ABSTRACT
This study aimed to describe the association of Borrelia burgdorferi s.s. with ixodid tick cell lines by flow cytometry and fluorescence and confocal microscopy. Spirochetes were stained with a fluorescent membrane marker (PKH67 or PKH26), inoculated into 8 different tick cell lines and incubated at 30°C for 24 h. PKH efficiently stained B. burgdorferi without affecting bacterial viability or motility. Among the tick cell lines tested, the Rhipicephalus appendiculatus cell line RA243 achieved the highest percentage of association/internalization, with both high (90%) and low (10%) concentrations of BSK-H medium in tick cell culture medium. Treatment with cytochalasin D dramatically reduced the average percentage of cells with internalized spirochetes, which passed through a dramatic morphological change during their internalization by the host cell as observed in time-lapse photography. Almost all of the fluorescent bacteria were seen to be inside the tick cells. PKH labeling of borreliae proved to be a reliable and valuable tool to analyze the association of spirochetes with host cells by flow cytometry, confocal and fluorescence microscopy.
Subject(s)
Animals , Borrelia burgdorferi , Staining and Labeling/methods , Ticks/cytology , Ticks/microbiology , Borrelia burgdorferi/isolation & purification , Cell Line , Cells, Cultured , Culture Media , Flow Cytometry/methods , Fluorescent Dyes , Microscopy, Confocal/methods , Organic Chemicals , Phagocytosis , Reproducibility of Results , Spirochaetales/isolation & purification , Tick-Borne Diseases/microbiology , Time FactorsABSTRACT
PURPOSE: To evaluate the nutritious state in children of low social-economic class in order to look over a possible corelationship among this status and the infections caused by enteroparasites. METHODS: 103 children were submitted to a nutritions evolution and to accomplish a exam of serial samples. The method employed searched protozoan cystos, helmints eggs and larvae and it counted helmint eggs when presented through 4 different methods of diagnosis. RESULTS: The results obtained show that the poor environmental and social-economic conditions helped create a high infectious frequency caused by enteroparasites, mainly by Trichuris trichiura and Ascaris lumbricoides, among the helminthes, and Endolimax nana and Giardia lamblia, among the protozoans. light malnutrition without protein deficit was found in 93.55% and moderate malnutrition in 6.45% of the children malnutrition (30.7%). CONCLUSION: Then, it is possible to suggest that children besides bearing many parasitosis were not weth their nutritional state deeply compromised.