Phylogenetic variants of Mycoplasma hominis from pregnant women and women presenting with infertility in Nnamdi Azikiwe University Teaching Hospital, Nnewi, Nigeria

Background: Much controversies have been associated with the pathogenicity of Mycoplasma hominis but little has been done to unravel the mystery behind the different views. This study aimed at investigating the genetic variants abounding within M. hominis and the distribution of the virulent genes among the variants. Methodology: Twenty (20) M. hominis isolates from high vaginal swabs of women (11 from pregnant women and 9 from women presenting with infertility) attending the Obstetrics and Gynaecology clinics of Nnamdi Azikiwe University Teaching Hospital (NAUTH), Nnewi, Nigeria, were sequenced using 16S rRNA universal gene target for the purpose of phylogenetic analysis and epidemiological typing. The isolates were also screened for the presence of M. hominis variable adherence antigen (vaa) and p120 virulent genes using primer constructs from the respective genes in a conventional PCR protocol. Results: Of the 20 M. hominis vaginal isolates, 4 phylogenetic strains were detected; strain MHS43 constituted 10/20 (50.0%) [2/9 (22.2%) from infertile women and 8/11 (72.7%) from pregnant women]; strain MHBS constituted 3/20 (15%) [3/9 (33.3%) from infertile women and 0/11 (0%) from pregnant women]; strain MHSWP2 constituted 4/20 (20.0%) [3/9 (33.3%) from infertile women and 1/11 (9.1%) from pregnant women]; while strain MHKC87 constituted 3/20 (15%) [1/9 (11.1%) from infertile women and 2/11 (18.2%) from pregnant women].Each of vaa and p120 genes was detected in 14 of 20 isolates, while 6 isolates did not carry the genes. A 2-way ANOVA test showed that none of the genes was significantly associated with a particular strain (p=0.8641). Conclusions: The different views regarding the pathogenicity of M. hominis may be linked to the heterogeneity within the species and lack of homogeneity in the virulent genes as witnessed both in the intra species and intra strain levels.

Prevalence of bacterial vaginosis in pregnant women attending Nnamdi Azikiwe University Teaching Hospital, Nnewi, Nigeria using the complete Amsel's diagnostic criteria

Background: Bacterial vaginosis (BV) in pregnant women remains a cause for clinical concern among clinicians and health care professionals. BV has been linked to prenatal, antenatal and postnatal challenges in pregnant women. Information on prevalence of BV across trimesters of pregnancy is expected to give better clinical insight into the pathophysiology of this polymicrobial disorder. This study was conducted to determine the prevalence of BV in pregnant women attending the Nnamdi Azikiwe University Teaching Hospital (NAUTH), Nnewi, Nigeria. Methodology: This was a cross-sectional study of 120 pregnant women (40 in each trimester of pregnancy) who had symptoms suggestive of BV, selected by systematic random sampling from among the women attending the Obstetrics and Gynaecology (O & G) clinic of NAUTH, Nnewi. Each subject participant was examined by the attending clinician, and high vaginal swab (HVS) sample was collected for diagnostic analysis of BV using with complete Amsel's clinical criteria, which consists of three of the four criteria; (i) adherent and homogenous vaginal discharge, (ii) vaginal pH > 4.5, (iii) detection of clue cells on saline wet mount, and (iv) amine odor after the addition of potassium hydroxide (positive Whiff test). Results: The mean age of the 120 selected participants was 27.25±6.09 years. The age groups 25-29 (36.7%) and 20-24 years (33.3%) constituted the largest proportion, while age groups <20 (5.0%) and 40-45 years (5.0%) constituted the least. Of the 120 participants, 26 (21.7%) were positive for BV by the Amsel's criteria. Pregnant women in age group <20 years had the highest prevalence of BV (100%, 6/6), followed by those in the age groups 20-24 (27.5%), 40-45 (16.7%), 25-29 (15.9%), 30-34 (9.1%) and 35-39 years (0%) (X 2=28.063, p=0.0001). Prevalence of BV was significantly higher in single (unmarried) pregnant women (45.5%, X 2=4.038, p=0.045), women with primary school education level (66.7%, X 2=14.530, p=0.001), unemployed women (36.1%, X 2=13.278, p=0.0013), and nulliparous women [36.4%, X 2 (for trend) = 4.805, p=0.0274), while there was no significant difference in the prevalence of BV with relation to trimester of pregnancy (X 2=2.750, p=0.253). Conclusion: This study reveals a relatively high prevalence of BV and significant association with factors such as age group, education and occupational status among pregnant women attending NAUTH Nnewi. Regular screening of women for BV prenatally may enable appropriate interventions to prevent adverse pregnancy outcomes

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.

Modulation of the immune response to Mycobacterium tuberculosis during malaria/M. tuberculosis co-infection.

Tuberculosis (TB) causes significant morbidity and mortality on a global scale. The African region has 24% of the world's TB cases. TB overlaps with other infectious diseases such as malaria and HIV, which are also highly prevalent in the African region. TB is a leading cause of death among HIV-positive patients and co-infection with HIV and TB has been described as a syndemic. In view of the overlapping epidemiology of these diseases, it is important to understand the dynamics of the immune response to TB in the context of co-infection. We investigated the cytokine response to purified protein derivative (PPD) in peripheral blood mononuclear cells from TB patients co-infected with HIV or malaria and compared it to that of malaria- and HIV-free TB patients. A total of 231 subjects were recruited for this study and classified into six groups; untreated TB-positive, TB positive subjects on TB drugs, TB- and HIV-positive, TB- and malaria-positive, latent TB and apparently healthy control subjects. Our results demonstrate maintenance of interferon (IFN)-γ production in HIV and malaria co-infected TB patients in spite of lower CD4 counts in the HIV-infected cohort. Malaria co-infection caused an increase in the production of the T helper type 2 (Th2)-associated cytokine interleukin (IL)-4 and the anti-inflammatory cytokine IL-10 in PPD-stimulated cultures. These results suggest that malaria co-infection diverts immune response against M. tuberculosis towards a Th-2/anti-inflammatory response which might have important consequences for disease progression.

Phytochemical and antifungal activities of Uvaria. chamae leaves and roots, Spondias mombin leaves and bark and Combretum racemosum leaves.

The effects of crude and dilutions of aqueous, methanolic and n-hexane extracts of Uvaria chamae (roots and leaves), Spondias mombin (leaves and bark) and Combretum racemosum (Leaves), on pathogenic candida albicans and aspergillus niger was studied. The aim was to contribute to the search for a cheaper, conventional cure for both fungi. Phytochemical analysis revealed varying degrees of alkaloids, glycosides, saponin, lipid and oil, tannins, flavonoids, terpenoids and acids. Agar diffusion method was used for anti fungal assay. Minimum inhibitory concentration (mic) used was 10 mg/ml of extract and dilutions of the non polar solvents of 10 (-1) and 10(-2) was used. Results showed that none of the plant parts was active against aspergillus niger. Combretum racemosum had no antifungal effect on tested organisms as well as the different dilutions. However crude methanolic extract of uvaria (roots and leaves), spondias ( bark and leaves ), and n-hexane extracts of uvaria (leaves and roots), produced anti candidal effects with diameters in this order 14.67 +/- 0.72mm, 10.67 +/- 0.52 mm, 11.00 +/- 0.47 mm, 15.00 +/- 0.47 mm, and 14.67 +/- 0.72 mm respectively. Some of the plant parts especially uvaria had zones of inhibition at a confidence limit comparable with control drug which is ketoconazole and it had inhibitory effects at a diameter of 20.06 +/- 0.40 mm.

Polymerase chain reaction assay of ureaplasma strains isolated from high vaginal swabs of women in Ibadan, Nigeria.

Human ureaplasma previously had one species known as Ureaplasma urealyticum but was recently separated into 2 species, U. urealyticum and U. parvum. This study was carried out to separate the ureaplasma strains isolated from women attending a tertiary-care hospital in Nigeria. Thirty (30) Ureaplasma strains isolated from the vaginal tracts of 13 pregnant and 17 non-pregnant women were assayed. The polymerase chain reaction (PCR) technique was employed using two primer pairs: UMS-170/UMA-263 specific for U. urealyticum and UMS-57/UMA-222 specific for U. parvum. The positivity bands of the primer pairs were 476 bp and 326 bp for U. urealyticum and U. parvum respectively. All isolates were found to be U. urealyticumn (100%). Eleven (84.6%) of the 13 U. urealyticum from pregnant women were from asymptomatic women while from the non-pregnant women; 6 (35.3%) were from women with complaint of infertility problems; 5 (29.4%) from those who complained of vaginal discharge, one (5.9%) was asymptomatic while the remaining 5 (29.4%) had various other complaints. U. urealyticum is thus the prevalent species of Ureaplasma among pregnant and non-pregnant women in the study population and this to the best of our knowledge is a pioneer study to speciate human ureaplasmas in this country.

Genital mycoplasmas in semen samples of males attending a tertiary care hospital in Nigeria: any role in sperm count reduction?

Semen samples from 54 married men attending the outpatient clinics for problems of infertility and routine semen analysis were examined for the presence of genital mycoplasmas. The mean age of the men was 36.1 years with a range of 25 55 years. Majority of the men 57.4% (31 of 54) were in their fourth decade of life (30 39 years). This age group also had the highest percentage 57.2% (8 of 14) of positive isolates of genital mycoplasmas on semen culture. A total of 21 organisms obtained from 14 (26.0%) positive samples were isolated. Mycoplasma and Ureaplasma spp. separately isolated from the samples yielded frequencies of 1 (1.9%) and 6 (11.1%) respectively and the remaining 7 (13.0%) samples were infected with both organisms. A breakdown of the mycoplasma species include 5 (23.8%) M. hominis, 2 (9.5%) M. fermentans and 1 (4.8%) M. penetrans. Apart from one isolate of M. hominis other Mycoplasma species were found in association with Ureaplasma species. Fifteen (71.4%) of the 21 isolates [8 (53.3%) ureaplasmas and 7 (46.7%) mycoplasmas] were isolated from samples with sperm counts less than 20 million/ml while the remaining 6 (21.6%) isolates [5 (83.3%) ureaplasmas and 1 (16.7) mycoplasma] were from samples with counts greater than 20 million/ml. This finding could indicate a possible influence of genital mycoplasmas especially mycoplasmas species on sperm count.

Presence of mycoplasma and ureaplasma species in the vagina of women of reproductive age.

OBJECTIVE: Against the background of genital mycoplasmas being implicated as pathogens in the genital tracts of adult humans, this study was carried out to determine the prevalence of genital mycoplasmas among women of reproductive age attending a tertiary-care hospital in Nigeria. METHODS: One hundred and sixty-eight high vaginal specimens from 114 pregnant and 54 non-pregnant women with or without complaints were studied for the presence of genital mycoplasmas using selective bacteriological culture techniques. Isolates were identified biochemically and serologically. RESULTS: Sixty (35.7 %) isolates of genital mycoplasmas were obtained from the study population. Identification showed 30 (17.9%) as Mycoplasma species, 13(7.7%) as Ureaplasma species and 17(10.1%) were positive for both organisms. Thirteen (11.4%) ureaplasma isolates were from pregnant women and 17 (31.5%) from non-pregnant women (p< 0.05). Thirty-eight (80.8%) of the Mycoplasma species were identified as Mycoplasma hominis, 6(12.8%) as M.fermentans and 3(6.4%) as M. penetrans. High prevalence of the Mycoplasma species were obtained from asymptomatic pregnant women (84.8%) while the Ureaplasma species were found more from symptomatic non-pregnant women- 35.3 % from infertility patients, 29.4% from those with vaginal discharge and 29.4% from those with other gynaecological complaints. CONCLUSION: These findings could be important since genital mycoplasmas have been associated with various adverse conditions especially in pregnant women and neonates while Ureaplasma species have been implicated in infertility. Their specific roles need further investigations.

Biochemical and serological characterization of mycoplasma strains isolated from the genital tracts of humans in Nigeria.

Fifty-five (55) Mycoplasma strains isolated from the genital tracts of humans were biochemically characterized using various biochemical tests and also serologically identified by growth inhibition technique using 5 mycoplasma antisera namely M. hominis PG2 1: M. genitalium G37: M. penetrans GTU54 and 2 strains of M. fermentans PG18 (HRC 6-62-S-170 and MB713-501-069). Biochemically, 43 (78.2%) strains were identified as Mycoplasma hominis, 8 (14.5%) strains as M. fermentans and 4 (7.3%) as M. penetrans. The M. hominis strains hydrolyzed only arginine while the M. fermentans and M. penetrans strains in addition to arginine hydrolysis also broke down glucose fermentatively and oxidatively. The M. fermentans strains showed varying reactions to phosphatase activity and to the reduction of tetrazolium chloride. Serologically, 4 (7.3%) mycoplasma strains were confirmed as M. penetrans GTU54 and of the 8 M. fermentans strains, 4 (7.3%) were identified as M. fermentans PG18 serotype HRC 6-62-S-170 and the other 4 (7.3%) as M. fermentans PG18 serotype MB 713-501-069. Only 13 (30.2%) of the 43 M. hominis strains were identified as M. hominis serotype PG2 1. None was identified as M. genitalium. The heterogeneity of the mycoplasma strains especially M. hominis was observed in this study and the need for the use of multiple antisera in growth inhibition test is hereby supported.