In vitro erythrophagocytosis by cultured macrophages stimulated with extraneous substances and those isolated from the blood, spleen and bone marrow of Boran and N'Dama cattle infected with Trypanosoma congolense and Trypanosoma vivax

A standard radioactive chromium (51Cr) release assay was used to assess the in vitro phagocytosis and lysis of bovine erythrocytes by cultured splenic, bone marrow and peripheral blood monocyte-derived (PBM) macrophages isolated from healthy and Trypanosoma congolense and T. vivax-infected cattle of the Boran and N'Dama breeds. Recombinant cytokines (rHuTNF-alpha and rBolFN-gamma) and non-acid-dialysed peripheral blood mononuclear cell (PBMNC) culture supernatants stimulated these PBM for enhanced activities. The stimulants caused increases in the rate of erythrocyte phagocytosis and lysis by cultured PBM in a concentration-dependent manner. But very high stimulant concentrations caused deceased in vitro erythrophagocytosis. However, bacterial lipopolysaccharide (LPS) and acid-dialysed PBMNC culture supernatants did not cause any increase in cultured PBM erythrophagocytosis. In vitro erythrocyte phagocytosis and lysis by splenic, bone marrow and peripheral blood monocyte (PBM)-derived macrophages of Boran breed of cattle infected with Trypanosoma congolense increased from 14 days post-infection (DPI) onwards and thereafter maintained at various levels above pre-infection. Cultured splenic macrophages showed the greatest erythrocyte destruction capability while PBM-derived macrophages was the least. The rates of in vitro erythrocyte phagocytosis and lysis were higher with the cultured PBM of the Boran than those of the N'Dama cattle during T. congolense infection. The rate of in vitro erythrocyte destruction was however, similar in both groups of cattle during T. vivax infection. These results correlated positively with the dynamics and degree of anaemia developed by these groups of animals during both T. congolense and T. vivax infections. Cattle infected with T. congolense and T. vivax developed varying degrees of normocytic normochromic anaemia during infection. Boran cattle developed a more severe anaemia, and had to be treated with diminazine aceturate, than N'Dama cattle during T. congolense infection. Both breeds of cattle developed a milder but similar degree of anaemia during T. vivax infection. None of the animals were treated. The results of this study indicated a role of in vivo macrophage stimulatory factors, notably cytokines such as TNF-alpha and IFN-gamma in host's serum, as well as parasite antigens, which may act singly or in concert, in the process of enhanced erythrocyte destruction, hence anaemia by the mononuclear phagocytic system (MPS) during bovine trypanosomosis.     

Non-immune control of trypanosomosis: in vitro oxidative burst of PMA- and trypanosome-stimulated neutrophils of Boran and N'Dama cattle

An in vitro assay that measures the generation of superoxide anions (O2-) was used to assess the level of oxidative burst of phorbol myristate acetate (PMA)- and trypanosome-stimulated neutrophils isolated from healthy Boran and N'Dama cattle, and those infected with Trypanosoma congolense. PMA stimulation of healthy bovine neutrophils resulted in between 300-400 % increase in O2- generation. Neutrophils of Boran cattle exhibited slightly higher but insignificant O2- generation capacity than those of the N'Dama breed. In vitro stimulation by trypanosomes of neutrophils isolated from Trypanosoma congolense-infected cattle caused significant increases in O2- generation, especially on days 14, 28 and 42 post-infection, of both breeds of cattle. No significant differences were observed in O2- generation capacity of the neutrophils of both breeds of infected cattle throughout the period of assay. The results of this study have shown that PMA and trypanosomes do cause an enhanced in vitro oxidative burst, hence trypanosome phagocytosis and killing activity of neutrophils. Neutrophils have been shown to play very significant roles in parasite clearance, hence reduction of trypanosome parasitaemia. The rates of both in vitro generation of O2- and trypanosome phagocytosis over time did not differ significantly between Boran and N'Dama breeds of cattle, even during T congolense infection in this study. Hence, it may be inferred that sustained and higher parasitaemia, more pronounced neutropenia, inadequate bone marrow response and less effective trypanosome-specific immune response, rather than defective neutrophil trypanosome destruction, may be the problem of trypanosusceptible cattle breeds.     

Responses of bovine chimaeras combining trypanosomosis resistant and susceptible genotypes to experimental infection with Trypanosoma congolense

West African N'Dama cattle have developed a genetic capacity to survive, reproduce and remain productive under trypanosomosis risk. The cellular and molecular bases of this so-called trypanotolerance are not known, but the trait is manifested by the N'Dama's greater capacity to control parasitaemia and anaemia development during an infection. In order to examine the role of the haematopoietic system in trypanotolerance, we have exploited the tendency for the placentas of bovine twin embryos to fuse. Placental fusion in cattle results in bone marrow chimaerism in twins. By comparison with the N'Dama, cattle of the East African Boran breed are relatively susceptible. We evaluated the role of the haemopoietic system in trypanotolerance by comparing the performance of five Chimaeric Boran/N'Dama twin calves with that of singletons of the two breeds. Chimaeric Boran/N'Dama pairs of twins were produced in recipient Boran cows by embryo transfer, and the majority of haemopoietic cells in all twinned individuals were of Boran origin. Thus, N'Dama chimaeras differed from N'Dama singletons in that the bulk of their haemopoietic system was derived from their susceptible Boran twins, while Boran chimaeras differed little from Boran control animals. All cattle became parasitaemic and developed anaemia. The N'Dama chimaeras did not manage their anaemia and white blood cell counts effectively. However, they were able to limit parasitaemia development. These results suggest that trypanotolerance is the result of two mechanisms, one that improves parasite control and is independent of the genetic origin of the haemopoietic tissue, and another that is influenced by haemopoietic tissue genotype and which improves control over anaemia. The capacity to maintain growth during infection was similarly dependent on the genetic origin of the haemopoietic tissue.     

Cytokine mRNA profiles in bovine macrophages stimulated with Trypanosoma congolense

It is known that different breeds of cattle display differential susceptibilities to Trypanosome congolense infections, and that N'Dama cattle remain more productive after infection than Boran cattle which are more susceptible to T. congolense. Macrophages from both breeds were cultured in vitro and the expressions of a number of cytokines and iNOS mRNA were analyzed using real time RT-PCR after stimulation with antibody-opsonized trypanosomes. No significant difference was seen between the responses of the two breeds. However, RNA levels of TNF-alpha in the IFN-gamma-primed macrophages were about 100-fold higher than those in the non-primed macrophages. A significant ten-fold decrease was seen for the anti-inflammatory cytokine IL-10. These results indicate that priming of the cells with IFN-gamma cause a serious shift toward an inflammatory response.     

Susceptibility of N'Dama cattle to experimental challenge and cross-species superchallenges with bloodstream forms of Trypanosoma congolense and T. vivax.

Susceptibility to Trypanosoma congolense, T. vivax challenge and cross species-superchallenges, and related effects on health and productivity were assessed in N'Dama cattle. Twenty-five N'Dama bulls aged 3-4 years and previously primed with trypanosome infections through natural tsetse exposure over more than one year were used. The experimental herd was divided in five groups each composed of five randomly selected animals. Group 1 was challenged with T. congolense, Group 2 with T. vivax, Group 3 was inoculated with T. congolense followed by a cross-superchallenge with T. vivax, Group 4 was inoculated with T. vivax followed by T. congolense cross-superchallenge. Animals in Group 5 were used as controls. Both T. vivax and T. congolense cross-superchallenges were carried out on Day 14 subsequent to respective initial T. congolense and T. vivax inoculations. All challenges were performed by intradermal needle inoculation of stocks of trypanosome bloodstream forms. In challenged animals (Group 1 to 4), parasitaemia profiles and packed red cell volumes (PCV) were measured for four months. Weight changes were recorded monthly and daily weight gain (DWG) computed. All cattle challenged with T. congolense became parasitaemic. Conversely, one animal in Group 2 and two in Group 3 never displayed patent T. vivax parasitaemia. Both in single (Group 1), initial (Group 3) and cross-superchallenged (Group 4) cattle higher percentage of positive blood samples and higher parasitaemia level were obtained following T. congolense than T. vivax inocula (Group 2, 3 and 4) (P<0.04 or greater). Overall the pre-challenge period, PCV values and DWGs were nearly identical in the five groups. Conversely, over the post-challenge period, cattle singly, initially and cross-superinoculated with T. congolense (Group 1, 3 and 4) displayed lower PCV values and DWGs in comparison with both control animals (Group 5) and with singly T. vivax challenged cattle (Group 2) (P<0.05 or greater). No difference in mean PCV levels and DWGs was found between animals in Group 2 and cattle in Group 5. It was concluded that trypanotolerant N'Dama cattle suffered more from T. congolense and mixed T. congolensel T. vivax infections, while pure T. vivax infection did not produce appreciable negative effects on their health and productivity. Therefore, considering that tsetse and trypanosomosis control campaigns are costly and are justified only when derived economic benefits exceed those of control, and also that an ample mosaic of farming systems exists in West Africa, species-specific trypanosome prevalence and relative impact should be assessed in various cattle populations and breeds differing in trypanosome susceptibility before advising any intervention. Moreover, virulence and related effects of T. congolense and T. vivax endemic stocks on health and productivity in local cattle populations should also be estimated in order to counsel appropriate economic protection measures against trypanosmosis, i.e. vector control and/or strategic use of trypanocidal drugs.     

Analysis of host genetic factors influencing African trypanosome species infection in a cohort of Tanzanian Bos indicus cattle

Trypanosomosis caused by infection with protozoan parasites of the genus Trypanosoma is a major health constraint to cattle production in many African countries. One hundred and seventy one Bos indicus cattle from traditional pastoral Maasai (87) and more intensively managed Boran (84) animals in Tanzania were screened by PCR for the presence of African animal trypanosomes (Trypanosoma congolense, Trypanosoma vivax and Trypanosoma brucei), using blood samples archived on FTA cards. All cattle screened for trypanosomes were also genotyped at the highly polymorphic major histocompatibility complex (MHC) class II DRB3 locus to investigate possible associations between host MHC and trypanosome infection. Overall, 23.4% of the 171 cattle tested positive for at least one of the three trypanosome species. The prevalence of individual trypanosome species was 8.8% (T. congolense), 4.7% (T. vivax) and 15.8% (T. brucei). The high prevalence of T. brucei compared with T. congolense and T. vivax was unexpected as this species has previously been considered to be of lesser importance in terms of African bovine trypanosomosis. Significantly higher numbers of Maasai cattle were infected with T. brucei (23.0%, p=0.009) and T. congolense (13.8%, p=0.019) compared with Boran cattle (8.3% and 3.6%, respectively). Analysis of BoLA-DRB3 diversity in this cohort identified extensive allelic diversity. Thirty-three BoLA-DRB3 PCR-RFLP defined alleles were identified. One allele (DRB3*15) was significantly associated with an increased risk (odds ratio, OR=2.71, p=0.034) of T. brucei infection and three alleles (DRB3*35, *16 and *23) were associated with increased risk of T. congolense infection. While further work is required to dissect the role of these alleles in susceptibility to T. brucei and T. congolense infections, this study demonstrates the utility of FTA archived blood samples in combined molecular analyses of both host and pathogen.     

A survival analysis of trypanosomosis diagnostic-test performance under natural infection challenge

Little is known about the time-to-first detection and the time difference (TD) between first parasitological and first serological diagnosis of Trypanosoma spp. infections under natural infection challenge in cattle. The objective of our study was to estimate these measures of "longitudinal aspects" of diagnostic performance and to investigate potential biological factors. Emphasis was on diagnosis at the genus level (Trypanosoma spp.). Twelve N'Dama, 12 Gobra zebu and 12N'DamaxGobra (F1) crossbred cattle (all animals non-infected at the start of the experiment, six male and six female animals in each cohort) were exposed to natural high tsetse challenge in the Niamina East area in The Gambia [Acta Trop. 71 (1998) 57]. The animals were investigated parasitologically (detection of trypanosomes by buffy-coat technique), serologically (detection of T. brucei, T. congolense and T. vivax antigen by enzyme-linked immunosorbent assay (ELISA)) and clinically (packed-cell volume, PCV) over a period of 180 days. The time-to-first detection of trypanosomes, trypanosomal antigen (cut-off as suggested by test supplier) and drop in PCV (subject-based cut-off values) were recorded as outcomes of interest. Thus, incidence was both parasitologically (I(p)), serologically (I(s)) and clinically (I(c)). Recurrent events were not considered. The TD between first parasitological and first serological detection was established as I(s) time minus I(p) time. The effect of breed and sex on the time-to-first detection and on TD was investigated using Cox (proportional hazard) regression and ANOVA, respectively.We found that time-to-first parasitological detection of trypanosomosis in N'Dama animals was significantly longer than in the two other breeds (Cox regression, P=0.002). A similar but less-strong (P=0.063) effect of breed on time-to-first detection of trypanosomal antigen was found, whereas no breed effect was observed for clinical detection (P=0.432). Sex had no effect in all detection systems. The TD varied between -56 and 115 (mean 28). Marked differences among breeds and between sexes were not observed (ANOVA, P=0.8). We suggest that incidence studies are more suitable for detecting risk factors for animal trypanosomosis than prevalence-based (cross-sectional) studies because the latter often result in misinterpretation of factors that increase the survival time with infection as risk factors.     

The distribution of tsetse (Diptera: Glossinidae) and bovine trypanosomosis in the Matutuine District, Maputo Province, Mozambique

A tsetse and bovine trypanosomosis survey was conducted during 1998 and 1999 in the Matutuine District of Maputo Province (Mozambique). A total of 59 Glossina brevipalpis and 17 Glossina austeni were captured throughout the district. Survey results suggest that Glossina brevipalpis is mainly concentrated in dense vegetation along the Maputo River and in the wetlands east of the river. G. austeni, on the other hand, was captured mainly in dense thickets in drier areas. Both tsetse species are suspected to be vectors of bovine trypanosomosis. Bovine trypanosomosis (75,5% Trypanosoma congolense) was diagnosed in 53 animals (13,9%) from seven sampling sites distributed throughout the district. The prevalence of cattle with anti-trypanosomal antibodies was high (29,9%).The incidence of trypanosomal infections in sentinel cattle was also high. The widespread distribution of bovine trypanosomosis and the high prevalence of infection are likely to have a significant impact on cattle production and, hence, the cattle restocking exercise in the district.     

Comparative studies on N''Dama and zebu cattle following repeated infections with Trypanosoma congolense

Twenty N'Dama and eight zebu cattle were inoculated intradermally with bloodstream forms of a cloned strain of Trypanosoma congolense originating from East Africa. All inoculated cattle became parasitaemic. Zebus showed consistently higher levels of parasitaemia and lower packed red cell volume (PCV) percentages than did N'Damas. Three of the eight zebus required treatment when high numbers of trypanosomes were present in the blood and PCV values dropped below 15 per cent. None of the N'Dama cattle needed treatment. Statistical analysis was performed on the data to assess the variability of parasitaemia and PCV levels before and during infection of the N'Dama cattle. The variation in PCV values was large between individuals during the early stages of the disease and diminished as infection continued. After trypanocidal drug treatment and a recovery period of 14 months, the same animals were inoculated intradermally with T congolense bloodstream forms isolated and cloned in The Gambia. Differences in susceptibility to the ensuing disease were apparent when comparing N'Dama and zebu cattle. Five zebu cattle needed trypanocidal drug treatment, while none of the N'Damas needed drug intervention. Ranking the 20 infected N'Damas according to average PCV levels revealed that the animals responded similarly to both infections.     

Implications of the re-invasion of Southeast Uganda by Glossina pallidipes on the epidemiology of bovine trypanosomosis

A study to assess the influence of re-invasion of Glossina pallidipes on the epidemiology of bovine trypanosomosis was conducted in Southeast Uganda. A total of 1,992 cattle were screened in villages, with (949) and without G. pallidipes (1043) for trypanosomosis using a combination of the BCT and HCT methods. The prevalence of trypanosomosis (15.5%), Trypanosoma brucei infection (1.4%), T. congolense infection (7.2%), T. vivax infection (5.3%) and mixed infection (1.6%) in cattle in villages with was significantly higher than in those without G. pallidipes: trypanosomosis (7.1%), T. brucei infection (0.6%), T. congolense infection (2.0%), T. vivax infection (3.3%) and mixed infection (1.2%) (overall trypanosome infection, chi2=35.5, d.f.=1, P<0.05; T. brucei infection, chi2=8.06, d.f.=1, P<0.05; T. congolense infection, chi2=22.8, d.f.=1, P<0.05 and T. vivax infection, chi2=6.4, d.f.=1, P<0.05). Infections of Trypanosoma congolense were predominant in cattle in villages with G. pallidipes, while T. vivax infections were predominant in cattle in villages without. In all villages, T. brucei infections were fewer than either T. congolense or T. vivax infections. The risk of transmission of T. brucei, T. congolense and T. vivax infections was 3, 2.7 and 1.6 times, respectively, higher in villages with G. pallidipes than in those without, despite the presence of G. f. fuscipes in either set of villages. The mean PCV (28.27+/-0.41, 95% CI) and mean herd size (3+/-0.46) of cattle in villages with G. pallidipes were significantly (P<0.05) lower than in those in villages without (mean PCV, 29.48+/-0.34; mean herd size, 4+/-0.72). It is evident that presence of G. pallidipes brings about an increase in the prevalence of T. congolense, which causes a more severe disease in cattle than other species of trypanosomes. This is a rare case of a re-invasion of a tsetse species whose disease transmission capability calls for refocusing of the traditional national tsetse and trypanosomosis control strategies to contain it.     

Relationships between trypanosome infection measured by antigen detection enzyme immunoassays, anaemia and growth in trypanotolerant N'Dama cattle.

Relationships were evaluated between trypanosome infection as measured by antigen detection enzyme immunoassays (antigen ELISA), anaemia as determined by average packed red cell volume (PCV), and animal performance as assessed by daily weight gain in 99 N'Dama cattle in Gabon exposed to natural tsetse challenge at 11.5 months of age and recorded 14 times over a 13 week period. Approximately half the animals were found to be infected for an average of five of the 14 times that they were examined: 38% with Trypanosoma congolense, 13% with Trypanosoma vivax and 49% with a mixed infection. Trypanosoma congolense infections had significant deleterious effects on animal growth, while T. vivax infections did not. Animals found on several occasions to be infected with T. congolense had significantly lower PCV values than those demonstrated to be infected on fewer occasions. No relationship was found between mean optical density (OD) values in antigen ELISA and PCV values. Animals capable of maintaining PCV values, even when antigen ELISA positive on a high number of occasions, grew at the same rate as uninfected animals. Animals that could not maintain PCV values when infected had poorer growth. Antigen ELISA has the potential to increase the efficiency of selection of trypanotolerant N'Dama cattle under tsetse challenge in the field, in three main ways. (1) Accurate identification of trypanosome species, especially in mixed species infections, clarifies relations between infection, anaemia and animal performance. (2) Detection of animals antigenaemic without patent parasitaemia could allow individuals with superior ability to control trypanosome infection to be identified. (3) More accurate measurement of the proportion of time an animal is infected allows more accurate evaluation of its anaemia control capability.     

Estimation of trypanosomal status by the buffy coat technique and an antibody ELISA for assessment of the impact of trypanosomosis on health and productivity of N'Dama cattle in The Gambia

The buffy coat/dark ground phase contrast technique (BCT) and an indirect antibody enzyme immunoassay (ELISA) were employed to assess the trypanosomal status of 32 N'Dama cattle, aged 19-28 months, exposed to natural challenge of Glossina morsitans submorsitans and G. palpalis gambiensis. Prior to the start of the investigation animals experienced 9-16 months of tsetse challenge in the study area. Blood and corresponding serum samples were examined monthly for a period of 8 months for patent parasitaemia by BCT and presence of Trypanosoma vivax and T. congolense antibodies by ELISA. In the ELISA, the reactivity of sera to anti-trypanosomal antibodies was expressed in percent positivity (pp). Packed cell volumes (PCV) and body weights were also recorded monthly, and daily weight gain (DWG) computed to assess the impact of trypanosomal status on health and productivity. During the study period, the overall parasitaemic trypanosome prevalence was 3% (6/199), while the serological prevalence was 54.7% (109/199). Both diagnostic tests revealed a predominance of T. vivax over T. congolense infections in N'Dama cattle. Sensitivity of the immunoassay was 83.3%. In T. vivax-parasitaemic cattle, antibodies persisted for 4-6 months after the parasite was detected by BCT. A significantly higher overall mean PCV level was observed in blood samples obtained from cattle found, in any particular month, negative by BCT and ELISA, compared with those blood samples from animals responding serologically positively for anti-trypanosome antibodies. Likewise, mean DWG was significantly higher in cattle found negative for both tests in comparison to animals presenting detectable anti-trypanosome antibodies and those detected positive by both tests. A significant negative relationship was observed between pp values and PCV levels in animals seropositive for T. vivax and/or T. congolense. Similarly, a negative relationship was observed between DWGs and pp values. PCV levels were significantly positively correlated with DWGs. It was concluded that serological screening could provide useful information complementary to that obtained by the use of BCT not only to assess more accurately the trypanosomal status of cattle populations, but also to evaluate the effects of trypanosome infection on animal health and productivity and estimate the trypanosomosis risk.     

Susceptibility and immune responses of zebu and taurine cattle of West Africa to infection with Trypanosoma congolense transmitted by Glossina morsitans centralis

Following tsetse-transmitted infection with Trypanosoma congolense, major differences in development of localised skin reactions, the ability to control parasitaemia, the degree of anaemia and in antibody response to trypanosomes were found between the reputedly trypanotolerant breeds of cattle (N'Dama, N'Dama/Baoule crosses, Baoule) and the trypanosusceptible West African Zebu. The local skin reactions that developed in the Zebu were large and severe while those that occurred in the other breeds were smaller and less severe or mild. The timing of appearance of parasitaemia and the height of the first peaks were similar in all the animals, but the Zebu were less able to control subsequent waves of parasitaemia. Possibly reflecting these events, it was only in the Zebu that significant anaemia developed. Neutralizing antibody against homologous metacyclic trypanosomes developed between 14 to 18 days after infection in all breeds of cattle; however, marked differences were found when antibody to trypanosomes derived from first peak parasitaemias were tested in the Zebu and Baoule. Neutralizing antibody against these parasites appeared in the Baoule on day 24 but were not detected in Zebu until day 51. Furthermore, the antibody titres were 3 log2 higher in the Baoule. It was concluded that the trypanotolerance exhibited by the West African taurine cattle might be related to a) their ability to control trypanosome numbers in the skin and in the bloodstream, an outcome that was possibly brought about by the earlier and superior immune response and b) failure to develop anaemia which might be associated with their capacity to control parasitaemia.     

Trypanosome-induced hypothyroidism in cattle.

Three Boran (Bos indicus) cattle infected with T. congolense IL 1180, and two uninfected control Boran cattle were used to study the effect of trypanosomiasis on the function of the thyroid gland. On a weekly basis, plasma thyroxine (T4) was measured by 125I-radioimmunoassay. Results indicated that T. congolense caused a significant decline in plasma T4 concentration in infected animals.     

Hypothalamic-pituitary-adrenal axis responsiveness to insulin-induced hypoglycaemia is modified by trypanosome infection in Boran (Bos indicus) cattle.

Ten Boran (Bos indicus) cattle were used to study the stress responsiveness of the hypothalamic-pituitary-adrenal (HPA) axis during trypanosome infection. Five cattle were infected with Trypanosoma congolense IL 1180 by tsetse challenge and five cattle served as controls. All infected animals developed acute trypanosomiasis. Insulin-induced hypoglycaemia (50 per cent of pre-insulin glucose concentration) was used as a stress factor. Acute hypoglycaemia was observed in three infected and three control animals after insulin challenge. Two animals from each group either did not respond or responded slowly. Hypoglycaemia in infected animals completely failed to induce an HPA axis response, while in control animals an HPA axis response was indicated by a significant increase in plasma adrenocorticotrophic hormone (ACTH) and cortisol concentrations (P less than 0.01). The results show that trypanosomiasis in Boran cattle can cause a decrease in the stress responsiveness of the HPA axis as indicated by a blunted ACTH/cortisol response to insulin-induced hypoglycaemia.     

Protective efficacy of isometamidium chloride and diminazene aceturate against natural Trypanosoma brucei, Trypanosoma congolense and Trypanosoma vivax infections in cattle under a suppressed tsetse population in Uganda

The protective efficacy of isometamidium chloride (ISMM) and diminazene aceturate (DIM) against Trypanosoma brucei, Trypanosoma congolense and Trypanosoma vivax infections in cattle under a suppressed tsetse population was assessed in southeast Uganda. A total of 66 and 57 trypanosome-infected cattle were treated with ISMM and DIM, respectively together with 177 trypanosome-free animals not treated were followed for 12 months, checked every 4 weeks. There was no statistical difference in the mean time to infection with any trypanosome species in animals treated with ISMM or DIM. However, the mean time to trypanosome infection was significantly longer for treated animals than controls. The mean time to infection with each of the three trypanosome species differed significantly, with the average time to T. vivax infection the lowest, followed by T. congolense and then T. brucei. The protective efficacy of DIM was as good as that of ISMM; implying curative treatments against trypanosomosis are sufficient for combination with tsetse control. Isometamidium chloride or DIM had the highest impact on T. brucei and T. congolense infections in cattle.     

The interaction of Trypanosoma congolense and Haemonchus contortus infections in trypanotolerant N'Dama cattle.

The interactions between Trypanosoma congolense and Haemonchus contortus infections were studied in N'Dama calves. A total of 38 N'Dama bulls was divided into four groups and each group infected either with H. contortus 1 week after infection with T. congolense or with T. congolense 4 weeks after infection with H. contortus, or with either infection singly. Parasitological (faecal egg counts, parasitaemia), haematological (packed cell volume, white blood cell counts, albumin) and clinical parameters (body weight change, mortality rate) were compared among the various groups. The results showed a reduced prepatent period and a markedly increased pathogenicity of H. contortus infections in animals with a concurrent T. congolense infection. The most harmful combination was a H. contortus infection 1 week after the T. congolense infection which resulted in a progressive and severe anaemia, accompanied by hypoalbuminaemia, increased weight loss and high mortality. The anaemia induced by dual infections showed a low responsiveness to chemotherapy and in several cases supportive treatment did not help recovery. The results also showed that animals with a concurrent T. congolense and H. contortus infection ran a higher risk of succumbing during the infection, and also during 10 weeks following treatment. Although infections with T. congolense alone produced no clinical signs, they were found to significantly reduce the ability of infected animals to mount a normal response to a subsequent H. contortus infection. It was concluded that the increased H. contortus egg excretion observed in animals infected with both parasites might significantly increase the risk of nematode infections and that the reduced prepatent period might necessitate more frequent anthelmintic treatments. These interactions should, therefore, be considered wherever attempts are made to control these two diseases.     

Parasitological prevalence of bovine trypanosomosis in Kindo Koisha district,Wollaita zone,south Ethiopia

A cross sectional survey to determine the distribution and prevalence of trypanosomosis was conducted in Kindo Koisha district, in the Wollaita zone in southern Ethiopia. A total of 1 008 adult cattle was examined at eight different localities. Dark field examination of the buffy coat, as well as stained thin blood film examination and packed cell volume (PCV) evaluation were the diagnostic techniques used. The overall prevalence of bovine trypanosomosis was 15 %. Among the positive animals, 108 (71.1%), 43 (28.4%) and 1 (0.6%) were due to Trypanosoma vivax, Trypanosoma congolense and mixed infection (T. vivax and T. congolense), respectively. The infection rate of T. vivax and T. congolense varied significantly (P < 0.01). The mean PCV of the positive and negative animals ranged between 18.3-32.1% and 26.8-33.4%, respectively. The mean PCV of negative animals (28 %) was significantly higher than the mean PCV of positive animals (22.3%) (P < 0.001). There was an inverse association of PCV with the prevalence of trypanosomosis (P > 0.05). The herd average PCV values of each site decreased with increasing proportion of the positive herds of that particular site. Of the diagnostic tests employed, the microhaematocrit buffy coat technique is relatively sensitive and it has an added advantage of indicating the general condition of the animal by haematocrit measurement. In view of the risk of trypanosomosis, a control intervention through the strategic application of appropriate trypanocidal drugs is recommended. A tsetse fly control scheme to reduce host-tsetse fly contact is equally as important as chemotherapy and chemoprophylaxis against trypanosomosis.     

Prevalence and source of trypanosome infections in field-captured vector flies (Glossina pallidipes) in southeastern Zambia

The prevalence of trypanosome infections in tsetse flies, Glossina pallidipes, collected from Chiawa and Chakwenga in Zambia with endemic trypanosomosis was assessed by polymerase chain reaction (PCR). Out of the 550 G. pallidipes, 58 (10.5%) flies were found to harbor trypanosome DNA. Infection rates of tsetse with Trypanosoma vivax universal, Trypanosoma congolense savannah, T. congolense forest and T. congolense kilifi were 4.2% (23/550), 4.7% (26/550), 1.1% (6/550) and 1.6% (9/550), respectively. To determine the mammalian hosts of T. congolense and T. vivax infections from the tsetse flies, mammalian mitochondrion DNA of blood meal in these flies were analyzed by PCR and subsequent gene sequence analysis of the amplicons. Sequence analysis showed the presence of cytochrome b gene (cyt b) of 7 different mammalian species such as human, elephant, buffalo, goat, warthog, greater kudu and cattle. Goats which were main livestock in these areas were further examined to know the extent of its contribution in spreading the infection. We examined the prevalence of trypanosome infections in the domestic goat population in 6 settlements in Chiawa alone. Of the 86 goats sampled, 4 (4.6%), 5 (5.8%), 4 (4.6%) and 4 (4.6%) were positive for T. vivax universal, T. congolense savannah, forest and kilifi, respectively. These findings showed that the host-source of trypanosome infections in vector fly give a vital information about spread of infection. The result of this study will certainly contribute in elucidating more the epidemiology of trypanosomosis.