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Parasites have been associated with possible anticancer activity, including Trypanosoma cruzi, which has been linked to inhibiting the growth of solid tumors. To better understand this antitumor effect, we investigated the association of anti-T. cruzi antibodies with B cells of the acute lymphoblastic leukemia (ALL) SUPB15 cell line. The antibodies were generated in rabbits. IgGs were purified by affinity chromatography. Two procedures (flow cytometry (CF) and Western blot(WB)) were employed to recognize anti-T. cruzi antibodies on SUPB15 cells. We also used CF to determine whether the anti-T. cruzi antibodies could suppress SUPB15 cells. The anti-T. cruzi antibodies recognized 35.5% of the surface antigens of SUPB15. The complement-dependent cytotoxicity (CDC) results demonstrate the cross-suppression of anti-T. cruzi antibodies on up to 8.4% of SUPB15 cells. For the WB analysis, a band at 100 kDa with high intensity was sequenced using mass spectrometry, identifying the protein as nucleolin. This protein may play a role in the antitumor effect on T. cruzi. The anti-T. cruzi antibodies represent promising polyclonal antibodies that have the effect of tumor-suppressive cross-linking on cancer cells, which should be further investigated.
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Anticorpos Antiprotozoários , Leucemia-Linfoma Linfoblástico de Células Precursoras , Trypanosoma cruzi , Trypanosoma cruzi/imunologia , Leucemia-Linfoma Linfoblástico de Células Precursoras/imunologia , Humanos , Linhagem Celular Tumoral , Animais , Coelhos , Anticorpos Antiprotozoários/imunologia , Proteínas de Ligação a RNA/imunologia , Proteínas de Ligação a RNA/metabolismo , Nucleolina , Fosfoproteínas/imunologia , Fosfoproteínas/metabolismoRESUMO
Background. Research studies indicate that immunization with protein extracts of Trypanosoma cruzi, the protozoan parasite that causes Chagas disease, prevents the appearance of tumors in 60% of mice injected with the murine lung carcinoma tumor line. The molecular basis of this process is unknown, although the presence of specific antigens in tumor cells and on the surface of T. cruzi suggests an antiparasitic immune response, with an effective cross-reaction against cancer cells, hence the importance to identify the antigens involved and determine their potential as target cells in anticancer therapy. Aim. This study aimed to determine the presence of antigenic proteins of T. cruzi shared with acute lymphoblastic leukemia and neuroblastoma cells. Material and methods. To achieve this, polyclonal antibodies against T. cruzi were developed in rabbits, and reactivity was determined with protein extracts of acute lymphoblastic leukemia cells and neuroblastoma. The immunodetection of five different strains of T. cruzi against anti-T. cruzi polyclonal antibodies was also performed. Conclusion. The study allows the knowledge of the immunological interactions between cancer and parasites to be expanded and, therefore, contributes to the design of more and better projects that improve the therapeutic strategies applied in oncology.
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Malaria is a parasitic disease of global importance due to its high annual death toll. The treatment for this infection is difficult for the increase in the populations of parasites resistant to the existing medicines, the development of new antimalarials is urgent needed. Several products developed for the control of malaria from herbalist have had a profound impact, for example, quinine obtained from the bark of the cinchona tree and recently those derived from artemisinin, whose discovery was the reason for the awarding of the 2015 Nobel Prize. The aim of the present study was to evaluate a compound named kramecyne extracted of "chayotillo" (Krameria cystisoides) plant used by the antiparasitic effect against some blood and intestinal protozoa (Giardia duodenalis y Trypanosoma cruzi). In addition is using for the treatment of inflammatory diseases. Measuring parasitaemia at different times, it was observed that in mice treated with kramecyne, it reached only 14% of parasitaemia at 7 days with a dose of 15 mg/kg, using chloroquine as a control drug, because it has not been demonstrated that parasites that infect rodents have developed resistance against this drug. Our results showed that kramecyne decreases the expression of parasite proteins that participate in biological processes, such as invasion, cytoadherence, pathogenicity and energy metabolism. With these results, it is proposed that this compound has repercussions on the metabolism of the parasite and could be useful for use as an antimalarial.
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Antimaláricos , Malária , Animais , Antimaláricos/farmacologia , Antimaláricos/uso terapêutico , Antiparasitários/farmacologia , Éteres Cíclicos , Malária/tratamento farmacológico , Malária/parasitologia , Camundongos , Peróxidos , Extratos Vegetais/farmacologia , Plasmodium berghei , Plasmodium falciparum , ProteômicaRESUMO
Congenital Chagas disease is considered a form of dispersion of Trypanosoma cruzi related to human migration from endemic, often rural to previously non-endemic urban areas. This fact increases the Chagas disease establishment risk inside of family members by vertical transmission pathway. Congenital Chagas disease cases in newborns could not identified by the health professional even in endemic regions. Here we present the first family cluster of Chagas disease cases from Chiapas: one of the most important endemic areas in South of Mexico, where vertical T. cruzi transmission incidence rate is ranged between 2% to 22% revealing an important public health problem. Two cases inside a family from Chiapas, México with positive antibodies against T. cruzi detected by ELISA are presented; one of them got the infection through vertical pathway. We think that congenital Chagas disease should not be ignored in a newborn born from an asymptomatic Chagas disease mother, who may transmit the parasite infection randomly.
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INTRODUCTION: Dengue, Zika and Chikungunya are RNA Arboviruses present in some areas of Mexico, mainly in the endemic state of Chiapas that is characterized by presence of the vector that transmit them and an ecology that favors high transmission. According to the national epidemiological surveillance system, Dengue has intensified since 2018 and outbreaks continue in various states while for Zika and Chikungunya a decrease in cases has been reported in recent years. The main objective of this study was to determine the incidence of Dengue, Zika and Chikungunya infections during pregnancy in the state of Chiapas. PRINCIPAL FINDINGS: The presence of previous and current infections and coinfections diagnosed by molecular (RT-PCR) and immunological (ELISA for IgG determination) techniques indicates a wide circulation of viruses in asymptomatic people, specifically in pregnant women showing that silent infections in dry season contributes to the preservation of viruses. CONCLUSIONS: From 136 studied samples, 27.7% tested positive for DENV, 8% for ZIKV and 24.1% for CHIKV by RTPCR and the values of IgG in sera show that 83.9% were positive for IgG antibodies against DENV, 65% against ZIKV and 59.1% against CHIKV. Results demonstrated presence of ZIKV and CHIKV, not detected by the epidemiological surveillance system, so the importance of establishing proactive epidemiological systems more strict, especially because these infections in pregnant women can cause severe health problems for newborn children.
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Febre de Chikungunya/complicações , Coinfecção , Dengue/complicações , Complicações Infecciosas na Gravidez/virologia , Infecção por Zika virus/complicações , Adulto , Anticorpos Antivirais/sangue , Febre de Chikungunya/epidemiologia , Dengue/epidemiologia , Doenças Endêmicas , Feminino , Humanos , Imunoglobulina G/sangue , Transmissão Vertical de Doenças Infecciosas , México/epidemiologia , Gravidez , Complicações Infecciosas na Gravidez/epidemiologia , RNA Viral/sangue , RNA Viral/isolamento & purificação , Infecção por Zika virus/epidemiologiaRESUMO
Giardia lamblia is one of the most important worldwide causes of intestinal infections, yet little is known about its cellular physiology, especially the diversity of ionic channels that this parasite expresses. In this work, we show that injection of mRNA isolated from trophozoites of Giardia, into Xenopus laevis oocytes, induces expression of three types of chloride currents (here referred to as ICl-G1, ICl-G2, and ICl-G3), which have different biophysical and pharmacological properties. ICl-G1 currents show inward rectification and voltage dependence are enhanced by hypotonicity, show a selectivity sequence of (I > Br > Cl > F), and are inhibited by NPPB, DIDS, SITS, 9AC, DPC, and Zinc. These findings suggest that ICl-G1 is the result of expression of chloride channels related to ClC2. ICl-G2 currents show outward rectification and are dependent of intracellular calcium, its selectivity sequence is (Cl > Br > I > F) and are inhibited by NPPB, DIDS, SITS, 9AC, DPC, niflumic acid, tannic acid, and benzbromarone. These findings suggest that they are produced by calcium dependent chloride channels (CaCC). The third type of currents (ICl-G3) appears only after a hypoosmotic challenge, and has similar properties to those described for ICl-swell, such as outward rectification, instant activation, and slow inactivation at large depolarizing voltages. They were blocked by NPPB, DIDS, 9AC, NIf, DCPIB, and tamoxifen. Our results indicate that Giardia intestinalis has at least three types of anion conductances.
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Canais de Cloreto/biossíntese , Giardia lamblia/genética , Oócitos/metabolismo , RNA Mensageiro/administração & dosagem , RNA de Protozoário/administração & dosagem , Trofozoítos/genética , Xenopus laevis/metabolismo , Animais , Cálcio/metabolismo , Canais de Cloreto/genética , Canais de Cloreto/metabolismo , Fenômenos Eletrofisiológicos , Feminino , Giardia lamblia/crescimento & desenvolvimento , Concentração de Íons de Hidrogênio , Injeções , Potenciais da Membrana , Oócitos/citologia , RNA Mensageiro/genética , RNA de Protozoário/genética , Trofozoítos/crescimento & desenvolvimento , Xenopus laevis/genéticaRESUMO
Resumen: Introducción: Trichinella spiralis es un nemátodo tisular que se aloja en el músculo esquelético de humanos y otros mamíferos y causa una serie de alteraciones fisiológicas. Las proteínas de los productos de excreción-secreción de T. spiralis juegan un papel importante en la aparición y regulación de estas alteraciones. Sin embargo, aún no se conoce el efecto de estos productos en la infección e invasión del parásito al hospedero. Métodos: Mediante un análisis electroforético en una dimensión, Western blot y espectrometría de masas, se evaluaron las diferencias y similitudes entre proteínas antigénicas y de superficie de cuatro aislados de T. spiralis obtenidos de hospederos accidentales (perros) y la cepa de referencia aislada de cerdos (MSUS/MEX/91/CM). Resultados: Utilizando ontología de genes, se encontraron cinco proteínas exclusivas de los hospederos accidentales. Después del análisis, se encontró que estas proteínas forman parte de la matriz extracelular del parásito, cuentan con actividad catalítica y están implicadas en la adhesión a las células del hospedero. La actividad antigénica de las cuatro cepas aisladas de hospederos accidentales es idéntica a la reportada para T. spiralis, visualizándose el triplete antigénico característico de 43, 45 y 47 kDa. Conclusiones: Las proteínas exclusivas de los hospederos accidentales proveen información para entender el mecanismo de acción de este parásito para penetrar el músculo y evadir la respuesta inmune en el hospedero.
Abstract: Background: Trichinella spiralis is an intestinal and tissue nematode specific for mammalian skeletal muscle, causing a series of physiological alterations. T. spiralis excretory-secretion products play an important role in the appearance and regulation of these alterations. However, the effect of these products on the infection and invasion of the parasite to the host is unknown. Methods: Differences and similarities between antigenic proteins and surface proteins of four accidental hosts isolates (dogs) of T. spiralis and the reference strain isolated from pigs (MSUS/MEX/91/CM) were assessed by electrophoresis, western blot and mass spectrometry. Results: Using gene ontology, five proteins exclusive to the accidental hosts were analyzed. The results showed that these proteins are part of the extracellular matrix of the parasite, present catalytic activity, and bind to host cells. The antigenic activity the four strains showed the antigenic triplet characteristic of T. spiralis of 43, 45 and 47 kDa. Conclusions: Five proteins exclusive to dog isolates provided information to understand the mechanism of action of this parasite to penetrate the muscle and evade the immune response in the host.
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Animais , Cães , Ratos , Triquinelose/parasitologia , Trichinella spiralis/metabolismo , Proteômica/métodos , Espectrometria de Massas , Suínos , Triquinelose/imunologia , Western Blotting , Trichinella spiralis/isolamento & purificação , Trichinella spiralis/imunologia , Ratos Wistar , Eletroforese , Antígenos de Helmintos/imunologiaRESUMO
BACKGROUND: Trichinella spiralis is an intestinal and tissue nematode specific for mammalian skeletal muscle, causing a series of physiological alterations. T. spiralis excretory-secretion products play an important role in the appearance and regulation of these alterations. However, the effect of these products on the infection and invasion of the parasite to the host is unknown. METHODS: Differences and similarities between antigenic proteins and surface proteins of four accidental hosts isolates (dogs) of T. spiralis and the reference strain isolated from pigs (MSUS/MEX/91/CM) were assessed by electrophoresis, western blot and mass spectrometry. RESULTS: Using gene ontology, five proteins exclusive to the accidental hosts were analyzed. The results showed that these proteins are part of the extracellular matrix of the parasite, present catalytic activity, and bind to host cells. The antigenic activity the four strains showed the antigenic triplet characteristic of T. spiralis of 43, 45 and 47 kDa. CONCLUSIONS: Five proteins exclusive to dog isolates provided information to understand the mechanism of action of this parasite to penetrate the muscle and evade the immune response in the host.
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Proteômica/métodos , Trichinella spiralis/metabolismo , Triquinelose/parasitologia , Animais , Antígenos de Helmintos/imunologia , Western Blotting , Cães , Eletroforese , Espectrometria de Massas , Ratos , Ratos Wistar , Suínos , Trichinella spiralis/imunologia , Trichinella spiralis/isolamento & purificação , Triquinelose/imunologiaRESUMO
A variety of drugs are used in giardiasis treatment with different levels of efficiency, presence of side effects, and even formation of resistant strains, so that it is important to search new only-one-dose treatments with high efficiency and less side effects. Kramecyne, an anti-inflammatory compound isolated from methanolic extract of Krameria cytisoides, does not present toxicity, even at doses of 5,000 mg/kg. The objective was to determine the antigiardial effect of kramecyne over Giardia intestinalis in vitro and in vivo and analyze the expression of genes ERK1, ERK2, and AK on kramecyne treated trophozoites by Real Time Polymerase Chain Reaction (RTPCR). The median lethal dose (LD50) was 40 µg/mL and no morphological changes were observed by staining with blue trypan and light microscopy; experimental gerbil infection was eliminated with 320 µg/Kg of weight. After treatment there were no differences between intestines from treated and untreated gerbils. Kramecyne did not have significant effect over ERK1 and AK, but there are differences in ERK2 expression (p = 0.04). Results show antigiardial activity of kramecyne; however the mode of action is still unclear and the evaluation of ultrastructural damage and expressed proteins is an alternative of study to understand the action mechanism.
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OBJECTIVE: To determine the Trypanosoma cruzi infection prevalence in 1125 pregnant women and the transmission frequency to their children from Tapachula and Palenque, Chiapas. MATERIALS AND METHODS: We determined the prevalence by serology tests and the transmission frequency by polymerase chain reaction (PCR) and T. cruzi reactivity capacity after 12 months. RESULTS: Total maternal infection prevalence were 23/1 125 (2.04%), 9/600 (1.5%) were from Tapachula and 14/525 (2.6%) from Palenque. The seropositive women were between 20 and 35 years old, 31.8% have Premature Rapture of Membrane and 9.1% have history of perinatal death. The total percentage of positive newborns by PCR was 9/23 (39.13%), out of those 2/9 (22.2%) are from Tapachula and 7/14 (50%) from Palenque. The Maternal Fetal transmission frequency was. 2/9 (22.2%) in Tapachula and 1/14 (7.14%) in Palenque, all positive infants were asynthomatic. CONCLUSION: The maternal-fetal transmission rate in Chiapas State is variable; the reason could be the maternal immunological status and T. cruzi strain.
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Doença de Chagas/transmissão , Transmissão Vertical de Doenças Infecciosas , Complicações Infecciosas na Gravidez/epidemiologia , Adulto , Doenças Assintomáticas , Doença de Chagas/epidemiologia , Feminino , Geografia Médica , Humanos , Lactente , Recém-Nascido , Transmissão Vertical de Doenças Infecciosas/estatística & dados numéricos , Masculino , México/epidemiologia , Gravidez , Resultado da Gravidez , Estudos Soroepidemiológicos , Trypanosoma cruzi/imunologia , Adulto JovemRESUMO
Resumen: Objetivo: Determinar la prevalencia de infección de Trypanosoma cruzi en 1 125 mujeres embarazadas y la frecuencia con que éstas la transmiten a sus hijos en Tapachula y Palenque, Chiapas. Material y métodos: Se determinó la prevalencia materna por serología, la frecuencia de transmisión por reacción en cadena de la polimerasa (PCR) y la capacidad reactiva a T. cruzi a los 12 meses de edad del niño. Resultados: La prevalencia materna total fue 23/1 125 (2.04%); 9/600 (1.5%) en Tapachula y 14/525 (2.6%) en Palenque. Las mujeres seropositivas tenían entre 20 y 35 años; 31.8% tuvo ruptura prematura de membrana (RPM) y 9.1% tuvo antecedentes de muerte perinatal. La frecuencia de transmisión materno-fetal en Tapachula fue de 2/9(22.2%) y en Palenque de 1/14(7.14%); todos los niños positivos eran asintomáticos. Conclusión: La tasa de transmisión materno-fetal en el estado de Chiapas es variable; la razón podría ser el estado inmunológico de la madre o la cepa de T. cruzi.
Abstract: Objective: To determine the Trypanosoma cruzi infection prevalence in 1125 pregnant women and the transmission frequency to their children from Tapachula and Palenque, Chiapas. Materials and methods: We determined the prevalence by serology tests and the transmission frequency by polymerase chain reaction (PCR) and T. cruzi reactivity capacity after 12 months. Results: Total maternal infection prevalence were 23/1 125 (2.04%), 9/600 (1.5%) were from Tapachula and 14/525 (2.6%) from Palenque. The seropositive women were between 20 and 35 years old, 31.8% have Premature Rapture of Membrane and 9.1% have history of perinatal death. The total percentage of positive newborns by PCR was 9/23 (39.13%), out of those 2/9 (22.2%) are from Tapachula and 7/14 (50%) from Palenque. The Maternal Fetal transmission frequency was. 2/9 (22.2%) in Tapachula and 1/14 (7.14%) in Palenque, all positive infants were asynthomatic. Conclusion: The maternal-fetal transmission rate in Chiapas State is variable; the reason could be the maternal immunological status and T. cruzi strain.
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Humanos , Masculino , Feminino , Gravidez , Recém-Nascido , Lactente , Adulto , Adulto Jovem , Complicações Infecciosas na Gravidez/epidemiologia , Doença de Chagas/transmissão , Transmissão Vertical de Doenças Infecciosas/estatística & dados numéricos , Trypanosoma cruzi/imunologia , Resultado da Gravidez , Estudos Soroepidemiológicos , Doença de Chagas/epidemiologia , Doenças Assintomáticas , Geografia Médica , México/epidemiologiaRESUMO
Introducción. El grupo Microsporidia incluye parásitos intracelulares obligados que afectan varios huéspedes. Los géneros que infectan humanos son Enterocytozoon y Encephalitozoon . Microsporidia es considerado un organismo oportunista en pacientes inmunocomprometidos e inmunocompetentes, que se ha convertido en un importante problema de salud pública. El objetivo del trabajo fue identificar Microsporidium spp en muestras fecales de pacientes con leucemia linfoblástica aguda, en diferente etapa de su tratamiento. Métodos. Se analizaron 77 muestras de niños con diagnóstico de leucemia linfoblástica aguda menores de 12 años, por análisis coproparasitoscópico de Faust. Las muestras se tiñeron con Ziehl Nielsen, tricrómica de Masson y calcoflúor. Finalmente se realizó la amplificación por PCR del gen ribosomal. Resultados. De los pacientes analizados, 16/77 (20.77%) resultaron positivos para Microsporidia, independientemente de que presentaran diarrea. El PCR fue más efectivo para la identificación que el análisis microscópico de las muestras teñidas. Conclusiones. Este trabajo enfatiza la importancia de la microsporidiosis como infección emergente en pacientes con leucemia linfoblástica aguda bajo tratamiento quimioterapéutico, incrementando las complicaciones clínicas adicionales a la leucemia.
Background. The phylum Microsporidia includes obligate intracellular parasites that affect several hosts. The most frequent genera to affect humans are Enterocytozoon and Encephalitozoon. Microsporidium is considered an opportunistic parasite in both immunocompromised and immunocompetent patients and have become an important health problem. The aim of this study was to identify Microsporidium spp in fecal samples of patients with acute lymphoblastic leukemia (ALL), with and without diarrhea, at different treatment stages. Methods. Seventy seven samples from children <12 years old with diagnosis of ALL were analyzed by the Faust coproparasitoscopic method, Ziehl-Nielsen, trichrome and calcofluor staining methods and polymerase chain reaction. Results. Results showed that 16/77 (20.77%) children presented Microsporidium and there was no relationship between microsporidial infection and diarrhea. Polymerase chain reaction was more effective than analysis by light microscopy of staining samples in the identification of the parasite. Conclusions. This work emphasizes the importance of microsporidiosis as an emerging infection in patients with ALL undergoing chemotherapy, increasing the additional clinical complications of leukemia.
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Despite its importance as a health problem issue, almost nothing is known about the membrane physiology of Giardia lamblia and practically there exist no information so far regarding the variety and properties of ion channels that this protozoan parasite possesses. To address this subject we resorted to an indirect method, consisting in the injection of mRNA and further characterization of ion currents in Xenopus oocytes. In this work, we show that oocytes injected with mRNA isolated from cultured trophozoites of G. lamblia, strain Portland-1 express novel potassium currents that appear over the second day after injection and show time- and voltage-dependent activation followed by a slow inactivation. They start activating at -90 mV, with V1/2 of -30 mV; its time constant of activation (at +60 mV) is 0.11 sec, whereas that of inactivation is 1.92 sec, V1/2 = -44.6 mV. Such K currents were effectively blocked by K channel blockers TEA and 4AP, as well as Ba(2+), quinine, quinidine, charybdotoxin, dendrotoxin-1, capsaicin, margatoxin, and diltiazem. These results suggest that such currents are the result of expression of Giardia's voltage-gated K channels heterologously expressed in Xenopus laevis oocytes.
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Four different isolates of Trichinella spp. (Z1, Z2, Z3, and Z4) obtained from the skeletal muscle of street dogs in the state of Zacatecas, Mexico were serial passaged in Wistar rats; infective larvae from the skeletal muscle of the rats were collected and frozen in liquid nitrogen. After centrifugation, DNA was extracted and the 5SRNAr and IsRNAr genes were amplified. The isolates were identified by the size of the amplified products from the 5SRNAr and IsRNAr genes (750 and 290 bp, respectively). The amplicons obtained by PCR were sequenced, aligned, and compared to the reference strain Trichinella spiralis MSUS/MEX/91//EM isolated from pigs. Based on our results, we determined that the Trichinella isolates from canine (Z1-Z4) belonged to the T. spiralis species and had 83% identity with the reference strain. The phylogenetic tree constructed from the sequences showed differences between the isolates from pig and dog. These genetic differences may be related to the immune response of the host or the pathogenicity of the isolates. Therefore, these findings have important epidemiological and public health implications.
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Doenças do Cão/parasitologia , Músculo Esquelético/parasitologia , Trichinella/genética , Triquinelose/veterinária , Animais , DNA de Helmintos/química , DNA Espaçador Ribossômico/química , Cães , Eletroforese em Gel de Ágar , México , Filogenia , RNA de Helmintos/genética , RNA Ribossômico 5S/genética , Ratos , Ratos Wistar , Alinhamento de Sequência , Trichinella/classificação , Trichinella/isolamento & purificação , Triquinelose/parasitologiaRESUMO
The assemblage of 37 Giardia intestinalis samples was determined, 19 obtained from puppy feces, 12 from stools of different human subjects under 3 years of age and 6 from axenic culture. The assemblages were classified according to the restriction pattern of beta-giardin gene with Hae III enzyme. Results showed that dog assemblages were grouped AI (52.6%) and AII (47.4%), while 41.7% human samples belonged to genotype AI and 58.3% to genotype AII. All axenic cultures belonged to assemblage AI; types AI and AII were both found in dog and human feces by Hae III restriction enzyme assay, suggesting a similarity between human and dog parasites. These results suggest that domestic animals infected with Giardia could produce cysts potentially infective for humans.
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Proteínas do Citoesqueleto/genética , Doenças do Cão/parasitologia , Giardia lamblia/classificação , Giardíase/veterinária , Proteínas de Protozoários/genética , Animais , Pré-Escolar , Desoxirribonucleases de Sítio Específico do Tipo II/metabolismo , Doenças do Cão/epidemiologia , Cães , Fezes/parasitologia , Feminino , Giardia lamblia/genética , Giardíase/parasitologia , Humanos , Lactente , Masculino , México/epidemiologiaRESUMO
Agarose gel electrophoresis of gdh gene fragments, amplified by Multiplex, was used to classify the assemblage of 24 Giardia isolates obtained from axenic cultures, children's stools, and feces of puppies from different dog breeds. Isolates were compared with seven reference strains of Giardia intestinalis. The results showed that 22/24 isolates (91%) belonged to assemblage A and could be further subclassified as assemblage A1 (18/22, 81%) and assemblage A2 (4/22, 19%). One sample revealed a mixture of A1/A2 genotypes, and another was assemblage G, indicating mixed infections by different strains in the same host, and an association with the assemblage reported in animals. The procedure described is useful to determine the Giardia genotype that parasitizes each host to conduct epidemiological studies assessing the close association between human- and animal-infecting strains and to monitor the adaptability of animal strains to humans.
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Giardia lamblia/classificação , Giardia lamblia/genética , Reação em Cadeia da Polimerase/métodos , Proteínas de Protozoários/genética , Desidrogenase do Álcool de Açúcar/genética , Animais , Criança , Pré-Escolar , Análise por Conglomerados , Doenças do Cão/parasitologia , Cães , Fezes/parasitologia , Genótipo , Giardia lamblia/isolamento & purificação , Giardíase/parasitologia , Giardíase/veterinária , Humanos , Epidemiologia Molecular , Homologia de SequênciaRESUMO
The purpose of the present paper was to analyse the association between sequences of Entamoeba histolytica and their relationship with the development of hepatic abscesses in hamsters, using a complementary DNA library for E. histolytica. From the sequences obtained, we designed oligonucleotides for amplification by PCR. Trophozoites were isolated from faeces of 11 patients in whom cysts from E. histolytica were identified, and these trophozoites were then subjected to monoaxenic culture. Then 1 x 10(5) trophozoites were inoculated into hamster liver, with three hamsters used for every culture. Sequences were obtained for ubiquitin, lectin surface precursor, replication factor MCM3 and surface antigen. The associations between sequences and hepatic abscesses were: 11/11 for ubiquitin, 9/11 for lectin precursor, 4/11 for replication factor and 1/11 for surface antigen. These results suggest that ubiquitin could be an important protein involved in the mechanism of E. histolytica invasion.
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Entamoeba histolytica/patogenicidade , Entamebíase/patologia , Abscesso Hepático Amebiano/patologia , Ubiquitina/genética , Animais , Sequência de Bases , Cricetinae , Entamoeba histolytica/genética , Entamoeba histolytica/crescimento & desenvolvimento , Entamoeba histolytica/isolamento & purificação , Entamebíase/parasitologia , Biblioteca Gênica , Humanos , Abscesso Hepático Amebiano/parasitologia , Dados de Sequência Molecular , Proteínas de Protozoários/química , Proteínas de Protozoários/genética , Ubiquitina/químicaRESUMO
The presence of human Giardia in several animals suggests a zoonotic transmission. We studied G. Intestinalis isolates obtained from: children with diarrhea (n=6), asymptomatic children (n=7), axenic cultures (n=7) and dogs (n=11). The sequence corresponding to 16 S rRNA was amplified by PCR, sequenced and compared with genotypes A, B and Dog sequences reported in the Gene Bank database. Results show that 9/20 (45%) of children isolates belonged to genotype A and 11/20 (55%) showed some variable sites, allowing classification in three arbitrary clusters: A1, A2 and A3. In addition 7/11 (63%) of dog isolates were genotype A, including those dogs that lived in the same locality as the children lived, while 4/11 (37%) belonged to an arbitrary A4 cluster living in a different locality. In this study, genotype A was associated with samples from children and dogs, and, therefore, we could infer zoonotic transmission as a way of getting the disease.
Assuntos
Doenças do Cão/parasitologia , Giardia lamblia/genética , Giardíase/parasitologia , Animais , Sequência de Bases , Criança , DNA de Protozoário/química , DNA de Protozoário/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Cães , Genes de RNAr/genética , Genótipo , Giardia lamblia/classificação , Giardia lamblia/isolamento & purificação , Giardíase/transmissão , Giardíase/veterinária , Humanos , Epidemiologia Molecular , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Alinhamento de Sequência , Análise de Sequência de DNA , ZoonosesRESUMO
The paper describes an introductory characterization of antigenic stimulation of excretion-secretion products (ESP) of Gnathostoma binucleatum advanced third-stage larvae cultured in vitro and proteinases present in this products. Excretory and secretory proteins were obtained after 10 larvae were maintained in 5% CO(2) RPMI medium. The supernatant was collected each week for two months. The proteins were dialyzed, concentrated, and separated in 10% SDS-PAGE gels under reducing conditions and transferred to nitrocellulose paper for immunoblot analyses. G. binucleatum immunized mice serum was used to determine protein antigenicity. Four proteins of 40, 80, 120, and 208 kDa persisted for two months and three proteins, 80, 120, and 208 kDa were recognized for antibodies of mice. In SDS-PAGE gelatin substrate gels ESP resolved as two proteins with molecular weight of 80 and 208 kDa that were sensitive to a metalloproteinase inhibitor, and thus it may be inferred that they might be used for diagnosis of gnathostomiasis.
Assuntos
Antígenos de Helmintos/análise , Gnathostoma/imunologia , Peptídeo Hidrolases/análise , Infecções por Spirurida/parasitologia , Animais , Anticorpos Anti-Helmínticos/imunologia , Antígenos de Helmintos/química , Antígenos de Helmintos/imunologia , Western Blotting , Cães , Eletroforese em Gel de Poliacrilamida , Feminino , Gnathostoma/enzimologia , Humanos , Soros Imunes/imunologia , Larva/enzimologia , Larva/imunologia , Camundongos , Peso Molecular , Peptídeo Hidrolases/química , Infecções por Spirurida/imunologiaRESUMO
Resistance is a practical problem associated to the use of benzimidazoles in the antigiardial therapy. Since benzimidazole-resistant strains of fungi have shown increased sensitivity to phenylcarbamates, in this study we synthesized and in vitro tested novel substituted phenylcarbamates against the protozoa Giardia intestinalis. IRE-6A and IRE-7B, two 4-R-ethyl-phenylcarbamates demonstrated an important antigiardial activity although that was modest when compared to albendazole in axenic cultures of Giardia intestinalis. Results of this study suggest a potential role of phenylcarbamates as alternative to benzimidazoles in the therapy of giardiasis.