A comparative study of the oral microbiome compositions of healthy postmenopausal, premenopausal, and prepubertal Nigerian females, using 16S rRNA metagenomics methods

Introduction: There is a paucity of information on the oral microbiome compositions of Nigerians, mostly due to lack of appropriate molecular techniques. In this pilot study, we sought to determine and characterize the oral bacterial compositions of “healthy” females. Materials and Methods: Oral samples were collected from three randomly selected females aged 56, 28, and 8 years. DNA was extracted and 16S rRNA V4 region was amplified using custom‑barcoded primers before sequencing with Illumina MiSeq platform. Quantitative Insights into Microbial Ecology pipeline was used for 16S rRNA recognition. Distribution of taxonomic categories at different levels of resolution was done using the ribosomal RNA similarities to entries in the REFseq protein database. Diversity score was calculated based on the inverse Simpson’s index. Results: The inverse Simpson’s diversity index for the postmenopausal, premenopausal, and prepubertal was 7.74, 6.95, and 7.42 respectively. A total of 12 phyla, 70 genera, and 85 species were detected. Firmicutes followed by Proteobacteria, Actinobacteria, Bacteroidetes, and Fusobacteria dominated the oral microbiome of the subjects. Streptococcus thermophilus (33.19%) was the most abundance species in subject 1, while subject 2 was highly predominated by Haemophilus parainfluenzae (80.65%), and subject 3 was predominated by Haemophilus influenzae (23.05%). Conclusion: The study has revealed that bacteria with varying diversities colonized the subjects and it highlighted the importance of metagenomics in deciphering the oral bacterial compositions from females of different age groups. More studies are needed using metagenomics approach, to appreciate these bacterial organisms that are associated with health and disease in our environment.Keywords: Metagenomics, Nigerian females, oral microbiom

Self-help treatment methods and aetiology of infertility among couples seeking in-vitro fertilization in a private fertility clinic, Awka, Nigeria

BACKGROUNDInfertility is considered a serious problem in low andmiddle-income countries due to the premium placedon childbearing. OBJECTIVESTo determine self-help treatment methods, aetiologyof infertility, sources of information for AssistedReproductive Technology and sources of verbal orphysical abuse of women seeking AssistedReproductive Technology. MATERIALS AND METHODSThis is a cross-sectional descriptive study. A total ofthirty-six couples were recruited for in-vitrofertilization using systematic sampling technique.Structured questionnaires were administered to allthe selected subjects in order to extract relevantbaseline information on self-help treatment methodsand socio-demographic data. Inclusion criteria weresubjects consenting to participate in this study, thosewithin age range between 25 and 60 years old, thosewhose infertility (primary or secondary) is persistinglonger than one year and those who have not beenreceiving antibiotics treatment before thecommencement of the study. Subjects were excludedon the basis of being under 25 years old, thosereceiving antibiotics treatment before the study andthose not visiting the clinics for assistedreproduction. Clinical and laboratory techniqueswere used to determine the causes of the subject’sinfertility. Sono saline hysterogram and laparoscopyand dye test were used to reveal the aetiology offemale infertility while semen analysis was used toassess the male subjects. RESULTSA total of thirty-six women were recruited for in-vitrofertilization and embryo transfer (IVF/ET) procedure.Majority of the subjects 15 (41.7%) had previouslyself-treated infections with herbal medications only.Fourteen women (38.9%) reported frequent abuse bytheir family relatives either verbally or physically fortheir infertility. Blocked fallopian tubes was thecommonest aetiology of infertility in females 15(41.7%) as revealed by laparoscopy and dye test. Itwas observed that the 21 couples (58.3%) who hadprevious history of sexually transmitted diseasesrecorded highest frequency in the distribution ofpotential risk factors associated with genitalbacterial infections. Twenty-three couples (63.9%)reported social media as the most frequent sourcesof information for Assisted Reproductive Technology(ART). CONCLUSIONSocial media has been reported as the major sourceof information concerning assisted reproductivetechnology among couples. Bilateral tubal blockageand poor semen quality are the commonestaetiology of infertility in this study. Majority of thesubjects had treated sexually transmitted infectionswith herbal medications. However, there is urgentneed for public education on the contribution ofcouples to infertility and current treatment methods

Vaginal bacteriome of Nigerian women in health and disease: A study with 16S rRNA metagenomics

Introduction: The argument on what bacteria make up healthy vagina and bacterial vaginosis (BV) remain unresolved. Black women most often are placed in grade IV vaginal communities as lacking Lactobacillus-dominated microbes. We sought to determine the vaginal microbiota compositions of healthy and those with BV using 16S rRNA metagenomics methods. Materials and Methods: Twenty-eight women provided vaginal swabs for Nugent scoring. Fifteen had BV (Nugent score 7–10), whereas 13 were normal (Nugent score 0–3). DNA was extracted and 16S rRNA V4 region amplified using custom bar-coded primers prior to sequencing with MiSeq platform. Sequence reads were imported into Illumina BaseSpace Metagenomics pipeline for 16S rRNA recognition. Distribution of taxonomic categories at different levels of resolution was done using Greengenes databases. Manhattan principal component analysis was used for similarity clustering. Results: Non-BV subjects were colonized by 12 taxonomic phyla that represent 182 genera and 357 species. Overall, 23 phyla representing 388 genera and 805 species were identified in BV subjects. Firmicutes represented 95% of the sequence reads in non-BV subjects with Lactobacillus-dominated genera and Lactobacillus crispatus–dominated species, followed by Proteobacteria (3.78%), Actinobacteria (0.74%), and Bacteriodetes (0.05%). In BV subjects, Firmicutes represented 59% of the classified sequence reads, followed by Bacteroidetes (19%), Actinobacteria (15.8%), Fusobacteria (4.08%), Proteobacteria (1.48%), and Tenericutes (1.25%). Conclusion: Non-BV healthy Black African, Nigerian women had Lactobacillus genera as the predominant microbiota, contrary to published reports. The study shows that BV subjects had varying proportions of diverse bacteria similar to studies from other parts of the world