RESUMO
Enteroparasites in children from three marginal urban districts of Trujillo (Peru) were studied to treat these children and to design a prevention and control programme. A total of 845 children were examined. The general prevalence of enteroparasites was of 66.3%, and 45.6% were multiparasitized. The pathogenic enteroparasite prevalence were 23.8% (Giardia lamblia), 4.6% (Iodamoeba buschlii), 2.6% (Cyclospora cayetanensis), 2.2% (Hymenolepis nana), and 2% (Cryptosporidium spp.). G. lamblia was the most frequent parasite both in diarrheic children (28.1%) as well as in nondiarrheic ones (19.5%). The G. lamblia genotypes were molecularly characterized by sequence analysis of the glutamate dehydrogenase (gdh) gene using PCR and RFLP. Sequence analysis revealed both Assemblage A (AI and AII) and Assemblage B (BIV), with the predominance of Assemblage AI. All the samples with Assemblage A were diarrheic but not those with Assemblage B. This is the first study of molecular characterization of G. lamblia in Peruvian children and confirms the importance of asymptomatic patients in the transmission of the giardiosis, especially in places with poor hygiene and sanitation.
Assuntos
Giardia lamblia/isolamento & purificação , Giardíase/epidemiologia , Enteropatias Parasitárias/epidemiologia , Enteropatias Parasitárias/parasitologia , Amoeba/isolamento & purificação , Animais , Criança , Pré-Escolar , Cyclospora/isolamento & purificação , DNA de Protozoário/análise , DNA de Protozoário/isolamento & purificação , Diarreia/epidemiologia , Diarreia/parasitologia , Genótipo , Giardia lamblia/classificação , Giardia lamblia/enzimologia , Giardia lamblia/genética , Giardíase/parasitologia , Glutamato Desidrogenase/genética , Humanos , Hymenolepis/isolamento & purificação , Enteropatias Parasitárias/fisiopatologia , Peru/epidemiologia , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Prevalência , População UrbanaRESUMO
The great difficulties in treating people and animals suffering from cryptosporidiosis have prompted the development of in vitro experimental models. Due to the models of in vitro culture, new extracellular stages of Cryptosporidium have been demonstrated. The development of these extracellular phases depends on the technique of in vitro culture and on the species and genotype of Cryptosporidium used. Here, we undertake the molecular characterization by polymerase chain reaction-restriction fragment length polymorphism of different Cryptosporidium isolates from calves, concluding that all are C. parvum of cattle genotype, although differing in the nucleotide at positions 472 and 498. Using these parasites, modified the in vitro culture technique for HCT-8 cells achieving greater multiplication of parasites. The HCT-8 cell cultures, for which the culture had not been renewed in seven days, were infected with C. parvum sporozoites in RPMI-1640 medium with 10% IFBS, CaCl2 and MgCl2 1 mM at pH 7.2. Percentages of cell parasitism were increased with respect to control cultures (71% at 48 h vs 14.5%), even after two weeks (47% vs 1.9%). Also, the percentage of extracellular stages augmented (25.3% vs 1.1% at 96 h). This new model of in vitro culture of C. parvum will enable easier study of the developmental phases of C. parvum in performing new chemotherapeutic assays.
Assuntos
Cryptosporidium/crescimento & desenvolvimento , Estágios do Ciclo de Vida/fisiologia , Animais , Sequência de Bases , Bovinos , Linhagem Celular Tumoral , Cryptosporidium/classificação , Cryptosporidium/genética , DNA de Protozoário/genética , DNA Ribossômico/genética , Genótipo , Estágios do Ciclo de Vida/genética , Masculino , Camundongos , Dados de Sequência Molecular , Oocistos/crescimento & desenvolvimento , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , RNA Ribossômico 18S/genética , Fatores de TempoRESUMO
The great difficulties in treating people and animals suffering from cryptosporidiosis have prompted the development of in vitro experimental models. Due to the models of in vitro culture, new extracellular stages of Cryptosporidium have been demonstrated. The development of these extracellular phases depends on the technique of in vitro culture and on the species and genotype of Cryptosporidium used. Here, we undertake the molecular characterization by polymerase chain reaction-restriction fragment lenght polymorphism of different Cryptosporidium isolates from calves, concluding that all are C. parvum of cattle genotype, although differing in the nucleotide at positions 472 and 498. Using these parasites, modified the in vitro culture technique for HCT-8 cells achieving greater multiplication of parasites. The HCT-8 cell cultures, for which the culture had not been renewed in seven days, were infected with C. parvum sporozoites in RPMI-1640 medium with 10 percent IFBS, CaCl2 and MgCl2 1 mM at pH 7.2. Percentages of cell parasitism were increased with respect to control cultures (71 percent at 48 h vs 14.5 percent), even after two weeks (47 percent vs 1.9 percent). Also, the percentage of extracellular stages augmented (25.3 percent vs 1.1 percent at 96 h). This new model of in vitro culture of C. parvum will enable easier study of the developmental phases of C. parvum in performing new chemotherapeutic assays.
Assuntos
Animais , Bovinos , Masculino , Camundongos , Cryptosporidium/crescimento & desenvolvimento , Estágios do Ciclo de Vida/fisiologia , Sequência de Bases , Linhagem Celular Tumoral , Cryptosporidium/classificação , Cryptosporidium/genética , DNA de Protozoário/genética , DNA Ribossômico/genética , Genótipo , Estágios do Ciclo de Vida/genética , Dados de Sequência Molecular , Oocistos/crescimento & desenvolvimento , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , /genética , Fatores de TempoRESUMO
Intestinal parasitism was studied in children of Trujillo (Peru) to create a prevention and control program. Fecal samples of 489 children were examined. The general prevalence of intestinal parasitosis was found to be 68%. The most frequent pathogenic enteroparasites were Giardia lamblia (26.4%), Cyclospora cayetanensis (13%), Hymenolepis nana (2%), Hymenolepis diminuta (1.6%), and Cryptosporidium spp. (1%). All these parasites appeared both in diarrheic and nondiarrheic children, except Cryptosporidium, which invariably caused diarrhea. Multiple parasitism was frequent, 45.6% of the children presenting two, three, or four intestinal parasites. Cryptosporidium was the only parasite that was not associated with the others. Only five children were affected of cryptosporidiosis, presenting explosive diarrhea, nausea, and vomiting. Cryptosporidium species and genotypes involved in the infantile cryptosporidiosis were determined by polymerase chain reaction-restriction fragment length polymorphism. Four children were parasitized by Cryptosporidium hominis and only one by Cryptosporidium parvum. Our results confirm that anthroponotic transmission of Cryptosporidium is predominant in Peru.