Justin Masumu

Five-fold increase in Trypanosoma congolense isolates resistant to diminazene aceturate over a seven-year period in Eastern Zambia.

Two groups of Trypanosoma congolense isolates collected from cattle in 1996 (n=39) and 2003 (n=38) in the Eastern Province of Zambia were analyzed by BclI-PCR-RFLP to assess the evolution of diminazene aceturate (DA) resistance over a period of seven years. The results show a significant increase of DA resistance in this relatively short period of time. In 1996, among the 39 isolates, 61.5% were found sensitive, 12.8% resistant and 25.7% had a mixed BclI-PCR-RFLP profile. In 2004, among the 38 isolates, 10.5% were found sensitive, 63.2% were resistant and 26.3% showed a mixed BclI-PCR-RFLP profile. In vivo tests in mice showed that isolates with a sensitive or mixed RFLP profile were sensitive to DA whereas isolates with a resistant RFLP profile were resistant. Since there are no indications that the drug pressure has increased between 1996 and 2003, it is suggested that genetic exchange of resistance genes might explain the increased frequency of resistance to DA.

Virulence in Trypanosoma congolense Savannah subgroup. A comparison between strains and transmission cycles

Trypanosoma congolense strains have been shown to differ in their virulence both between subgroups and within the Savannah subgroup between strains. This review revisits these findings and complements them with information on the virulence of T. congolense Savannah subgroup strains isolated from cattle (domestic transmission cycle) in different geographical areas and of strains isolated in protected areas where trypanotolerant wildlife species are the reservoir of the trypanosomes (sylvatic transmission cycle). The virulence of a total of 62 T. congolense Savannah subgroup strains (50 domestic and 12 sylvatic), determined using a standard protocol in mice, was compared. Virulence varied substantially between strains with, depending on the strain, the median survival time of infected mice varying from five to more than sixty days. The proportion of highly virulent strains (median survival time <10 days) was significantly (P = 0·005) higher in strains from the sylvatic transmission cycle. The analysis highlights repercussions of the domestication of the trypanosomiasis transmission cycle that may have to be taken in consideration in the development of trypanosomiasis control strategies.

Comparison of the transmissibility of Trypanosoma congolense strains, isolated in a trypanosomiasis endemic area of eastern Zambia, by Glossina morsitans morsitans

Transmission experiments were conducted to compare the transmissibility of genetically different Trypanosoma congolense (Savannah subgroup) strains isolated from cattle in a trypanosomiasis endemic area of eastern Zambia. A total of 17 strains were compared. Three strains were extremely virulent with a short pre-patent period, high parasitaemia and a short median survival time (between 5 and 9 days) in mice. The remainder of the strains belonged to the moderate (6 strains) or low (8 strains) virulence categories with median survival times between 10 and 30 days and >30 days, respectively. Batches of 40 teneral Glossina morsitans morsitans (Diptera: Glossinidae) were offered a single bloodmeal on mice infected with one of those strains. Flies were dissected to determine their infection status 21 days later. The proportion of flies with procyclic and metacyclic infections differed significantly between trypanosome strains and were significantly higher in flies infected with extremely virulent strains (P=0.033 and P=0.016 for the differences in the procyclic infection rate of strains with moderate and low virulence, respectively and P=0.005 and P=0.019 for the differences in the metacyclic infection rate of strains with moderate and low virulence, respectively). On the other hand, moderately virulent strains had, in general, higher procyclic and metacyclic infection rates compared to low virulent strains. But the differences were not significant (P>0.05). The outcome of those experiments shows clear differences in transmissibility of trypanosome strains associated with their virulence. This observation confirms the theory for the evolution and maintenance of virulence in a parasite population and may explain the persistence of virulent trypanosome strains in a susceptible host population.

Vector competence of Glossina austeni and Glossina brevipalpis for Trypanosoma congolense in KwaZulu-Natal, South Africa

Tsetse-transmitted trypanosomosis (nagana) has been the cause of stock losses in the recent past and still presents a major problem to livestock owners in certain areas of KwaZulu- Natal, South Africa. Over 10 000 cattle mortalities were reported in the 1990 nagana outbreak. Although information on the distribution and abundance of the tsetse flies Glossina brevipalpis and Glossina austeni in KwaZulu-Natal exists, data on their vector competence are lacking. This study aimed to determine the rate of natural Trypanosoma congolense infection by field-collected as well as colony-reared flies of these species. A total of 442 field-collected G. brevipalpis and 40 G. austeni flies were dissected immediately after collection to determine their infection rates, whilst 699 G. brevipalpis and 49 G. austeni flies were fed on susceptible animals in 10 and four batches, respectively, for use in xenodiagnosis experiments. Teneral colony flies were fed on infected animals and dissected 21 days post infection to confirm their infectivity testing. Glossina austeni harboured 8% immature and mature infections. In G. brevipalpis, the infection with the immature stages was lower (1%) and no mature infections were observed. Although all four batches of G. austeni transmitted T. congolense to four susceptible animals, no transmission resulted from 10 batches of G. brevipalpis fed on susceptible cattle. Colony-derived G. austeni (534) and G. brevipalpis (882) were fed on four bovines infected with different T. congolense isolates. Both G. austeni and G. brevipalpis acquired trypanosome infection from the bovines, with immature infection ranges of 20% - 33% and 1% - 4%, respectively. Parasites, however, only matured in G. austeni (average = 4%). Glossina austeni plays a larger role in the epidemiology of animal trypanosomosis in KwaZulu-Natal than G. brevipalpis and therefore more focus should be aimed at the former when control measures are implemented.

Cross-protection between Trypanosoma congolense strains of low and high virulence.

The aim of this study was to assess the existence of possible cross-protection between Trypanosoma congolense strains of low and extreme virulence circulating in the same trypanosomiasis focus. Groups of six mice were infected using one of three strains of low virulence and challenged with one of three strains of extreme virulence. A group of six mice was used as control for each strain of low and extreme virulence. The results showed that mice infected with one of the strains of extreme virulence developed high parasitaemia and a significant drop of the PCV compared to mice infected with a strain of low virulence and challenged with one of the strains of extreme virulence. With an exception of one strain of extreme virulence (strain F), the survival time of mice infected with the strains of extreme virulence was shorter compared to mice infected with strains of low virulence and subsequently challenged with a strain of extreme virulence. These results suggest that in an area where trypanosomes of various virulence profiles circulate, livestock infected with T. congolense strains of low virulence can be protected against the adverse effects of extremely virulent T. congolense strains.

Endemic type of animal trypanosomiasis is not associated with lower genotype variability of Trypanosoma congolense isolates circulating in livestock

In order to verify whether the low impact on livestock production in endemic areas is related to a low number of trypanosome strains circulating in livestock, 37 Trypanosoma congolense isolates collected from cattle in 11 sites in an endemic trypanosomiasis area in Eastern Zambia were characterised for genotype variability using a modified amplified fragment length polymorphism technique (AFLP). Isolates were further cloned to evaluate the occurrence of mixed infections in individuals. The results obtained revealed a high genotype diversity (94.6%) among these isolates. Apart from one site, all isolates gave different AFLP profiles in each of the sites. When clones were compared, three (8%) of the 37 isolates had mixed infections. These results indicate the circulation of a high number of strains in this trypanosomiasis endemic area despite the low impact the disease has on livestock production.

Transmissibility, by Glossina morsitans morsitans, of Trypanosoma congolense strains during the acute and chronic phases of infection.

In order to verify whether chronic trypanosomal infections can affect the transmissibility of Trypanosoma congolense by tsetse flies, batches of Glossina morsitans morsitans were fed on mice infected with the same level of parasitemia (10(8.1)trypanosomes/ml of blood) of two cloned low virulent T. congolense strains during the acute and the chronic phases of infection. Results showed that the proportions of procyclic infections in flies that were fed during the acute phase (32.6% and 45.4% for isolates 1 and 2, respectively) were significantly higher (chi(2)=4.7, P<0.05 and chi(2)=23.7, P<0.0001, respectively) compared to the proportions of procyclic infections of flies fed during the chronic phase of infection (18.8% and 14.9% for isolates 1 and 2, respectively). Similarly the proportions of metacyclic infections in flies fed during the acute phase (32.6% and 45.4% for isolates 1 and 2, respectively) were significantly higher (chi(2)=6.3, P<0.05 and chi(2)=23.7, P<0.0001, respectively) compared to the proportions of metacyclic infections in flies fed during the chronic phase of infection (16.8% and 14.9% for isolates 1 and 2, respectively). No significant difference was found in the maturation rate of both strains during the acute phase compared to the chronic phase of infection (P>0.05). The results of this study suggest that T. congolense loses part of its transmissiblity by tsetse flies during the chronic phase of infection.