Browsing by Author "Jaspan, Heather B"

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    Alterations in preconception, antenatal, and postnatal maternal gut microbiota influence offspring intestinal microbiota and immunity
    (2017) Nyangahu, Donald D; Jaspan, Heather B; Horsnell, William
    Maternal microbiota during pregnancy, as well as maternal disease state, may impact offspring gut bacterial colonisation. Here, we explore the impact of maternal antibiotics during gestation and/or nursing on offspring gut microbiota. Further, we investigate the effect of preconception helminth infections on maternal and infant gut microbiota. For maternal antibiotic experiments, dams were fed vancomycin, polymyxin B, or both, in drinking water during gestation, nursing or gestation plus nursing, and their offspring microbiota analysed at 14 days of life, alongside immunity in the spleens. Offspring born to vancomycin treated mothers had significantly higher relative abundance of Proteobacteria and Tenericutes while maternal oral polymyxin B led to significantly lower abundance of Proteobacteria and Deferribacteres in infants. Maternal oral vancomycin led to significant reduction in proportions of infant central memory CD4+ T cells (CD4+CD44hiCD62Lhi) regardless of antibiotic timing. Effector memory CD4+ T cells were significantly lower in pups born to dams treated with polymyxin B while nursing while proportions of central memory CD4 T cells were significantly increased in gestation only or gestation plus nursing pups. In addition, oral vancomycin in dams during nursing resulted in significantly reduced proportions of both total and follicular B cells in offspring born to antibiotic treated dams. Pups born to Vancomycin treated mothers had a significant delay in growth when infected with Respiratory Syncytial Virus (RSV). On the other hand, pups born to mothers treated with Polymyxin B during gestation or gestation plus nursing were susceptible to Nippostrongylus brasiliensis (Nb) infection. In the second study, we infected female BALB/c mice with 500Nb L3 three weeks prior to mating and examined the effect of preconception helminth infection on offspring microbiota and immunity. Preconception Nb infections led to alterations of maternal gut microbiota during pregnancy. In addition, we observed dramatic differences in offspring microbiota in pups born to previously helminth infected dams. Coriobacteriaceae were predominant in pups born to previously Nb infected dams when compared to uninfected dams. Overall, manipulation of maternal microbiota during gestation or lactation profoundly impacts offspring growth, intestinal microbiota and immunity to RSV and helminths.
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    Bacterial disease and antimicrobial susceptibility patterns in HIV-infected, hospitalized children: a retrospective cohort study
    (Public Library of Science, 2008) Jaspan, Heather B; Huang, Lyen C; Cotton, Mark F; Whitelaw, Andrew; Myer, Landon
    BACKGROUND: Serious bacterial infections are a major source of morbidity and mortality in HIV-infected children. The spectrum of disease is wide, and responsible organisms vary according to setting. The use of antibiotic prophylaxis and the emergence of multi-drug resistant bacteria necessitate examination of responsible organisms and their antibiotic susceptibility. METHODOLOGY/PRINCIPAL FINDINGS: A retrospective cohort study of all HIV-positive pediatric admissions at an urban public sector hospital in Cape Town between January 2002 and June 2006 was conducted. Children between the ages of one month and nine years with laboratory confirmed HIV status, serious bacterial infection, and a hospital length of stay of 5 days or more, were eligible for inclusion. Organisms isolated from blood, urine, and cerebral spinal fluid cultures and their antimicrobial susceptibility were examined, and compared according to timing of isolation to distinguish nosocomial versus community-acquired. One hundred and forty-one children were identified (median age 1.2 years), 39% of whom were on antiretrovirals started before or during this hospitalization. Bacterial infections involved all organ systems, however pneumonia was most common (67%). S. pneumoniae and S. aureus were the most common gram positive and K. pneumoniae was the most common gram negative organism. K pneumoniae isolates were resistant to many first and second line antibiotics, and were all considered nosocomial. All S. aureus isolates were methicillin resistant, some of which were community-acquired. Conclusions/Significance Bacterial infections are an important source of co-morbidity in HIV-infected children in resource-limited settings. Clinicians should have a low threshold to initiate antibiotics in children requiring hospitalization. Broad-spectrum antibiotics should be used judiciously. Clinicians caring for HIV-infected children should be cognizant of the most common organisms affecting such children, and of their local antimicrobial susceptibilities, when treating empirically for serious bacterial infections.
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    CD4 and CD8 T-cell responses to acellular pertussis and rotavirus vaccination in breast-fed HIV exposed, uninfected infants
    (2017) Nundalall, Trishana; Jaspan, Heather B; Gray, Clive M
    Introduction: Vaccination is one of the most efficient ways to prevent infectious diseases, however due to the naivety and regulation of immunity found in infants, induction of vaccine-mediated immunity is challenging. Respiratory and diarrheal diseases are major contributors to infant mortality. Additionally, Human Immunodeficiency Virus-1 (HIV) infections increase the risk of mortality. Current advances in Prevention of Mother-to-Child Transmission (PMTCT) have prevented HIV infections in almost 97% of infants being born to HIV-infected mothers. As a result there is an increasing number of HIV exposed, uninfected infants (HEU). HEU infants have a higher rate of infectious disease related mortality and morbidity compared to unexposed infants, the underling causes of these differences are still not understood. In this dissertation, responses to two childhood vaccines, live, attenuated rotaviral vaccine (Rotarix) and acellular Pertussis (aP), were analyzed in HEU infants, with specific focus on T-cell responses to Rotarix and aP, due to the current lack of published data on T-cell responses. Additionally, the influence of feeding mode, that is breast or formula feeding, was also assessed as it is well established that breast fed infants contract fewer infections compared to formula fed infants. Methods: This dissertation included infants from a larger cohort which includes three groups of infants; HIV unexposed breast fed (UBF), HIV exposed breast-fed (EBF) and HIV exposed formula fed (EFF) infants. Infants were recruited at birth and followed up until 36 weeks of age. As no Rotavirus vaccine T-cell assay was previously published, multiple techniques were utilized to attempt to optimize an assay capable of detecting Rotavirus (RV) vaccine-specific T-cell responses. To determine T-cell responses to Bordertella pertussis (BP), blood was collected from infants at each time-point and 200ul was stimulated with BP antigen in a 12-hour whole blood assay. Cells from all assays were fixed and stained for flow cytometric analysis of CD4 and CD8 T-cell responses. The markers used included live/ dead, CD3, CD4 and CD8 for identification of T-cell populations, IFNγ, IL-2 and TNFα cytokines, HLA-DR and Ki67 for activation and proliferation, and CD45RA and CD27 memory differentiation. Data analysis was then completed using Microsoft Excel, Flow.Jo V9, GraphPad prism V6, Pestle 1.7 and Spice V5.33 software packages. Results: Despite multiple attemps it was not possible to optimise an assay capable of consistently detecting Rotavirus vaccine specific responses. This was partly due to interferance from contaminating agents in the protein antigens used, and difficulty in culturing and purification of whole virus. Assessment of aP spcific CD4+ T-cell memory demonstrated an overall increase in terminally differentiated (TD) memory cells accross time. This mirrored the ontogeny of the total T cell pool which showed an overall decrease in naïve T-cell frequencies with a consequent increase in late and terminally differentiated CD4 and CD8 T-cell populations over time through the first months of life. Both total and aP specific CD4+ early differentiated (ED) memory T-cells remained unchanged over time. ED CD8+ memory T-cells peaked at week 15 in EBF infants. A similar observation was found in UBF infants but at a non-significant level. EFF infants had no significant changes in CD8+ naive, ED and late differentiated (LD) memory populations over time. Additionally all infants demonstrated high levels of Ki67 expression at D4-7, which is prior to vaccination and maintained this level of proliferation after vaccination. HEU infants had higher levels of activation compared to HU infants in the first week of life but this normalised to HU infant levels by week 7. Furthermore EFF infants had peak T-cell activation at week 7 as compared to week 15 in EBF infants. In addition HU infants had better cytokine responses than HEU infants at week 7 but similar responses at week 15 and 36. In Addition, EFF infants also had increased vaccine specific CD4+ responses at week 7 and week 36 compared to EBF infants. This was true for overall cytokine expressing CD4 T-cells and single TNFα expressing CD4+ T-cells. Disscussion: Given the important role T-cells play in the clearance of Rotavirus, it is important that an assay capable of detecting RV vaccine specific T-cell responses be developed. Furthermore, T cells play a role in providing help for antibody responses to BP and for killing of intracellular bacteria. Our findings regarding immunity to aP suggest that all infants, regardless of HIV exposure status and feeding mode, are able to mount a T cell response to aP vaccination. However the differing ontogeny of responses seen in all three groups of infants lends some insight on the complex determinants of vaccine T -cell immunogenicity. In this case, age since vaccination, HIV exposure, and feeding mode resulted in apparent changes in vaccine responses as well as T cell differetiation and activation.
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    Correction to: Microbial function and genital inflammation in young South African women at high risk of HIV infection
    (2022-03-09) Alisoltani, Arghavan; Manhanzva, Monalisa T; Potgieter, Matthys; Balle, Christina; Bell, Liam; Ross, Elizabeth; Iranzadeh, Arash; du Plessis, Michelle; Radzey, Nina; McDonald, Zac; Calder, Bridget; Allali, Imane; Mulder, Nicola; Dabee, Smritee; Barnabas, Shaun; Gamieldien, Hoyam; Godzik, Adam; Blackburn, Jonathan M; Tabb, David L; Bekker, Linda-Gail; Jaspan, Heather B; Passmore, Jo-Ann S; Masson, Lindi
    Correction to: Microbiome 8, 165 (2020) https://doi.org/10.1186/s40168-020-00932-8
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    Disruption of maternal gut microbiota during gestation alters offspring microbiota and immunity
    (BioMed Central, 2018-07-07) Nyangahu, Donald D; Lennard, Katie S; Brown, Bryan P; Darby, Matthew G; Wendoh, Jerome M; Havyarimana, Enock; Smith, Peter; Butcher, James; Stintzi, Alain; Mulder, Nicola; Horsnell, William; Jaspan, Heather B
    Background: Early life microbiota is an important determinant of immune and metabolic development and may have lasting consequences. The maternal gut microbiota during pregnancy or breastfeeding is important for defining infant gut microbiota. We hypothesized that maternal gut microbiota during pregnancy and breastfeeding is a critical determinant of infant immunity. To test this, pregnant BALB/c dams were fed vancomycin for 5 days prior to delivery (gestation; Mg), 14 days postpartum during nursing (Mn), or during gestation and nursing (Mgn), or no vancomycin (Mc). We analyzed adaptive immunity and gut microbiota in dams and pups at various times after delivery. Results In addition to direct alterations to maternal gut microbial composition, pup gut microbiota displayed lower α-diversity and distinct community clusters according to timing of maternal vancomycin. Vancomycin was undetectable in maternal and offspring sera, therefore the observed changes in the microbiota of stomach contents (as a proxy for breastmilk) and pup gut signify an indirect mechanism through which maternal intestinal microbiota influences extra-intestinal and neonatal commensal colonization. These effects on microbiota influenced both maternal and offspring immunity. Maternal immunity was altered, as demonstrated by significantly higher levels of both total IgG and IgM in Mgn and Mn breastmilk when compared to Mc. In pups, lymphocyte numbers in the spleens of Pg and Pn were significantly increased compared to Pc. This increase in cellularity was in part attributable to elevated numbers of both CD4+ T cells and B cells, most notable Follicular B cells. Conclusion Our results indicate that perturbations to maternal gut microbiota dictate neonatal adaptive immunity.
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    The effect of HIV-exposure on immune responses to expanded programme on immunization vaccines and antigens
    (2013) Kidzeru, Elvis Banboye; Jaspan, Heather B; Gray, Clive M
    Immunization against vaccine-preventable infections is essential to reducing childhood morbidity and mortality. The immaturity and tolerogenicity of the immune system of infants renders them susceptible to infectious diseases and makes induction of protective immunity via vaccines a challenge. HIV-exposed infants are HIV uninfected and born to HIV-infected mothers and have increased morbidity and mortality of unknown aetiology. We hypothesise that T cells of HIV-exposed uninfected (HEU) infants have impaired proliferative ability and cytokine production in response to vaccine antigens than HIV unexposed (HU) infants.
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    Effects of delayed BCG vaccination on cellular immune responses in HIV-exposed infants
    (2014) Tchakoute, Christophe Toukam; Jaspan, Heather B; Gray, Clive
    Includes abstract. Includes bibliographical references.
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    The impact of myeloid derived suppressor cells on vaccine immunogenicity in South African HIV-infected and uninfected mothers and their infants
    (2016) Kidzeru, Elvis Banboye; Jaspan, Heather B
    BACKGROUND: Each year over 4 million infants die from infections, of which many are vaccinepreventable. Young infants respond poorly to vaccines, but the basis of reduced immunity is controversial. We hypothesized that myeloid-derived suppressor cells (MDSC) that might be induced during gestation, would persist at birth leading to active suppression of infant-immune responses. OBJECTIVE: We evaluated the ontogeny of MDSC and the effect of MDSC on vaccine immunogenicity during early life in South African infants and mothers, and in HIVexposed uninfected (HEU) infants and HIV+ mothers. METHODS: HIV-infected and uninfected mothers and their infants were recruited from Khayelitsha, Cape Town and followed-up for one year. In whole PBMC and after MDSC (CD15+) depleted, we measured BCG, Hepatitis B, Tetanus toxoid and Bordetella pertussis vaccine-specific CD4+ T cell proliferation by CFSE and IFN-γ responses using ELISpot assay.
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    In Silico Characterisation of Putative Prophages in Lactobacillaceae Used in Probiotics for Vaginal Health
    (2022-01-20) Happel, Anna-Ursula; Kullin, Brian R; Gamieldien, Hoyam; Jaspan, Heather B; Varsani, Arvind; Martin, Darren; Passmore, Jo-Ann S; Froissart, Rémy
    While live biotherapeutics offer a promising approach to optimizing vaginal microbiota, the presence of functional prophages within introduced Lactobacillaceae strains could impact their safety and efficacy. We evaluated the presence of prophages in 895 publicly available Lactobacillaceae genomes using Phaster, Phigaro, Phispy, Prophet and Virsorter. Prophages were identified according to stringent (detected by ≥4 methods) or lenient criteria (detected by ≥2 methods), both with >80% reciprocal sequence overlap. The stringent approach identified 448 prophages within 359 genomes, with 40.1% genomes harbouring at least one prophage, while the lenient approach identified 1671 prophages within 83.7% of the genomes. To confirm our in silico estimates in vitro, we tested for inducible prophages in 57 vaginally-derived and commercial Lactobacillaceae isolates and found inducible prophages in 61.4% of the isolates. We characterised the in silico predicted prophages based on weighted gene repertoire relatedness and found that most belonged to the Siphoviridae or Myoviridae families. ResFam and eggNOG identified four potential antimicrobial resistance genes within the predicted prophages. Our results suggest that while Lactobacillaceae prophages seldomly carry clinically concerning genes and thus unlikely a pose a direct risk to human vaginal microbiomes, their high prevalence warrants the characterisation of Lactobacillaceae prophages in live biotherapeutics.
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    Intra-and inter-clade cross-reactivity by HIV-1 Gag specific T-cells reveals exclusive and commonly targeted regions: implications for current vaccine trials
    (Public Library of Science, 2011) Zembe, Lycias; Burgers, Wendy A; Jaspan, Heather B; Bekker, Linda-Gail; Bredell, Helba; Stevens, Gwynneth; Gilmour, Jill; Cox, Josephine H; Fast, Patricia; Hayes, Peter
    The genetic diversity of HIV-1 across the globe is a major challenge for developing an HIV vaccine. To facilitate immunogen design, it is important to characterize clusters of commonly targeted T-cell epitopes across different HIV clades. To address this, we examined 39 HIV-1 clade C infected individuals for IFN-γ Gag-specific T-cell responses using five sets of overlapping peptides, two sets matching clade C vaccine candidates derived from strains from South Africa and China, and three peptide sets corresponding to consensus clades A, B, and D sequences. The magnitude and breadth of T-cell responses against the two clade C peptide sets did not differ, however clade C peptides were preferentially recognized compared to the other peptide sets. A total of 84 peptides were recognized, of which 19 were exclusively from clade C, 8 exclusively from clade B, one peptide each from A and D and 17 were commonly recognized by clade A, B, C and D. The entropy of the exclusively recognized peptides was significantly higher than that of commonly recognized peptides (p = 0.0128) and the median peptide processing scores were significantly higher for the peptide variants recognized versus those not recognized (p = 0.0001). Consistent with these results, the predicted Major Histocompatibility Complex Class I IC 50 values were significantly lower for the recognized peptide variants compared to those not recognized in the ELISPOT assay (p<0.0001), suggesting that peptide variation between clades, resulting in lack of cross-clade recognition, has been shaped by host immune selection pressure. Overall, our study shows that clade C infected individuals recognize clade C peptides with greater frequency and higher magnitude than other clades, and that a selection of highly conserved epitope regions within Gag are commonly recognized and give rise to cross-clade reactivities.
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    Probiotics for vaginal health in South Africa: what is on retailers’ shelves?
    (BioMed Central, 2017-01-19) Happel, Anna-Ursula; Jaumdally, Shameem Z; Pidwell, Tanya; Cornelius, Tracy; Jaspan, Heather B; Froissart, Remy; Barnabas, Shaun L; Passmore, Jo-Ann S
    Background: Probiotics are widely used to improve gastrointestinal (GI) health, but they may also be useful to prevent or treat gynaecological disorders, including bacterial vaginosis (BV) and candidiasis. BV prevalence is high in South Africa and is associated with increased HIV risk and pregnancy complications. We aimed to assess the availability of probiotics for vaginal health in retail stores (pharmacies, supermarkets and health stores) in two major cities in South Africa. Methods: A two-stage cluster sampling strategy was used in the Durban and Cape Town metropoles. Instructions for use, microbial composition, dose, storage and manufacturers’ details were recorded. Results: A total of 104 unique probiotics were identified in Cape Town and Durban (66.4% manufactured locally). Cape Town had more products than Durban (94 versus 59 probiotics), although 47% were common between cities (49/104). Only four products were explicitly for vaginal health. The remainder were for GI health in adults (51.0%) or infants (17.3%). The predominant species seen overall included Lactobacillus acidophilus (53.5%), L. rhamnosus (37.6%), Bifidobacterium longum ssp. longum (35.6%) and B. animalis ssp. lactis (33.7%). Products for vaginal health contained only common GI probiotic species, with a combination of L. acidophilus/B. longum ssp. longum/B. bifidum, L. rhamnosus/L. reuteri or L. rhamnosus alone, despite L. crispatus, L. gasseri, and L. jensenii being the most common commensals found in the lower female reproductive tract. Conclusion: This survey highlights the paucity of vaginal probiotics available in South Africa, where vaginal dysbiosis is common. Most vaginal products contained organisms other than female genital tract commensals
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    The role of polyclonal intravenous immunoglobulin in treating HIV-infected children with severe bacterial infections: A retrospective cohort study
    (BioMed Central Ltd, 2008) Huang, Lyen C; Myer, Landon; Jaspan, Heather B
    BACKGROUND:Mortality among HIV-infected children in developing countries remains high after serious bacterial infections despite the use of antibiotics. Intravenous immunoglobulin (IVIG) has been used as an adjuvant therapy to treat these infections, but little data exists regarding its efficacy, and previous studies have focused on IVIG as a prophylactic agent. We examined the impact of IVIG as an adjuvant therapy in reducing mortality and length of hospital stay in HIV-infected children with serious bacterial infections. METHODS: This retrospective study focused on pediatric admissions at a large urban hospital between 2002 and 2006. Children between the ages of one month and nine years of age with laboratory confirmed HIV-status, serious bacterial infection, no prior exposure to IVIG, and a hospital length of stay of 5 days or more, were eligible for inclusion. RESULTS: A total of 140 children (median age 1.2 years) met inclusion criteria; lower respiratory tract infection was diagnosed in 94 (67%) of the children, while 74 (53%) had bacterial sepsis. Fifty-four (39%) children were receiving antiretroviral therapy and 39 (28%) were receiving tuberculosis treatment. Overall 73 (52%) were treated with IVIG, with the majority (74%) of children receiving a single dose. Thirteen (9%) died during their hospital admission. In crude analysis IVIG was significantly associated with increased mortality was (Odds Ratio (OR): 5.8; 95% Confidence Interval (CI): 1.2-27.1) and this association was weakened by adjustment for other predictors of mortality (OR 4.3, 95% CI 0.7-27.9, p = 0.123). IVIG use was also associated with longer hospital stays. CONCLUSION: Administration of one to three doses of IVIG during the acute phase of illness does not appear to reduce mortality or the length of hospital stays in HIV-infected children with serious bacterial infections. However, the retrospective nature of this study makes confounding by indication difficult to control and further studies regarding the timing, dosing, and method of administration are required. Nonetheless the routine use of IVIG in resource-limited settings should be carefully considered given its high cost.
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    Vaginal microbial diversity of the genital tract of South African adolescent females
    (2017) Breetzke, Aerin Olivia; Passmore, Jo-Ann; Jaspan, Heather B; Lennard, Katie
    Young, reproductive-aged women are at highest risk of acquiring human-immunodeficiency virus (HIV). The Women's Initiative in Sexual Health (WISH) study was designed to investigate potential biological reasons for this high risk in HIV negative, South African adolescent females. Little is known about the 'normal' microbiome of this population. As such, the aim of this substudy was to quantify specific bacterial species (L. crispatus, L. jensenii, L. gasseri, L. iners, G. vaginalis and P. bivia) by quantitative real time PCR (qPCR) from adolescent female lateral vaginal wall swabs, and to assess associations between the quantities of these bacteria and bacterial vaginosis (BV) status, inflammation levels, age, hormonal contraceptive usage, and sexually transmitted infections (STIs). Samples were collected from 143 participant adolescent females in total, aged between 16 and 22 years of age, with a median of 18 years of age, from the Masiphumelele Youth Clinic in Cape Town, South Africa. Bacterial DNA was extracted from lateral vaginal wall swabs using the MoBio Powersoil® DNA Isolation Kit after enzymatic digestion. Positive bacterial reference strains were cultured in MRS buffer and Schwedler's broth, after which the DNA was extracted using the Qiagen Blood and Tissue DNA Maxi Extraction Kit. The quality and concentration of the DNA was confirmed using Qubit technology. The positive control DNA was amplified with PCR using species specific primers and the product run on an agarose gel to confirm primer specificity. The positive control DNA was serially diluted from 106 to 10-2 copies/μL to form a standard curve for absolute quantification through qPCR. Multiple steps were taken in order to optimize the qPCR experiments in terms of protocols, initial denaturation and annealing temperatures, cycle length and number, primers, and serial dilutions of the positive control DNA. The optimization for the P. bivia qPCR protocol presented the most issues, with the final quantification results being unreliable and requiring further work. Once the qPCR conditions were optimized for each bacterium; all samples, non-template control and standards were run in triplicate to quantify the number of bacterial copies per ng of DNA for each participant. The average of the three values were used as the final quantities and then used for downstream analyses. The bacterium L. crispatus, L. jensenii and L. gasseri, had median readings of 3.957 copies/ng, 1.568 copies/ng, and 17.58 copies/ng, respectively, with increased L. iners (2807 copies/ng) and G. vaginalis (8540 copies/ng). BV negative participants had increased levels of L. crispatus (p=0.0004, p=0.0002) and L. gasseri (p=0.0016, p<0.0001) in comparison to both BV intermediate and BV positive participants. L. jensenii (p<0.0001) and L. iners (p=0.0461) readings were increased in BV negative participants compared with BV positive and BV intermediate participants, respectively. BV positive participants had increased levels of G. vaginalis in comparison with both BV intermediate (p=0.0059) and BV negative (p<0.0001) adolescents. The 47 immunological factors, assessed via luminex, were categorized into high and low genital inflammation based on the unsupervised analysis by partitioning around medoids (PAM) using an R package 'cluster' with a k-value of 2. The inflammation-low group had increased levels of L. crispatus (p=0.0005), L. gasseri (p=0.033) and L. jensenii (p=0.0046) in comparison to the genital inflammation-high group. In participants with two viral STIs (Herpes Simplex Virus 2 and Human Papilloma Virus), there were increased copies/ng of G. vaginalis in comparison with participants with none (p=0.0098) or one viral STI (p=0.0324). Participants with high-risk HPV subtypes had significantly higher copy numbers of L. crispatus in comparison to the participants with low risk HPV subtypes (p=0.0181). Further, the only association demonstrated between the qPCR-based bacterial levels and the hormonal contraceptive prescribed was indicated by L. jensenii (ANOVA p=0.0222), possibly due to the low copy number readings. In conclusion, BV status, low levels of genital inflammation and the presence of two viral STIs indicate an association with bacterial copy numbers reported in this study, with increased median levels of L. iners and G. vaginalis across all adolescent participants compared to the other reported bacterial copy numbers. This indicates a possible alternate 'normal' microbiota profile of the FGT in adolescents in Masiphumelele.