Microbiological examinations were performed with use of classic methods employed in microbiological diagnostics. Material collected from the patients was cultured on proper growth media for proliferation and subsequent isolation of pure cultures. Aerobic bacteria were proliferated on Columbia agar solid medium with 5% sheep blood at 37°C. Anaerobic bacteria were proliferated on Schaedler K3 solid medium with 5% sheep blood at 37°C in anaerobic conditions obtained with Biomerieux GENbag anaer generators (Marcy-l'Étoile, France). Species identification was performed after isolation and proliferation of cultured microorganism strains with use of the following reagent sets: ENTEROtest 24N, NEFERMtest 24N, STREPTOtest 24, STAPHYtest, ANAEROtest 23, OXItest, PYRAtest, and TNW_lite 6.5 computer program for species identification of microorganisms (Erba-Lachema, Brno, Czech Republic). Also the following Biomerieux (Marcyl'Etoile, France) biochemical tests were used: Katalaza, Slidex Staph Kit, and API Candida. Performance and interpretation of results of the tests was carried out according to the manufacturer's recommendations with diagnostic reagent sets.
Api candida
Manufactured by bioMérieux
Sourced in France
The API Candida is a laboratory product developed by bioMérieux. It is designed to identify and differentiate between various Candida species. The product provides a standardized identification method for clinical microbiology laboratories.
Automatically generated - may contain errors
Lab products found in correlation
Slidex staph kit, by bioMérieux (3 mentions)
Columbia agar, by bioMérieux (3 mentions)
Chromid candida, by bioMérieux (2 mentions)
Genbag anaer kits, by bioMérieux (2 mentions)
Schaedler k3 agar, by bioMérieux (2 mentions)
Sabouraud agar, by bioMérieux (2 mentions)
Genbag anaer, by bioMérieux (2 mentions)
Api 20ne, by bioMérieux (1 mentions)
Api 20e, by bioMérieux (1 mentions)
Chromid candida agar, by bioMérieux (1 mentions)
9 protocols using api candida
1
Comprehensive Microbial Identification Protocols
2
Microbiological Identification Protocol
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Microbiological tests were carried out using classical methods routinely used in microbiological diagnostics. The material collected from the examined patients was cultured on appropriate culture media to multiply and then isolate pure microbial cultures. Aerobic bacteria were multiplied on solid medium Columbia agar with a 5% addition of sheep blood at 37°C. Anaerobic bacteria were multiplied on Schaedler K3 solid medium with a 5% addition of sheep blood at 37°C in an-aerobic conditions obtained using Biomerieux Genbag Anaer kits (Marcy l’Etoile, France). Candida fungi were multiplied and initially identified using chromogenic medium ChromID Candida from Biomerieux (Marcy l’Etoile, France).
After isolation and multiplication of the cultured strains of microorganisms, their species identification was carried out using the following sets of reagents: ENTEROtest 24 N, NEFERMtest 24 N, STREPTOtest 24, STAPHYtest 24, ANAEROtest 23, OXItest, PYRAtest, and the computer program TNW_lite 6.5 for species identification of Erba-Lachema microorganisms (Brno, Czech Republic). The following biochemical tests from Biomerieux (Marcy l’Etoile, France) were also used: Catalysis, Slidex Staph Kit, and API Candida. Execution, reading, and interpretation of the test results were performed in accordance with the recommendations of manufacturers of diagnostic reagent kits.
After isolation and multiplication of the cultured strains of microorganisms, their species identification was carried out using the following sets of reagents: ENTEROtest 24 N, NEFERMtest 24 N, STREPTOtest 24, STAPHYtest 24, ANAEROtest 23, OXItest, PYRAtest, and the computer program TNW_lite 6.5 for species identification of Erba-Lachema microorganisms (Brno, Czech Republic). The following biochemical tests from Biomerieux (Marcy l’Etoile, France) were also used: Catalysis, Slidex Staph Kit, and API Candida. Execution, reading, and interpretation of the test results were performed in accordance with the recommendations of manufacturers of diagnostic reagent kits.
3
Comprehensive Oral Microbial Profiling
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Oral microbiological examination was performed by taking a swab from the bottom of the mouth, using sterile swabs, in tubes with AMIES transport medium, with charcoal (DELTALAB, Rubi, Spain). The following media from Biomerieux (Marcy l’Etoile, France) were used to cultivate microorganisms: Columbia agar, Schaedler K3, ChromID Candida and Genbag anaer kits for the cultivation of anaerobic bacteria. Species identification of the cultured microorganisms was carried out using the following sets of reagents: ENTEROtest 24N, NEFERMtest 24N, STREPTOtest 24, STAPHYtest, ANAEROtest 23, OXItest, PYRAtest and the computer program TNW lite 6.5 for the identification of microorganisms by Erba-Lachema (Brno, Czech Republic). Biochemical tests by Biomerieux (Mercy l’ Etoile, France) were also used: Katalase, Slidex Staph Kit, and API Candida.
4
Candida Infection Assessment Protocol
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I. An assessment of their clinical history: Previous urologic problems, operation, terminal haematuria, dysuria and taking of antibiotics, or any other medications.
II. Clinical and radiological examination; III. Laboratory investigations of urine samples: Including Mid-stream urine samples or catheterized urine samples were obtained under aseptic conditions in sterile screw-capped wide mouth containers. Samples were subjected to microscopic examination and culture. Identification of Candida species by Gram-stain, Germ Tube test, subculture on chromID™ Candida Agar (CAN2): (bioMérieux) and biochemically by API Candida (bioMérieux).
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5
Microbiological Testing of Clinical Samples
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Microbiological tests were performed by the Department of Microbiology and Immunology in Zabrze of the Medical University of Silesia in Katowice. The samples were inoculated on suitable culture media (Columbia agar, Schaedler K3 agar, and Sabouraud agar) from bioMerieux (Marcy l'Etoile, France). Aerobic bacteria were propagated on Columbia agar solid medium with 5% sheep blood at 37°C. Anaerobic bacteria were propagated on Schaedler K3 solid medium with 5% sheep blood at 37°C under anaerobic conditions using a GENbag Anaer (bioMerieux, Marcy l'Etoile, France). Candida fungi were propagated on selective Sabouraud agar solid medium at 35°C under aerobic conditions. Upon isolation and further culture of each microorganism, their species were identified by the following tests: Api 20 E, Api 20 NE, and Api Candida (bioMerieux, Marcy l'Etoile, France) and ENTEROtest 24 N, NEFERMtest 24 N, STREPTOtest 24, STAPHYtest 24, and ANAEROtest 23 (Erba-Lachema, Brno, Czech Republic).
The data from individual patients were treated as confidential and were not identifiable in any documentation that emerged in relation to the examination. The study represented a separate part of the main research project at the Medical University of Silesia supported by the Grant KNW-2-102/10 and was performed following the guidelines of the Declaration of Helsinki.
The data from individual patients were treated as confidential and were not identifiable in any documentation that emerged in relation to the examination. The study represented a separate part of the main research project at the Medical University of Silesia supported by the Grant KNW-2-102/10 and was performed following the guidelines of the Declaration of Helsinki.
6
Microbiological Identification of Aerobic, Anaerobic, and Fungal Isolates
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Microbiological investigations were performed in the Department of Microbiology and Immunology in Zabrze, Medical University of Silesia in Katowice. The material for microbiological testing was inoculated on suitable culture media (Columbia agar, Schaedler K3 agar, and Sabouraud agar) from Biomerieux (Marcy-l'Etoile, France). Aerobic bacteria were propagated on Columbia agar medium with 5% sheep blood at 37°C. Anaerobic bacteria were propagated on Schaedler K3 medium with 5% sheep blood at 37°C in anaerobic conditions using Genbag anaer (Biomerieux, Marcy-l'Etoile, France). Candida fungi were propagated on selective Sabouraud agar medium at 35°C in aerobic conditions. Upon isolation and further culture of each microorganism, their species were identified with the help of the following reagent sets: ENTEROtest 24N, NEFERMtest 24N, STREPTOtest 24, STAPHYtest 24, ANAEROtest 23 (Erba-Lachema, Brno, Czech Republic), and Api Candida (Biomerieux, Marcy-l'Etoile, France).
7
Candida Identification Protocol
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All vaginal swabs were streaked onto Sabouraud Dextrose Agar (SDA) supplemented with chloramphenicol (REF 610103 Liofilchem R© srl Italy) and incubated at 37 °C/24–48 h. Once cultures positives, the strains were purified and identified using chromatophilic medium ChromID® Candida Agar (REF 43 639, BioMérieux, Marcy l’Etoile, France), API® Candida (REF 10 500, BioMérieux, Marcy l’Etoile, France), Apiweb Standalone version 1.3.2. and the Germ tube test [10 (link), 11 (link)].
8
Microscopic Detection of Fungal BLOs
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This study was approved by the ethics committee of the University of Concepción, under number E-224-18. Prior to the sample collection, each participant received information, oral and written, about the importance of Detection of bacterium-like organelles (BLOs) on the inside of fungal cells by optical microscopy.
We carried out a wet examination in yeast colonies randomly chosen in search of BLOs inside the fungal cells, using as a negative control C. albicans ATCC 90028. Each colony was placed in 20 µL of saline solution and observed using an optical microscope, with a 100X objective lens.
Tests for the identification of Candida spp. We took an inoculum from the crops of pure yeast in Sabouraud Agar, which was then sown on CHRO-Magar Candida (Difco, USA) and incubated under aerobic conditions at 37°C for 24-48 hours. The interpretation was performed according to the manufacturer's instructions. Subsequently, the results were confirmed using the identification system API®Candida, following the manufacturer's recommendations (BIOMÉRIEUX, France).
We carried out a wet examination in yeast colonies randomly chosen in search of BLOs inside the fungal cells, using as a negative control C. albicans ATCC 90028. Each colony was placed in 20 µL of saline solution and observed using an optical microscope, with a 100X objective lens.
Tests for the identification of Candida spp. We took an inoculum from the crops of pure yeast in Sabouraud Agar, which was then sown on CHRO-Magar Candida (Difco, USA) and incubated under aerobic conditions at 37°C for 24-48 hours. The interpretation was performed according to the manufacturer's instructions. Subsequently, the results were confirmed using the identification system API®Candida, following the manufacturer's recommendations (BIOMÉRIEUX, France).
9
Diagnosing Vulvovaginal Candidiasis
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The case definition of VVC included the presence of symptoms, the demonstration of blastospores and pseudohyphae in a wet vaginal smear that had been treated with 10% potassium hydroxide, and a positive fungal culture. All strains were identified using the API Candida (bioMérieux, Marcy l'Etoile, France).
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