The FAO/IAEA DGIS Coordinated research programmes on trypanosomiasis diagnosis and animal production in Africa.

During the period 1987-1992, Professor Dick Zwart played a key role in guiding the work of two African research networks established by the Joint FAO/IAEA Division and supported financially by the Netherlands Directorate General of Development Co-operation. These networks brought together staff from African institutes involved in the diagnosis and control of trypanosomiasis and in animal production, with scientists from International Agricultural Research Centres and other advanced research institutions in the Netherlands and elsewhere. Through this approach, the value of recently-developed immunoassay methods was assessed for diagnosing trypanosomiasis and monitoring control programmes, and similar methods were used to monitor animal productivity and ways of improving this on small-holder farms. Furthermore it strengthened the portion together with the functioning of the national agriculture research institutions. This paper outlines the main technical and other accomplishments of these networks.

An ELISA for the diagnosis of Babesia ovis infection utilizing a synthetic, Babesia bovis-derived antigen.

The development of an enzyme-linked immunosorbent assay (ELISA) for the detection of Babesia ovis antibodies is described. In an initial study, a crude Babesia bovis antigen and a synthetic B. bovis-derived antigen (designated 11C5) were used to screen 46 B. ovis-positive and 55 negative sheep sera. A 95% correlation between the two antigenic preparations was found with the positive sera; no negative sera gave positive reactions. The synthetic antigen was then used in the screening of 1466 sera collected from sheep from 18 regions of Turkey. A high incidence of B. ovis-positive reactions was found from all regions (60-80%) in sheep over 1 year old, while from two smaller samples the incidence in young sheep was much less (28 and 52%). This test is superior to existing ones because the synthetic antigen can be produced in a highly reproducible state, is specific and is stable over extended periods of time.

The pathogenesis of experimental Schistosoma bovis infections in Sudanese sheep and goats.

The pathogenic effects of experimental Schistosoma bovis infection in Sudanese sheep and goats were investigated by a variety of clinical, parasitological, physiological and histopathological techniques; uninfected animals of each species were used as controls. Infected animals of both species lost or failed to gain weight and developed a haemorrhagic diarrhoea, inappetence, marked anaemia, hypoalbuminaemia, hyperglobulinaemia, hyperproteinaemia and eosinophilia. These changes first became noticeable around the time of onset of oviposition and their severity was generally related to faecal egg counts. Red cell breakdown and albumin catabolism were much higher in infected than in control animals of the same species, and it was concluded that these changes were due to haemorrhage resulting from the extrusion of large numbers of eggs through the intestinal mucosa. Although all the animals were infected with the same number of cercariae, both the number of worms reaching maturity and the tissue egg counts tended to be higher in sheep than in goats. On the other hand, goats had significantly higher faecal egg counts than sheep and it is suggested that this was the reason for the generally more severe disease in the former species.

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. 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.

Schistosoma bovis in calves: the development and clinical pathology of primary infections.

The clinico-pathological effects of Schistosoma bovis were monitored in zebu calves for a year after exposure to 100 or 200 cercariae/kg body weight and were related to the number and reproductive activities of the parasites present. The disease was characterised by diarrhoea, weight loss or poor weight gain, anaemia, serum protein changes and eosinophilia. These changes were broadly related to the level of infection and were most prominent during the two months following patency when faecal egg counts were highest. Subsequently, the condition of most animals improved. This was associated with a marked reduction in faecal egg excretion which in turn was related to worm deaths, reduced worm fecundity and egg retention in the tissues.

Pathophysiological changes associated with Parascaris equorum infection in the foal.

The mechanisms involved in the pathophysiological disturbances associated with the presence of mature Parascaris equorum in the small intestine were investigated with radioisotopic techniques. The results suggested that, compared with worm-free controls, infected foals had a reduction in gut motility, an increase in the body solids ratio, a lowering of the body pool of albumin and a decreased ability to incorporate dietary methionine into plasma protein.

Bovine trypanosomiasis: the red cell kinetics of ndama and Zebu cattle infected with Trypanosoma congolense.

The responses of susceptible Ndama and Zebu cattle to needle challenge with Trypanosoma congolense were followed using parasitological, haematological and radio-isotopic methods and compared with those of corresponding uninfected animals. In both breeds, infection became patent at the same time but peak parasitaemias were significantly lower, were attained later and were of short duration in the Ndama. All infected animals became anaemic, the severity of which correlated with the level and duration of parasitaemia. However, even when parasites could no longer be detected in the blood, packed cell volumes showed little tendency to recover. The anaemia was due to increased intravascular red cell destruction and was more pronounced in the Zebu. Haemodilution was not a feature. Increased red cell syntheisis occurred in infected animals of both breeds but particularly in the Zebu; this accounted for the capacity to maintain packed cell volume levels following the initial drop associated with parasitaemia. However, in most cases red cell synthesis was less than expected from the degree of anaemia, suggesting impairment of bone marrow function. Measurement of red cell iron utilization indicated that this was due to effective from re-utilization from degraded red cells arising from reticulo-endothelial blockade. It is concluded that the anaemia in this disease and its underlying processes are broadly in line with the number of parasites in the blood and that the superior resistance of the Ndama cattle lies in their ability to control parasitaemia rather than their capacity to mount a more efficient erythropoietic response.

The pathophysiology of ovine fascioliasis: studies on the feed intake and digestibility, body weight and nitrogen balance of sheep given rations of hay or hay plus a pelleted supplement.

The effect of Fasciola hepatica on feed intake and digestibility, body weight and nitrogen balance was measured during the course of experimental infections in sheep given a diet of hay or hay with 'concentrate'. A system of paired feeding was used to allow comparisons between infected and control animals. After the sixth week the appetites and body weights of all infected animals declined but both features were more prominent in the group given the diet of hay alone. By week 14 these animals became moribund and were necropsied whereas those given hay with concentrate which had the same fluke burdens, survived until week 20. At necropsy, the loss of weight in both groups was comparable but greater than in their pair-fed uninfected controls, suggesting that inappetence was not solely responsible. Measurements of feed digestibility revealed few differences between infected and control animals on either diet, but nitrogen retention was markedly lower in the infected animals after the eighth week, and sufficient to account for their inferior weight performance. Reduced nitrogen retention was primarily a reflection of a high urinary excretion in the infected animals, faecal excretion remaining unaltered despite the loss of substantial amounts of nitrogen into the gut as a result of the parasites' haematophagic activities.

The blood volumes and erythrokinetics of Ndama and Zebu cattle experimentally infected with Trypanosoma brucei.

The responses of susceptible Ndama and Zebu cattle to experimental infection with Trypanosoma brucei were compared using haematological, parasitological and radioisotopic methods. Animals of both breeds became anaemic, but this was more severe in the Zebu cattle, one of which died. Although the prepatent period was the same in animals of both breeds, the levels of the first and subsequent peaks of parasitaemia were higher in the Zebu. The anaemia was due to an accelerated rate of red cell break-down which was more marked in the Zebu cattle. Haemodilution was not a feature. There was no evidence of dyshaemopoiesis but iron reutilisation from degraded erythrocytes was impaired. The greater resistance of the Ndama to T brucei infection could not be attributed to the capacity of this breed to mount a more effective erythropoietic response than the Zebu.

Genetic resistance to helminths. The influence of breed and haemoglobin type on the response of sheep to re-infection with Haemonchus contortus.

The influence of genetic factors on acquired resistance to Haemonchus contortus infections in sheep was investigated. Animals whose primary infections were terminated with an anthelmintic failed to develop any immunity against subsequent challenge as judged by worm numbers. Nevertheless, all were better able to retard the development and reduce the fecundity and haematophagic activities of their parasite populations than animals undergoing primary infections. High levels of resistance, as judged by all these parameters, were observed in most animals when the challenge larvae were superimposed on existing worm populations. The patterns of worm establishment and disease indicated that genetic factors operated in determining resistance, since fewer worms became established and less severe clinical and pathophysiological changes were observed in Scottish Blackface than in Finn Dorset sheep with the same haemoglobin type. Similar advantages were displayed by animals with haemoglobin AA and to a lesser extent those with haemoglobin AB over haemoglobin BB types. The importance of breed was further indicated by the occurrence of 'self-cure' in the majority of the Scottish Blackfaces but in only one Finn Dorset. There was no evidence that this reaction was associated with haemoglobin type.

Genetic resistance to helminths: a possible relationship between haemoglobin type and the immune responses of sheep to non-parasitic antigens.

The results of some preliminary studies on a possible association between haemoglobin type and the antibody responses of sheep to a variety of non-parasitic antigens are described. Sheep with haemoglobin AA showed a significantly better response to human serum albumin than animals of the same breed with haemoglobin BB. There was also suggestive evidence of a better response by haemoglobin AA types to rabbit red cells and horse gamma globulin. These results indicate that the greater resistance of these sheep to gastrointestinal nematodes is reflection of their superior immunological competence.

Genetic resistance to helminths. Comparison of the development of Ostertagia circumcincta infections in Scottish Blackface sheep of different haemoglobin type.

Over the years a number of investigators have reported that sheep with haemoglobin A are more resistant to Haemonchus contortus than animals of the same breed with haemoglobin B. The experiment described here was an attempt to ascertain whether a similar association might exist between haemoglobin type and resistance to non-haematophagic parasites. The results indicate that such a relationship might exist, since Scottish Blackface sheep with HbA showed milder biochemical and pathophysiological changes than their HbB counterparts and at the same time harboured smaller numbers of adult worms and more inhibited larvae when necropsied 16 days after a primary infection with 100,000 Ostertagia circumcincta third-stage larvae.