axillary microbiota compositions from men and women in a ...1 1 axillary microbiota compositions...

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Axillary Microbiota Compositions from Men and Women in a Tertiary 1

Institution-South East Nigeria: Effects of Deodorants/Antiperspirants on 2

Bacterial Communities. 3

Kingsley C Anukam1*, 2, 3, Victoria Nmewurum1, Nneka R Agbakoba1 4

1Department of Medical Laboratory Science, Faculty of Health Sciences & Technology, 5

Nnamdi Azikiwe University, Nnewi Campus, Anambra State, Nigeria. 6

2Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Nnamdi Azikiwe 7

University, Anambra State, Nigeria. 8

3Uzobiogene Genomics, London, Ontario, Canada. 9

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*Correspondence: Dr. Kingsley C Anukam: [email protected]; 11

[email protected] 12

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ABSTRACT 14

The axillary skin microbiota compositions of African populations that live in warm climate is not 15

well studied with modern next-generation sequencing methods. To assess the microbiota 16

compositions of the axillary region of healthy male and female students, we used 16S rRNA 17

metagenomics method and clustered the microbial communities between those students that 18

reported regular use of deodorants/antiperspirants and those that do not. Axillary skin swab was 19

self-collected by 38 male and 35 females following uBiome sample collection instructions. 20

Amplification of the V4 region of the 16S rRNA genes was performed and sequencing done in a 21

pair-end set-up on the Illumina NextSeq 500 platform rendering 2 x 150 base pair. Microbial 22

taxonomy to species level was generated using the Illumina Greengenes database. 26 phyla were 23

identified in males with Actinobacteria as the most abundant (60%), followed by Firmicutes 24

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(31.53%), Proteobacteria (5.03%), Bacteroidetes (2.86%) and others. Similarly, 25 phyla were 25

identified in females and Actinobacteria was the most abundant (59.28%), followed by Firmicutes 26

(34.28%), Proteobacteria (5.91%), Bacteroidetes (0.45%) and others. A total of 747 genera were 27

identified, out of which 556 (74.4%) were common to both males and females and 163 (21.8%) 28

were exclusive to males while 28 (3.8%) were exclusive to females. Corynebacterium (53.89% vs 29

50.17%) was the most relative abundant genera in both male and female subjects, followed by 30

Staphylococcus (19.66% vs 20.90%), Anaerococcus (4.91% vs 7.51%), Propionibacterium 31

(1.21% vs 1.84%). There was a significant difference (P=0.0075) between those males that 32

reported regular use of antiperspirant/deodorants and those that reported non-use of 33

antiperspirants/deodorants in the relative abundance of Corynebacterium (68.06% vs 42.40%). 34

Higher proportion of Corynebacterium was observed in male subjects than females, while more 35

relative abundance of Staphylococcus was found in females than males. This study detected 36

Lactobacilli in the axilla of over 82% of female and over 81% of male subjects, though in low 37

relative abundance which suggests that Lactobacillus taxa might be considered as part of the 38

normal axillary bacterial community. The study also revealed that the relative abundance of 39

Corynebacterium (68.06% vs 42.40%) was higher in those that reported regular use of 40

deodorants/antiperspirants. 41

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Keywords: axilla, microbiome, microbiota, skin, Africa, deodorants, antiperspirants, students. 43

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INTRODUCTION 45

The bacterial microbiota compositions of the axillary skin of African people is less well studied 46

with the modern next-generation sequencing technology resulting in little or poor knowledge on 47

the microbial communities that could be mined for diagnostic and therapeutic purposes. Previous 48


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studies on the axillary microbiota have relied on culture-dependent methods whereby 49

Staphylococci or Corynebacteria genera have consistently been incriminated (Jackman, 1983; 50

Taylor, et al., 2003). This is due to the fact that culture methods utilize artificial media that can 51

support the growth of these bacteria leading to underestimation of other microbes present on the 52

body site (Kong and Segre, 2012). One culture-dependent study that was conducted in people 53

affected by Albinism and those with normal pigmented skin in Northern Tanzania showed that 54

Staphylococcus was the commonest microorganism isolated in over 90% of the samples (Kiprono 55

et al., 2012). In the last decade, the use of next-generation sequencing approach has revealed an 56

avalanche of microbial communities that inhabit the axillary region showing the predominance of 57

Staphylococci, Corynebacteria, Anaerococcus and Peptoniphilus (Egert et al., 2011; Callewaert et 58

al., 2013; Troccaz et al., 2015). 59

Body malodour is the most common reason human adults generally use deodorants or 60

antiperspirants in order to obtain an appealing body odour or to mask and reduce sweat from the 61

apocrine glands. Bacteria present in the skin are responsible for body odour, whereby sweat 62

components which are odourless are broken down to odour-causing substances such as steroid 63

derivatives, short volatile branched-chain fatty acids and sulphanylalkanols. In the underarm or 64

axilla, malodour arises due to biotransformation by the microbiota of dipeptide-conjugated 65

thioalcohols, particularly S-[1-(2-hydroxyethyl)-1-methylbutyl]-(l)-cysteinylglycine (Cys-Gly-66

3M3SH) (Bawdon et al., 2015). Most students in tertiary institutions around the world are 67

conscious of body odour and application of deodorants have recorded corresponding influence on 68

the species diversities of the axillary microbiome (Callewaert et al 2013). In Western societies, 69

over 95% of the young adult population are concerned about their personal hygiene and are less 70

tolerant toward unpleasant body odour and they make use of underarm deodorants and 71


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antiperspirants(Callewaert et al., 2014). In the same way, the adult population in Africa and 72

particularly students, utilize deodorants with the perception of increased social confidence and 73

improvement in the quality of life (Pierard et al., 2003). We do not have documented information 74

on the social predictors that motivate university students in Nigeria on the regular use of 75

antiperspirants/deodorants but it is believed to be a common phenomenon as marketers advertise 76

such products with hype, brazenly. In this study we hypothesized that the relative abundance of 77

bacterial communities in adult male students may be different from adult female students. The 78

objectives of this study are two folds: first to determine the microbiota compositions of the axillary 79

region of healthy male and female students using 16S rRNA metagenomics method and second to 80

separate the microbial communities between those students that reported regular use of 81

deodorants/antiperspirants and those that do not. 82

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MATERIALS AND METHODS 85

Ethics Review Committee Approval 86

This study was carried out in accordance with the recommendations of the ethic review committee 87

of the Faculty of Health Sciences, Nnamdi Azikiwe University. All subjects gave written informed 88

consent in accordance with the Declaration of Helsinki. 89

Study Participants and Sample Collections 90

A total of 100 participants comprising of 50 male and 50 female students from the Faculty of 91

Health Sciences & Technology , Nnamdi Azikiwe University, Nnewi Campus were recruited in 92

the study. The selection criteria involved those with no history of dermatological disorders or other 93

chronic medical disorders and with no current skin infections. Participants were between the ages 94


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of 17 years old to 35 years old. They provided signed informed consents. Socio-demographic data, 95

skin health or disease history and regular use of deodorant/antiperspirants were obtained from the 96

participants through the administered questionnaires. Skin (Axilla) sample was self-collected 97

following uBiome® sample collection instructions. A moistened sterile cotton swab (uBiome) was 98

thoroughly swabbed for 20 seconds in the axillary region to detach and absorb the microorganisms, 99

and it was vigorously agitated for 20 seconds in a sterilized reaction vial or tube containing a lysis 100

and stabilization buffer that preserves the DNA for transport at ambient temperatures. The tubes 101

were sent to uBiome Inc. in California, United States America for DNA extraction and sequencing. 102

Sequencing results were analyzed with bioinformatic tools at Uzobiogene Genomics, London, 103

Ontario, Canada. 104

DNA Extraction and Sequencing of the 16S rRNA V4 region 105

Bacterial DNA was extracted from the axilla swabs using an in-house protocol developed by 106

uBiome Inc. Briefly, samples were lysed using bead-beating, and DNA was extracted in a class 107

1000 clean room by a guanidine thiocyanate silica column-based purification method using a 108

liquid-handling robot. PCR amplification of the 16S rRNA genes was performed with primers 109

containing universal primers amplifying the V4 region (515F: GTGCCAGCMGCCGCGGTAA 110

and 806R: GGACTACHVGGGTWTCTAAT) as previously described (Caporaso et al, 2011). In 111

addition, the primers contained Illumina tags and barcodes. DNA samples were barcoded with a 112

unique combination of forward and reverse indexes allowing for simultaneous processing of 113

multiple samples. PCR products were pooled, column-purified, and size-selected through 114

microfluidic DNA fractionation. Consolidated libraries were quantified by quantitative real-time 115

PCR using the Kapa Bio-Rad iCycler qPCR kit on a BioRad MyiQ before loading into the 116

sequencer. Sequencing was performed in a pair-end modality on the Illumina NextSeq 500 117


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platform rendering 2 x 150 bp pair-end sequences. The sequencer has a flow cell with four lanes. 118

This means that each sample was read in four different lanes (L001 to L004), and each produced 119

forward (R1) and reverse (R2) reads. 120

Metagenomics Sequence Analysis 121

Raw sequence reads were demultiplexed using Illumina’s BCL2FASTQ algorithm. Reads were 122

filtered using an average Q-score > 30. The 8 paired-end sequence FASTQ reads for each sample 123

were imported into MG-RAST pipeline for quality check (QC). Artificial replicate sequences 124

produced by sequencing artifacts were removed following Gomez-Alvarez, et al., (2009) protocol. 125

Any human host specific species sequences were removed using DNA level matching with bowtie 126

(Langmead et al. 2009) and low-quality sequences were removed using a modified DynamicTrim 127

method by Cox et al. (2011). Quantitative Insights into Microbial Ecology (QIIME) pipeline was 128

used for 16S rRNA recognition. Sequences were pre-screened using QIIMEUCLUST algorithms 129

for at least 97% identity to ribosomal sequences from the RNA databases. Reads passing all above 130

filters were aligned to the database of 16S rRNA gene sequences. Microbial taxonomy to species 131

level was generated using the Illumina BaseSpace Greengenes database. 132

133

RESULTS 134

We hereby present the 16S rRNA dataset of the axillary skin microbiome compositions from the 135

students. Out of 100 axillary swab samples that were collected from male and female students, 38 136

male and 35 female samples passed quality check and were analyzed with bioinformatics tools. 137

On average the base pair count contains 29,129,902bp of DNA sequence and the sequence count 138

contains 194,736 sequences ranging from 32bp to 151bp and averaging 149bp in length 139


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(std.deviation from average length 8.777). All of the sequences have unique identifications. The 140

average GC-content is 55.770% (std.deviation 2.740) and GC-ratio 0.802 (std.deviation 0.095). 141

Distribution of the taxonomic categories shows that the axilla of the male subjects had phyla that 142

ranged from 5-22, Class (10-38), Order (14-80), Family (30-163), Genus (42-326), and Species 143

(35-565) as shown in Figure 1. 144

In contrast, the female subjects had phyla (6-18), Class (11-32), Order (17-69), Family (31-148), 145

Genus (47-292), and Species (79-566) presented in Figure 2. 146

26 phyla were identified in males with Actinobacteria as the most abundant (60%), followed by 147

Firmicutes (31.53%), Proteobacteria (5.03%), Bacteroidetes (2.86%) and others. Two phyla, 148

Fibrobacteres and Nitrospirae appeared exclusive to the males. Similarly, 25 phyla were 149

identified in females and Actinobacteria was the most abundant (59.28%), followed by Firmicutes 150

(34.28%), Proteobacteria (5.91%), Bacteroidetes (0.45%) and others as shown in Figure 3. 151

Caldithrix, occurred exclusively in the female samples. 152

153

Figure 3: Phyla relative abundance (%) in both male and female subjects 154

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At the Family taxonomic level, 257 families were identified, out of which 211 were common to 156

both male and females and 40 were exclusive to males, while 6 families were exclusive to females. 157

Three common families including Corynebacteriaceae (56%/52%), Staphylococcaceae 158

(20%/22%) and Clostridiaceae (6.8%/10.6%) appeared as the most abundant families in both 159

males and females respectively. Among the exclusive families identified in females were 160

Caldithrixaceae, Sporichthyaceae, Halothiobacillaceae, Cohaesibacteraceae, Nannocystaceae and 161

Sulfolobaceae. 162


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At the genera taxonomic level, a total of 747 genera were identified, out of which 556 (74.4%) 163

were common to both males and females and 163 (21.8%) were exclusive to males while 28 (3.8%) 164

were exclusive to females. Corynebacterium (53.89%) was the most relative abundant genera in 165

males, followed by Staphylococcus (19.66%), Anaerococcus (4.91%), Propionibacterium 166

(1.21%), Bacteroides (1.14%), Kaistella (0.85%), Faecalibacterium (0.79%), Blautia (0.76%), 167

Acinetobacter (0.69%) and others. Similarly, Corynebacterium (50.17%) was the most relative 168

abundant genera in females, followed by Staphylococcus (20.90%), Anaerococcus (7.51%), 169

Acinetobacter (2.79%), Propionibacterium (1.84%), Enhydrobacter (1.68%), Micrococcus 170

(1.64%), Finegoldia (1.47%), Peptoniphilus (1.08%), Exiguobacterium (1.03%), Mycobacterium 171

(0.41%), Pseudoclavibacter (0.28%) and others as shown in Figure 4. 172

Figure 4: Comparative relative abundance (%) of taxonomic genera in male and female subjects 173

174

Comparatively, there was a significant difference between male and female on the relative 175

abundance of Corynebacterium (P=0.016), Acinetobacter (P=0.050), Enhydrobacter (P= 0.0001), 176

Finegoldia (P=0.000013), Micrococcus (P=0.0005), and Kaistella (P=0.0145). The proportion of 177

Lactobacillus genera found in 29/35 females was higher (0.02%) compared to 0.01% found in 31/38 of 178

males. 179

At the species taxonomic level, a total of 1994 species were identified of which 1134 species were 180

common to both male and female subjects, while 612 species were exclusively found in males 181

(Supplementary Table 1) and 248 species were identified exclusively in females (Supplementary 182

Table 2). Among the male subjects, Corynebacterium appendicis (19.86%) was the most abundant 183

species, followed by Corynebacterium glaucum (6.35%), Corynebacterium sundsvallense 184

(6.18%), Corynebacterium coyleae (5.65%), Corynebacterium tuberculostearicum (5.18%), 185

Corynebacterium tuscaniense (4.41%), Corynebacterium riegelii (4.05%), Corynebacterium 186


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imitans (4.02%), Anaerococcus octavius (3.82%), Staphylococcus haemolyticus (2.82%) and 187

others represented in Figure 5. 188

In contrast, among the female subjects, Corynebacterium tuberculostearicum (20.80%) was the 189

most abundant species identified in all females subjects followed by Corynebacterium coyleae 190

(9.75%), Corynebacterium appendicis (5.41%), Corynebacterium glaucum (5.04%), 191

Anaerococcus octavius (4.39%), Corynebacterium mucifaciens (3.38%), Staphylococcus 192

haemolyticus (3.04%), Corynebacterium kroppenstedtii (2.86%), Finegoldia magna (2.80%), 193

Micrococcus yunnanensis (2.55%), Staphylococcus aureus (2.51%) and others as shown in Figure 194

6. Comparative relative abundance of Corynebacterium species is presented in Figure 7. 195

Interestingly, 62 Corynebacterium species were found in males, with Corynebacterium auriscanis 196

and Corynebacterium renale as exclusive, while 63 Corynebacterium species were identified in 197

females with Corynebacterium casei, Corynebacterium glucuronolyticum and Corynebacterium 198

pseudodiphtheriticum as exclusive. 199

The axillae of the subjects were also colonized by Lactobacillus species found in 29/35 of female 200

subjects. Among the 29 Lactobacillus species present in female subjects, Lactobacillus equi, 201

Lactobacillus equicursoris, Lactobacillus plantarum, Lactobacillus fabifermentans, Lactobacillus 202

pantheris and Lactobacillus oris occurred exclusively. The male subjects (31/38) had 203

Lactobacillus ruminis, Lactobacillus paracasei, Lactobacillus acidifarinae, Lactobacillus casei, 204

Lactobacillus versmoldensis, and Lactobacillus hayakitensis as exclusive (Figure 8). 205

Staphylococcus species appears to be the second most abundant in both genders, however 34 206

species were identified in males with Staphylococcus haemolyticus (2.82%) as the most abundant 207

species followed by Staphylococcus aureus (1.78%), Staphylococcus gallinarum (0.80%), 208


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Staphylococcus caprae (0.73%), Staphylococcus epidermidis (0.23%) and others as shown in 209

Figure 9. 210

In females, 33 species were found showing Staphylococcus haemolyticus (3.04%) as the most 211

relative abundant species followed by Staphylococcus aureus (2.51%), Staphylococcus caprae 212

(0.99%), Staphylococcus auricularis (0.80%), Staphylococcus gallinarum (0.46%), 213

Staphylococcus epidermidis (0.32%) and others. 214

The use of antiperspirant/deodorants was reported by 23 males while 15 male subjects stated that 215

they do not use such products. At the genera taxonomic level, there was a significant difference 216

(P=0.0075) between those males that reported regular use of antiperspirant/deodorants and those 217

that reported non-use of antiperspirants/deodorants in the relative abundance of Corynebacterium 218

(68.06% vs 42.40%). In contrast, a reverse trend was observed in the relative abundance of Staphylococcus 219

(P = 0.047) (2.25% vs 45.10%) as shown in Figure 10. 220

At the species taxonomic level, males that reported non-use of antiperspirant/deodorants had more relative 221

abundance of Corynebacterium appendicis (22.69% vs 14.30%), Corynebacterium glaucum (7.43% vs 222

4.24%), Corynebacterium tuscaniense (6.55% vs 0.18%), Corynebacterium coyleae (6.27% vs 4.43%), 223

Corynebacterium imitans (5.56% vs 0.99%), Corynebacterium riegelii (4.88% vs 2.41%) and others 224

represented in Figure 11. 225

Conversely, male subjects that reported regular use of antiperspirants/deodorants had more relative 226

abundance of Staphylococcus species than male subjects that reported non-use of 227

antiperspirants/deodorants. For example, Staphylococcus aureus (4.76% vs 0.26%), Staphylococcus 228

gallinarum (1.85% vs 0.26%), Staphylococcus haemolyticus (8.06% vs 0.16%), Staphylococcus kloosii 229

(0.13% vs 0.07%), Staphylococcus caprae (2.11% vs 0.036%), Staphylococcus epidermidis (0.66% vs 230

0.02%), Staphylococcus hominis (0.084% vs 0.003%) and others shown in Figure 12. 231

232

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Interestingly, male subjects that reported non-use of antiperspirants/deodorants had more Lactobacillus 234

species in their axilla. Out of 30 Lactobacillus species identified, 23 Lactobacillus were present in non-use 235

of antiperspirants compared with 14 Lactobacillus species found in male subjects that reported regular use 236

of antiperspirants/deodorants. 237

Although not surprising, out of the 35 female subjects, only 2 reported non-use of 238

antiperspirants/deodorants, while 33 stated regular use of antiperspirants/deodorants. At the genera 239

taxonomic level, Corynebacterium (52.26%) was the most relative abundance in the female subjects that 240

reported regular use of antiperspirants/deodorants, followed by Staphylococcus (23.10%), Anaerococcus 241

(7.25%), Acinetobacter (3.11%), Propionibacterium (2.04%), Enhydrobacter (1.87%), Micrococcus 242

(1.83%), Finegoldia (1.58%), Exiguobacterium (1.15%), Peptoniphilus (1.09%). Comparatively, at the 243

species taxonomic level, the relative abundance of the species that occurred 1.0% and above in both male 244

and female subjects that reported regular use of antiperspirants/deodorants is represented in Figure 13. 245

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DISCUSSIONS 247

In this study, for the first time in Nigeria, we led an elaborate determination of, and obtained detailed insight 248

into the axillary bacterial communities from both healthy adult male and female students using next 249

generation high throughput sequencing approach. Based on the 16S rRNA dataset obtained, over 99.39% 250

(in males) and 99.92% (in females) of the total sequence reads were assigned to four out of 26 phyla 251

representing Actinobacteria, Firmicutes Proteobacteria and Bacteroidetes. Our study is in line 252

with the findings of other studies in Europe by Grice et al., (2009a, 2009b) and Costelo et al., 253

(2009). It is noteworthy that out of 747 genera, only three genera Corynebacterium, 254

Staphylococcus, and Anaerococcus constituted 78.46% of the total reads, which shows that 255

Corynebacterium and Staphylococcus occupy an importance niche in the human axilla. Similar 256

finding was reported by Callewaert et al (2013). 257


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Our hypothesis appears to be supported by the results obtained at the genera taxonomic level as 258

74.4% bacterial communities were common to both males and females and 21.8% were 259

exclusively identified in males while 3.8% were exclusive to females. However, it remains to be 260

determined whether the exclusive bacterial communities in males or females confer any 261

differential health benefit, physiological role or pathogenic potential. A relatively higher 262

proportion of Corynebacterium was observed in male subjects than females, probably suggesting 263

that gender may play a role. Previous study by Fierer et al (2008) found that Corynebacterium tend 264

to colonize male skin especially the hand, more than females. Other studies by Zeeuwen et al 265

(2012) showed that there are differences in the pattern of Corynebacterium colonization in the 266

upper buttocks of males and females. It has been postulated that due to anatomical and 267

physiological differences between male and female subjects, especially in hair growth, skin 268

thickness, sex hormones, sweat and sebum production, may be responsible for these microbial 269

differences in the axilla (Giacomoni et al., 2009). In contrast, there are more relative abundance of 270

Staphylococcus in females than males, similar to the study conducted on the axillae of adult 271

Belgians (Callewaert et al., 2013). It should be noted that previous studies that utilized culture-272

dependent methods never reported Lactobacilli as being part of the skin and or axillary microbiota. 273

For the fact that in this study, we detected Lactobacilli in the axilla of over 82% of females and 274

over 81% of male subjects, though in low relative abundance compared with Corynebacteria and 275

Staphylococci, indicates that Lactobacillus taxa should be considered as part of the normal axillary 276

bacterial community. The female subjects had more relative abundance of Lactobacillus taxa than 277

male subjects, which is consistent with the findings of Lebeer et al., (2019) that found a 10-fold 278

higher relative abundance in women than men. 279


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At the species taxonomic level, the proportion of Corynebacterium appendicis was more 280

pronounced in males than females in the ratio of 19.86% vs 5.41%, while the reverse was the case for 281

Corynebacterium tuberculostearicum found in higher proportion in females than males in the ratio 282

of 20.80% vs 5.18%. The axillary physiological role of this difference remains to be determined, 283

but previous study by Bawdon et al (2015) revealed that Corynebacterium tuberculostearicum is 284

a low producer of malodour precursor, a dipeptide-conjugated thioalcohol, particularly S-[1-(2-285

hydroxyethyl)-1-methylbutyl]-(l)-cysteinylglycine (Cys-Gly-3M3SH). In another study, 286

individuals with higher odour intensities had a greater proportion of Corynebacterium 287

tuberculostearicum (Troccaz et al., 2015). 288

This study revealed that the dominant Staphylococcus species in the sampled population in both 289

male and female subjects were Staphylococcus haemolyticus and Staphylococcus aureus. This is 290

inconsistent with the study by Egert et al. (2011) that showed the dominant Staphylococcus species 291

in the axilla were Staphylococcus epidermidis and Staphylococcus hominis. 292

By clustering the bacterial communities from males that reported regular use of 293

antiperspirant/deodorants and those that reported non-use of antiperspirants/deodorants, we 294

observed that the relative abundance of Corynebacterium (68.06% vs 42.40%) was higher in those 295

reported regular use of deodorants. The implication of this is that the use of deodorants/antiperspirants 296

facilitates the proliferation of Corynebacterium species as high levels of strong body odour were observed 297

by Taylor et al (2003) in individuals with a microbiota dominated by Corynebacterium. Interestingly, in 298

this study, those that reported non-use of deodorants/antiperspirants, the axillae were dominated by 299

Staphylococci, as staphylococci-dominated axillae revealed low levels of odour (Taylor et al 2003). We 300

found out a significant reduction in the species richness and diversities of Lactobacillus taxa from those 301

that reported regular use of deodorants/antiperspirants than those that do not , thus suggesting that beneficial 302

bacteria such as Lactobacilli are impacted negatively by the use of these products. 303


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In this study, Staphylococcus haemolyticus (8.06% vs 0.16%), and Staphylococcus hominis (0.084% vs 304

0.003%) were more in male subjects that reported regular use of deodorants/antiperspirants, which suggests 305

that they may be having more malodour as Staphylococcus hominis and Staphylococcus haemolyticus have 306

been identified as efficient bio-transformers of Cys-Gly-3M3SH (Bawdon et al., 2015). 307

The limitations associated with this study especially on the use of antiperspirants/deodorants verges on the 308

inability to collect information on the exact regular products used. The subjects that used deodorants and 309

or antiperspirants may have different levels of bacterial communities. 310

311

CONCLUSION 312

We have shown in this study that the axilla of the sampled students is composed of bacterial 313

communities that largely represented Actinobacteria, Firmicutes Proteobacteria and 314

Bacteroidetes. A relatively higher proportion of Corynebacterium was observed in male subjects 315

than females, while more relative abundance of Staphylococcus was found in females than males. 316

This study detected Lactobacilli in the axilla of over 82% of female and over 81% of male subjects, 317

though in low relative abundance which suggests that Lactobacillus taxa might be considered as 318

part of the normal axillary bacterial community. The study also revealed that the relative 319

abundance of Corynebacterium (68.06% vs 42.40%) was higher in those reported regular use of 320

deodorants. The implication of this is that the use of deodorants/antiperspirants may facilitate the 321

proliferation of malodour-producing Corynebacterium and Staphylococcus species, while decreasing 322

beneficial bacteria such as Lactobacilli in the axilla. 323

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Figures and Figure legends 328

329

Figure 1: Taxonomic distribution categories in male subjects 330

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Figure 2: Taxonomic distribution categories in female subjects 333


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334

Figure 3: Phyla relative abundance (%) in both male and female subjects 335

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Figure 4: Comparative relative abundance (%) of taxonomic genera in male and female subjects 339


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340

Figure 5: The most relative abundant (%) species in the axilla of male subjects 341

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Figure 6: The most relative abundant (%) species in the axilla of female subjects 346


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347

Figure 7: Comparative relative abundance (%) of Corynebacterium species in the axilla of 348

the subjects 349

350

351

Figure 8: Lactobacillus species identified in the axillary skin of female and male subjects 352


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353

Figure 9: Comparative relative abundance (%) of Staphylococcus species in the axilla of the subjects 354

355

Figure 10: Comparative relative abundance (%) of genera in male subjects 356


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357

358

Figure 11: Comparative relative abundance of Corynebacterium species in male subjects that 359 reported regular use of deodorants/antiperspirants and those that don’t. 360

361


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362

Figure 12: Relative abundance (%) of Staphylococcus species in male subjects that reported 363

regular use of deodorants/antiperspirants and those that don’t. 364

365

366

Figure 13: Relative abundance (%) of species from female and male subjects that reported 367 use of antiperspirants/deodorants. 368


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369

Supplementary Tables: 370

Supplementary Table 1: Bacterial species exclusively identified in male subjects 371

Supplementary Table 2: Bacterial species exclusively identified in the axilla of female subjects 372

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453

AUTHOR CONTRIBUTIONS 454

KCA and NRA conceived and designed the study. KCA sourced for funding, wrote the protocol, 455

did literature search, did bioinformatics analysis, interpreted the data and wrote the final 456

manuscript. VM did the survey experiments, collected the samples, did initial literature searches, 457


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taxonomic data organization & statistical analysis, wrote the draft manuscript and recruited the 458

subjects. KCA and NRA supervised the study and approved the submitted manuscript. 459

460

CONFLICT OF INTEREST: The authors declare that there are no personal, professional or 461

financial relationships that could potentially be construed as a conflict of interest. 462

463

ACKNOWLEDGMENTS 464

We sincerely thank uBiome Inc, San Francisco, California, USA (uBiome has been liquidated and 465

bought over by a Korean company) for awarding a grant-in-kind to Dr. Kingsley Anukam and for 466

carrying out the metagenomics sequencing. We gratefully acknowledge the student volunteers who 467

freely participated in the study. KCA is a visiting reader to the Departments of Medical Laboratory 468

Science and Pharmaceutical Microbiology, Nnamdi Azikiwe University. 469

470


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