The objective of this study was to perform a preliminary characterization of the microbial populations of the normal human vulva. Genomic DNA was isolated from samples of the labia majora and labia minora from four healthy women, and sequences of bacterial 16S rRNA genes in each were determined. The sequences were compared with those of known bacterial species to classify the numerically abundant populations in these communities. Even among this limited number of individuals, the microbiota of the human vulva was found to be quite diverse. Each woman had a distinctive microbiota and no single species was common to all women. The microbiota of the labia majora and labia minora differed, although both had appreciable numbers of lactobacilli and strict anaerobes. A greater diversity of populations inhabited the labia majora compared with the labia minora. The results indicated that the microbiota of the vulva includes populations known to be commensals of the microbiota of the skin, colon and vagina, and is much more complex than previously thought, suggesting that more extensive investigations are warranted. INTRODUCTIONThe human vulva consists of several distinct ecotopes that are defined by their physical and chemical characteristics. The physical characteristics include distinct histological architectures and anatomical proximity to the vagina, urethra, anus and Bartholin's gland, whilst the chemical factors include the composition and amount of vaginal secretions, as well as contamination by urine and faecal material. As a result, the microbiota of the human vulva is complex and unique (Elsner & Maibach, 1990). Cultivationdependent methods have been used previously to study the microbiology of the labia majora. The microbiota of the labia majora is characterized by micro-organisms that are either related to vaginal and urethral inhabitants (such as Lactobacillus spp.) or are common for intertriginous skin, including Gram-negative rods, non-pathogenic Neisseria, Gardnerella vaginalis and Staphylococcus aureus (Aly et al., 1979;Elsner & Maibach, 1990). Technical difficulties in the isolation of bacteria have confounded efforts to obtain a comprehensive understanding of the organisms present in various anatomical locations on the human body (Berg, 1999;Henderson et al., 1998; Hooper et al., 2001;Pace, 1997;Rappe & Giovannoni, 2003;Tlaskalová-Hogenová et al., 2004). This raises the possibility that organisms that normally reside in the human vulva were overlooked in previous studies that employed cultivation-dependent methods.Micro-organisms present on the vulva of healthy women may have clinical significance because their presence may affect the proliferation of non-indigenous populations (Atassi et al., 2006;Reid & Burton, 2002), including those that play a crucial role in triggering various common diseases such as vulvovaginitis (Wilson, 2005), urinary tract infection (Reid, 1999), bartholinitis (Nakatsu et al., 1998; Sing et al., 1998) and abscesses of Bartholin's gland (Tanaka et al., 2005). Vulvovaginitis ...

Isolates of anaerobic fungi obtained from the rumen, duodenum and faeces of sheep were identified as Piromyces mae based on their morphological characteristics observed using light microscopy. There was no significant morphological variation among the isolates of P. mae from the rumen, duodenum and faeces. Isozymes of 12 isolates of P. mae (one each from the rumen, duodenum and faeces from 4 different sheep) were analysed by PAGE. A total of 12 isozymes were studied and 5 isozyme loci were successfully typed. They were malic enzyme, malate dehydrogenase, shikimate dehydrogenase, alpha-esterase and beta-esterase. All the isolates of P. mae regardless of whether they were from the rumen, duodenum or faeces or from different animals produced very similar isozyme banding patterns for each of the enzyme systems. The similar isozyme profiles of the isolates indicate that they are of the same species although they exist in different regions of the alimentary tract.