INTRAGASTRIC INFECTION OF CONVENTIONAL AND GERM-FREE MICE WITH GIARDIA-LAMBLIA

The effects of experimental infection with Giardia lamblia were studied in 30-day old conventional and germfree CFW mice (7 animals in each group) of both sexes. Cysts were observed in the feces of both groups 6 to 7 days after intragastric infection of each animal with about 2.5 x 10(5) G. lamblia trophozoites. Fecal cyst level was statistically higher in germfree mice (about 10(5) cysts/g feces) when compared with the conventional group (about 10(4) cysts/g feces). The peak of infection in the conventional group apparently occurred on the 10th day after infection as indicated by an increase of fecal weight and by histopathological examination. Intense infiltration of the lamina propria and high reactional hyperplasia of the lymphoid component were observed in the conventional group. There was no infiltration or hyperplasia in germfree infected mice and fecal weight was relatively constant throughout the experiment. These results suggest that, as is the case for other intestinal pathogenic protozoa, the intestinal microflora is indispensable for the expression of the pathogenicity but not for the multiplication of G. lamblia.

EFFECTS OF PROTEIN-DEFICIENCY AND NATURAL INTESTINAL INFECTION WITH GIARDIA-LAMBLIA ON JEJUNAL INTRAEPITHELIAL LYMPHOCYTES IN RATS OF DIFFERENT AGES

1. The interaction between experimental protein deprivation and natural intestinal infection by Giardia lamblia was studied in terms of its effects on the intraepithelial lymphocyte (IEL) population and morphology of the jejunal mucosa of rats of different ages.2. Young, adult and old male Wistar rats received a protein-deficient diet (2% casein) or a control diet (20% casein) for 42 days. Mucosal height and the number of lymphocytes located among 500 consecutive epithelial cells (EC) along the villi or crossing the basement membrane were determined in PAS-stained jejunal fragments.3. The number of IEL increased progressively with animal age, from 14 to 25 per 100 epithelial cells, with significant differences between age ranges. However, the number of IEL did not differ between control and protein-deficient rats in any of the age groups. The proportion of lymphocytes crossing the basement membrane was approximately two-fold greater in young (2.8/100 EC) and adult (5.8/100 EC) protein-deficient animals than in their respective controls (1.6 and 2.8/100 EC). The intensity of parasite colonization was moderate, from 3 to 5/100 EC and did not differ between groups. The pattern of morphologic changes of jejunal mucosa in protozoal infection did not differ between control and protein-deficient animals in any of the three age groups.4. We conclude that intestinal infection with Giardia lamblia probably stimulated the local immune response, masking the reduction of the IEL population induced by protein deficiency. The increase in lymphocyte numbers with age may be related to prolonged antigenic stimulation promoted by infection.

Protease activity in extracellular products secreted in vitro by trophozoites of Giardia duodenalis

There are evidences that Giardia trophozoites contain and/or release proteolytic enzymes that may be implicated in pathogenesis of giardiasis. This report describes a preliminary characterization of the proteolytic activity in excretory/secretory (E/S) products of Giardia duodenalis trophozoites of an axenic Brazilian strain (BTU-11) and the reference strain Portland 1 (P1). The protease activity of E/S products in conditioned medium by trophozoites of each strain was analyzed using substrate (gelatin and collagen) impregnated SDS-PAGE and hemoglobin assay. The protease characterization was based on inhibition assays including synthetic inhibitors. Proteolytic products were detected in the conditioned medium by trophozoites of both assayed strains. In the gels containing copolymerized gelatin and collagen, E/S products promoted degradation of the substrates and the most evident proteolysis zones were distributed in the migration regions of 77 to 18 kDa and 145 to 18 kDa, respectively, in the patterns of gelatinolytic and collagenolytic activities. Degradation of hemoglobin was also observed, and the pattern of hydrolysis was similar in both E/S products assayed. Inhibition assays showed that the main proteolytic activity in both E/S products is due to cysteine proteases although the presence of serine proteases was also indicated, mainly in the hydrolysis of hemoglobin.

Uptake of bacteria by trophozoites of Giardia duodenalis (Say)

Rod-like bacteria were found in the cytoplasm of trophozoites of Giardia duodenalis (Say) in domestic rats (Rattus rattus). These structures were always in phagocytic vacuoles without signs of bacteria digestion or degradation of the trophozoite cytoplasm. The uptake of the bacteria was observed from their attachment to the trophozoite membrane until their total incorporation by phagocytosis.

Invited review Giardia duodenalis: Inter-strain variability of proteins, antigens, proteases, isoenzymes and nucleic acids

Giardia duodenalis isolates from asymptomatic or symptomatic patients and from animals present similarities and differences in the protein composition, antigenic profile, pattern of proteases and isoenzymes, as well as in nucleic acids analysis. In the present overview, these differences and similarities are reviewed with emphasis in the host-parasite interplay and possible mechanisms of virulence of the protozoon.

Avaliação de diferentes métodos no diagnóstico laboratorial de Giardia lamblia

The protozoan parasite Giardia lamblia is responsible for a common intestinal infection in all regions of the world. In this study, four laboratory tests were evaluated for diagnostic reproducibility in the detection of this infection: Coprotest®, Direct modified method, Faust method and iron-hematoxylin staining. Positive diagnoses were tested for association with factors such as the age group and gender of the subject and the month when the sample was taken. Feces of 200 children in the Araraquara region (SP, Brazil) were examined by all four methods and the results compared. G. lamblia was found to be the most frequent parasite and 8.0% of the children showed giardiasis. There was no apparent correlation with gender, but most of the parasites were found in three- to five-year-olds. The highest frequency of infection occurred in January. The most reliable diagnostic results for G. lamblia were achieved by combining at least two methods of good reproducibility, i.e. Coprotest-Faust, Direct-Faust or Coprotest-Direct (A > 0.81).

Comparação de quatro métodos laboratoriais para o diagnóstico da Giardia lamblia em fezes de crianças da região de Araraquara-SP

Pós-graduação em Biociências e Biotecnologia Aplicadas à Farmácia - FCFAR

Prevalência de parasitas intestinais e caracterização genotípica de Giardia duodenalis em creche do município de Pratânia, estado de São Paulo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Identificação e caracterização dos produtos de excreção/secreção de trofozóitos de Giardia duodenalis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diagnóstico e caracterização molecular de Giardia duodenalis e Cryptosporidium spp. em amostras fecais de bovinos e ovinos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ocorrência de Criptosporidium spp. (Tyzzer, 1907) e Giardia spp. (Kunstler, 1882) em leitões ao desmame

Pós-graduação em Ciência Animal - FMVA

Estudo clássico e molecular de Giardia lamblia isolada de uma população infantil da região de Presidente Prudente-SP/Brasil

Pós-graduação em Biociências e Biotecnologia Aplicadas à Farmácia - FCFAR

Efeito in vitro de inibidores de proteases sobre trofozoítos de Giardia duodenalis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)