Morphology, antibacterial, and molecular analysis using the RAPD method on bamboo in Bengkulu, Indonesia

Event Abstract Back to Event Isolation and antimicrobial activity of probiotic strains for caries prevention Nor Zaihana Abdul Rahman1*, Rohazila Mohd Hanafiah1, Siti Aisyah Abdul Ghafar1 and Nurul Iffah Abdul Rashid1 1 Department of Basic Sciences & Oral Biology, Faculty of Dentistry, Universiti Sains Islam Malaysia, Malaysia Background Probiotics can be described as live microorganism which when consumed in adequate amounts, it will confer health benefits to the host. The benefits arising from probiotics has been mostly shown on gut health. However, its function to oral health is still poorly investigated. The aim of this study was to isolate lactic acid bacteria (LAB) from local fermented food and to determine its antibacterial activity, in order to select candidate probiotic strains for preventing caries. Methods The probiotic strains were isolated from five local fermented food which are tapai ubi, tapai pulut, rebung, thosai and taucu. Ten-fold serial dilutions of the fermented food samples were made in sterile peptone water before plating on de Man Rogosa and Sharpe (MRS) agar. The pure cultures were then randomly picked and biochemically identified. Then, the antibacterial activity of LAB against Streptococcus mutans were assessed by using disk diffusion method. Results A total of 120 LAB were isolated from five different fermented foods. The morphology of the isolates were circular, convex, dull opaque white or translucent white. Gram-staining identification showed that the isolates were Gram-positive rods. Of the 120 LAB isolates, 6 strains displayed moderate to strong antibacterial activity against S. mutans with the inhibition zones ranging from 7-12 mm. Conclusion The antibacterial activity demonstrated from LAB isolated from local fermented foods suggests that the isolates from local fermented foods possess antibacterial properties against pathogen responsible for causing dental caries. These strains are currently investigated in depth to assess whether they can be fully characterized as probiotics. Keywords: Probiotics, Lactic acid bacteria, Dental Caries, Streptococcus mutans, Oral Health Conference: International Conference on Drug Discovery and Translational Medicine 2018 (ICDDTM '18) “Seizing Opportunities and Addressing Challenges of Precision Medicine”, Putrajaya, Malaysia, 3 Dec - 5 Feb, 2019. Presentation Type: Poster Presentation Topic: Miscellaneous Citation: Abdul Rahman N, Mohd Hanafiah R, Abdul Ghafar S and Abdul Rashid N (2019). Isolation and antimicrobial activity of probiotic strains for caries prevention. Front. Pharmacol. Conference Abstract: International Conference on Drug Discovery and Translational Medicine 2018 (ICDDTM '18) “Seizing Opportunities and Addressing Challenges of Precision Medicine”. doi: 10.3389/conf.fphar.2018.63.00103 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 19 Oct 2018; Published Online: 17 Jan 2019. * Correspondence: Dr. Nor Zaihana Abdul Rahman, Department of Basic Sciences & Oral Biology, Faculty of Dentistry, Universiti Sains Islam Malaysia, Nilai, Malaysia, zaihana@usim.edu.my Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Nor Zaihana Abdul Rahman Rohazila Mohd Hanafiah Siti Aisyah Abdul Ghafar Nurul Iffah Abdul Rashid Google Nor Zaihana Abdul Rahman Rohazila Mohd Hanafiah Siti Aisyah Abdul Ghafar Nurul Iffah Abdul Rashid Google Scholar Nor Zaihana Abdul Rahman Rohazila Mohd Hanafiah Siti Aisyah Abdul Ghafar Nurul Iffah Abdul Rashid PubMed Nor Zaihana Abdul Rahman Rohazila Mohd Hanafiah Siti Aisyah Abdul Ghafar Nurul Iffah Abdul Rashid Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

<em><span>Data collected by WHO in 2019 shows an estimated 17.9 million (equivalent to 32% of global death). The total number is due to cardiovascular disease, 85% were due to heart attacks and strokes. One of the basic pathophysiologies of stroke is atero-thromboembolic (thrombosis). The accumulation of clots formed by fibrin in blood vessels can inhibit several processes in the body system, such as blood flow, oxygen transportation, and transportation of nutritional and non-nutritional compounds needed. Fibrinolytic enzymes from food microbes have attracted attention for further research as thrombolytic agents. The Bacillus genus from fermented foods can produce strong fibrinolytic enzymes, such as Bacillus natto from natto (Japan), Bacillus subtilis KCK-7 from Chungkook-jang (Korea), Bacillus weihenstephanensis from shrimp paste (Vietnam), Bacillus amyloliquefaciens MH18B1 from Hawaijar (India), Rhizopus chinensis 12 from Chinese wine (China), and Bacillus pumillus and Bacillus subtilis K2 from fermented soybean food products such as tempe gembus, red oncom, and moromi (Indonesia). The high protein content in soybeans can be used as a medium for the growth of proteolytic and fibrinolytic microorganisms. Studies in Asia regarding fibrinolytic enzymes from fermented foods are still limited, even though the Asian region comprises many countries rich in fermented foods. This review is relevant to healthy food ingredients from local fermented foods. Recently, microbial fibrinolytic enzymes from fermented foods have received tremendous interest in the medical field. Many local fermented foods are a potential source of fibrin-degrading enzymes, which can be further developed to treat diseases like stroke and heart attack. This article aims to review microbial fibrinolytic enzymes from various fermented foods and their impact on health, especially thrombosis. This manuscript review uses the narrative review method. From the overall discussion, it can be concluded that fermented foods have properties as fibrinolytic agents.</span></em>

Probiotics are live microorganisms acclaimed to provide health benefits when consumed and are generally considered safe for consumption. The lactic acid bacteria (LAB) are common groups of bacteria with certain health benefits, including the ability of some strains to play probiotic roles. Some indigenous fermented foods localized in Ado-Ekiti carry LAB that may have some probiotic properties. The aim of this study was to determine the probiotic potential of some LAB isolated from some local fermented foods in Ado- Ekiti. Samples of locally fermented foods: “garri”, “iru”, “ogi”, and “fura de nono” were obtained from the local market in Ado-Ekiti for isolation of LAB. All samples were cultured using standard methods. A total of sixteen (16) isolates were recovered from the samples and screened for their antibacterial activity against four human pathogenic bacterial strains which were Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, and Staphylococcus aureus. The LAB with enhanced antibacterial activity were further screened for potential probiotic characteristics using pH tolerance test, bile salt tolerance test, NaCl tolerance test, temperature sensitivity, and lactose utilization. The LAB isolates identified from the local food samples included: Lactobacillus fermentum, Acetobacter spp., Bacillus subtilis, Lactobacillus plantarum, and Lactococcus lactis. Out of the 16 strains obtained, five strains that showed considerable antibacterial activity were evaluated for their probiotic potential. The five strains were susceptible to conventional antibiotics. It was also shown that the five strains were able to grow at the selected range of pH 4.0 – 7.0, they were able to survive at the temperature ranges between 20 oC and 45 oC, they were lactose-tolerant, and they were able to tolerate 1 – 5% sodium chloride (NaCl) concentration. Some isolated LAB exhibited excellent probiotic characteristics and thus can be recommended as a potential use as probiotics. The findings of this study suggest that popular local foods in Ado Ekiti, such as “garri”, “iru”, “ogi”, and “fura de nono”, harbor some LAB with potential probiotic properties, and can be explored for possible health benefits.

Probiotic microorganisms, offering health benefits when consumed in sufficient quantities, are gaining recognition for their potential role in promoting wellness. This study focuses on isolating and characterising potential probiotic yeast strains sourced from fermented food products. This research evaluates the gastrointestinal tolerance and antimicrobial activity of isolated yeast strains, with the potential application in probiotic supplements and functional foods. Yeast strains were isolated from fermented food sources and identified using morphological analysis, PCR, gene sequencing, and genetic identification. Gastrointestinal tolerance was assessed through simulated gastric fluid (SGF) exposure, and antimicrobial activity was tested against foodborne pathogens. Six yeast strains (Diutina mesorugosa, Pichia kudriavzevii, Candida mesorugosa, Candida sp) were identified. They exhibited varying resistance to low pH in SGF, suggesting survivability in the stomach. Some strains selectively inhibited specific Gram-negative pathogens like Pseudomonas aeruginosa and Salmonella sp. These findings suggest the isolated yeast strains may serve as probiotics, promoting digestive health and food safety. They are potentially used in probiotic supplements and functional foods, promising improved overall well-being.

Aging has become one of the most important health considerations. Aging is caused by the generation of reactive oxygen species and the activity of free radicals which degenerate the cells in the human body. Aging is the prime factor for different types of diseases like cancer, cardiovascular problems, type II diabetics, hypertension, imbalance of important gut microbiota, and other diseases. Probiotics from fermented foods aid in health promoting functionalities as a function of antioxidant activity and promoting longevity which allows them to be a potential source of supplement. These factors open an interesting prospect of screening well known probiotics, especially lactic acid bacteria (LAB), which containing antioxidant activity from various Thai local fermented foods. This study aimed to screen targeted lactic acid bacteria (LAB) which is well known bacteria group, from 31 plant and animal based Thai local fermented foods in different regions (central, north, west, and north-east) by CaCO3 containing MRS agar and analyzed its probiotic properties and antioxidant activities. It was found that all 18 isolates considered as potential LAB as it had shown clear zone on CaCO3 containing MRS agar, and most isolates exhibited over 70% scavenging of DPPH in both supernatant and pellet except two isolates from plasom and kung jom having 49.12% for supernatant and 58.08% for pellet respectively. But only one isolate, called NYC8, from plant based fermented food showed significant potential probiotic properties, including hydrophobicity, acid-bile tolerance, and antimicrobial activity. Additionally, NYC8 isolate showed reducing ability equivalent to L-cysteine 195.91 µM along as well as lipid peroxidation inhibition capacity 44.34% for pellet and 4.34% for supernatant, and it was the one that had the one of the highest DPPH scavenging activities. After doing 16S rDNA sequencing, it was found that NYC8 isolate had 100% similarity with Lactiplantibacillus plantarum ATCC 14917T (ACGZ01000098) and Lactiplantibacillus argentoratensis 6365T (CP032751). This isolated strain might be further analyzed and developed as a potential longevity promoting supplement product

The present review explores recent scientific evidence on in situ folate fortification through African staple foods fermentation. Specifically, it highlights, folate-producing microorganisms isolated from these foods, conditions influencing folate production, and key factors determining the success or failure of this approach. Relevant studies were identified by searching literature in Web of Science, Scopus, PubMed, and Google Scholar in order of preference. Fourteen studies on microbial folate synthesis with promising results were reported for five fermented cereal-based products from Africa: bensaalga, injera, ogi, motoho, and togwa. Since these fermented foods contained an interesting level of folate, their consumption could contribute to folate intake. In these studies, 29 microbial isolates, including 14 lactic acid bacteria strains and 15 yeast strains were identified as the most promising folate producers. Yeast strains produced varying amounts of folate, with differences observed between strains. Findings suggest that optimizing folate production in fermented foods requires cautious microbial strain selection, strategic strain combination, and optimum monitoring of parameters such as temperature and incubation time. Understanding these factors may help improve the effectiveness of microbial folate enrichment in local fermented foods, which could support increased dietary folate intake in African populations.

Dendrobium is one of the largest genera in the Orchidaceae family. Information on the genetic diversity and relationships among species and hybrids is important for breeding purposes and species conservation. The objectives of this study were to assess genetic relatedness and to determine whether morphological, molecular, or combined analysis can discriminate among Dendrobium species, commercial hybrids, and interspecific hybrids. A total of 81 Dendrobium accessions were characterized with 12 amplified fragment length polymorphism (AFLP) primer pairs and 21 morphological characters. Mean genetic relatedness for morphological characters, AFLP analysis, and combined analysis were 0.61, 0.37, and 0.43, respectively. Dendrograms were generated using an unweighted pair group method with arithmetic averages (UPGMA); the analysis was performed on a Jaccard similarity coefficient matrix. The data from morphological characters revealed that the Jaccard similarity coefficient ranged from 0.20 to 1.0, where the tested 81 Dendrobium accessions could be grouped into four clusters. For the AFLP analysis, the number of polymorphic fragments for each primer varied from 80 to 284 with 78% average percentage of polymorphic loci and the similarity coefficient ranging from 0.125 to 1.0 with Dendrobium accessions grouped into three clusters. The similarity coefficients estimated through a combined analysis of morphological and AFLP data ranged from 0.21 to 1.0 and the Dendrobium accessions appeared clustered into two groups. The results revealed some similarities among the three data sets. The combined data set was the most useful in discriminating Dendrobium accessions based on species sections and relationship among species and their hybrids. The correlation between the AFLP data and the combined data was highly significant ( r = 0.98, P > 0.001), indicating the usefulness of AFLP data for species discrimination and hybrid identity in the absence of floral morphological characters.

In fermented foods, Enterococcus faecium, a bacterium with probiotic potential and uses in the food and health sectors, contributes to the production of bioactive metabolites and the preservation of microbial balance. Isolating and characterizing E. faecium from a variety of traditional Iraqi fermented foods, such as red pickles, Arab cheese, and red fermented sauerkraut, was the main objective of this study. Using 16S rRNA gene sequencing, three isolates (S2, AB1, and L1) were shown to be 99–100% comparable to reference strains (GenBank Accession Nos. MF582727.1, PQ590703.1, and ON631310.1). Biochemical profiling, including as motility, urease, oxidase, catalase, and IMVC assays, was used in conjunction with molecular identification. All of the isolates had comparable phenotypic traits and were non-motile, Gram-positive cocci that were negative for urease, indole, methyl red, catalase and oxidase, and VP and citrate. Their varied food sources were reflected in slight sequencing differences. Crucially, in well diffusion studies, all three isolates demonstrated antimicrobial action against Escherichia coli and produced distinct inhibitory zones, suggesting that they are capable of synthesizing bioactive substances including hydrogen peroxide, organic acids, and maybe bacteriocins. These results support the isolates' safe use in food preservation and human health by highlighting their dual roles as possible probiotic strains with antibacterial qualities and as functional microorganisms in local fermented foods.

Phenotypic and genotypic analyses of lactic acid bacteria in local fermented food, breast milk and faeces of mothers and their babies

Since the beneficial effects of viable probiotic bacteria as dietary supplements have gained huge research interest, Lactobacillus spp. with probiotic characteristics are widely used to prepare fermented dairy products such as yoghurts, milk-shakes etc. In this study, eight (08) homemade yoghurt samples were collected from different regions in the country for isolation of probiotic Lactobacillus spp. Among the samples, four (04) isolates were identified as Lactobacillus plantarum based on their growth and biochemical characteristics. The Isolates were resistant to NaCl (1-9%) and bilesalt (0.05-0.3%) and showed good growth in the acidic condition, while maximum growth was observed at pH around 6.0. The isolates were examined for their antibacterial activity against nine (09) different test pathogens and found all pathogens are inhibited their growth to some extent but maximum zone of inhibition was observed against Bacillus cereus (53.20 mm) and minimum was against Staphylococcus aureus (19 mm) after 72 hour incubation. The results of the present study indicate that, homemade yoghurts in Bangladesh are potential source of probiotic Lactobacillus spp. Further extensive research on isolation and characterization of probiotic organisms from local fermented foods and their growth optimization might be required for development of probiotic enriched food supplements in our country.

Traditional or local fermented foods have been the favourite sources for Lactic acid bacteria (LAB) used for starter cultures. Traditional food such as fermented cassava ‘tapai ubi’, fermented glutinous rice ‘tapai pulut’ and fermented shrimp paste ‘belacan’ have been local heritage consumed as Malay delicacies. In this work, 33 LAB samples were isolated from tapai ubi, tapai pulut and belacan and out of these only 5 LAB isolates (PG, PH, BG, UG and UL) showed inhibitory properties against selected indicator organisms (Bacillus subtilis, Escherichia coli, Salmonella typhimurium, and Staphylococcus aureus). Morphologically, isolate PG, PH, BG are cocci, UL is rod and UG is coccobacillus. Biochemically, isolates (PG, PH, BG UL and UG) are found to be Gram positive, non motile, lactose fermenter and catalase negative. The 16s Ribosomal RNA gene sequencing was carried out and each was identified with an accession number (PB: MT645488, PH: MT645489; BG: MT645490 UG: MT645491 and UL MT645492). Isolates PG and PH from tapai pulut belonged to Pediococcus pentosaceus (at 99% and 98%, respectively). Meanwhile, isolate from Belacan BG belonged to Enterococcus faecium (99%), and those from fermented shrimp, UG and UL belonged to Weissella confusa (99%), and Lactobacillus fermentum (98%), respectively. Majority of the isolates demonstrated broad spectrum inhibition against both Gram positive and negative indicator strains. Compared to the rest of isolates, PH exhibited the highest antibacterial activity against Bacillus subtilis. These results suggested that isolate PH are the most potent isolates which is producing antimicrobial agent with potential as food preservatives.

Nowadays, people prefer to consume foods with better nutrition to enjoy life more healthily. However, Muslims must also eat halal food following the Islamic way of life; eating nutritious food alone is insufficient. Indonesia, the largest Muslim population in the world, presents a market potential for halal food manufacturers and consumers worldwide. Several varieties of regional fermented foods were listed on the halal-positive food list and free from the requirement of halal certification, as per Indonesian Halal rules. The review of Indonesian fermented foods listed in the category of positive halal foods is the main topic of this essay. The procedures used to review the chosen themes include finding the literature in online sources, screening for topic inclusion, and evaluating, extracting, and discussing the accessible data found in the publications. The potential functional local fermented food varieties are included on the positive halal list: Tape (sticky rice and cassava), Dadih, and Tempeh. These fermented foods' microbiological and chemical characteristics were discussed. Therefore, the functional value of these fermented foods is rich in phytochemicals, anti-cancer (β-glucan), antioxidant compounds (isoflavones) and probiotic components (lactic acid bacteria and yeasts).

Bioethanol is a liquid chemical produced from sugar-, starch-or lignocellulosic-based biomass through fermentation by ethanol-producing microbes. Ethanol-producing yeast generally has limited tolerance to ethanol and has limitation to high temperatures above 40°C. High-temperature tolerant yeast is required because it potentially reduces the risk of contamination and it also reduces the cost of the cooling process. This study aims to determine ethanol-producing yeasts that have tolerance to ethanol and high temperatures from local fermented food products. This study uses a descriptive method conducted in three stages. Isolation and selection of yeast were performed from 18 local fermented foods in Indonesia. Temperature and ethanol tolerance of selected yeast were performed by using a spot test method. The ethanol content was tested using Gas Chromatography (GC). The results exhibited that isolate F08b had the highest tolerance to ethanol and temperature. The isolate was able to grow up to a temperature of 50°C and a concentration of 18% ethanol. Meanwhile, isolate F10 was able to produce the highest ethanol concentration at 3.37% (v/v) in 48th-hour fermentation.

Carbon nanotubes (CNT) have potential biomedical applications, and investigations are shifting towards the production of such nanotubes using renewable natural sources. CNTs were synthesized at various temperatures of 700, 750, 800, 850 and 900 °C, respectively, using a local fermented food known as “tapai ubi” or fermented tapioca as a precursor. The liquid part of this fermented food was heated separately at 80°C and channeled directly into the furnace system that employs the thermal chemical vapor deposition (CVD) method. Ferrocene, which was the catalyst was placed in furnace 1 in the thermal CVD process. The resulting CNTs produced from the process were studied using field emission scanning electron microscopy (FESEM) and raman spectroscopy. The FESEM images showed the growth morphology of the CNTs at the different temperatures employed. It was observed that the higher the synthesis temperature up to a point, the diameter of CNTs produced, after which the diameter increased. CNTs with helical structures were observed at 700 °C with a diameter range of 111 - 143 nm. A more straightened structure was observed at 750 °C with a diameter range of 59 - 121 nm. From 800 °C onwards, the diameters of the CNTs were less than 60 nm. Raman analysis revealed the present of D, G and G’ peak were observed at 1227-1358, 1565-1582, and 2678-2695 cm−1, respectively. The highest degree of crystallity of the carbon nanotubes synthesized were obtained at 800 °C. The radial breathing mode (RBM) were in range between 212-220 and 279-292 cm−1. Carbon nanotubes also being functionalized with Polyethylene bis(amine) Mw2000 (PEG 2000-NH2) and showed highly cells viability compared to non-functionalized CNT. The nanotubes synthesized will be applied as drug delivery in future study.

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