Analysis of microtubule cytoskeleton distribution using a fluorescent taxoid in two trichomonadid protozoa: Trichomonas gallinae and Tritrichomonas foetus.

Trichomonas gallinae and Tritrichomonas foetus are flagellated parasitic protozoa of the upper digestive tract of birds and the urogenital tract of cattle, respectively. Both of these species are important in the veterinary field, due to the fact that they cause significant economic losses. Therefore, we investigated the morphology of these parasites by studying microtubule cytoskeleton organization. FLUTAX-2, an active fluorescent derivative of Taxol, was used in this study. This fluorescent taxoid binds to polymerized alphabeta-tubulin dimers. Our results showed that FLUTAX-2 was able to bind to and stabilize microtubules of intact T. gallinae and T. foetus trophozoites, allowing the microtubular cytoskeleton to be easily observed by fluorescence microscopy. T. foetus and T. gallinae had no differences in their FLUTAX-2 binding profiles. Further studies may allow this technique to be improved, and it may possibly be used as a routine laboratory method for the diagnosis of avian and bovine trichomonosis.

Characterisation of an ATP diphosphohydrolase (Apyrase, EC 3.6.1.5) activity in Trichomonas vaginalis.

In the present report the enzymatic properties of an ATP diphosphohydrolase (apyrase, EC 3.6.1.5) in Trichomonas vaginalis were determined. The enzyme hydrolyses purine and pyrimidine nucleoside 5'-di- and 5'-triphosphates in an optimum pH range of 6.0--8.0. It is Ca(2+)-dependent and is insensitive to classical ATPase inhibitors, such as ouabain (1 mM), N-ethylmaleimide (0.1 mM), orthovanadate (0.1 mM) and sodium azide (5 mM). A significant inhibition of ADP hydrolysis (37%) was observed in the presence of 20 mM sodium azide, an inhibitor of ATP diphosphohydrolase. Levamisole, a specific inhibitor of alkaline phosphatase, and P(1), P(5)-di (adenosine 5'-) pentaphosphate, a specific inhibitor of adenylate kinase, did not inhibit the enzyme activity. The enzyme has apparent K(m) (Michaelis Constant) values of 49.2+/-2.8 and 49.9+/-10.4 microM and V(max) (maximum velocity) values of 49.4+/-7.1 and 48.3+/-6.9 nmol of inorganic phosphate x min(-1) x mg of protein(-1) for ATP and ADP, respectively. The parallel behaviour of ATPase and ADPase activities and the competition plot suggest that ATP and ADP hydrolysis occur at the same active site. The presence of an ATP diphosphohydrolase activity in T. vaginalis may be important for the modulation of nucleotide concentration in the extracellular space, protecting the parasite from the cytolytic effects of the nucleotides, mainly ATP.

[Serologic screening of infectious diseases in blood banks as an indicator of morbidity in the population]. / Rastreamento sorológico para doenças infecciosas em banco de sangue como indicador de morbidade populacional.

Screening tests of 62,814 blood donations carried out between October 85 and October 87 in all five blood banks of Goiânia, Central Brazil, were analysed. The private institutions (3 blood banks) collected 80% of all donations and the remaining were collected by public blood banks. Seroprevalence of HIV, HBsAg, Chagas' disease and syphilis were compared with data obtained in the surveillance system and from previous surveys in an attempt to validate this source of health information. A seroprevalence of 0.07% for AIDS (one ELISA test) was calculated, which may suggest the presence of 1900 infected individuals, in the population of the State of Goiás as a whole. This figure seems reasonably close to the official data obtained by compulsory notification. For Chagas' disease, hepatitis B and syphilis seroprevalence of 3.3%, 1.3% and 4.1% were obtained, respectively. The potential usefulness and the limitations of screening in blood banks as a source of reliable morbidity indicator is discussed.