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At least 19 recordsLinked to original sources
PubMed1997

A comparison of the enzymatic properties of the major cysteine proteinases from Trypanosoma congolense and Trypanosoma cruzi.

Congopain and cruzipain, the major cysteine proteinases from Trypanosoma congolense and Trypanosoma cruzi, were compared for their activities towards a series of new, sensitive fluorogenic substrates of the papain family of cysteine proteinases and for their sensitivity to inhibition by cystatins and related biotinylated peptidyl diazomethanes. Low Ki values, in the 10 pM range, were found for the interaction of both proteinases with natural cystatin inhibitors. The kinetic constants for the hydrolysis of cystatin-derived substrates, and the inhibition by related diazomethanes were essentially identical. Unlike cathepsins B and L, the related mammal papain family proteinases, congopain and cruzipain accomodate a prolyl residue in P2'. Substrates having the sequence VGGP from P2 to P2' were hydrolysed by both congopain and cruzipain with a k(cat)/Km greater than 4.10(3) mM(-1) s(-1). Irreversible diazomethane inhibitors, deduced from the unprime sequence of cystatin-derived substrates, inhibited the two parasite proteinases. N-terminal labelling of diazomethanes with a biotin group did not alter the rate of inhibition significantly, which provides a useful tool for examining the distribution of these enzymes in the parasite and in the host. Despite their similar activities on cystatin-derived substrates, congopain and cruzipain had significantly different pH-activity profiles when assayed with a cystatin-derived substrate. They were correlated with structural differences, especially at the presumed S2 subsites.

Animals↗
PubMed1988

Endocytosis and intracellular occurrence of the variant surface glycoprotein in Trypanosoma congolense.

Trypanosoma congolense bloodstream forms were examined for binding sites of polyclonal anti-variant surface glycoprotein (VSG) antibodies using immunoelectron microscopy. Besides the surface, the antibodies labeled intracellular vesicles, the tubular membrane system, secondary lysosomes, and the digestive vacuole. Protein A gold (PAG), peroxidase gold (POG), anti-VSG antibodies preincubated with PAG, ferritin, concanavalin A-ferritin, and microperoxidase were examined for their suitability as endocytosis tracers in combination with immunoelectron microscopy. Endocytosis of PAG and POG was most effective and was mediated by vesicles transporting the tracer to secondary lysosomes. Gold particles eventually accumulated in the digestive vacuole. Apparantly only low amounts of VSG were internalized during endocytosis. VSG export from the cell interior to the flagellar pocket was not observed during excessive endocytosis of PAG, whereas after incubation with substances causing the formation of filopodia by binding to the surface coat, VSG-labeled vesicles were present near the flagellar pocket.

Animals↗
PubMed1999

Purification and characterisation of a trypsin-like serine oligopeptidase from Trypanosoma congolense.

Trypanosoma brucei contain a serine oligopeptidase (OP-Tb) that is released into (and remains active in) the blood of trypanosome-infected animals. Here a similar enzyme from Trypanosoma congolense is described. This oligopeptidase, called OP-Tc, was purified using three-phase partitioning, and ion-exchange and affinity chromatography. OP-Tc is inhibited by alkylating agents, by serine peptidase-specific inhibitors including 3,4-dichloroisocoumarin, 4-(2-aminoethyl)benzenesulfonylfluoride and diispropylfluoro-phosphate and by other peptidase inhibitors including leupeptin, antipain and peptidyl chloromethyl ketones. Reducing agents such as dithiothreitol enhanced activity as did heparin, spermine and spermidine. The enzyme has trypsin-like specificity since it cleaved fluorogenic peptides that have basic amino acid residues (Arg or Lys) in the P1 position. Potential substrates without a basic residue in P1 were not hydrolysed. Although OP-Tc has weak arginine aminopeptidase activity, the enzyme clearly preferred substrates that had amino acids in the P2 and P3 positions. Overall, OP-Tc appears to be less efficient than OP-Tb because it usually displayed lower k(cat)/Km values for the substrates tested. However, like OP-Tb, the best substrate for OP-Tc was Cbz-Arg-Arg-AMC (Km = 0.72 microM, k(cat) = 96 s(-1)). OP-Tc preference for amino acids in the P2 position was (Gly,Lys,Arg) > Phe > Leu > Pro. The results also suggest that the P3-binding site has hydrophobic characteristics. OP-Tc may not be a naturally immunodominant molecule because neither IgG nor IgM anti- OP-Tc antibodies were detected in the blood of experimentally infected cattle.

Animals↗
PubMed2004

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.

Animals↗
PubMed2002

Comparative evaluation of the antibody-detection ELISA technique using microplates precoated with denatured crude antigens from Trypanosoma congolense or Trypanosoma vivax.

Two FAO/IAEA indirect enzyme-linked immunosorbent assays (ELISA), which use microplates precoated with denatured crude Trypanosoma congolense or Trypanosoma vivax antigen for detecting anti-trypanosomal antibodies in bovine sera, were evaluated for their sensitivity, specificity and positive and negative predictive values, using 320 Ugandan field samples (known negative sera, n = 80; known positive sera, n = 80; cattle herds where control of tsetse and trypanosomosis was practiced, n = 80; and cattle herds where there was no such control, n = 80). Cut-off points of 30% and 25% positivity were determined for the T. congolense and T. vivax assays, respectively, using a modified ROC (receiver operating characteristic) analysis. The T. congolense assay had estimated diagnostic sensitivity and specificity of 63.7% and 57.5%, respectively, while the T. vivax assay had estimated diagnostic sensitivity and specificity of 81.3% and 81.3%, respectively. The two assays conducted in parallel had estimated diagnostic sensitivity and specificity of 82.5% and 88.7%, respectively. Using the sera from the cattle in the area with control (detected prevalence of trypanosomosis 0%), both the T. congolense and T. vivax assays had negative and positive predictive values of 100% and 0%, respectively. Using the sera from the cattle in the area without control (detected prevalence of trypanosomosis 15%), the T. congolense assay had negative and positive predictive values of 91% and 33%, respectively, and the T. vivax assay had negative and positive predictive values of 93% and 27%, respectively. The T. congolense assay was in fair agreement with the buffy coat technique (BCT) (kappa = 0.25), while the T. vivax assay was in substantial agreement with the BCT (kappa = 0.625), and both assays conducted in parallel were in substantial agreement with the BCT (kappa = 0.708). Both assays were found to be proficient and suitable for the diagnosis of bovine trypanosomosis, especially when used in parallel.

Animals↗
PubMed1976

The generation of toxic activity from trypanosoma congolense.

Trypanosoma congolense organism, on incubation at 20 degrees C for 91/2h, were found to generate phospholipase like activity which was capable of mediating lysis of both nucleated cells and erythrocytes as well as acute inflammatory response on intradermal inoculation.

Animals↗
PubMedDate not supplied

Diminazene aceturate residues in the tissues of healthy, Trypanosoma congolense and Trypanosoma brucei brucei infected dogs.

The tissue distribution and residue profile of diminazene aceturate was investigated in healthy dogs and in dogs infected with Trypanosoma congolense and Trypanosoma brucei brucei. The drug was administered at 3.5 mg/kg i.m. and tissue samples were taken post mortem from the animals at 48, 72, 120, 168 and 240 h after injection. The drug was distributed to various organs and tissues of the body with the highest concentrations occurring in liver and kidney. Higher drug levels were obtained in the tissues of healthy dogs compared with trypanosome infected animals except in the brain. The levels of residues in the healthy animals were significantly different (P less than 0.05) from those of the infected dogs. The drug residues were still detectable in the tissues of the animals 10 days after drug administration.

Animals↗
PubMed1978

Mechanisms of the anemia in trypanosomiasis: studies on the role of the hemolytic fatty acids derived from Trypanosoma congolense.

Trypanosoma congolense releases a number of hemolytic free fatty acids on autolysis of which the most potent is linoleic acid. These fatty acids can lyse washed rat and bovine erythrocytes in vitro. Autolysed T. congoleuse will cause increased fragility of erythrocytes in whole rat blood but not in whole bovine blood. The lack of effect of these fatty acids on bovine erythrocytes tends to be confirmed by the failure of this material to influence the survival of these cells in vivo. Calculations based on the amount of free fatty acids required to cause hemolysis in vivo also confirm that this mechanism of erythrocyte destruction is probably unimportant in bovine trypanosomiasis.

Anemia↗
PubMed2000

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.

Anemia↗
PubMed2006

The pathogenesis of anaemia in goats experimentally infected with Trypanosoma congolense or Trypanosoma brucei: use of the myeloid:erythroid ratio.

The present study examined the development of anaemia in Small East African goats experimentally infected with Trypanosoma congolense or Trypanosoma brucei. Experimental goats received a primary trypanosome challenge on day 0, treated with diminazene aceturate on day 49 and received a secondary trypanosome challenge on day 77 of the 136-day experiment. Both primary and secondary challenges were characterised by reduced peripheral erythrocyte counts, fall in packed cell volume (PCV), hypohaemoglobinaemia and reductions in the myeloid:erythroid ratios (M:E) compared with the uninfected goats. The progressive reduction in the M:E ratios denoted increased erythrogenesis in response to increased destruction of erythrocytes in blood by infecting trypanosomes or their products. The more rapid fall in M:E ratio in T. congolense infections shows that this parasite causes more severe clinical pathological effects in goats than T. brucei.

Anemia↗
PubMed1998

Sensitivity and specificity of antigen-capture ELISAs for diagnosis of Trypanosoma congolense and Trypanosoma vivax infections in cattle.

Sensitivity and specificity of the FAO/IAEA antigen-ELISA kits for diagnosis of bovine trypanosomosis were investigated using sera from experimental cattle infected by tsetse challenge with cloned populations of Trypanosoma congolense (three populations) or T. vivax (one population). The kits are based on monoclonal antibodies that recognise internal antigens of tsetse-transmitted trypanosomes. Ten cattle were infected with each trypanosome population for at least 60 days, and in combination with uninfected cohorts (n = 16) were used in a double-blind study design. Sensitivity and specificity of the tests depended on the choice of positive-negative thresholds expressed as percent positivity with respect to the median OD of four replicates of the strong positive reference serum provided with the kit. In general, while overall specificities were high, sensitivities of the antigen-ELISAs were poor. For example, at a cut-off of 5% positivity, the sensitivities of the antigen-ELISAs were 11% for samples (n = 1162) from T. congolense infected cattle (n = 30), and 24% for samples (n = 283) from T. vivax infected cattle (n = 10). The corresponding specificity values were 95% and 79%, respectively. At a cut-off of 2.5% positivity sensitivity for T. congolense was 25%, and for T. vivax 35%; corresponding specificity values were 85% and 63% respectively. There were no values of the positive-negative threshold at which both sensitivity and specificity were satisfactory. Restricting the analyses to samples taken more than 2 weeks after tsetse challenge did little to improve sensitivity estimates. Trypanosome species specificities of the antigen-ELISAs were also poor. Sensitivity and species specificity of the antigen-ELISA for Trypanosoma brucei infections were not investigated. In contrast to the antigen-ELISA, the sensitivity of the buffy-coat technique when applied to the same experimental animals was fairly high at 67% for T. congolense infections and 60% for T. vivax infections. For samples taken more than 2 weeks after tsetse challenge, high sensitivity estimates of 96% for T. congolense and 76% for T. vivax infections were obtained.

Africa South of the Sahara↗
PubMed1987

Lectin mediated establishment of midgut infections of Trypanosoma congolense and Trypanosoma brucei in Glossina morsitans.

D+Glucosamine, which has been shown in vitro to specifically inhibit tsetse midgut lectin activity, when fed to Glossina morsitans morsitans with the infective feed significantly increased midgut infection rates of Trypanosoma congolense and T. brucei rhodesiense. All flies infected with T.b. rhodesiense and maintained on a diet of blood with D+Glucosamine throughout their lives developed midgut infections. Midgut extracts from flies bred for refractoriness to infection with trypanosomes showed significantly greater erythrocyte agglutinating activity and were more trypanocidal in vitro than flies bred for susceptibility to trypanosome infection. It is concluded that susceptibility to trypanosome infection in tsetse is mediated through midgut lectins.

Agglutination Tests↗
PubMed1992

Sequential infection of tsetse flies with Trypanosoma congolense and Trypanosoma brucei.

The question whether tsetse flies can be experimentally infected with more than one trypanosome species or strain by sequential feeding was investigated using DNA probe technology to identify directly the small numbers of trypanosomes in the fly gut. Bloodstream form trypanosomes of Trypanosoma congolense or T. brucei ssp. were used for initial infection, followed by sequential feeds using either T. congolense or T. brucei ssp. Midgut trypanosome populations were subsequently analysed by hybridising dot blots with species-specific DNA probes. Two different T. brucei stocks were also fed in succession and the midgut trypanosome populations analysed by molecular karyotype. Contrary to expectations from previous reports, it was comparatively easy to superinfect flies with a second trypanosome species or stock, although the presence of trypanosomes already in the gut did not aid establishment of those incoming. Thus, to develop a mixed infection, a prerequisite for trypanosome mating, flies do not necessarily have to pick up both parental trypanosomes on their first feed.

Animals↗
PubMed1989

Detection of Trypanosoma congolense and Trypanosoma brucei subspecies by DNA amplification using the polymerase chain reaction.

The nuclear DNA of Trypanosoma congolense contains a family of highly conserved 369 base pair (bp) repeats. The sequences of three cloned copies of these repeats were determined. An unrelated family of 177 bp repeats has previously been shown to occur in the nuclear DNA of Trypanosoma brucei brucei (Sloof et al. 1983a). Oligonucleotides were synthesized which prime the specific amplification of each of these repetitive DNAs by the polymerase chain reaction (PCR). Amplification of 10% of the DNA in a single parasite of T. congolense or T. brucei spp. produced sufficient amplified product to be visible as a band in an agarose gel stained with ethidium bromide. This level of detection, which does not depend on the use of radioactivity, is about 100 times more sensitive than previous detection methods based on radioactive DNA probes. The oligonucleotides did not prime the amplification of DNA sequences in other trypanosome species nor in Leishmania, mouse or human DNAs. Amplification of DNA from the blood of animals infected with T. congolense and/or T. brucei spp. permitted the identification of parasite levels far below that detectable by microscopic inspection. Since PCR amplification can be conducted on a large number of samples simultaneously, it is ideally suited for large-scale studies on the prevalence of African trypanosomes in both mammalian blood and insect vectors.

Animals↗
PubMed1979

Non-specific induction of increased resistance in mice to Trypanosoma congolense and Trypanosoma brucei by immunostimulants.

Administration of the immunostimulants Corynebacterium parvum, Bacillus Calmette-Guérin (BCG) or Bordetella pertussis prior to, or at the same time as, challenge with Trypanosoma congolense significantly increased survival times in mice, both of trypano-susceptible (A/J) and trypano-resistant (C57Bl) strains. The increased survival time was associated with significant alterations in parasitaemia, which included lengthening of the pre-patent period, a delay in the time taken to reach the first peak of parasitaemia and a reduction in the level of parasitaemia. Similar results were obtained when these strains of mice were challenged with Trypanosoma brucei following pre-treatment with C. parvum. Thus, by the use of immunostimulants it was possible to reduce the susceptibility of mice to trypanosomiasis and the hope is that this can also be achieved with domestic livestock.

Animals↗
PubMed1980

Hemolytic complement and serum C3 levels in Zebu cattle infected with Trypanosoma congolense and Trypanosoma vivax and the effect of trypanocidal treatment.

Total hemolytic complement and C3 levels were found to drop to 6.25% and 50% of preinfection levels, respectively, during trypanosome infections. Chemotherapeutic elimination of the trypanosomes with Berenil led to recovery of preinfection levels within 7 days and 11 days when cattle infected with Trypanosome congolense and Trypanosoma vivax, respectively, were treated 37 days after onset of infection. Recovery was slower in T. vivax-infected cattle treated on day 50. Berenil treatment had no effect on complement levels in control animals. The possible causes and implications of these low complement levels in bovine trypanosomiasis are discussed.

Amidines↗