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Abstract
Human hairs have been known to be easily contaminated with microorganisms. This study was performed in order to measure what bacterial species and how much microorganisms contaminate human hairs in specific place. Virgin human hairs were left at 6 positions in inside corner and beside window in a laboratory for 7 days. The number of viable bacterial cells, which were determined by most probable number method, contaminating the human hairs was measured at a maximum of 106/g hair and a minimum of 103/g hair in inside corner and maximum of 106/g hair and a minimum of 103/g hair beside window. The bacterial cells-contaminating human hairs were observed via fluorescence light microscopy after 4′,6-diamino-2-phenylindole (DAPI) staining. The bacterial community contaminating human hairs was analyzed via the thermal gradient gel electrophoresis (TGGE) technique, based on the diversity of the 16S-rDNA variable region. In total, approximately 20 bacterial species were detected from 12 groups of hair samples. In this study, general experimental methods-fluorescence staining, TGGE and MPN-were combined to develop new method for observation and estimation of bacteria contaminating human hairs.
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Figure 1.
Fixation of hair bundle in an opened square box, by which human hairs can be naturally moved and contact with air (A). Six of them were placed in inside corner and another six were placed beside window (B).
Figure 2.
Hair contaminated with various fungal hyphae and conidia (A, B and C), as observed under bright-field light microscopy (× 400). Bacterial cells at grooves on the hair surfaces were not noted, but round spots were seen on the hair cuticles (D); however, these stains cannot be conclusively identified as fungal cells.
Figure 4.
Microscopic image of human hair surface contaminated with microorganisms. Bacterial cells (A), fungal (yeast) cells (B, arrow marks) and communities of microorganisms (C) made fluorescent by DAPI staining.
Figure 5.
TGGE patterns for bacterial communities contaminating the surfaces of human hairs left in inside corners (lane 1 to 6) and beside windows (lane 7 to 12) for 7 days. Sequencing of amplicons generated from DNAs extracted from the numbered bands showed more than 90% identity with the followings: 1. Uncultured bacterium (GQ111303); 2. Janthinobacterium sp. (FJ605429); 3. Bacillus sp. (FJ941090); 4, 25. Stenotrophomonas sp. (FJ897489); 5, 27. Erwinia sp. (FJ816023); 6. Bacillus sp. (AM292066); 7. Oxalobacter sp. (AV367035); 8, 26. Naxibacter sp. (FJ621304); 9, 22. Deinococcus sp. (DQ223543); 10. Pseudomonas sp. (AB461633); 11. Uncultured soil bacterium FACE.R5 (FJ621196); 12. Uncultured Stenotrophomonas sp. (FN428834); 13. Bacillus sp. (FJ981907); 14. Uncultured soil bacterium Kei21 (FJ184368); 15. uncultured Comamonas sp. (AM711890); 16. Uncultured bacterium clone (GQ101918); 17, 30. Pantoea ananat (AF491932); 18. Pantoea eucalypti (GQ169378); 19. Uncultured bacterium sp. (GQ114033); 20, 29. Endophytic bacterium C01 (FJ205678); 21. Uncultured bacterium sp. (FQ111303); 23. Uncultured bacterium (EU438875); 24. Uncultured Actinobacterium sp. (EU810962).
Table 1.
Most probable number (MPN) of bacteria contaminated on hair surface. Hairs were left in inside corner or beside windows in a laboratory for 7 days
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