given only two injections of GST (total 0.48 mg protein); a third group of calves was vaccinated twice with a total of 2.0 mg KLH in PBS. All three vaccination schedules induced specific antibody. Both GST vaccination schedules induced significant reductions in faecal egg counts compared to non-vaccinated controls and in this experiment tissue egg densities were also significantly reduced. A striking finding, however, was that adult worm counts were not reduced by vaccination. An essentially similar outcome resulted from KLH vaccination, since there were significant reductions in faecal and tissue egg counts in the absence of a reduction in adult worm numbers.(ABSTRACT TRUNCATED AT 250 WORDS)

Protective effect of irradiated metacercariae of Fasciola gigantica and irradiated cercariae of Schistosoma bovis against fascioliasis in goats.

Sensitisation of goats for eight weeks with metacercariae of Fasciola gigantica gamma-irradiated at 3 kr resulted in significant resistance to an homologous challenge with normal metacercariae. However, serum sorbitol dehydrogenase assay suggested that, whereas little damage was produced by immunising infections, considerable hepatic damage was caused by flukes remaining from challenge infections. On the other hand sensitisation with cercariae of Schistosoma bovis gamma-irradiated at 3 kr did not stimulate any resistance against heterologous challenge with F. gigantica.

Studies on heterologous resistance between Schistosoma bovis and Fasciola gigantica in Sudanese cattle.

Using the local strains of Schistosoma bovis and Fasciola gigantica, it was shown that Sudanese zebu calves with mature primary infections of F. gigantica were highly resistant to challenge with S. bovis cercariae, and vice versa. Liver enzyme tests showed that, in both cases, the primary infections had caused some liver damage. Primary infection with irradiated S. bovis cercariae, which did not cause significant liver damage, did not protect significantly against challenge with F. gigantica.

Erythrokinetics and albumin metabolism in primary experimental Schistosoma bovis infections in Zebu calves.

Red cell kinetics and albumin metabolism were studied in calves infected with either 100 or 200 Schistosoma bovis cercariae per kg body weight, by the use of 59Fe-labelled transferrin, 51Cr-labelled erythrocytes and 125I-labelled albumin; a third group of worm-free animals acted as controls. The anaemia which developed in the infected calves was shown to be due basically to an accelerated rate of red cell loss from the circulation, and became evident around the seventh week of infection, increased in severity during the following two months, and subsequently subsided. In view of its close similarity to the pattern of faecal egg excretion, it was concluded that haemorrhage into the intestine caused by the exit of eggs was the principal aetiological factor: haemolysis was excluded by the absence of both splenomegaly and hyperferraemia . Erythropoiesis was also accelerated in infected animals, but could not keep pace with the rate of red cell breakdown to which the animals were concurrently subjected. Haemodilution was involved, but not to a significant extent. The hypoalbuminaemia associated with infection was caused by an increased rate of albumin catabolism, and a plasma volume expansion, and was accompanied by marked depletion of all albumin pools, but particularly the extravascular pool. The pattern of albumin catabolism closely followed that of red cell loss, suggesting that passage of plasma as whole blood into the intestine was the basic cause of hypoalbuminaemia. Red cell losses and albumin hypercatabolism were more severe in the more heavily infected group, and although both subsided as egg counts fell, they remained evident even 1 year after infection. This may partly explain the failure of infected animals to regain the weight lost during earlier stages of disease.

Observations on cattle schistosomiasis in the Sudan, a study in comparative medicine. V. The effect of praziquantel therapy on naturally acquired resistance to Schistosoma bovis.

Studies in the White Nile area of the Sudan have shown that Zebu cattle acquire a high degree of resistance to Schistosoma bovis as a result of repeated natural exposures without, however, being able to eliminate their populations of adult schistosomes, although these do show greatly suppressed fecundity. To test whether these adult worms are necessary for the maintenance of resistance we cured six "naturally resistant" cattle (TC group) with a double treatment of 25 mg/kg praziquantel and compared their response to a 70,000 cercariae challenge with groups of "naturally resistant" but untreated cattle (UC group) and with previously unexposed, challenged cattle (CC group). Challenge was carried out 7 weeks after the second dose of praziquantel. The results confirmed that untreated cattle are "naturally resistant" and also showed that resistance was not abrogated by cure of the naturally-acquired infections. Thus, fecal egg counts after challenge reached mean maxima of 2,432 eggs per gram (epg) in the CC, but only 5 epg and 28 epg in the TC and UC groups, respectively. Similarly, mean worm counts were 85% and 69% lower in the TC and UC groups, respectively, and mean tissue egg densities were reduced by 72-99%, and 56-80%. Histopathologically, the TC and UC groups were also far less affected than the CC. Effective praziquantel treatment does therefore not destroy naturally acquired resistance to S. bovis, and may benefit infected livestock even in the absence of transmission control. The situation in human schistosomiasis is less clear, but there are several epidemiological and experimental indications of a similar conclusion for S. mansoni.

Observations on cattle schistosomiasis in the Sudan, a study in comparative medicine. VI. Demonstration of resistance to Schistosoma bovis challenge after a single exposure to normal cercariae or to transplanted adult worms.

Calves were immunized with Schistosoma bovis by a single experimental exposure to 10,000 normal cercariae. Some of these calves were perfused 14 weeks later, and a part of their worm loads was surgically transplanted into groups of normal recipient calves: "WPR" group calves received 500 pairs of worms; "MR" group calves received between 650 and 1,000 male worms alone. All three groups were subsequently challenged 10 weeks after surgery with 20,000 cercariae, as were a previously unexposed group of controls ("CC"). Mean post-challenge fecal egg counts in the animals immunized with cercariae ("PC" group) rose to a maximum of only 60 eggs per gram (e.p.g.), compared to 376 e.p.g. in the CC, and maximum fecal egg counts in the WPR and MR groups were also somewhat lower than in the CC, at 152 and 250 e.p.g., respectively. In spite of the much lower fecal egg counts in the PC than in the CC group, calculated adult "challenge" worm recoveries were only reduced by 11%, but PC group tissue egg densities derived from the challenge were 78-100% lower than in the CC. The WPR and MR groups had 43% and 37%, respectively, fewer worms than the CC, and mean tissue egg densities were lower by 39-63% and 63-76%, respectively, though in most cases there were no statistically significant differences from the CC.(ABSTRACT TRUNCATED AT 250 WORDS)

Observations on cattle schistosomiasis in the Sudan, a study in comparative medicine. IV. Preliminary observations on the mechanism of naturally acquired resistance.

Suppression of egg production is the main parasitological manifestation of naturally acquired resistance to Schistosoma bovis in Sudanese cattle. In preliminary investigations on the mechanisms involved, 700-4,000 "suppressed" adult worms were surgically transplanted from six "resistant" donor cattle with very low fecal egg counts (0-8 eggs/g, epg) into six normal recipients. After transplantation, large numbers of eggs were excreted in the feces of the recipient cattle, beginning at between 5 and 16 days after operation, and reaching counts of 55-405 epg at between 6 and 20 days post transplantation. In the cattle with the highest egg counts, egg counts soon fell sharply from peak levels, whereas in cattle with lower peak counts, more steady counts were maintained. All the recipients were perfused at days 46-56, when between 0.1% and 78.5% of the transplanted worms were recovered. In the second experiment, 1,000-ml quantities of pooled sera from "resistant" donors were injected intraperitoneally into each of four normal recipient calves, while another four were injected with pooled sera from uninfected cattle. All the calves were challenged percutaneously the next day with 7,500 cercariae each, and the course of infection was followed by parasitological and clinical measurements until perfusion 18 weeks later. The results showed that the "immune" sera had a negligible effect on the numbers of worms which developed, and had no significant effect on the fecal egg counts or clinical parameters studied. There was, however, some evidence from the tissue egg counts of a reduction in the fecundity of the worms in calves injected with "immune" sera.(ABSTRACT TRUNCATED AT 250 WORDS)

Clinical diagnosis of schistosomiasis in Sudanese cattle.

This study was done in the White Nile Province to characterise the history and signs of naturally occurring Schistosoma bovis infection in cattle (Gorag). Necropsy and laboratory examinations were performed on 10 animals six to 30 months of age which were in poor condition. They were selected because of a history suggestive of schistosomiasis. All the animals showed some degree of S. bovis infection; eight had a moderate or heavy degree of infection. Also all had liver damage due to either past or active Fasciola gigantica infection. Although concurrent infection with these two trematodes is common an owner who diagnoses Gorag is most likely referring to the syndrome caused by S. bovis as being the major cause of the poor performance observed. Fasciolicide treatment may eliminate active fascioliasis as the principal damaging agent. Also differentiating signs of haemorrhagic diarrhoea, severely sunken eyed appearance and only moderate inappetence are common in animals with acute schistosomiasis.

Effects of praziquantel and metrifonate on Schistosoma bovis infections in Sudanese cattle.

Twelve nine-month-old zebu calves were each experimentally infected with 10,000 Schistosoma bovis cercariae. Four were treated orally with 20 mg/kg praziquantel at weeks 9 and 14 after infection, and four were treated orally three times with metrifonate at week 11 (25 mg/kg) and again at week 14 (50 mg/kg). Praziquantel proved to be highly effective, reducing faecal egg counts near to zero; the mean live worm count in the treated calves at week 16 was only 32, compared to 2850 in the untreated group, a reduction of 98.9 per cent. In contrast, metrifonate treatment caused only a very short-lived, partial reduction in faecal egg counts, and no reduction in live worm counts.

Observations on cattle schistosomiasis in the Sudan, a study in comparative medicine. I. Epizootiological observations on Schistosoma bovis in the White Nile Province.

A 2-year epizootiological study was carried out on Schistosoma bovis in cattle in an enzootic area of the Sudan. The prevalence of infection, as judged by the Pitchford fecal egg counting technique, was very high, approaching 90% in 1.5-year-old animals. There was, however, a lower prevalence in older cattle, and this trend was also seen with the fecal egg counts. This suggests that the cattle gradually acquire resistance to reinfection, particularly since there seems to be little age-related variation in the amount of water contact. Monthly incidence rates, estimated from fecal examinations of initially uninfected "tracer" calves showed a marked seasonal pattern, being much higher in the hot summer months, and snail infection rates showed a similar seasonal pattern. In addition, both the monthly incidence measurements and the snail infection rates showed that transmission was much heavier in 1976 than in 1977. Epizootics such as those recorded previously in this area presumably arise from exceptionally intense transmission years.

Observations on cattle schistosomiasis in the Sudan, a study in comparative medicine. II. Experimental demonstration of naturally acquired resistance to Schistosoma bovis.

Epizootiological observations on Schistosoma bovis in cattle at Kosti, Sudan, showed a significant fall in age-specific prevalence and intensity with age, based on fecal egg count. To test the possibility that this is due to acquired resistance, Kosti cattle and a control group of cattle of similiar breed and age from a nonenzootic area were experimentally challenged with 70,000 S. bovis cercariae. Clinical observations showed very clearly that the Kosti cattle were able to withstand almost completely the effects of the challenge, whereas the controls developed lethal infections. Resistance was further demonstrated by clear differences between the two groups in terms of their body weights, hematological measurements, histopathological and pathophysiological responses, and worm and egg counts. The data suggested that the main basis of the resistance was a suppression of egg production by the worms from the challenge, rather than absolute prevention of their maturation. There was also evidence of a suppression of the fecundity of worms in the naturally infected Kosti cattle.

Observations on cattle schistosomiasis in the Sudan, a study in comparative medicine. III. Field testing of an irradiated Schistosoma bovis vaccine.

Previous work has shown that cattle can acquire a strong resistance to Schistosoma bovis infection following repeated natural exposure. Partial resistance to a laboratory challenge with S. bovis has also been demonstrated in calves after immunization with an irradiated schistosomular or cercarial vaccine. The aim of the present study was to see whether this type of vaccine could protect calves under the very different conditions of natural exposure to S. bovis in the field. Thirty 6- to 9-month-old calves were each immunized with 10,000 irradiated S. bovis schistosomula by intramuscular injection and 8 weeks later were released into an enzootic area along with 30 unvaccinated animals. The calves were followed up for 10 months, during which period protection was evidenced by a lower mortality rate, a slower rate of acquisition of infection, and lower fecal egg counts in the vaccinated calves. Necropsy of the survivors showed 60--70% reductions in worm and tissue egg counts of the vaccinated calves as compared to those not vaccinated.

The pathology of experimental Schistosoma bovis infection in sheep.

Five desert sheep were exposed to 5000 or 10000 Schistosoma bovis cercariae each, and the parasitological clinical and pathological parameters were recorded. The pre-patent period was approximately 7 weeks and from that time onwards, the animals became progressively ill and emaciated. Following necropsy at the 12th week, a percentage schistosome recovery ranging from 36%--63.6% was found, and there were high egg densities in the intestinal tract and the liver. These organs revealed severe pathological lesions which were described in detail along with the changes occurring in other tissues.