c.i.-fluorescent-brightening-agent-28 and Diarrhea

Microsporidia, obligate intracellular parasites, were first defined by Nageli in 1857. Microsporidia phylum consists of 200 genus and 1500 species. They have a wide host spectrum including insects, fish, and mammals. It has been shown that they may also infect humans and may be existed both in symptomatic and asymptomatic forms. There are eight species infecting humans, which include Anncaliia (Brachiola, Nosema), Encephalitozoon, Entrocytozoon, Microsporidium, Nosema, Pleistophora, Trachipleistophor, and Vittaforma. The species most commonly infect humans are Encephalitozoon intestinalis and Enterocytozoon bieneusi. The aim of this study was to determine the prevalence of Microsporidia by using two different chemiluminescence stains, namely uvitex 2B and calcoflour and detect species by molecular analysis in diarrheal patients. For this purpose, we studied stool samples of 200 patients with diarrhea sent to Gazi University Health Practice and Research Hospital, Microbiology Laboratory and Ankara Numune Training and Research Hospital Microbiology Laboratory between 2012-2013. The stool samples were stained with chemiluminescent stains uvitex 2B and calcoflour methods; the Microsporidia prevalence was found to be 38% (77/200) by fluorescent microscopic examination. Statistically an excellent consistency was found between the chemiluminescent stains uvitex 2B and calcoflour (Cohen's kappa= 0.881). A statistical analysis for the consistency of uvitex 2B and calcoflour in terms of numerical density (low, high) and luminescence of spores (dim, bright) showed a moderate consistency between the two stains with respect to determining numerical density of spores (Cohen's kappa= 0.354), while there was no consistency in terms of luminescence of spores (Cohen's Kappa= 0.001). No significant difference was found between the Microsporidia prevalence with respect to age group or clinics (p > 0.05). A sex-based analysis showed that Microsporidia prevalence was more common in women (50%) than men (30.8%) (p< 0.05). In 77 samples that were detected positive for Microsporidia with uvitex 2B and calcoflour stains determination of genus and species level were done by using multiplex nested polymerase chain reaction (PCR) method. With this technique, seven (9.1%) of 77 isolates were detected as E.bieneusi, and 70 (90.9%) as Encephalitozoon spp. When the Microsporidia genus was considered, the Microsporidia prevalence did not show differences with respect to age, sex, and refe

Topics: Animals; Benzenesulfonates; Diarrhea; Encephalitozoon; Feces; Female; Genes, Fungal; Humans; Luminescence; Male; Microsporidia; Microsporidiosis; Polymerase Chain Reaction; Prevalence; Staining and Labeling

To evaluate three fluorescent chitin stains for detecting microsporidia spores in specimens from acquired immunodeficiency syndrome (AIDS) patients with chronic diarrhea.. We compared the Fungifluor, Calcofluor White, and Fungiqual A fluorochrome stains for identifying Enterocytozoon bieneusi and Septata intestinalis spores in stool, intestinal fluid, biopsy imprints, and paraffin biopsy sections. The modified chromotrope trichrome stain was used as the standard light microscopic technique for stool and fluid specimens. Stained and unstained paraffin sections and fluid preparations were also evaluated. Multiple specimens from 50 consecutive symptomatic AIDS patients and archival material from known microsporidia-positive AIDS patients were analyzed.. Spores of E bieneusi and S intestinalis fluoresce brightly with all three fluorochrome stains in all of the types of diagnostic specimens. Fluorescing debris and the much larger fungal forms were readily distinguished. Spores were equally well detected in unfixed and formalin-fixed stool specimens, but were not as well detected after sodium acetate-acetic acid, polyvinyl acetate, and ethanol fixation. Bouin's tissue fixative gave a higher background staining than formalin. Spores were readily detected in archival paraffin sections and stool preparations, even when the specimens had been stained previously. Repeat fluorochrome staining was possible. The methods also could detect extraintestinal parasites in paraffin sections.. The three fluorescent chitin stains are sensitive and rapid methods for detecting microsporidia spores in stool, intestinal fluid, biopsy imprint, and tissue specimens, even from archived material.

Topics: Acquired Immunodeficiency Syndrome; Animals; Benzenesulfonates; Biopsy; Body Fluids; Chitin; Diarrhea; Feces; Fixatives; Fluorescent Dyes; Humans; Intestines; Microscopy, Fluorescence; Microsporida; Microsporidiosis; Pilot Projects; Stilbenes; Triazines