Hydrolyzable tannins from Poincianella (Caesalpinia) microphylla fruits: Metabolite profiling and anti-Trichomonas vaginalis activity.

Poincianella (Caesalpinia) microphylla fractions were analyzed by LC-DAD-MS and by statistical tools to determine the possible bioactive compounds against T. vaginalis. Tannins enriched-fractions toxicity (M5 and M10) were analyzed in in vitro against human red cells and in in vivo model of Galleria mellonella. Twenty-six compounds were detected from the P. microphylla fractions. The main compounds identified were hydrolyzable tannins (gallotanins and ellagitannis), such as O-digalloyl hexoside, O-digalloyl HHDP -hexoside, tri-O-galloyl HHDP-hexoside, O-galloyl HHDP-DHHDP-hexoside and their isomers. In addition, procyanidin dimer, epicatechin, ellagic acid and O-(digalloyl) quinic acid were also identified. Based on univariate statistical analyses, stronger correlations with the anti-T. vaginalis properties were observed for the compounds 7 (O-galloyl HHDP-DHHDP-hexoside - Geraniin isomer), 3 (O-digalloyl HHDP -hexoside - mallotinic acid), 2 (O-digalloyl) quinic acid), 1 (O-digalloyl hexoside) and 9 (unknown). Tannins enriched-fractions (M5 and M10) presented anti-Trichomonas activity (IC50 70.41 µg/mL and 142.1 µg/mL, respectively) and no toxicity in the in vivo model of G. mellonella. This innovative approach allowed us to identify likely bioactive compounds in the extracts, although the mechanism(s) underlying anti-trichomonal activity encompass a complex trait.

Interações entre antiparasitários e alimentos / Interactions between antiparasitic drugs and food

O uso, por vezes indiscriminado, de antiparasitários pode levar a consequências importantes na saúde do indivíduo, principalmente relacionadas a alterações no sistema gastrointestinal. As interações entre fármacos e nutrientes podem ocorrer quando um alimento ou nutriente altera a resposta esperada de um medicamento ou quando este interfere sobre o estado nutricional do indivíduo. Essa alteração da eficácia pode resultar em falha no tratamento ou até mesmo na toxicidade do fármaco. A fim de conhecer, identificar e prevenir interações indesejáveis entre antiparasitários e alimentos, o presente artigo de revisão aborda os principais fármacos e alimentos ou nutrientes envolvidos, bem como as consequências que tais interações podem acarretar ao indivíduo. O estado nutricional é de suma importância nas infecções parasitárias, pois é o determinante entre uma maior carga de parasitos ou a resistência total contra a infecção. De forma geral, indivíduos parasitados são inapetentes e emagrecidos, o que em crianças pode comprometer o desenvolvimento físico e intelectual. Portanto, o conhecimento e uma correta orientação médica, farmacêutica e nutricional levam a um tratamento bem sucedido...

The often indiscriminate use of antiparasitic drugs can lead to serious consequences for the health of the individual, mainly related to alterations in the gastrointestinal system. Interactions between drugs and nutrients may occur when a food or nutrient changes the expected response of a drug or when a medicine interferes with the individual's nutritional status. These changes may result in treatment failure or even toxicity of the drug. As an aid to the detection, identification and prevention of undesirable interactions between foods and antiparasite drugs, in this review we discuss the major drugs and foods or nutrients involved and the consequences that these interactions can have for the individual. Nutritional status is important in parasitic infections, as it is a factor determining the parasite load level or the total resistance against infection. Generally, individuals infected by parasites have a poor appetiteand are emaciated, which in children can damage physical and intellectual development. Therefore, knowledge of the parasite and proper medical, pharmaceutical and nutritional guidance lead to a more successful treatment...

Cattle tick-associated bacteria exert anti-biofilm and anti-Tritrichomonas foetus activities.

Research on microbiota in cattle tick and the evaluation of its activity against other microorganisms can contribute to identify new molecules potentially useful to control infections caused by bacteria and protozoa. Biofilms pose increasing problems worldwide, mainly due to their resistance to antimicrobial therapies and host immune response. In this study we investigate the ability Rhipicephalus (Boophilus) microplus-associated bacteria may exhibit to produce anti-biofilm and trichomonicidal compounds. Gut, ovary, salivary glands, and Gené organ were collected from engorged R. microplus female. Homogenates of each tissue were inoculated onto 15 distinct culture media. Anti-biofilm and trichomonicidal activities were analyzed by culturing each bacterium isolated in a liquid medium. Results showed that R. microplus cattle tick microflora varies for different tissues. Bacteria belonging to different genera (Aeromonas, Bacillus, Brevibacillus, Castelaniella, Comamonas, Kocuria, and Microbacterium) were identified. Interestingly, all bacterial species found displayed pronounced activity against Staphylococcus epidermidis and Pseudomonas aeruginosa biofilms, and also against the cattle pathogen Tritrichomonas foetus, confirming the hypothesis that cattle tick could be a source of bacteria active against pathogens. This is the first study showing that bacteria isolated from a tick exert anti-biofilm and trichomonicidal activities.

Anti-protozoal activity of diamine derivatives.

Ten N-monoalkylated diamines were synthesized and evaluated for in vitro activities against Trichomonas vaginalis and Giardia lamblia. Several compounds displayed a good inhibition of parasite growth, with MIC less or equal to 20µg/mL. N-hexadecil-1,4-butanediamine was found to be the most active compound in vitro against T. vaginalis with MIC of 2.5µg/mL, twice more active in comparison to the reference drug metronidazole (MTZ). Seven of the studied compounds showed a better anti-G. lamblia activity than MTZ.

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.

Heterogeneity in extracellular nucleotide hydrolysis among clinical isolates of Trichomonas vaginalis.

Trichomonas vaginalis is a parasitic protozoan that causes trichomonosis, a sexually-transmitted disease, with serious sequelae to women and men. As the host-parasite relationship is complex, it is important to investigate biochemical aspects of the parasite that contribute to our understanding of trichomonal biology and pathogenesis. Nucleoside triphosphate diphosphohydrolase 1 (NTPDase 1), which hydrolyses extracellular ATP and ADP, and ecto-5'-nucleotidase, which hyrolyses AMP, have been characterized in laboratory isolates of T. vaginalis. Here we show that the extracellular ATP: ADP hydrolysis ratio varies among fresh clinical isolates, which presented higher ATPase and ADPase activities than long-term-grown isolates. Growth of parasites in iron-replete and iron-depleted medium resulted in different, albeit minor, patterns in extracellular ATP and ADP hydrolysis among isolates. Importantly, some isolates had low or absent ecto-5'-nucleotidase activity, regardless of environmental conditions tested. For isolates with ecto-5'-nucleotidase activity, high- and low-iron trichomonads had increased and decreased levels of activity, respectively, compared to organisms grown in normal TYM-serum medium. This suggests a regulation in expression of either the enzyme amounts and/or activity under the control of iron. Finally, we found no correlation between the presence or absence of dsRNA virus infection among trichomonad isolates and NTPDase and ecto-5'-nucleotidase activities.

Growth kinetic study of Tetratrichomonas didelphidis isolated from opossum Lutreolina crassicaudata and interaction with a prokaryotic cell.

Tetratrichomonas didelphidis is a flagellate protozoan found in the intestine, cecum and colon of opossums, Didelphis marsupialis. This work reports the occurrence of T. didelphidis in another opossum species, Lutreolina crassicaudata. The strain was cultivated in monoxenic culture with Escherichia coli in Diamond (TYM) medium without maltose and with starch solution (trypticase-yeast extract-starch), pH 7.5 at 28 degrees C. The growth kinetic study of T. didelphidis showed a longer time of growth and a higher number of trophozoites when inoculated with E. coli than in axenic cultures, in aerobiosis as well as under anaerobic conditions. Scanning electron microscopy showed that the bacteria adhered throughout the protozoan body and probably evoked endocytic channels, strongly suggesting the existence of endocytosis of rods by T. didelphidis. Our preliminary results suggest that the in vitro culture of T. didelphidis depends on E. coli as a growth-promoting partner, and requires monoxenic cultivation.

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.

Morphologic aspects of Tetratrichomonas didelphidis isolated from opossums Didelphis marsupialis and Lutreolina crassicaudata.

Tetratrichomonas didelphidis (Hegner & Ratcliffe, 1927) Andersen & Reilly, 1965 is a flagellate protozoan found in the intestine, cecum, and colon of Didelphis marsupialis. The parasitic protozoa used in this study was found and isolated in the intestine of opossums in Pavlova starch-containing medium in Florianópolis, State of Santa Catarina, Brazil, from D. marsupialis and Lutreolina crassicaudata. The strains were cultivated in Diamond medium without maltose and with starch solution, pH 7.5 at 28 degrees C. The specimens were stained by the Giemsa method and Heidenhain's iron hematoxylin. The light microscopy study of the trophozoites revealed the same morphologic characteristics as specimens previously described.

Hemolytic activity of Monocercomonas spp.

The hemolytic activity of an isolate of Monocercomonas spp. from Tropidophis melanurus (snake: Boidae) was investigated. The isolate was tested against human erythrocytes of groups A, B, AB and O and against erythrocytes of six adult animals of different species (rabbit, rat, chicken, horse, bovine, and sheep). Results show that Monocercomonas spp. exerted an hemolytic activity against all erythrocytes tested.