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Axillary Microbiota Compositions from Men and Women in a Tertiary

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Axillary Microbiota Compositions from Men and Women in a Tertiary bioRxiv preprint doi: https://doi.org/10.1101/2020.03.11.986364; this version posted March 12, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Axillary Microbiota Compositions from Men and Women in a Tertiary 2 Institution-South East Nigeria: Effects of Deodorants/Antiperspirants on 3 Bacterial Communities. 4 Kingsley C Anukam1*, 2, 3, Victoria Nmewurum1, Nneka R Agbakoba1 5 1Department of Medical Laboratory Science, Faculty of Health Sciences & Technology, 6 Nnamdi Azikiwe University, Nnewi Campus, Anambra State, Nigeria. 7 2Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Nnamdi Azikiwe 8 University, Anambra State, Nigeria. 9 3Uzobiogene Genomics, London, Ontario, Canada. 10 11 *Correspondence: Dr. Kingsley C Anukam: [email protected]; 12 [email protected] 13 14 ABSTRACT 15 The axillary skin microbiota compositions of African populations that live in warm climate is not 16 well studied with modern next-generation sequencing methods. To assess the microbiota 17 compositions of the axillary region of healthy male and female students, we used 16S rRNA 18 metagenomics method and clustered the microbial communities between those students that 19 reported regular use of deodorants/antiperspirants and those that do not. Axillary skin swab was 20 self-collected by 38 male and 35 females following uBiome sample collection instructions. 21 Amplification of the V4 region of the 16S rRNA genes was performed and sequencing done in a 22 pair-end set-up on the Illumina NextSeq 500 platform rendering 2 x 150 base pair. Microbial 23 taxonomy to species level was generated using the Illumina Greengenes database. 26 phyla were 24 identified in males with Actinobacteria as the most abundant (60%), followed by Firmicutes 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.03.11.986364; this version posted March 12, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 25 (31.53%), Proteobacteria (5.03%), Bacteroidetes (2.86%) and others. Similarly, 25 phyla were 26 identified in females and Actinobacteria was the most abundant (59.28%), followed by Firmicutes 27 (34.28%), Proteobacteria (5.91%), Bacteroidetes (0.45%) and others. A total of 747 genera were 28 identified, out of which 556 (74.4%) were common to both males and females and 163 (21.8%) 29 were exclusive to males while 28 (3.8%) were exclusive to females. Corynebacterium (53.89% vs 30 50.17%) was the most relative abundant genera in both male and female subjects, followed by 31 Staphylococcus (19.66% vs 20.90%), Anaerococcus (4.91% vs 7.51%), Propionibacterium 32 (1.21% vs 1.84%). There was a significant difference (P=0.0075) between those males that 33 reported regular use of antiperspirant/deodorants and those that reported non-use of 34 antiperspirants/deodorants in the relative abundance of Corynebacterium (68.06% vs 42.40%). 35 Higher proportion of Corynebacterium was observed in male subjects than females, while more 36 relative abundance of Staphylococcus was found in females than males. This study detected 37 Lactobacilli in the axilla of over 82% of female and over 81% of male subjects, though in low 38 relative abundance which suggests that Lactobacillus taxa might be considered as part of the 39 normal axillary bacterial community. The study also revealed that the relative abundance of 40 Corynebacterium (68.06% vs 42.40%) was higher in those that reported regular use of 41 deodorants/antiperspirants. 42 43 Keywords: axilla, microbiome, microbiota, skin, Africa, deodorants, antiperspirants, students. 44 45 INTRODUCTION 46 The bacterial microbiota compositions of the axillary skin of African people is less well studied 47 with the modern next-generation sequencing technology resulting in little or poor knowledge on 48 the microbial communities that could be mined for diagnostic and therapeutic purposes. Previous 2 bioRxiv preprint doi: https://doi.org/10.1101/2020.03.11.986364; this version posted March 12, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 49 studies on the axillary microbiota have relied on culture-dependent methods whereby 50 Staphylococci or Corynebacteria genera have consistently been incriminated (Jackman, 1983; 51 Taylor, et al., 2003). This is due to the fact that culture methods utilize artificial media that can 52 support the growth of these bacteria leading to underestimation of other microbes present on the 53 body site (Kong and Segre, 2012). One culture-dependent study that was conducted in people 54 affected by Albinism and those with normal pigmented skin in Northern Tanzania showed that 55 Staphylococcus was the commonest microorganism isolated in over 90% of the samples (Kiprono 56 et al., 2012). In the last decade, the use of next-generation sequencing approach has revealed an 57 avalanche of microbial communities that inhabit the axillary region showing the predominance of 58 Staphylococci, Corynebacteria, Anaerococcus and Peptoniphilus (Egert et al., 2011; Callewaert et 59 al., 2013; Troccaz et al., 2015). 60 Body malodour is the most common reason human adults generally use deodorants or 61 antiperspirants in order to obtain an appealing body odour or to mask and reduce sweat from the 62 apocrine glands. Bacteria present in the skin are responsible for body odour, whereby sweat 63 components which are odourless are broken down to odour-causing substances such as steroid 64 derivatives, short volatile branched-chain fatty acids and sulphanylalkanols. In the underarm or 65 axilla, malodour arises due to biotransformation by the microbiota of dipeptide-conjugated 66 thioalcohols, particularly S-[1-(2-hydroxyethyl)-1-methylbutyl]-(l)-cysteinylglycine (Cys-Gly- 67 3M3SH) (Bawdon et al., 2015). Most students in tertiary institutions around the world are 68 conscious of body odour and application of deodorants have recorded corresponding influence on 69 the species diversities of the axillary microbiome (Callewaert et al 2013). In Western societies, 70 over 95% of the young adult population are concerned about their personal hygiene and are less 71 tolerant toward unpleasant body odour and they make use of underarm deodorants and 3 bioRxiv preprint doi: https://doi.org/10.1101/2020.03.11.986364; this version posted March 12, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 72 antiperspirants(Callewaert et al., 2014). In the same way, the adult population in Africa and 73 particularly students, utilize deodorants with the perception of increased social confidence and 74 improvement in the quality of life (Pierard et al., 2003). We do not have documented information 75 on the social predictors that motivate university students in Nigeria on the regular use of 76 antiperspirants/deodorants but it is believed to be a common phenomenon as marketers advertise 77 such products with hype, brazenly. In this study we hypothesized that the relative abundance of 78 bacterial communities in adult male students may be different from adult female students. The 79 objectives of this study are two folds: first to determine the microbiota compositions of the axillary 80 region of healthy male and female students using 16S rRNA metagenomics method and second to 81 separate the microbial communities between those students that reported regular use of 82 deodorants/antiperspirants and those that do not. 83 84 85 MATERIALS AND METHODS 86 Ethics Review Committee Approval 87 This study was carried out in accordance with the recommendations of the ethic review committee 88 of the Faculty of Health Sciences, Nnamdi Azikiwe University. All subjects gave written informed 89 consent in accordance with the Declaration of Helsinki. 90 Study Participants and Sample Collections 91 A total of 100 participants comprising of 50 male and 50 female students from the Faculty of 92 Health Sciences & Technology , Nnamdi Azikiwe University, Nnewi Campus were recruited in 93 the study. The selection criteria involved those with no history of dermatological disorders or other 94 chronic medical disorders and with no current skin infections. Participants were between the ages 4 bioRxiv preprint doi: https://doi.org/10.1101/2020.03.11.986364; this version posted March 12, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 95 of 17 years old to 35 years old. They provided signed informed consents. Socio-demographic data, 96 skin health or disease history and regular use of deodorant/antiperspirants were obtained from the 97 participants through the administered questionnaires. Skin (Axilla) sample was self-collected 98 following uBiome® sample collection instructions. A moistened sterile cotton swab (uBiome) was 99 thoroughly swabbed for 20 seconds in the axillary region to detach and absorb the microorganisms, 100 and it was vigorously agitated for 20 seconds in a sterilized reaction vial or tube containing a lysis 101 and stabilization buffer that preserves the DNA for transport at ambient temperatures. The tubes 102 were sent to uBiome Inc. in California, United States America for DNA extraction and sequencing.
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