Intestinal and haematic parasitism in the birds of the Almuñecar (Granada, Spain) ornithological garden.

Birds from the Almuñecar ornithological garden (Granada, Spain) were surveyed from June 2006 to May 2007 to establish programmes to prevent, control, and treat intestinal and haematic parasites. A total of 984 faecal samples and 41 samples of blood were collected from Psittacidae, Cacatuidae, Phasianidae, and Anatidae. One or more intestinal parasites were identified in 51.6% of the samples. Blood parasites were found in 26.8% of the birds examined. The most frequent pathogenic endoparasites were coccidians, such as Cyclospora sp. (4.5%), Eimeria sp. (4.1%) and Isospora sp. (2%) and helminths such as Capillaria sp. (10. 1%), Ascaridia sp. (4.9%) and Heterakis gallinarum (4.9%). All the parasites varied with season but the most were found year round. Multiple parasitic infections by intestinal parasites were common, with 196 of 984 faecal samples having 2-5 intestinal parasites. The most frequent cases of multiple parasitism were Blastocystis plus Entamoeba sp. and Blastocystis plus Cyclospora sp. The haematic protozoa detected were Haemoproteus sp. (17%) and Plasmodium sp. (7.3%). Multiple parasitism by Haemoproteus sp. and Plasmodium sp. was detected in 1 sample of Gallus gallus. After each sampling, some of the affected animals were treated according to our results, and the corresponding programmes of prevention and control were designed.

Prevalence of enteroparasites and genotyping of Giardia lamblia in Peruvian children.

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.

More productive in vitro culture of Cryptosporidium parvum for better study of the intra- and extracellular phases.

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.

More productive in vitro culture of Cryptosporidium parvum for better study of the intra- and extracellular phases

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.

Intestinal parasitism in Peruvian children and molecular characterization of Cryptosporidium species.

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.