Background: Studies on the impact of the skin microbiota on human health have been gaining more attention. Bacteria are associated with various diseases, although certain strains of bacteria, which are known as probiotics, are considered beneficial. Mixtures of several bacteria (bacterial cocktail) isolated from targeted organs have shown promising modulatory activities for use in skin therapeutics. The objectives of this study were to determine and identify the microbial communities on the skin that can potentially be used as probiotics, as determined by bacterial isolation and cultivation, followed by next-generation sequencing (NGS). Results: Samples were collected by swabbing on forehead and cheek skin. Genomic DNA from bacterial swab samples were directly extracted to be further processed into NGS. Cultivation of skin bacteria was carried out in subsequent medium. Thus, around twenty bacterial isolates with different characteristics were selected and identified by both culture-based method and 16sRNA sequencing. We found that Actinobacteria and Firmicutes are the most abundant phylum present on the skin as presented by NGS data, which constitute to 67% and 28.59% of the whole bacterial population, consecutively. However, Staphylococcus hominis, Staphylococcus warneri, and Micrococcus luteus (AN MK968325.1; AN MK968315.1; and MK968318.1 respectively) were able to be obtained in the samples of cultivable, and could be potentially developed as probiotics in skin microbiome therapeutic as well as for postbiotic formulation. Conclusion: Skin microbiome is considered to provide several probiotics for skin therapeutic. However, some opportunistic pathogens were discovered in this study population. Thus, the promising formula of bacterial cocktail for skin microbiome therapeutic must be thoroughly elucidated to avoid unwanted species. Our study is the first human skin microbiome profile of Indonesia resulted from a Next Generation Sequencing as an effort to show a representative of tropical country profile.
Objective: This study was conducted to optimize the genomic deoxyribonucleic acid (DNA) based molecular detection of gelatin derived from porcine by performing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and duplex PCR method employing cyt B gene.Methods: Optimization was carried out for DNA extraction, PCR conditions, and the sensitivity of the PCR-RFLP method. Due to the very low DNA trace in gelatin after the various manufacturing process, the extraction was optimized to obtain sufficient DNA which was visible on the agarose gel. PCR-RFLP was carried out using universal primers and BsaJI restriction enzyme, and duplex PCR was carried out using two sets of porcine-specific primers. Porcine and bovine DNA were mixed in various concentration to confirm sensitivity of both methods, i.e. 100%, 50%, 10%, 1%, 0.5%, 0.1%, 0.05%, and 0.01%Results: Both methods, PCR-RFLP, and Duplex PCR, were able to detect as low as 0.01% porcine DNA, indicated by the presence of porcine DNA amplicon bands (131 bp and 228 bp for PCR-RFLP, 212 bp and 398 bp for duplex PCR). Although DNA bands presented in low intensity, identification of porcine and bovine species and estimation of DNA quantities were possible.Conclusion: Both conventional PCR methods, i.e. PCR-RFLP and Duplex PCR, were sensitive, specific, and suitable as a rapid initial detection method for molecular detection of porcine in gelatin capsule shells.
Background: Postbiotic fractions of several lactic acid bacteria have potential as microbial therapeutics for skin health and may also appeal to consumers who wish to avoid animal-based products. We aim to establish the optimum plant-peptone fermentation of Streptococcus macedonicus MBF10-2, which possess Bacteriocin Like-Inhibitory Substance activity in our previous study, to produce bacterial bioactive fractions. We evaluate their potential antibacterial and antioxidant actions, and as well assess the preliminary safety for human skin application. Methods: Fermentation was carried out by using plant peptone modified MRS, i.e., soy peptone and Vegitone, a non-animal-carbon sources that substitute proteose peptone in MRS medium. Fractions of MBF10-2 lysate and cell-free supernatant were collected and processed as follows, i.e. cell disruption, fraction separation and fractions freeze-drying. Fractions were confirm for antibacterial properties by the agar well diffusion method and assess for antioxidant activity using DPPH, while safety assessment was carried-out by skin patch assay. Result: Maximum growth of MBF10-2 achieved by fermentation in soy peptone- and in Vegitone-modified media was 9.00 and 7.99 g total cell mass, respectively. The antibacterial property of fractions was most effective against Micrococcus luteus T18. The lysate fraction exhibited a mild antioxidant potency (IC50 840 µg/mL), and all bioactive fractions were proven safe and non-allergenic for human skins. Conclusion: Strep. macedonicus MBF10-2 postbiotics bioactive fractions were indicated as being safe for topical application. This is the first report on the production of a safe Strep. macedonicus bioactive postbiotic possessing mild antibacterial and mild-to-weak antioxidant. Keywords: antibacterial; antioxidant; lysate; soy peptone; Streptococcus macedonicus MBF 10-2
Studies on the impact of skin microbiota on human health have been gaining more attention. The skin microbiome is considered to provide several probiotics for skin therapeutic. Beneficial bacteria mixtures (bacterial cocktail) isolated from targeted organs have shown promising modulatory activities for use in skin therapeutics. The objectives of this study were to determine and identify the microbial communities on the skin that can be potentially used as probiotics-postbiotics. Determination and identification of skin microbiota were carried out simultaneously by employing next-generation sequencing (NGS) of direct sampling, as well as by bacterial cultivation; twenty bacterial isolates with different characteristics were selected and identified by both culture-based methods and 16sRNA sequencing. We found that Actinobacteria and Firmicutes are the most abundant phylum present on the skin as presented by NGS data, which constitute to 67% and 28.59% of the whole bacterial population, consecutively. Three strains, i.e., Staphylococcus hominis (AN MK968325.1), Staphylococcus warneri (AN MK968315.1), and Micrococcus luteus (MK968318.1), were obtained from cultivable samples. They were potential to be developed further as probiotics in skin microbiome therapeutic as well as for postbiotic formulation. However, the promising formula of bacterial cocktail for skin microbiome therapeutic must be elucidated thoroughly to avoid unwanted effects by performing visual observation on agar plate and by molecular approach, q-PCR.
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