RESUMO
Trypanosoma cruzi, the causal agent of American trypanosomiasis, and Leishmania spp., the causal agents of Leishmaniasis, are prevalent in more than 20 American countries, including Mexico. Dogs have been reported as incidental hosts for both parasites and may be helpful as transmission sentinels. We surveyed the dog population in a rural locality of the Merida municipality in Yucatan, Mexico, to evaluate the seroreactivity against T. cruzi and Leishmania spp. using two antigens, parasite homogenate (H) and iron superoxide dismutase extract (FeSODe), with two serological techniques (ELISA and Western Blot). Our study found that 3.33% of the tested dogs were seroreactive to T. cruzi using ELISA-H, and 29.5% were seroreactive to FeSODe antigen, with a 94.4% consistency between the two tests. Similarly, for L. mexicana, 1.6% were seroreactive using ELISA-H, and 9.8% were seroreactive using ELISA-FeSODe, with an 83.3% consistency between tests. For L. braziliensis, no dogs were seroreactive using ELISA-H, but 16.4% were seroreactive using ELISA-FeSODe, with a 90% consistency between tests. Finally, for L. infantum, 4.9% were seropositive using ELISA-H, and 6.6% were seropositive using ELISA-FeSODe, with a 75% consistency between tests. These results show noticeable evidence of exposure of dogs to trypanosomatid parasites and highlight the potential disease risk for the people and their companion animals in the region.
Assuntos
Doença de Chagas , Leishmania , Parasitos , Trypanosoma cruzi , Animais , México/epidemiologia , Doença de Chagas/epidemiologia , Doença de Chagas/veterináriaRESUMO
Trypanosomatidae is a family of unicellular parasites belonging to the phylum Euglenozoa, which are causative agents in high impact human diseases such as Leishmaniasis, Chagas disease and African sleeping sickness. The impact on human health and local economies, together with a lack of satisfactory chemotherapeutic treatments and effective vaccines, justifies stringent research efforts to search for new disease therapies. Here, we present in vitro trypanocidal activity data and mode of action data, repositioning leishmanicidal [1,2,3]Triazolo[1,5-a]pyridinium salts against Trypanosoma cruzi, the aetiological agent of Chagas disease. This disease is one of the most neglected tropical diseases and is a major public health issue in Central and South America. The disease affects approximately 6-7 million people and is widespread due to increased migratory movements. We screened a suite of leishmanicidal [1,2,3]Triazolo[1,5-a]pyridinium salt compounds, of which compounds 13, 20 and 21 were identified as trypanocidal drugs. These compounds caused cell death in a mitochondrion-dependent manner through a bioenergetic collapse. Moreover, compounds 13 and 20 showed a remarkable inhibition of iron superoxide dismutase activity of T. cruzi, a key enzyme in the protection from the damage produced by oxidative stress.
Assuntos
Doença de Chagas/tratamento farmacológico , Compostos de Piridínio/farmacologia , Tripanossomicidas/farmacologia , Trypanosoma cruzi/efeitos dos fármacos , Animais , Morte Celular/efeitos dos fármacos , Reposicionamento de Medicamentos , Humanos , Leishmaniose/tratamento farmacológico , Membranas Mitocondriais/metabolismo , Estresse Oxidativo/efeitos dos fármacos , América do Sul , Superóxido Dismutase/metabolismo , Tripanossomíase Africana/tratamento farmacológicoRESUMO
BACKGROUND: Chagas disease caused by Trypanosoma cruzi is endemic in Latin America. Human infection is mainly spread by Triatominae insects. Other forms of transmission are congenital, blood transfusion and organ transplantation. METHODS: Anti-T. cruzi antibodies were determined by enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) in 155 serum samples from mothers and their babies. Indirect immunofluorescence (IFA) and a commercial test were used to validate efficacy of a specific ELISA-iron-excreted superoxide dismutase assay. Sera from babies were collected at 6 and 12 months, whereas maternal samples were obtained after delivery. Calostrum and umbilical cord samples were simultaneously obtained. RESULTS: Anti-T. cruzi antibodies were detected in 8 (5.16%) mothers by ELISA-WB, in 7 (4.51%) using IFA and in 1 (0.64%) by a commercial kit. Nine (5.80%) 6-month-old children were positive by ELISA-WB and 7 (4.51%) by IFA; negative results were obtained when the commercial kit was used. At 12 month of age, 15 (9.67%) children were positive by ELISA-WB, 13 (8.38%) by IFA and 1 (0.64%) by the commercial test. Antibodies were detected in 4 mothers whose children were serologically negative. Four other mothers and their children were positive, but only one of them had detected antibodies in umbilical cord up to 12 months, thus assuming vertical transmission. CONCLUSIONS: The use of iron-excreted superoxide dismutase as antigen in serologic tests for detection of T. cruzi yielded promising results as diagnostic procedure.
Assuntos
Doença de Chagas/congênito , Doença de Chagas/diagnóstico , Complicações Infecciosas na Gravidez/parasitologia , Superóxido Dismutase/sangue , Trypanosoma cruzi/imunologia , Adulto , Anticorpos Antiprotozoários/sangue , Anticorpos Antiprotozoários/imunologia , Antígenos de Protozoários/imunologia , Western Blotting/métodos , Doença de Chagas/sangue , Doença de Chagas/enzimologia , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Técnica Indireta de Fluorescência para Anticorpo/métodos , Humanos , Lactente , Transmissão Vertical de Doenças Infecciosas , Mães , Gravidez , Complicações Infecciosas na Gravidez/sangue , Complicações Infecciosas na Gravidez/imunologia , Complicações Infecciosas na Gravidez/fisiopatologia , Superóxido Dismutase/imunologia , Trypanosoma cruzi/isolamento & purificação , Adulto JovemRESUMO
Clinical diagnosis of leishmaniasis is highly complex, presenting a wide range of clinical manifestations, sometimes non-specific, and thus the epidemiological study and diagnostic need specific molecular markers for each Leishmania species. Leishmania spp. posses different Fe-SOD isoforms, one of which is excreted into the external milieu and, presenting immunogenic characteristics, is a very reliable molecular marker. Superoxide dismutases (SODs) are antioxidant metal-enzymes responsible for the dismutation of superoxide ion into hydrogen peroxide and molecular oxygen, and it is considered an important virulence factor. In this manuscript we have purified the iron(Fe)-SOD excreted by Leishmania braziliensis using ion-exchange and molecular-sieve chromatography and we have studied it as an antigen in serodiagnostic analyses in ELISA and Western blot techniques, testing 213 human sera from Mexico. Indeed, L. braziliensis Fe-SODe has been purified 123.26 times with a specific activity of about 893.66 U/mg of protein. Applying the purified enzymes in serological tests we found 17.84% sera positive. We have demonstrated that the purified enzyme is more sensitive than the non-purified ones and we also demonstrated, for the first time, the presence of antibodies against L. braziliensis, not the main species in the country, in human population from Hidalgo and Nuevo Leon States.
RESUMO
OBJECTIVE: To classify 21 new isolates of Trypanosoma cruzi (T. cruzi) according to the Discrete Typing Unit (DTU) which they belong to, as well as tune up a new pair of primers designed to detect the parasite in biological samples. METHODS: Strains were isolated, DNA extracted, and classified by using three Polymerase Chain Reactions (PCR). Subsequently this DNA was used along with other isolates of various biological samples, for a new PCR using primers designed. Finally, the amplified fragments were sequenced. RESULTS: It was observed the predominance of DTU I in Colombia, as well as the specificity of our primers for detection of T. cruzi, while no band was obtained when other species were used. CONCLUSIONS: This work reveals the genetic variability of 21 new isolates of T. cruzi in Colombia.Our primers confirmed their specificity for detecting the presence of T. cruzi.
RESUMO
Leishmania sp. survival in the vertebrate host depends on the host macrophage immune response as well as on the parasite's defense against free radicals. Iron-superoxide dismutase (Fe-SOD) is a key antioxidant enzyme that contributes to radical superoxide dismutation, preventing the disease from surging and propagating itself. Leishmania sp. has various Fe-SOD isoforms, one of which (Fe-SODe) is excreted into the medium and, being highly immunogenic, can be considered a very good molecular marker. In this work, we purified the Fe-SOD enzymes excreted by L. peruviana and L. amazonensis and studied them as antigens in serodiagnosis. We used ELISA and Western blot techniques to test 51 human cutaneous leishmaniasis sera from Colombia. All 51 patients presented with dermal injuries caused by unknown Leishmania species. The results observed with the purified proteins were compared with those obtained when total soluble lysate and unpurified Fe-SODe were used as the antigen fraction. Thus, we conclude that the purified enzymes are more sensitive and specific than their unpurified counterparts and that there is no cross-reactivity between them.
Assuntos
Anticorpos , Leishmaniose Cutânea/diagnóstico , Superóxido Dismutase/imunologia , Superóxido Dismutase/isolamento & purificação , Anticorpos/imunologia , Especificidade de Anticorpos , Colômbia , Radicais Livres/metabolismo , Humanos , Leishmania/enzimologia , Leishmania/imunologia , Leishmaniose Cutânea/imunologiaRESUMO
The activity of a family scorpiand-like azamacrocycles against Leishmania infantum and Leishmania braziliensis was studied using promastigotes, axenic and intracellular amastigotes forms. All the compounds are more active and less toxic than meglumine antimoniate (Glucantime). Moreover, the data on infection rates and amastigotes showed that compounds P2Py, PN and P3Py are the most active against both species of Leishmania. On the other hand, studies on the inhibitory effect of these compounds on SOD enzymes showed that while the inhibition of the Fe-SOD enzyme of the promastigote forms of the parasites is remarkable, the inhibition of human CuZn-SOD and Mn-SOD from Escherichia coli is negligible. The ultrastructural alterations observed in treated promastigote forms confirmed that the compounds having the highest activity were those causing the largest cell damage. The modifications observed by (1)H NMR, and the amounts of catabolites excreted by the parasites after treatment with the compounds, suggested that the catabolic mechanism could depend on the structure of the side chains linked to the aza-scorpiand macrocycles.
Assuntos
Antiprotozoários/farmacologia , Compostos Aza/farmacologia , Leishmania braziliensis/efeitos dos fármacos , Leishmania infantum/efeitos dos fármacos , Compostos Macrocíclicos/farmacologia , Antiprotozoários/síntese química , Antiprotozoários/química , Compostos Aza/síntese química , Compostos Aza/química , Relação Dose-Resposta a Droga , Compostos Macrocíclicos/síntese química , Compostos Macrocíclicos/química , Estrutura Molecular , Testes de Sensibilidade Parasitária , Relação Estrutura-AtividadeRESUMO
OBJECTIVES: To evaluate the potential of iron-oxide dismutase excreted (SODeCRU) by T. cruzi as the antigen fraction in the serodiagnosis of Chagas disease and compile new epidemiological data on the seroprevalence of this disease in the suburban population of the city of Santiago de Querétaro (Mexico). DESIGN AND METHODS: 258 human sera were analyzed by the techniques of ELISA and Western blot and using the homogenate and the SODeCRU. RESULTS: A total of 31 sera were positive against ELISA/SODeCRU (12.4%), while 30 sera proved positive by WB/SODeCRU (11.6%). The comparison between the technique of ELISA and WB showed a sensitivity of 93%, and a specificity of 99%. The positive predictive value was 93% and the negative predictive value was 99%, with a Kappa (κ) value of 1. CONCLUSIONS: These preliminary data reveal the degree of infection of nonrural areas of Mexico and demonstrated that SODeCRU is an antigen useful to diagnose Chagas disease.
Assuntos
Doença de Chagas/sangue , Doença de Chagas/epidemiologia , Trypanosoma cruzi/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antiprotozoários/sangue , Western Blotting , Doença de Chagas/parasitologia , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Masculino , México , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Valor Preditivo dos Testes , Projetos de Pesquisa , Sensibilidade e Especificidade , Estudos Soroepidemiológicos , Testes Sorológicos , População Suburbana , TemperaturaRESUMO
Studies on the anti-proliferative activity in vitro of seven ternary nickel (II) complexes with a triazolopyrimidine derivative and different aliphatic or aromatic amines as auxiliary ligands against promastigote and amastigote forms of Leishmania infantum and Leishmania braziliensis have been carried out. These compounds are not toxic for the host cells and two of them are effective at lower concentrations than the reference drug used in the present study (Glucantime). In general, the in vitro growth rate of Leishmania spp. was reduced, its capacity to infect cells was negatively affected and the multiplication of the amastigotes decreased. Ultrastructural analysis and metabolism excretion studies were executed in order to propose a possible mechanism for the action of the assayed compounds. Our results show that the potential mechanism is at the level of organelles membranes, either by direct action on the microtubules or by their disorganization, leading to vacuolization, degradation and ultimately cell death.
Assuntos
Antiprotozoários/farmacologia , Complexos de Coordenação/farmacologia , Leishmania braziliensis/efeitos dos fármacos , Leishmania infantum/efeitos dos fármacos , Níquel/farmacologia , Organelas/ultraestrutura , Pirimidinas/farmacologia , Animais , Antiprotozoários/síntese química , Antiprotozoários/química , Cátions Bivalentes/farmacologia , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Complexos de Coordenação/síntese química , Complexos de Coordenação/química , Feminino , Leishmania braziliensis/metabolismo , Leishmania infantum/metabolismo , Ligantes , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/parasitologia , Camundongos , Camundongos Endogâmicos BALB C , Modelos Moleculares , Níquel/química , Organelas/efeitos dos fármacos , Testes de Sensibilidade Parasitária , Triazóis/farmacologiaRESUMO
Although human leishmaniasis has been reported in 20 states in Mexico, no case of leishmaniasis has been reported in cats to date. In the Yucatan Peninsula, it has been found that dogs may act as reservoirs for at least three Leishmania species (Leishmania mexicana, Leishmania braziliensis, and Leishmania panamensis). In this study we identified specific antibodies against these three Leishmania spp. and Trypanosoma cruzi in the sera from 95 cats from two States on the Yucatan Peninsula, namely Quintana Roo and Yucatan, by ELISA and Western blot techniques using whole extract and an iron superoxide dismutase excreted by the parasites as antigens. As well as demonstrating the presence of trypanosomatid antibodies in the feline population on the Yucatan Peninsula, we were also able to confirm the high sensitivity and specificity of the iron superoxide dismutase antigen secreted by them, which may prove to be very useful in epidemiological studies.
Assuntos
Anticorpos Antiprotozoários/sangue , Doenças do Gato/imunologia , Doença de Chagas/veterinária , Leishmania/imunologia , Leishmaniose/veterinária , Superóxido Dismutase/imunologia , Trypanosoma cruzi/imunologia , Animais , Anticorpos Antiprotozoários/imunologia , Antígenos de Protozoários/imunologia , Western Blotting , Doenças do Gato/epidemiologia , Doenças do Gato/parasitologia , Gatos , Doença de Chagas/epidemiologia , Doença de Chagas/imunologia , Doença de Chagas/parasitologia , Ensaio de Imunoadsorção Enzimática , Leishmania/isolamento & purificação , Leishmania/patogenicidade , Leishmaniose/epidemiologia , Leishmaniose/imunologia , Leishmaniose/parasitologia , México/epidemiologia , Carga Parasitária , Prevalência , Sensibilidade e Especificidade , Trypanosoma cruzi/isolamento & purificação , Trypanosoma cruzi/patogenicidadeRESUMO
The activity of five (1-5) abietane phenol derivatives against Leishmania infantum and Leishmania braziliensis was studied using promastigotes and axenic and intracellular amastigotes. Infectivity and cytotoxicity tests were performed with J774.2 macrophage cells using Glucantime as a reference drug. The mechanisms of action were analysed by performing metabolite excretion and transmission electron microscopy ultrastructural studies. Compounds 1-5 were more active and less toxic than Glucantime. The infection rates and mean number of parasites per cell observed in amastigote experiments showed that derivatives 2, 4 and 5 were the most effective against both L. infantum and L. braziliensis. The ultrastructural changes observed in the treated promastigote forms confirmed that the greatest cell damage was caused by the most active compound (4). Only compound 5 caused changes in the nature and amounts of catabolites excreted by the parasites, as measured by ¹H nuclear magnetic resonance spectroscopy. All of the assayed compounds were active against the two Leishmania species in vitro and were less toxic in mammalian cells than the reference drug.
Assuntos
Animais , Feminino , Camundongos , Antiprotozoários/farmacologia , Leishmania braziliensis/efeitos dos fármacos , Leishmania infantum/efeitos dos fármacos , Macrófagos/parasitologia , Terpenos/farmacologia , Antiprotozoários/química , Leishmania braziliensis/ultraestrutura , Leishmania infantum/ultraestrutura , Espectroscopia de Ressonância Magnética , Camundongos Endogâmicos BALB C , Microscopia Eletrônica de Transmissão , Testes de Sensibilidade Parasitária , Terpenos/químicaRESUMO
The activity of five (1-5) abietane phenol derivatives against Leishmania infantum and Leishmania braziliensis was studied using promastigotes and axenic and intracellular amastigotes. Infectivity and cytotoxicity tests were performed with J774.2 macrophage cells using Glucantime as a reference drug. The mechanisms of action were analysed by performing metabolite excretion and transmission electron microscopy ultrastructural studies. Compounds 1-5 were more active and less toxic than Glucantime. The infection rates and mean number of parasites per cell observed in amastigote experiments showed that derivatives 2, 4 and 5 were the most effective against both L. infantum and L. braziliensis. The ultrastructural changes observed in the treated promastigote forms confirmed that the greatest cell damage was caused by the most active compound (4). Only compound 5 caused changes in the nature and amounts of catabolites excreted by the parasites, as measured by ¹H nuclear magnetic resonance spectroscopy. All of the assayed compounds were active against the two Leishmania species in vitro and were less toxic in mammalian cells than the reference drug.
Assuntos
Antiprotozoários/farmacologia , Leishmania braziliensis/efeitos dos fármacos , Leishmania infantum/efeitos dos fármacos , Macrófagos/parasitologia , Terpenos/farmacologia , Animais , Antiprotozoários/química , Feminino , Concentração Inibidora 50 , Leishmania braziliensis/ultraestrutura , Leishmania infantum/ultraestrutura , Espectroscopia de Ressonância Magnética , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Eletrônica de Transmissão , Testes de Sensibilidade Parasitária , Terpenos/químicaRESUMO
Leishmania spp. has been recorded in humans and in dogs, and numerous studies have demonstrated that dogs act as reservoirs for visceral leishmaniasis. The objective of this study was to determine the prevalence of three species of the Leishmania genus and possible associated factors in sera of 218 dogs from two different populations in Mérida, Yucatán (Mexico). The sera were analyzed to detect antibodies against L. mexicana, L. braziliensis, and L. infantum using the superoxide dismutase- enzyme-linked immunosorbent assay (SOD-ELISA) and Western blot as confirmation. The Fe-SOD excreted was used as the antigenic fraction for the three Leishmania species. The prevalence values found were 30.2% (L. mexicana), 8.2% (L. braziliensis), and 11.9% (L. infantum), with L. mexicana seroprevalence being greater than L. braziliensis and L. infantum (p<0.05). Five percent (11/218) of the dogs showed antibodies against L. mexicana/L. braziliensis, 5.5% (12/218) with L. mexicana/L. infantum and 1.8% (4/218) with L. mexicana/L. braziliensis/L. infantum. No relationship (p>0.05) was found between antibodies against L. mexicana and breed, age, physical condition, or cutaneous lesions in dogs. This study provides evidence of antibodies against L. mexicana, L. braziliensis and L. infantum in dog populations from Mérida, Mexico.
Assuntos
Anticorpos Antiprotozoários/sangue , Doenças do Cão/epidemiologia , Doenças do Cão/imunologia , Leishmania braziliensis/imunologia , Leishmania infantum/imunologia , Leishmania mexicana/imunologia , Leishmaniose/veterinária , Animais , Western Blotting , Cães , Ensaio de Imunoadsorção Enzimática , Leishmaniose/imunologia , Masculino , México/epidemiologia , PrevalênciaRESUMO
OBJECTIVES: To evaluate the in vitro leishmanicidal activity of imidazole-based (1-4) and pyrazole-based (5-6) benzo[g]phthalazine derivatives against Leishmania infantum and Leishmania braziliensis. METHODS: The in vitro activity of compounds 1-6 was assayed on extracellular promastigote and axenic amastigote forms, and on intracellular amastigote forms of the parasites. Infectivity and cytotoxicity tests were performed on J774.2 macrophage cells using meglumine antimoniate (Glucantime) as the reference drug. The mechanisms of action were analysed by iron superoxide dismutase (Fe-SOD) and copper/zinc superoxide dismutase (CuZn-SOD) inhibition, metabolite excretion and transmission electronic microscopy (TEM). RESULTS: Compounds 1-6 were more active and less toxic than meglumine antimoniate. Data on infection rates and amastigote mean numbers showed that 2, 4 and 6 were more active than 1, 3 and 5 in both L. infantum and L. braziliensis. The inhibitory effect of these compounds on the antioxidant enzyme Fe-SOD of promastigote forms of the parasites was remarkable, whereas inhibition of human CuZn-SOD was negligible. The ultrastructural alterations observed in treated promastigote forms confirmed the greater cell damage caused by the most active compounds 2, 4 and 6. The modifications observed by (1)H-NMR in the nature and amounts of catabolites excreted by the parasites after treatment with 1-6 suggested that the catabolic mechanisms could depend on the structure of the side chains linked to the benzo[g]phthalazine moiety. CONCLUSIONS: All the compounds assayed were active in vitro against the two Leishmania species and were less toxic against mammalian cells than the reference drug, but the monosubstituted compounds were significantly more effective and less toxic than their disubstituted counterparts.
Assuntos
Antiprotozoários/farmacologia , Imidazóis/farmacologia , Leishmania braziliensis/efeitos dos fármacos , Leishmania infantum/efeitos dos fármacos , Ftalazinas/farmacologia , Pirazóis/farmacologia , Animais , Antiprotozoários/química , Antiprotozoários/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Feminino , Humanos , Imidazóis/química , Imidazóis/toxicidade , Macrófagos/parasitologia , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Eletrônica de Transmissão , Testes de Sensibilidade Parasitária , Ftalazinas/química , Ftalazinas/toxicidade , Pirazóis/química , Pirazóis/toxicidade , Relação Estrutura-Atividade , Superóxido Dismutase/metabolismoRESUMO
An increasing number of studies have reported high infection rates for American cutaneous leishmaniasis in dogs, which have thus been proposed as the reservoir host. Canine leishmaniasis is widespread in different states in Mexico, where a number of Leishmania species have been isolated from dogs. In the present study, the detection of different Leishmania species is described in stray dogs from two localities, namely Tulum and Celestún on the Yucatan Peninsula (Mexico). The use of iron-superoxide dismutase excreted by the parasites as the antigen fraction and enzyme-linked immunosorbent assay and western blot tests allowed us to confirm the presence of at least three species of Leishmania (Le. mexicana, Le. braziliensis, and Le. panamensis), some of which are reported for the first time in this species. In addition to a high prevalence of Le. mexicana and Le. braziliensis, and to a lesser degree, Le. panamensis, there is a significant prevalence of Trypanosoma cruzi, suggesting that the dog may be a source of transmission of trypanosomiasis. However, a more thorough epidemiological study on the dog population, both wild as well as urban, of the Yucatan Peninsula will be required to design a control strategy for these diseases, paying particular attention to the population affected and even broadening the study to other Mexican states as well as neighboring countries. These results again confirm that iron-superoxide dismutase excreted by the different trypanosomatid species constitutes a good source of antigen for serodiagnosis in epidemiological studies.
Assuntos
Antígenos de Protozoários , Doenças do Cão/diagnóstico , Doenças do Cão/parasitologia , Leishmania/imunologia , Leishmania/metabolismo , Leishmaniose Cutânea/veterinária , Animais , Western Blotting/veterinária , Vetores de Doenças , Doenças do Cão/epidemiologia , Doenças do Cão/transmissão , Cães , Ensaio de Imunoadsorção Enzimática , Leishmania/classificação , Leishmaniose Cutânea/diagnóstico , Leishmaniose Cutânea/epidemiologia , Leishmaniose Cutânea/transmissão , México/epidemiologia , Superóxidos , Trypanosoma cruzi/isolamento & purificação , Tripanossomíase/epidemiologia , Tripanossomíase/veterináriaRESUMO
A superoxide dismutase excreted by promastigote forms of L. (Viannia) peruviana (SODe-Lp), L. (Viannia) brazilensis (SODe-Lb), and L. (L.) amazonensis (SODe-La) is tested to evaluate its potential value as a diagnostic tool of mucocutaneous and Andean cutaneous leishmaniasis. We used 45 sera with mucocutaneous leishmaniasis (SL) and 68 with Andean cutaneous leishmaniasis (ACL). SODe-Lp antigen was recognized by 94% of the serum from ACL patients, and the SODe-Lb antigen was recognized by 93% of the serum from SL patients. Meanwhile, the result for SL and ACL patients with SODe-La antigen was 69% and 43% and SODe-Li was 11% and 9%, respectively. This suggest that antibodies to SODe-Lp undergo further response in patients with ACL and the antibodies to SODe-Lb do so preferentially in patients with SL. The SODe ELISA may be useful in endemic areas for discriminative assays between patients with different forms of leishmaniases and those with other clinical conditions.
Assuntos
Antígenos de Protozoários/análise , Leishmaniose Cutânea/diagnóstico , Superóxido Dismutase/análise , Animais , Antígenos de Protozoários/sangue , Ensaio de Imunoadsorção Enzimática , Humanos , Focalização Isoelétrica , Leishmania braziliensis/enzimologia , Leishmania infantum/enzimologia , Leishmaniose Cutânea/sangue , Peru , Superóxido Dismutase/sangueRESUMO
Chagas disease is present in Mexico but data on the presence of its vectors are not known for all the states. We conducted an epidemiological study to ascertain the presence of bedbugs in 23 communities of seven municipalities in the state of Querétaro, Mexico. Sampling was performed within and outside dwellings, using the technique of one person searching per hour per house. Four triatomine species were found: Triatoma mexicana (94%), Tri. dimidiata (3%), Tri. gerstaeckeri (2%) and Tri. pallidipennis (1%). Triatoma mexicana was found in six of the seven municipalities but has previously been found only in the state of Hidalgo, where it is considered the most important vector species of Trypanosoma cruzi. This is the first time that the presence of Tri. gerstaeckeri or Tri. dimidiata has been reported in the state of Querétaro. Tolimán municipality had the highest entomological indices: infestation index 73%; infection index 34.5%; crowding index 9.2%; density index 6.7%; and colonization index 20.6%. Although the prevalence and colonization indices were low, the infestation and infection indices indicate that different Triatoma species are becoming established in Querétaro. This work represents the first entomological study in this Mexican state and suggests that some triatomine vectors are extraordinarily mobile.
Assuntos
Doença de Chagas/parasitologia , Ectoparasitoses/transmissão , Insetos Vetores , Reduviidae , Triatoma , Trypanosoma cruzi/isolamento & purificação , Animais , Doença de Chagas/transmissão , Ectoparasitoses/epidemiologia , Entomologia/métodos , Hemípteros , Humanos , MéxicoRESUMO
Four superoxide dismutase (SOD) activities (SOD I, II, III, and IV) have been characterized in the epimastigote form of Trypanosoma cruzi. The total extract was subjected to two successive ammonium sulphate additions between 35 and 85%, and the resulting fraction was purified using two continuous chromatography processes (ion exchange and filtration). Enzymes were insensitive to cyanide but sensitive to hydrogen peroxide, properties characteristic of iron-containing SODs. The molecular masses of the different SODs were 20 kDa (SOD I), 60 kDa (SOD II), 50 kDa (SOD III) and 25 kDa (SOD IV), whereas the isoelectric points were 6.9, 6.8, 5.2 and 3.8, respectively. Subcellular location and digitonin experiments have shown that these SODs are mainly cytosolic, with small amounts in the low-mass organelles (SOD II and SOD I) and the mitochondrion (SOD III), where these enzymes play an important role in minimizing oxidative damage.
Assuntos
Superóxido Dismutase/isolamento & purificação , Trypanosoma cruzi/enzimologia , Animais , Superóxido Dismutase/classificaçãoRESUMO
Four superoxide dismutase (SOD) activities (SOD I, II, III, and IV) have been characterized in the epimastigote form of Trypanosoma cruzi. The total extract was subjected to two successive ammonium sulphate additions between 35 and 85 percent, and the resulting fraction was purified using two continuous chromatography processes (ion exchange and filtration). Enzymes were insensitive to cyanide but sensitive to hydrogen peroxide, properties characteristic of iron-containing SODs. The molecular masses of the different SODs were 20 kDa (SOD I), 60 kDa (SOD II), 50 kDa (SOD III) and 25 kDa (SOD IV), whereas the isoelectric points were 6.9, 6.8, 5.2 and 3.8, respectively. Subcellular location and digitonin experiments have shown that these SODs are mainly cytosolic, with small amounts in the low-mass organelles (SOD II and SOD I) and the mitochondrion (SOD III), where these enzymes play an important role in minimizing oxidative damage.
Assuntos
Animais , Superóxido Dismutase/isolamento & purificação , Trypanosoma cruzi/enzimologia , Superóxido Dismutase/classificaçãoRESUMO
We have characterized diverse strains or species of Leishmania isolated in humans that are currently circulating throughout Peru, by means of isoenzymatic characterization, kDNA analysis by restriction enzymes, and multiplex PCR assay. The cluster analysis gave five groups. Cluster 1 includes L. (L.) donovani together with the isolates LP4 and LP7, forming the donovani complex. Thus, this complex corresponds to the New World visceral form, L. (L.) chagasi. Cluster 2 is formed by the isolates LP1-LP3, LP6, LP10, LP9, and LP11, phylogenetically intermediate between Cluster 1 and Cluster 3, or they can be treated as hybrids. Cluster 3 is divided into two subgroups: one formed by L. (V.) peruviana, together with the isolates LP14 and LP5, and the second one formed by L. (V.) brazilensis and the isolate LP8. These two subgroups form part of the brazilensis complex. The three strains of L. (L.) infantum [L. (L.) infantum I and II and la LSI] make up Cluster 4. In Cluster 5, we include the three Mexican strains (LM1-LM3) forming one subgroup while we would place L. (L.) amazonensis in another subgroup. These two subgroups would comprise the complex mexicana.