Trichomonas vaginalis and Leishmania spp. are protozoal species responsible for millions of cases of parasitic diseases worldwide. Considering the potential of natural products and the need for more effective and less toxic alternatives to treat trichomoniasis and leishmaniasis, this study aimed to evaluate the effect of two series of triterpenes derivatives with different modifications at C-3 and C-28 positions of the ursolic acid (UA) and betulinic acid (BA) against trophozoites of Trichomonas vaginalis and promastigotes forms of Leishmania (L.) amazonensis. The compounds modified just at C-3 were the most active. The 3β-acetyl betulinic acid (1b) reduced the trophozoites viability of T. vaginalis at 74%, followed by the 3-oxo ursolic acid and 3-oxo betulinic acid (3a and 3b) compounds (55% of reduction). The compound 3β-isobutyl ursolic acid (7a) inhibited the viability of L. amazonensis promastigotes by 55%. Therefore, analyzing the structure-activity relationship and the data of literature, it is possible to suppose that the inclusion of polar groups in the skeletons could improve the antiprotozoal activity. Overall, further studies are necessary to develop triterpenic derivatives with more powerful trichomonicidal and leishmanicidal properties.

Trichomonas vaginalis is the etiologic agent of trichomoniasis, the most common non-viral sexually transmitted disease (STD) in the world. The diagnosis is based on wet mount preparation and direct microscopy on fixed and stained clinical specimens. The aim of this study was to compare the performance of different fixing and staining techniques used in the detection of T. vaginalis in urine. The smears were fixed and submitted to different methods of permanent staining and then, the morphological aspects of the parasites were analyzed and compared. The Papanicolaou staining with ethanol as the fixative solution showed to be the best method of permanent staining. Our data suggest that staining techniques in association with wet mount examination of fresh specimens contribute to increase the sensitivity in the diagnosis of trichomoniasis.

Trichomonas vaginalis is a flagellate protozoan that parasitises the urogenital human tract and causes trichomoniasis. During the infection, the acquisition of nutrients, such as iron and purine and pyrimidine nucleosides, is essential for the survival of the parasite. The enzymes for purinergic signalling, including adenosine deaminase (ADA), which degrades adenosine to inosine, have been characterised in T. vaginalis. In the evaluation of the ADA profile in different T. vaginalisisolates treated with different iron sources or with limited iron availability, a decrease in activity and an increase in ADA gene expression after iron limitation by 2,2-bipyridyl and ferrozine chelators were observed. This supported the hypothesis that iron can modulate the activity of the enzymes involved in purinergic signalling. Under bovine serum limitation conditions, no significant differences were observed. The results obtained in this study allow for the assessment of important aspects of ADA and contribute to a better understanding of the purinergic system in T. vaginalis and the role of iron in establishing infection and parasite survival.

Extracellular ATP may act as a danger signalling molecule, inducing inflammation and immune responses in infection sites. The ectonucleotidases NTPDase and ecto-5’-nucleotidase are enzymes that modulate extracellular nucleotide levels; these enzymes have been previously characterised in Trichomonas vaginalis. Iron plays an important role in the complex trichomonal pathogenesis. Herein, the effects of iron on growth, nucleotide hydrolysis and NTPDase gene expression in T. vaginalisisolates from female and male patients were evaluated. Iron from different sources sustainedT. vaginalis growth. Importantly, iron from haemoglobin (HB) and haemin (HM) enhanced NTPDase activity in isolates from female patients and conversely reduced the enzyme activity in isolates from male patients. Iron treatments could not alter the NTPDase transcript levels in T. vaginalis. Furthermore, our results reveal a distinct ATP, ADP and AMP hydrolysis profile between isolates from female and male patients influenced by iron from HB and HM. Our data indicate the participation of NTPDase and ecto-5’-nucleotidase in the establishment of trichomonas infection through ATP degradation and adenosine production influenced by iron.

Dados preocupantes demonstram a prevalência de parasitos intestinais e sua relação com fatores socioambientais entre indígenas Mbyá-Guarani. A prevalência foi determinada pelas técnicas de sedimentação espontânea em água, centrífugo-flutuação e Kato-Katz. Para cada família, um questionário socioeconômico foi aplicado. A prevalência geral de parasitos intestinais foi de 88,7% e 45,5% estavam poliparasitados. A prevalência em crianças (1-12 anos) foi 90,5% e em indivíduos entre 13-65 anos foi 85%, indicando que ambas as faixas etárias são amplamente parasitadas. A carga parasitária foi baixa a moderada para geohelmintos e 75% das famílias não possuem banheiro, sendo a prática da defecação feita ao ar livre. Estes achados sugerem que o poliparasitismo intestinal na comunidade Mbyá-Guarani é altíssimo, a ponto de ser a regra, e que se refere essencialmente ao estilo de vida tradicional e hábitos de saúde. Há a necessidade de aplicar urgentemente a associação de tratamento antiparasitário às melhorias sanitárias. Isto deve ser feito simultaneamente com atividades de educação em saúde para esta população.

Disturbing data reveal the prevalence of intestinal parasites and their relationship with socio-environmental factors among Mbyá-Guarani Indians. The prevalence was determined by spontaneous sedimentation in water, centrifugation-floatation, and Kato-Katz. A socioeconomic questionnaire was submitted to each family. The overall prevalence of intestinal parasites was 88.7%, and 45.5% were polyparasitized. There was 90.5% prevalence of enteric parasites in children (1-12- year-old), and 85% among 13-65-year-old individuals, indicating that both age groups are extensively parasitized. The parasite load was low to moderate for geohelminths and 75% of the families did not have latrine, thus the practice of defecation occurred outdoors. These findings suggest that the multiple intestinal parasitism in the Mbyá-Guarani community is high to the point of being the rule, and that it relates essentially to the traditional lifestyle and health habits. It is urgently necessary to implement the association of anti-parasitic treatment with sanitation improvement. This should be done simultaneously with health education activities for this population.

Trichomonas vaginalis is a parasite of the human urogenital tract that causes trichomonosis, the most prevalent non-viral sexually transmitted disease. Ectonucleoside triphosphate diphosphohydrolase (NTPDase) family members, which hydrolyse extracellular ATP and ADP and ecto-5′-nucleotidase, which hydrolyses AMP, have been characterised in T. vaginalis. For trichomonad culture, the growth medium is supplemented with 10% serum, which is an important source of nutrients, such as adenosine. Here, we investigated the ATP metabolism of T. vaginalis trophozoites from long-term cultures and clinical isolates under limited bovine serum conditions (1% serum). The specific enzymatic activities were expressed as nmol inorganic phosphate (Pi) released/min/mg protein, the gene expression patterns were determined by reverse transcriptase-polymerase chain reaction, the extracellular adenine nucleotide hydrolysis was analysed by high performance liquid chromatography and the cell cycle analysis was assessed by flow cytometry. Serum limitation led to the profound activation of NTPDase and ecto-5'-nucleotidase activities. Furthermore, the levels of NTPDase A and B transcripts increased and extracellular ATP metabolism was activated, which led to enhanced ATP hydrolysis and the formation of ADP and AMP. Moreover, the cell cycle was arrested at the G0/G1 stage, which suggested adenosine uptake. Our data suggest that under conditions of serum limitation, NTPDase and ecto-5'-nucleotidase play a role in providing the adenosine required for T. vaginalis growth and that this process contributes to the establishment of parasitism.

Trichomonas vaginalis é o agente etiológico da tricomoníase, a doença sexualmente transmissível (DST) não-viral mais comum no mundo. Esse protozoário flagelado atinge o parasitismo com sucesso em um ambiente hostil através dos vários mecanismos pelos quais estabelece sua patogenicidade e também por sua capacidade de evadir a resposta imune do hospedeiro. A infecção apresenta uma ampla variedade de manifestações clínicas, desde quadro assintomático até severa vaginite. A tricomoníase tem sido associada à transmissão do vírus da imunodeficiência humana (HIV), à doença inflamatória pélvica, ao câncer cervical, à infertilidade, ao parto prematuro e ao baixo peso de bebês nascidos de mães infectadas. A investigação laboratorial é essencial na diagnose dessa patogenia, uma vez que leva ao tratamento apropriado e facilita o controle da propagação da infecção. A prevalência mundial anual da tricomoníase é de 180 milhões de casos, e na Europa é responsável por 41% dos casos de vaginite. A terapia da tricomoníase inclui as mesmas medidas profiláticas destinadas às outras DSTs, como prática de sexo seguro e uso de preservativos. O metronidazol é o medicamento de escolha no tratamento da tricomoníase, entretanto, devido à ineficácia dos tratamentos de dose única e ao iminente surgimento de cepas resistentes, outras alternativas terapêuticas estão sendo investigadas.

Trichomonas vaginalis is the aethiologic agent of trichomoniasis, the sexually transmitted disease (STD) non-viral most common in the world. This flagellate protozoan successfully reaches the parasitism in a hostile environment through some mechanisms which establish its pathogenicity and also through its capacity to evade the host immune response. Trichomoniasis presents a large variety of clinical manifestations, from a totally asymptomatic infection to severe vaginitis. It has been associated to the increase in transmission of the human immunodeficiency virus (HIV), pelvic inflammatory disease, cervical cancer, infertility, and premature delivery and low birth weight of children born from infected mothers. The laboratorial inquiry is essential in diagnosis of this STD, leads to the appropriate treatment and facilitates the control of the spread of T. vaginalis infection. The annual world prevalence of trichomoniasis is 180 million cases and in Europe it is responsible for 41% of vaginitis cases. The treatment of trichomoniasis includes the same profilatic means devoted to others STDs, such as secure sex practice and preservative use. Metronidazole is the choose for the treatment of trichomoniasis, however, due to the fail in the single dose treatment and the imminent appearance of resistant strains, other therapeutic alternatives are been investigated.

Tetratrichomonas didelphidis is a flagellated protozoan found in the intestine of opossums. The specimens were stained by the Giemsa method and by FLUTAX-2, an active fluorescent derivative of Taxol which binds to the ab-tubulin polimerized of microtubules of cells. Giemsa stain revealed the morphological features of trichomonads such as four anterior flagella, undulating membrane, axostyle and posterior flagellum. An intense fluorescence was observed in living trophozoites of T. didelphidis and Trichomonas vaginalis (used as control), incubated with FLUTAX-2. An analysis of the composition of the cytoskeleton of T. didelphidis will contribute to understanding the cellular morphology of the parasites.

Several diagnostic techniques have been employed for the detection of Trichomonas vaginalis. Microtubules constitute the cytoskeleton in eukaryotic cells and are sensitive to antimitotic drugs, such as Taxol (paclitaxel). We used FLUTAX a fluorescent taxoid - to analyze the microtubule distribution in living trophozoites of T. vaginalis in urine and in vaginal discharge. A high intensity of fluorescence was observed in living T. vaginalis, epithelial cells and leukocytes present in urine and vaginal discharge. Our preliminary results show the perspective of a new diagnostic technique for trichomonosis and will contribute to the understanding of the cytoskeleton of T. vaginalis.

Shape variations of strains of Trichomonas vaginalis were demonstrated by scanning electron microscopy (SEM) following inoculation on McCoy cell monolayers and after interaction with human erythrocytes. The parasites adhered to both cells and presented amoeboid forms, with pseudopodia-like extensions. Several amoeboid organisms swam freely over the cell monolayers and produced aggregates. These unusual forms of the urogenital flagellate may be a response to the composition of the culture medium and may play an important role in the pathogenic process of T. vaginalis.

Tetratrichomonas didelphidis (Hegner and Ratcliffe 1927) Andersen and Reilly 1965 is a flagellate protozoan from the intestine, cecum and colon of Didelphis marsupialis Linnaeus, 1758. The prevalence of T. didelphidis in opossums D. albiventris was studied in the Botanical Garden, Porto Alegre City in the southernmost Brazilian State, Rio Grande do Sul, Brazil. T. didelphidis was found in seven of eight cultures of swabbed rectums, representing a prevalence of 87.5% in D. albiventris. In the present investigation it was observed that the T. didelphidis found in the intestine content of D. albiventris had the same morphological characteristics as those previously described by other authors in the D. marsupialis, being probably the same protozoan species in both host species.

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°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.

In in vitro assay the hemolytic activity of live isolates and clones of Trichomonas gallinae was investigated. The fresh isolates and clones were 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 showed that each of the isolates and clones tested lysed all human blood groups, as well as rabbit, rat, chicken, horse, bovine and sheep erythrocytes. No hemolysin released by the parasites could be identified. Parasite culture supernatants of hemolysis assays, tested in the presence of human erythrocytes did not induce any hemolytic activity. Trichomonads culture supernatants from 18, 24, and 48 hr extracts of sonicated parasites or cold killed organisms tested with human erythrocytes did not exhibit hemolysis. Although an hemolytic activity of T. gallinae was clearly demonstrated, a relationship between hemolysis and cell pathogenicity is not yet clearly established.

Tetratrichomonas didelphidis (Hegner and Ratcliffe, 1927) Andersen and Reilly, 1965 is a flagellate protozoan from the intestine, cecum, and colon of Didelphis marsupialis Linnaeus, 1758. The parasite was found and isolated in the rectal glands in Pavlova starch-containing media in Florianópolis, SC, Brasil from D.

ABSTRACT Trichomonas gallinae (Rivolta, 1878) is a parasitic protozoan of the upper digestive tract and various organs of different avian groups, being common among the Columbids, was isolated described and its prevalence studies in the Southern Brazilian State, Rio Grande do Sul. The morphology study of the live specimens done by examination of fresh and stained specimens, showed that the T. gallinae isolated has the same morphological characteristics as the upper digestive tract trichomonads previously described by other authors