Behavior of alfa-tomatine and tomatidine against several genera of trypanosomatids from insects and plants and Trypanosoma cruzi / Comportamento de alfa-tomatina e tomatidina contra vários gêneros de tripanossomatídeos de insetos e plantas e Trypanosoma cruzi

Glycoalkaloids are important secondary metabolites accumulated by plants as protection against pathogens. One of them, α-tomatine, is found in high concentrations in green tomato fruits, while in the ripe fruits, its aglycone form, tomatidine, does not present a protective effect, and it is usual to find parasites of tomatoes like Phytomonas serpens in these ripe fruits. To investigate the sensitivity of trypanosomatids to the action of α-tomatine, we used logarithmic growth phase culture of 20 trypanosomatids from insects and plants and Trypanosoma cruzi. The lethal dose 50% (LD50) was determined by mixing 107 cells of the different isolates with α-tomatine at concentrations ranging from 10-3 to 10-8 M for 30 min at room temperature. The same tests performed with the tomatidine as a control showed no detectable toxicity against the same trypanosomatid cultures. The tests involved determination of the percentage (%) survival of the protozoan cultures in a Neubauer chamber using optical microscopy. The LD50 values varied from 10-4 to 10-6 M α-tomatine. Slight differences were detected among the LD50 values of the analyzed samples, and none of them showed evidence of resistance to the action of tomatinase, as shown by some pathogenic fungi.

Os glicoalcaloides são metabólitos secundários importantes produzidos pelas plantas e estão envolvidos em sua proteção contra agentes patogênicos. Um deles, α-tomatina, é encontrado em altas concentrações em frutos de tomate verde, enquanto que, nos frutos maduros, sua forma aglicona, tomatidina, não apresenta um efeito protetor, sendo comum encontrar parasitas de tomates como Phytomonas serpens nesses frutos maduros. Para investigar a sensibilidade dos tripanossomatídeos à ação da α-tomatina, utilizamos formas de cultura em fase logarítmica de 20 tripanossomatídeos de plantas e insetos e Trypanosoma cruzi. A dose letal 50% (DL50) foi determinada, misturando 107 células das formas de cultura com concentrações de 10-3 a 10-8 M de α-tomatina durante trinta minutos a temperatura ambiente. Testes realizados com a tomatidina como controle não mostraram toxicidade detectável contra os mesmos tripanossomatídeos. Os testes foram avaliados pela porcentagem (%) de sobrevivência das formas de cultura dos protozoários observados por microscopia óptica em câmara de Neubauer. Os resultados da determinação de DL50 mostraram que esta variou entre 10-4 a 10-6 M de α-tomatina. Pequenas diferenças foram observadas entre os valores de DL50 das amostras analisadas, e nenhuma delas mostrou evidência de resistência pela ação da tomatinidase, como demonstrado em alguns fungos patogênicos.

Behavior of alfa-tomatine and tomatidine against several genera of trypanosomatids from insects and plants and Trypanosoma cruzi / Comportamento de alfa-tomatina e tomatidina contra vários gêneros de tripanossomatídeos de insetos e plantas e Trypanosoma cruzi

Glycoalkaloids are important secondary metabolites accumulated by plants as protection against pathogens. One of them, α-tomatine, is found in high concentrations in green tomato fruits, while in the ripe fruits, its aglycone form, tomatidine, does not present a protective effect, and it is usual to find parasites of tomatoes like Phytomonas serpens in these ripe fruits. To investigate the sensitivity of trypanosomatids to the action of α-tomatine, we used logarithmic growth phase culture of 20 trypanosomatids from insects and plants and Trypanosoma cruzi. The lethal dose 50% (LD50) was determined by mixing 107 cells of the different isolates with α-tomatine at concentrations ranging from 10-3 to 10-8 M for 30 min at room temperature. The same tests performed with the tomatidine as a control showed no detectable toxicity against the same trypanosomatid cultures. The tests involved determination of the percentage (%) survival of the protozoan cultures in a Neubauer chamber using optical microscopy. The LD50 values varied from 10-4 to 10-6 M α-tomatine. Slight differences were detected among the LD50 values of the analyzed samples, and none of them showed evidence of resistance to the action of tomatinase, as shown by some pathogenic fungi.(AU)

Os glicoalcaloides são metabólitos secundários importantes produzidos pelas plantas e estão envolvidos em sua proteção contra agentes patogênicos. Um deles, α-tomatina, é encontrado em altas concentrações em frutos de tomate verde, enquanto que, nos frutos maduros, sua forma aglicona, tomatidina, não apresenta um efeito protetor, sendo comum encontrar parasitas de tomates como Phytomonas serpens nesses frutos maduros. Para investigar a sensibilidade dos tripanossomatídeos à ação da α-tomatina, utilizamos formas de cultura em fase logarítmica de 20 tripanossomatídeos de plantas e insetos e Trypanosoma cruzi. A dose letal 50% (DL50) foi determinada, misturando 107 células das formas de cultura com concentrações de 10-3 a 10-8 M de α-tomatina durante trinta minutos a temperatura ambiente. Testes realizados com a tomatidina como controle não mostraram toxicidade detectável contra os mesmos tripanossomatídeos. Os testes foram avaliados pela porcentagem (%) de sobrevivência das formas de cultura dos protozoários observados por microscopia óptica em câmara de Neubauer. Os resultados da determinação de DL50 mostraram que esta variou entre 10-4 a 10-6 M de α-tomatina. Pequenas diferenças foram observadas entre os valores de DL50 das amostras analisadas, e nenhuma delas mostrou evidência de resistência pela ação da tomatinidase, como demonstrado em alguns fungos patogênicos.(AU)

Susceptibility of Trypanosoma evansi in the copaiba oil: in vitro test and in mice experimentally infected with the parasite / Susceptibility of Trypanosoma evansi in the copaiba oil: in vitro test and in mice experimentally infected with the parasite

Background: Trypanosoma evansi is a flagellate which belongs to the salivate section, commonly observed parasitizing blood of equines, ruminants, pigs, dogs and wild animals in different regions of the world. It causes many losses to farmers due to death of animals and drug spending in endemic areas. The treatment of this disease in Brazil is only performed with diminazene aceturate; however it has been ineffective for many animals. During the lats years many studies have been carried out with natural products such as the essential oils. Copaiba oil stands out due some properties described as anti-inflammatory, healing, antiedematogenic, antitumor, parasitic and antibacterial. Therefore, this study aimed to test, in vitro and in vivo, the susceptibility of T. evansi to copaiba oil. Materials, Methods & Results: The oils used in this study were obtained from Copaifera reticulata and Copaifera duckei trees, commonly found in the Tapajos National Forest. The procedure received authorization of IBAMA due the scientific purposes. This study identified three oils identified as copaiba 4-C (C. reticulata), copaiba 5-C (C. duckei) and copaiba 8-C (C. reticulata). The bioassay was performed in vitro using specific culture medium for T. evansi, previously described by Baltz, 1985. Copaiba oil was dissolved in dimethyl sulfoxide (DMSO) and tested in three concentrations (0.5, 1.0 and

Background: Trypanosoma evansi is a flagellate which belongs to the salivate section, commonly observed parasitizing blood of equines, ruminants, pigs, dogs and wild animals in different regions of the world. It causes many losses to farmers due to death of animals and drug spending in endemic areas. The treatment of this disease in Brazil is only performed with diminazene aceturate; however it has been ineffective for many animals. During the lats years many studies have been carried out with natural products such as the essential oils. Copaiba oil stands out due some properties described as anti-inflammatory, healing, antiedematogenic, antitumor, parasitic and antibacterial. Therefore, this study aimed to test, in vitro and in vivo, the susceptibility of T. evansi to copaiba oil. Materials, Methods & Results: The oils used in this study were obtained from Copaifera reticulata and Copaifera duckei trees, commonly found in the Tapajos National Forest. The procedure received authorization of IBAMA due the scientific purposes. This study identified three oils identified as copaiba 4-C (C. reticulata), copaiba 5-C (C. duckei) and copaiba 8-C (C. reticulata). The bioassay was performed in vitro using specific culture medium for T. evansi, previously described by Baltz, 1985. Copaiba oil was dissolved in dimethyl sulfoxide (DMSO) and tested in three concentrations (0.5, 1.0 and

Susceptibility of Trypanosoma evansi in the copaiba oil: in vitro test and in mice experimentally infected with the parasite

Background: Trypanosoma evansi is a flagellate which belongs to the salivate section, commonly observed parasitizing blood of equines, ruminants, pigs, dogs and wild animals in different regions of the world. It causes many losses to farmers due to death of animals and drug spending in endemic areas. The treatment of this disease in Brazil is only performed with diminazene aceturate; however it has been ineffective for many animals. During the last years many studies have been carried out with natural products such as the essential oils. Copaiba oil stands out due some properties described as anti-inflammatory, healing, antiedematogenic, antitumor, parasitic and antibacterial. Therefore, this study aimed to test, in vitro and in vivo, the susceptibility of T. evansi to copaiba oil. Materials, Methods & Results: The oils used in this study were obtained from Copaifera reticulata and Copaifera duckei trees, commonly found in the Tapajos National Forest. The procedure received authorization of IBAMA due the scientific purposes. This study identified three oils identified as copaiba 4-C (C. reticulata), copaiba 5-C (C. duckei) and copaiba 8-C (C. reticulata). The bioassay was performed in vitro using specific culture medium for T. evansi, previously described by Baltz, 1985. Copaiba oil was dissolved in dimethyl sulfoxide (DMSO) and tested in three concentrations (0.5, 1.0 and 2.0%) in culture medium containing the parasite. To the control test (without oil) the same volume of DMSO (10 µL) was added. Dimnazene aceturate was also used as a positive control at 0.5% of concentration. The counting of trypanosomes was performed in triplicate in a Neubauer chamber after 1, 3, and 6 hours after the experiment onset. For the tests in vivo, mice were infected (n = 40) and divided in 5 groups of 8 animals each. Group A consisted of healthy animals and Group B comprised animals infected with T. evansi and untreated. The other groups were infected and treated orally with oil of copaíba 4-C (Group C), copaiba 5-C (Group D) and copaiba 8-C (Group E), using a dose of 0.63 mL/kg/day for five consecutive days at intervals of 24 hours. In vitro tests with copaiba oil showed a reduction in the number of alive trypanosomes for the three tested concentrations, when compared to the control test after 1 and 3 h, similar to what occurred with testing aceturate. At 6 hours, it was not observed alive parasites in the test groups, differently from the control group which had an increase of trypanosomes compared to the time zero. The trypanocidal activity had a dose-dependent effect. In the in vivo experiment, the oil of copaiba administered orally had no curative efficacy for any of the groups; however group D treated with C. duckei showed prolonged longevity of the mice when compared to the groups B and C. Discussion: Trypanosoma cruzi and Leishmania amazonensis have been challenged on their susceptibility to oil of copaiba, with obtention of trypanocidal and leishmanicidal effects, similar process thet occurred in this study with T. evansi. According to scientific literature the copaíba oil increases the membrane permeability, as well as provides the depolarization of the mitochondrial membrane in parasite cells. A group of mice in this study showed prolonged longevity, showing that the variation of their compositions influence the trypanocidal effect. Based on these results it was concluded that the T. evansi may is susceptible to the oil of copaiba. Therefore, it can be natural product used as a new alternative and supplementar activity in the treatment of this protozoan, as have been suggested for leishmaniasis.

Susceptibility of Trypanosoma evansi in the copaiba oil: in vitro test and in mice experimentally infected with the parasite / Susceptibility of Trypanosoma evansi in the copaiba oil: in vitro test and in mice experimentally infected with the parasite

Background: Trypanosoma evansi is a flagellate which belongs to the salivate section, commonly observed parasitizing blood of equines, ruminants, pigs, dogs and wild animals in different regions of the world. It causes many losses to farmers due to death of animals and drug spending in endemic areas. The treatment of this disease in Brazil is only performed with diminazene aceturate; however it has been ineffective for many animals. During the lats years many studies have been carried out with natural products such as the essential oils. Copaiba oil stands out due some properties described as anti-inflammatory, healing, antiedematogenic, antitumor, parasitic and antibacterial. Therefore, this study aimed to test, in vitro and in vivo, the susceptibility of T. evansi to copaiba oil. Materials, Methods & Results: The oils used in this study were obtained from Copaifera reticulata and Copaifera duckei trees, commonly found in the Tapajos National Forest. The procedure received authorization of IBAMA due the scientific purposes. This study identified three oils identified as copaiba 4-C (C. reticulata), copaiba 5-C (C. duckei) and copaiba 8-C (C. reticulata). The bioassay was performed in vitro using specific culture medium for T. evansi, previously described by Baltz, 1985. Copaiba oil was dissolved in dimethyl sulfoxide (DMSO) and tested in three concentrations (0.5, 1.0 and

Background: Trypanosoma evansi is a flagellate which belongs to the salivate section, commonly observed parasitizing blood of equines, ruminants, pigs, dogs and wild animals in different regions of the world. It causes many losses to farmers due to death of animals and drug spending in endemic areas. The treatment of this disease in Brazil is only performed with diminazene aceturate; however it has been ineffective for many animals. During the lats years many studies have been carried out with natural products such as the essential oils. Copaiba oil stands out due some properties described as anti-inflammatory, healing, antiedematogenic, antitumor, parasitic and antibacterial. Therefore, this study aimed to test, in vitro and in vivo, the susceptibility of T. evansi to copaiba oil. Materials, Methods & Results: The oils used in this study were obtained from Copaifera reticulata and Copaifera duckei trees, commonly found in the Tapajos National Forest. The procedure received authorization of IBAMA due the scientific purposes. This study identified three oils identified as copaiba 4-C (C. reticulata), copaiba 5-C (C. duckei) and copaiba 8-C (C. reticulata). The bioassay was performed in vitro using specific culture medium for T. evansi, previously described by Baltz, 1985. Copaiba oil was dissolved in dimethyl sulfoxide (DMSO) and tested in three concentrations (0.5, 1.0 and