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 Table of Contents  
ORIGINAL ARTICLE
Year : 2020  |  Volume : 8  |  Issue : 1  |  Page : 6-19

Lactobacillus pentosus KCA1 Decreases Vaginal and Gut Microbiota Associated with Bacterial Vaginosis


1 Uzobiogene Genomics, London, Ontario, Canada; Department of Medical Laboratory Sciences, Faculty of Health Sciences and Technology, College of Health Sciences, Nnamdi Azikiwe University, Nnewi, Anambra State, Nigeria
2 Uzobiogene Genomics, London, Ontario, Canada
3 Department of Obstetrics and Gynaecology, Nnamdi Azikiwe University Teaching Hospital, Nnewi, Anambra State, Nigeria

Date of Submission15-Nov-2019
Date of Decision04-Dec-2019
Date of Acceptance10-May-2020
Date of Web Publication31-Jul-2020

Correspondence Address:
Dr. Kingsley Chidozie Anukam
Uzobiogene Genomics, London, Ontario, Canada

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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/njecp.njecp_31_19

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  Abstract 


Introduction: Bacterial vaginosis (BV) which affects 14%–50% of reproductive-aged women in Nigeria is misdiagnosed and underreported. Treatment option is antibiotics, which leads to recurrent infections. The objectives of this study are three folds: first, to determine effects of oral feeding of Lactobacillus pentosus KCA1 on the vaginal and gut microbiota of women diagnosed with BV; second, to measure the level of two pro-inflammatory cytokines interleukin-1 (IL-1)-beta and IL-6 before and after KCA1 consumption; and third, to determine the relative abundance of bacterial metabolic genes. Materials and Methods: Seven women diagnosed with BV by Nugent score (7–10) were recruited to provide vaginal and gut sample before and after 14-day oral intake of 3 g of L. pentosus KCA1. The DNA from the swabs was processed for 16S rRNA metagenomics using Illumina MiSeq platform. The paired-end sequence FASTQ reads were imported into Illumina Basespace pipeline for quality check. In addition, EzBioCloud pipeline was used for alpha- and beta-diversity estimation using PKSSU4.0 version and open reference UCLUST_MC2 for operational taxonomic units picking at 97% cutoff. Blood samples were analyzed using ELISA technique. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States was used to predict the metabolic functions from the 16S rRNA gene dataset. Results: On average, there was no significant difference at P = 0.05 in the alpha-indices typified by Shannon index. The beta-diversity showed different clustering positions with principal coordinate analysis. However, at individual taxonomic categories, there was a significant decrease in the relative abundance of some genera associated with BV after KCA1 feeding with a corresponding increase of Lactobacillus genus. Bacterial genes related to defense systems were upregulated in the vagina. There was a two-fold downregulation of IL-1-beta after consumption of KCA1. Conclusion: Our findings suggest that L. pentosus KCA1 taken orally lowers pro-inflammatory cytokine and IL-1-beta and decreases the relative abundance of BV-associated bacteria.

Keywords: Bacterial vaginosis, gut, Lactobacillus pentosus KCA1, microbiome, probiotics, vagina


How to cite this article:
Anukam KC, Agbakoba NR, Ejike CE, Oguejiofor CB. Lactobacillus pentosus KCA1 Decreases Vaginal and Gut Microbiota Associated with Bacterial Vaginosis . Niger J Exp Clin Biosci 2020;8:6-19

How to cite this URL:
Anukam KC, Agbakoba NR, Ejike CE, Oguejiofor CB. Lactobacillus pentosus KCA1 Decreases Vaginal and Gut Microbiota Associated with Bacterial Vaginosis . Niger J Exp Clin Biosci [serial online] 2020 [cited 2020 Aug 8];8:6-19. Available from: http://www.njecbonline.org/text.asp?2020/8/1/6/291198




  Introduction Top


Bacterial vaginosis (BV) is underreported, misdiagnosed, and inappropriately treated in Nigeria.[1] Our recent study reported that a large proportion of women (45.1%) assumed that vaginal discharge is the most common symptom associated with BV and 41.7% stated that BV recurrence is a common denominator in their lives. On the social burden of BV, 53.7% of women indicated that BV makes them avoid having sex while 29.5% avoid closeness with people, yet Nigerian clinicians still rely on antibiotics.[2] Antibiotics kills both the healthy bacteria, especially lactobacilli and those associated with BV, thus altering the vaginal and gut microecology.[3] It is widely acknowledged that lactobacilli and other genera that produce some metabolites such as lactic acid which contributes to vaginal health by maintaining the low pH of the vagina.[4] Although the etiology of BV is still unresolved, a recent metagenomics study has provided an insight into the phylogenetic diversity and species richness of the vaginal and gut microbiome of reproductive-aged women with BV in some selected Nigerian women.[5] The polymicrobial nature of BV is also believed to induce a pro-inflammatory environment, consisting of cytokines and toll-like receptor ligands.[6] In Nigeria, clinicians treat BV with antibiotic combinations, especially metronidazole or clindamycin. However, the recurrence rate remains high. Failure of the antibiotics and recurrences common in BV are attributed to various reasons such as drug resistance and failure of the agents to penetrate and eradicate biofilms formed by BV-associated organisms.[7] The concept of replenishing the vaginal microbiota can be traced back to antiquity, but it was reinvigorated four decades ago by delivering lactobacilli orally to repopulate the vagina.[8] A potential probiotic organism, Lactobacillus pentosus strain KCA1, was isolated for the first time from the vagina of a healthy Nigerian woman. Some microbiologists have scaled up their decades of research on few vaginal Lactobacillus strains as probiotics; thus, L. pentosus KCAI was shown to produce biosurfactants and hydrogen peroxide (H2O2) and inhibit the intestinal and urogenital pathogens[9] as well as exhibit varying degrees of acid bile tolerant.[10]

In order to establish the genetic capability and functional potentials, the full-genome sequence of L. pentosus KCA1 was determined in our previous study.[11]L. pentosus KCA1 was found to dedicate 121 genes to metabolism of cofactors and vitamins including five genes for biotin biosynthesis. The L. pentosus KCA1 genome also encodes putative phage defense systems including clustered regularly interspaced short palindromic repeats (CRISPRs) and abortive infection, novel toxin–antitoxin systems, and biosynthesis of an antibacterial peptide, a Class V cyclic bacteriocin precursor, designated as pentocin KCA1 (penA). There are five complete two-component systems in KCA1 which includes histidine kinase hpk1 (KCA1_0030) and response regulator rrp1 (KCA1_0029) and two-component response regulator TrxR (KCA1_2843), a transcriptional regulator of the AraC family.

There have been multiple studies showing significant improvements in treating vaginal infections with probiotic versus traditional antibiotic treatments, but so far, only a few strains have been clinically proven to be effective in particular to prevent BV, and no study has been done to determine the impact of a single Lactobacillus strain on the vaginal and gut microbiome. It is important to note that Lactobacillus strains are specific in their action according to Shanahan[12] and Van Barlen et al.,[13] denoting that one should not utilize data from one strain to infer to another untested strain that will provide the same benefit. It is critically important that L. pentosus KCA1, which has been characterized at molecular level, should be tested clinically for probiotic use.

The genome provides a basis for its application to maintain a balanced urogenital health, especially for women diagnosed with BV. The objectives of this study are three folds: first, to determine the effects before and after 14 days oral feeding of L. pentosus KCA1 on the vaginal and gut microbiota compositions of women diagnosed with BV; second, to measure the level of two pro-inflammatory cytokines interleukin-1 (IL-1) beta and IL-6 before and after KCA1 consumption; and third, to determine in silico, the relative abundance of microbial genes involved in metabolic functions using the 16S rRNA dataset.


  Materials and Methods Top


Ethical approval

Ethical approval was obtained from the Research Ethics Review Board of the Faculty of Health Science and Technology, Nnamdi Azikiwe University and General Hospital, Onitsha, in accordance with the Helsinki Declaration of 1975, as revised in 2000 (available at http://www.wma.net/e/policy/17-c_e.html).

Study location and population

This study was conducted at General Hospital Onitsha, Anambra State. This hospital is the only government hospital at Onitsha and serves the neighboring communities. One hundred and twenty women of reproductive-aged group who visited a clinic at General Hospital Onitsha (GHO) were recruited for this study. The individuals were randomly selected after informed consent.

Inclusion criteria are individuals with age range 14–49 years, not on antibiotic therapy within the last 1 month, and not menstruating at the time of inclusion. Individuals were excluded if found to be pregnant, use of antibiotic medication, declined informed consent, menstruating, and those that used douches, sprays, and spermicides in the last 48 h.

Collection of specimen

Two high vaginal swabs were collected by a qualified gynecologist with a nonlubricated sterile disposable plastic speculum, and samples were collected from the posterior vaginal fornix using a Dacron swab stick.

Microscopy and Nugent score

BV was determined using Gram staining technique and Nugent criteria. One set of the high vaginal swab was used to prepare a dry vaginal smear which was stained by the Gram staining technique. The slides were examined using the × 100 (oil immersion objective) and scored for BV using Nugent criteria. The slides were observed for the presence of four different bacterial morphotypes: large Gram-positive rods (Lactobacillus spp. morphotypes), small Gram-variable rods (Gardnerella vaginalis morphotypes), small Gram-negative rods (Bacteroides spp. morphotypes), and curved Gram-variable rods (Mobiluncus spp. morphotypes).

For Lactobacillus, scores range from 0 to 4, where 0 indicates that 30 or more organisms are identified and 4 indicates that no organisms are identified in the sample. For Gardnerella, however, a score of 0 indicates that no organisms are identified and the highest score of 4 indicates that 30 or more organisms are identified. For Mobiluncus, scores range from 0 to 2 with a score of 2, indicating that 5 or more organisms are identified in the sample. At the end of scoring process, total scores ranging from 0 to 10 was computed. A total score of 7 or more was termed a case of BV; a score of 4–6 was considered intermediate while a score of 0–3 was considered normal as standardized by Nugent et al.[14]

Oral administration of Lactobacillus pentosus KCA1

Seven women diagnosed with BV by Nugent score (7–10) were recruited to provide vaginal and gut sample before and after 14-day oral intake of 3 g (2.5×108 cfu/g) of L. pentosus KCA1 (Winclove Probiotics, Amsterdam, The Netherlands) suspended in lukewarm water or dairy and taken daily for 14 days. Blood samples were also collected before and after 14 days of oral consumption of L. pentosus KCA1.

Extraction of bacterial DNA from vaginal swabs/stool samples and sequencing of the amplified 16S rRNA region

Bacterial DNA was extracted from the vaginal swabs using an in-house protocol developed by uBiome Inc. Briefly, samples were lysed using bead beating, and DNA was extracted in a class 1000 cleanroom by a guanidine thiocyanate silica column-based purification method using a liquid-handling robot. Polymerase chain reaction (PCR) amplification of the 16S rRNA genes was performed with primers containing universal primers amplifying the V4 region (515F: GTGCCAGCMGCCGCGGTAA and 806R: GGACTACHVGGGTWTCTAAT). In addition, the primers contained Illumina tags and barcodes. Samples were barcoded with a unique combination of forward and reverse indexes allowing for simultaneous processing of multiple samples. PCR products were pooled, column-purified, and size-selected through microfluidic DNA fractionation. Consolidated libraries were quantified by quantitative real-time PCR using the Kapa Bio-Rad iCycler qPCR kit on a BioRad MyiQ before loading into the sequencer. Sequencing was performed in a pair-end modality on the Illumina NextSeq 500 platform rendering 2 × 150 bp pair-end sequences.

Metagenomics sequence analysis

Raw sequence reads were demultiplexed using Illumina's BCL2FASTQ algorithm. Reads were filtered using an average Q-score >30. The paired-end sequence FASTQ reads were imported into MG-RAST pipeline for quality check. EzBioCloud microbiome taxonomic profile (MTP) pipeline[15] was employed for alpha- and beta-diversity estimation using PKSSU4.0 version database and open reference UCLUST_MC2 for operational taxonomic units (OTUs) picking at 97% cutoff. Sequences were prescreened using QIIME-UCLUST algorithms for at least 97% identity to ribosomal sequences from the RNA databases.[16] Rarefication to 1000 reads per sample was employed to calculate microbial diversity. Alpha-diversity was calculated for species richness by Abundance Coverage Estimate (ACE), Chao1, and Jackknife methods, while diversity indexes were calculated by Shannon, nonparametric Shannon, and Simpson indexes. Principal coordinate analysis with Jensen–Shannon divergence distance metrics were used to evaluate beta-diversity between vaginal and gut samples.[17] Linear discriminant analysis effect size (LEfSe)[18] was used to identify biologically and statistically significant differences in the OTU relative abundance. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) was used to predict the metabolic function of the metagenomes from the 16S rRNA gene dataset,[19] with reference to the Kyoto Encyclopedia of Genes and Genomes ortholog categorizations.[20] Blood samples for cytokines were also collected before and after KCA1 consumption and processed with ELISA technique.

Statistical analysis

Student's t-test as obtained in Excel 2019 Office was used to test differences at P < 0.05 in the proportion of microbiota before and after oral intake of L. pentosus KCA1.


  Results Top


Vaginal Microbiome composition before and after consumption of Lactobacillus pentosus KCA1

The 16S rRNA data sets comprising data before and after consumption of L. pentosus KCA1 were compared using the Greengenes Version 13_8 (ftp://greengenes.microbio.me/greengenes_release/gg_13_5/gg_13_8_otus.tar.gz) in Illumina Basespace algorithm, as shown in [Figure 1]. At the phylum taxonomic categories, before consumption of L. pentosus KCA1, 22 phyla were identified. Firmicutes (47.60%) were the most abundant, followed by Bacteroidetes (27.39%), Actinobacteria (17.10%), Fusobacteria (6.03%), Tenericutes (1.38%), Proteobacteria (0.44%), Acidobacteria (0.01%), and others, as shown [Figure 2]. After 14-day consumption of L. pentosus KCA1, 22 phyla were also identified with obliteration of two phyla (Armatimonadetes and Planctomycetes) and introduction of two new phyla (Elusimicrobia and Nitrospirae). Firmicutes (46.68%) were the most abundant, followed by Bacteroidetes (26.31%), Actinobacteria (18.01%), Fusobacteria (7.29%), Tenericutes (0.97%), Proteobacteria (0.66%), Acidobacteria (0.01%), and others, as presented in [Figure 2].
Figure 1: Distribution of total taxonomic categories in bacterial vaginosis patients before and after KCA1 consumption

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Figure 2: Distribution of phylum taxonomic categories identified in bacterial vaginosis patients before and after Lactobacillus pentosus KCA1 consumption

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In addition, EzBioCloud MTP was used to determine alpha-diversity including species richness and diversity index. On average, there was no significant difference (P = 0.749) in the species richness-alpha indices typified by ACE, Chao1, Jackknife, and number of OTUs. Diversity indices such as NPShannon and Shannon (P = 0.749) were not significant. Similarly, Simpson and phylogenetic diversities (P = 0.565) were not significant before and after KCA1 consumption [Figure 3]. The beta-diversity showed different clustering positions with Bray–Curtis principal coordinate analysis [Figure 4]. However, at individual taxonomic categories, there was a significant difference in the relative abundance of the phylaProteobacteria and Actinobacteria and some genera associated with BV observed before and after KCA1 feeding. For example, there was a 23.29% decrease in the genus Gardnerella, Porphyromonas (26.39%), Prevotella (16.1%), Gemella (52.07%), Veillonella (54.17%), Atopobium (43.91%), Ureaplasma (56.11%), and Peptostreptococcus decreased by 59.72% with a corresponding 56.09% increase of Lactobacillus genus [Figure 5].
Figure 3: Alpha diversity index showing the species richness as determined by EzBioCloud microbiome taxonomic profile pipeline

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Figure 4: Beta-diversity showing different clustering positions before and after Lactobacillus pentosus KCA1 with Bray–Curtis principal coordinate analysis

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Figure 5: Vaginal bacteriome genus compositions before and after oral consumption of Lactobacillus pentosus KCA1

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At the species taxonomic level, some bacterial organisms associated with BV decreased with higher proportions, as shown in [Figure 6]. Interestingly, 113 (37%) species were exclusively identified in the vagina of the participants after KCA1 consumption. Notable species found after KCA1 were L. pentosus, Lactobacillus reuteri, Lactobacillus crispatus, Lactobacillus gasseri, Blautia wexlerae, Roseburia cecicola, Roseburia inulinivorans, and Faecalibacterium spp.
Figure 6: Selected vaginal bacteriome species before and after oral consumption of Lactobacillus pentosus KCA1

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Gut microbiome composition before and after consumption of Lactobacillus pentosus KCA1

The Gut microbiome before and after oral feeding of KCA1 rarefaction curve is presented in [Figure 7]. Microbiota composition at the genus taxonomic level shows that L. pentosus KCA1 led to an increase in the proportion of some genera. For example, there was a 41.32% increase in Faecalibacterium, Bacteroides (29.94%),  Escherichia More Details (64.36%), Clostridium (3.93%), Parasutterella (3.97%), Ruminococcus (1.86%), Streptococcus (2.69%), Lactobacillus (3.20%), Enterococcus (0.22%), Akkermansia (0.17%), and Butyricimonas (0.74%). Conversely, there was a decrease in the relative abundance of some genera associated with BV [Figure 8]. Alloprevotella decreased by 18.05%, Prevotella (1.6%), Haemophilus (8.08%), Intestinibacter (2.63%), Clostridium_g24 (0.19%), Coprococcus (5.13%), Prevotellaceae_uc (0.64%), Fusicatenibacter (8.33%), Megasphaera (0.54%), Porphyromonas (0.16%), Veillonella (0.35%), Actinomyces (0.02%), Peptostreptococcus (0.12%), Streptococcaceae_uc (0.15%), Acinetobacter (2.8%), Anaerococcus (0.04%), Aeromonas (0.17%), Eggerthella (0.05%),  Neisseria More Details (0.05%), Campylobacter (0.02%), Staphylococcus (0.02%), Clostridium_g21 (0.08%), Atopobium (0.02%), and Gardnerella (0.09%). Some genera that were identified before oral intake of KCA1 appears to be absent after KCA1 consumption. The Venn diagram plot shows that 67.2% of the genera were commonly present before and after, while 18.7% were exclusively found after KCA1 oral intake and 14.2% were exclusively found pre-KCA1 consumption [Figure 9]. Such genera include but not limited to Achromobacter, Actinobacillus_g1, Actinobaculum, Aggregatibacter, Alcaligenes, Alishewanella, Anaerofustis, Anaerotaenia, Arcobacter, Burkholderia, Mobiluncus, Pediococcus, Pelomonas, Peptococcus, Slackia, Sphingobacterium, Ureaplasma, Vallotia, and Vibrio. Some genera that occurred below 1% relative abundance are presented in [Figure 10].
Figure 7: Rarefaction curve showing the number of reads from gut samples associated with the number of operational taxonomic units

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Figure 8: Gut relative abundance genus (above 1%) before and after consumption of Lactobacillus pentosus KCA1

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Figure 9: Venn diagram showing the number (%) of genus and species identified in the gut that were common and exclusive before and after oral intake of Lactobacillus pentosus KCA1

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Figure 10: Gut relative abundance genus (below 1% cutoff) before and after consumption of Lactobacillus pentosus KCA1

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Metabolic functional prediction using the 16S rRNA marker genes

The results from PICRUSt indicated that certain bacterial metabolic functional genes were upregulated in the vagina after consumption of L. pentosus KCA1. For example, metabolic pathways for amino acid, alanine, were significantly upregulated (P = 0.008809) with LEfSe of 3.445098. Similarly, polar amino acid transport system substrate-binding protein with gene ortholog K02030 was significantly increased (LEfSe = 2.364761 and P = 0.035006) [Figure 11]. There was a significant difference in the predicted relative abundance of microbial gene pathways related to degradation of aromatic compounds (ko01220), with LEfSe = 2.571179 and P = 0.047645. There was also a high abundance of two-component system sensor histidine kinase; two-component system, OmpR family, lantibiotic biosynthesis sensor histidine kinase NisK/SpaK, and redox-sensing transcriptional regulator. There were acquisitions of bacterial metabolic genes related to defense systems such as CRISPR-associated protein Cmr1, and DNA mismatch repair protein MSH3, after consumption of KCA1 in the vagina [Table 1].
Figure 11: Bacterial metabolic functional genes upregulated in the vagina after oral intake of Lactobacillus pentosus KCA1

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Table 1: Bacterial metabolic functional genes predicted with Phylogenetic Investigation of Communities by Reconstruction of Unobserved States using EzBioCloud pipeline

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Several bacterial metabolic functions in the gut were similarly affected. IL-15 receptor-alpha was downregulated significantly at P = 0.025347, and methicillin resistance protein was also downregulated (P = 0.047645). Conversely, several microbial functional cofactors such as menaquinone biosynthesis pathway, Vitamin B6 (ko00750) metabolism, biotin (ko00780) metabolism, propanoate (ko00640), and lipoic acid (ko00785) metabolisms were upregulated [Figure 12].
Figure 12: Bacterial metabolic functional genes upregulated in the gut after oral intake of Lactobacillus pentosus KCA1

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Interleukin-1-beta

There was a two-fold downregulation of IL-1-beta in contrast to IL-6 after consumption of KCA1 [Figure 13].
Figure 13: Box plot of the serum interleukin-1-beta and interleukin-6 before and after 14 days oral intake of Lactobacillus pentosus KCA1

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  Discussions Top


In this study, we determined the composition of the vaginal and gut microbiota of women diagnosed with BV before and after 14-day consumption of L. pentosus KCA1. In our previous study, we used metagenomics approach to decipher the composition of vaginal and gut microbiota of a small cohort of women diagnosed with BV. The study revealed the polymicrobial nature, phylogenetic diversity, and species richness of the vaginal and gut microbiome of reproductive-aged women diagnosed with BV.[5] This present study is consistent with our previous study as the vaginal samples indicated fewer phyla in contrast to the gut. The consumption of KCA1 did not induce any significant difference in the species richness-alpha indices typified by ACE, Chao1, Jackknife, and number of OTUs. However, at individual taxonomic categories, L. pentosus KCA1 led to a decrease in the proportion of bacterial organisms associated with BV with a corresponding improvement in Nugent score typified by increase in lactobacilli. It should be noted that in healthy women, the vagina is colonized predominantly by lactobacilli as we demonstrated in our previous study whereby the vagina of healthy, non-BV Nigerian women was colonized by Lactobacillus-dominated bacterial communities, in addition to the presence of other lactic acid-producing bacteria as revealed by 16S rRNA metagenomics.[21] In a meta-analysis review, there was a statistically significant beneficial effect of probiotics observed in European populations after short-term follow-up days.[22] The decrease of most of the bacteria associated with BV such as G. vaginalis in the vagina after consumption of KCA1 and the corresponding increase of the genera lactobacilli suggest that KCA1 may be disrupting the bacterial biofilm associated with G. vaginalis survival.[23],[24]L. pentosus KCA1 is known to produce lactic acid and genes for the production of H2O2 and bacteriocins are encoded in the genome,[11] of which several studies have shown to affect G. vaginalis.[25] A recent study demonstrated the use of Lactobacillus strains against two different pathogens (G. vaginalis and Atopobium vaginae) responsible for BV.[26] Restoration of vaginal microbiota after insertion of a vaginal tablet has been reported.[27],[28] The decrease of the BV-associated genera (Porphyromonas, Prevotella, Gemella, Veillonella, Atopobium, Ureaplasma, and Peptostreptococcus) suggests that L. pentosus KCA1 influences multigenus metabolic activities. For example, a study reported that certain strains of Lactobacillus in the vagina were found to suppress the epithelial inflammatory response to Atopobium, linked to BV.[29] The identification of some species not found before L. pentosus KCA1 may indicate the possibility of cellular crosstalk and quorum sensing, thereby stimulating other related dormant cells to proliferate.[30]

While L. pentosus KCA1 led to decrease in BV-associated organisms in the vagina, it also decreased some BV-associated organisms in the gut such as Prevotella. Conversely, it stimulated the proliferation of some bacteria known to be associated with health. For example, there was a 41.32% increase in Faecalibacterium, especially Faecalibacterium prausnitzii, which has been found to produce high amounts of butyrate and anti-inflammatory compounds.[31] In the same vein, Akkermansia muciniphila proportion was elevated. It should be noted that Akkermansia is involved in metabolic functions and immune regulations and has been proposed as a promising probiotic.[32]

The in silico metabolic functional predictions indicated upregulation of some functions suggesting that L. pentosus KCA1 impacted the microbial activities. Intriguingly, the acquisition of microbial defense systems after consumption of KCA1 such as CRISPRs is an indication that KCA1 can modulate other bacterial functions. The genome of L. pentosus KCA1 encodes six CRISPR arrays with direct repeat length consensus exhibiting repeat polymorphisms.[11],[33]

The downregulation of IL-15 by L. pentosus KCA1 in the gut microbes is noteworthy, in that the cytokine correlates with adiposity and markers of the metabolic syndrome. Genome-wide association has identified the gene encoding this receptor, IL15RA, which resides also on human chromosome 10p, a location linked to obesity and type 2 diabetes.[34] The upregulation of menaquinone biosynthesis pathway and Vitamin B6 suggest that the consumption of L. pentosus KCA1 has health attributes beyond the vagina. In addition to these attributes, KCA1 has a direct effect on the human host as it downregulated serum cytokine IL-1-beta, such suggesting modulation of inflammatory processes.


  Conclusion Top


Our findings suggest that L. pentosus KCA1, taken orally, contributes positively to lowering pro-inflammatory cytokines, especially IL-1-beta, with a significant decrease in the relative abundance of BV-associated bacteria and a corresponding increase of Lactobacillus genus. Increased metabolism of Vitamin B6, biotin, lipoic acid, and propanoate was observed in the gut. Bacterial genes related to defense systems were upregulated in the vagina. The predicted metabolic functions indicate that some pathways in the vaginal microbial genes such as glycerol phosphoryl diester phosphodiesterase (K01126), mRNA interferase MazF (K07171), two-component system sensor histidine kinase, disease resistance protein (K13446), and transposase IS30 family (K07482) were significantly upregulated after consumption of L. pentosus KCA1.

Financial support and sponsorship

We sincerely thank uBiome Inc., San Francisco, California, USA, for awarding a grant-in-kind to Dr. Kingsley Anukam and for carrying out the metagenomics sequencing. We also thank Winclove Probiotics, Amsterdam, The Netherlands, for providing Lactobacillus pentosus KCA1 in powder form used for this study.

Conflicts of interest

Dr. Kingsley C Anukam isolated and sequenced the full genome of Lactobacillus pentosus KCA1 but has licensed the strain to Winclove Probiotics in The Netherlands.



 
  References Top

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