Limosilactobacillus reuteri DSM 17938 Inhibition of Biofilm Formation by Prevotella intermedia and Fusobacterium nucleatum Across Salivary pH: An In Vitro Study.

OBJECTIVES: This study aims to investigate Limosilactobacillus reuteri DSM 17938's antibiofilm effects on Prevotella intermedia and Fusobacterium nucleatum, common causes of alveolar osteitis. It seeks topical alternatives to prevent this condition posttooth extraction. The secondary objective is to assess these effects under different pH conditions (pH 4.5 and pH 7), mimicking oral cavity saliva pH dynamics. MATERIALS AND METHODS: Ethical approval was secured for the saliva collection process involving five healthy adult participants who had undergone wisdom tooth extraction. Saliva samples were diligently collected on the 7th day post-surgery. The unstimulated saliva underwent a series of treatments, including the addition of phenylmethylsulfonyl fluoride (PMSF), pH adjustments, centrifugation, and filtration. The pH levels were re-measured, and subsequent adjustments were made to achieve pH values of 4.5 or 7. Limosilactobacillus reuteri DSM 17938, with a concentration of 1×108 colony-forming units (CFU) per 5 drops, was utilized in the study. Biofilm testing involved incubating saliva samples with varying pH (4.5 or 7) alongside bacterial suspensions (Prevotella intermedia, Fusobacterium nucleatum, or a mixed species). The Interlac suspension was introduced, and plates were anaerobically incubated for 24 hours. Biofilm results were obtained using a spectrometer. The test is conducted in triplicate. STATISTICAL ANALYSIS: To scrutinize the impact of pH on biofilm development, the acquired data underwent a two-way ANOVA test in SPSS as part of the statistical analysis. A significance level of p<0.05 was used to determine statistical significance. RESULTS: Limosilactobacillus reuteri DSM 17938 significantly reduced biofilm formation across bacterial strains (p = 0.000). Statistical analysis indicated a significant impact of pH on biofilm development (p = 0.000) compared to no saliva samples, with higher formation observed under acidic conditions (pH 4.5). However, the pH levels of 4.5 and 7 did not result in significantly different bacterial biofilm formation (p = 0.529). CONCLUSION: This research highlights Limosilactobacillus reuteri DSM 17938's potency in inhibiting biofilm formation of Prevotella intermedia and Fusobacterium nucleatum. Salivary pH variations significantly influence biofilm development, emphasizing the need to consider pH when assessing probiotic effectiveness. Despite limitations in saliva sample sterilization, this study provides valuable insights into alternative approaches for preventing alveolar osteitis. Further research should explore clinical applications and refine sterilization methods for more accurate results.

Immunoinformatic of novel self-amplifying mRNA vaccine lipid nanoparticle against SARS-CoV-2.

We developed innovative self-amplifying mRNA (sa-mRNA) vaccine based on the derivative of S and Nsp3 proteins, which are considered crucial adhering to human host cells. We performed B-cell, Major histocompatibility complex (MHC) I, and II epitope which were merged with the KK and GPGPG linker. We also incorporated 5' cap sequence, Kozak sequence, replicase sequence, 3'/5' UTR, and poly A tail within the vaccine structure. The vaccine structure was subsequently docked and run the molecular dynamic simulation with TLR7 molecules. As the results of immune response simulation, the immune response was accelerated drastically up to >10-fold for immunoglobulin, interferon-γ, interleukin-2, immunoglobulin M (IgM) + immunoglobulin G (IgG) isotype, IgM isotype, and IgG1 isotype in secondary and tertiary dose, whereas natural killer cells, macrophages, and dendritic cells showed relatively high concentrations after the first dose. As our finding, the IgM + IgG, IgG1 + IgG2, and IgM level (induced by sa-mRNA vaccine) ensued three times with two-fold increase in days 25, and 50, then decreased after days 70-150. However, 150-350 days demonstrated constantly in the range of 20,000-21,000.

Evaluation of IL-1ß and CRP mRNA expression levels by RT-PCR in postorthodontic treatment patients with temporomandibular joint disorders: a cross-sectional Study.

Aim: Temporomandibular joint disorder (TMD), which affects the masticatory muscles, temporomandibular joint, and surrounding tissues, can manifest as inflammation. This study aims to explore the expression levels of the inflammatory biomarkers, interleukin (IL)-1ß and C-reactive protein (CRP), in TMD patients who have undergone orthodontic treatment. Materials and Methods: Buccal swabs from 105 postorthodontic treatment patients were analyzed using real-time polymerase chain reaction to assess the expression levels of IL-1ß and CRP in each group after messenger ribonucleic acid extraction. Patients were also examined using the Diagnostic Criteria for TMD (DC/TMD) to determine if they met the criteria for a TMD diagnosis. The TMD group was subdivided into three categories based on the DC/TMD. Results: The study included 37 patients who did not develop TMD (group 0) and 68 participants who developed TMD after orthodontic treatment, including 17 with pain-related TMDs (group 1), 29 with intra-articular TMDs (Group 2), and 22 with combined pain-related and intra-articular TMDs (group 3). CRP expression was higher than IL-1ß in groups 1 and 2, and IL-1ß expression was higher than CRP in group 3. The Kruskal-Wallis test showed that IL-1ß and CRP expression levels in groups 1, 2, and 3 were not statistically different. Sex and adult age had considerable effects on the occurrence of TMD in patients after orthodontic treatment. Conclusions: Higher IL-1ß expression was found in postorthodontic treatment patients with more complex TMD. This study strengthens the evidence of inflammation through IL-1ß and CRP expression in individuals with TMD, especially after orthodontic treatment.

Corrigendum to "The role of NMT induction on odontogenic proliferation and differentiation of dental pulp stem cells".

[This corrects the article DOI: 10.1016/j.heliyon.2021.e06598.].

Hydrogel-Based Biomaterial as a Scaffold for Gingival Regeneration: A Systematic Review of In Vitro Studies.

BACKGROUND: Hydrogel is considered a promising scaffold biomaterial for gingival regeneration. In vitro experiments were carried out to test new potential biomaterials for future clinical practice. The systematic review of such in vitro studies could synthesize evidence of the characteristics of the developing biomaterials. This systematic review aimed to identify and synthesize in vitro studies that assessed the hydrogel scaffold for gingival regeneration. METHODS: Data on experimental studies on the physical and biological properties of hydrogel were synthesized. A systematic review of the PubMed, Embase, ScienceDirect, and Scopus databases was conducted according to the Preferred Reporting System for Systematic Reviews and Meta-Analyses (PRISMA) 2020 statement guidelines. In total, 12 original articles on the physical and biological properties of hydrogels for gingival regeneration, published in the last 10 years, were identified. RESULTS: One study only performed physical property analyses, two studies only performed biological property analyses, and nine studies performed both physical and biological property analyses. The incorporation of various natural polymers such as collagen, chitosan, and hyaluronic acids improved the biomaterial characteristics. The use of synthetic polymers faced some drawbacks in their physical and biological properties. Peptides, such as growth factors and arginine-glycine-aspartic acid (RGD), can be used to enhance cell adhesion and migration. Based on the available primary studies, all studies successfully present the potential of hydrogel characteristics in vitro and highlight the essential biomaterial properties for future periodontal regenerative treatment.

The antimicrobial effect of Limosilactobacillus reuteri as probiotic on oral bacteria: A scoping review.

Dysbiosis among oral microbial community in the oral cavity can lead to several oral diseases. Probiotic therapy is known to correct these imbalances. Limosilactobacillus reuteri is one of the most studied strains of probiotics and can control oral microbiota through reuterin, a wide-spectrum antimicrobial agent. The objective of this review was to evaluate the effect of the antimicrobial activity of Limosilactobacillus reuteri on the oral bacteria of humans. This review used PubMed, Scopus, EMBASE, ScienceDirect, and Google Scholar databases as bibliographic resources. Studies with matching keywords were analyzed and screened with PRISMA-ScR recommendations. Sixteen articles were selected for this review, which included a total of 832 patients. Based on this review, Limosilactobacillus reuteri has a strong antibacterial effect against Streptococcus mutans in healthy individuals but is not effective against Lactobacillus. Additionally, it has a significant antibacterial effect against Porphiromonas gingivalis in patients with periodontitis, although its effectiveness is not stable in patients with peri-implant infections. Furthermore, Limosilactobacillus reuterihas varying results against other bacteria, indicating the need for further extensive research to ensure its efficacy.

Relationships between Solobacterium moorei and Prevotella intermedia in subgingival microbiota of periodontitis patients with halitosis: A preliminary study using qPCR.

Objective: Solobacterium moorei is suggested to be associated with the production of volatile sulphur compounds (VSCs) and can be found in subgingival plaques of deep periodontal pockets. We examined whether this bacterium's count was reduced in periodontitis patients with halitosis following non-surgical periodontal treatment, while the bacterial count of Prevotella intermedia was measured simultaneously as a control. Material & methods: This clinical study included 20 adults with chronic periodontitis who complained of halitosis. The bacterial relationship in the subgingival plaque sample was measured after 8 weeks post-treatment, including the probing pocket depth (PPD). Quantitative real-time PCR (qPCR) was used to measure the proportion of S. moorei, while the concentrations of H2S and CH3SH were determined using oral ChromaTM. Results: The presence of S. moorei was consistently observed in participants with periodontitis before and after non-surgical periodontal treatment and consistent showed a significantly lower proportion compared with P. intermedia. Solobacterium moorei showed a strong positive correlation with H2S and CH3SH concentrations, but a negative correlation with deep periodontal pocket measurements. Conversely, reduced P. intermedia may be more associated with a deep pocket, independent of the concentration of CH3SH. Conclusion: The study data showed that the proportion of S. moorei in the subgingival biofilm can be related to halitosis in periodontitis patients.

Quantity of the antigens of Streptococcus mutans serotype e and Candida albicans and its correlation with the salivary flow rate in early childhood caries.

Background: Streptococcus mutans involved in caries pathogenesis is classified into four serotypes, namely serotypes c, e, f, and k. Candida albicans can be found in the plaque of children with early childhood caries (ECC). Aims: The aim of this study was to analyze the quantity of the antigens of S. mutans serotype e and C. albicans and its correlation with the salivary flow rate in ECC. Materials and Methods: The antigen quantities of caries plaque samples and caries-free were determined using an enzyme-linked immunoassay with 450-nm optical density. Results: There was a significant difference between the quantity of S. mutans serotype e and C. albicans antigens in each salivary flow rate category (P < 0.05). The relationship between the antigen quantity of S. mutans serotype e and C. albicans was r = 0.624 (P > 0.05) for caries plaque samples and r = 0.628 (P > 0.05) for caries-free samples. Conclusion: the antigen quantities of S. mutans serotype e and C. albicans and the salivary flow rate might correlate to the pathogenesis of ECC.

The Discrepancy between Clove and Non-Clove Cigarette Smoke-Promoted Candida albicans Biofilm Formation with Precoating RNA-aptamer.

This study explores the influence of precoating aptamer (Ca-apt1) on C. albicans viability while the fungus was growing in the presence of exposing condensed cigarette smoke (CSC), prepared from clove (CCSC) and non-clove (NCSC) cigarettes, for 48 h. Using qPCR, we found that mRNA expression of adhesion-associated genes ( ALS3 and HWP1) was impaired by precoating C. albicans yeast cells with the aptamer. Conversely, the gene transcription was upregulated when aptamer-uncoated yeast was pre-treated with either CSC. In addition, by analysing the result of MTT ([3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] assay, we found that the presence of added CCSC or NCSC in growth medium for 48 h was significantly enhanced C. albicans biofilm development. However, the presence of precoated aptamer was significantly impaired biofilm development accelerated by the NCSC. The inhibitory effect of the Ca-apt1 was not dependent on the precoated aptamer (1 and 10%). Interestingly, we noted that the enhancer effect of treated CCSC was no longer effective when the yeast had been precoated with 10% aptamer tested. Additionally, light microscopy analysis revealed that precoating aptamer alleviates morphological changes of C. albicans (from yeast to hypha formation) that are enhanced by adding CCSC or NCSC in the growth medium. In conclusion, these results suggest that administration on Ca-ap1 exhibits a significant protective effect on CSC-induced biofilm formation by C. albicans.

The role of NMT induction on odontogenic proliferation and differentiation of dental pulp stem cells.

This study was conducted to investigate the odontogenic proliferation and differentiation of dental pulp stem cells (DPSCs) after induction by nanoparticle mineral trioxide (NMT). DPSCs were isolated from permanent teeth and placed in tubes containing Dulbecco's modified Eagle's medium, followed by immunocytochemistry analysis. The viability of DPSCs exposed to NMT was measured using MTT assay with trypan blue dye exclusion. Alkaline phosphatase (ALP) activity was evaluated using ALP colorimetric reactions by reacting NMT supernatants with fluorescent-specific ALP substrates. The concentration of osteocalcin was determined using an instant human osteocalcin enzyme-linked immunosorbent assay (ELISA) kit. A human dentin sialophosphoprotein (DSPP) ELISA kit coated with anti-human DSPP antibody was employed to measure DSPP levels. There was a significant difference between ALP activity after exposing the cells to NMT and trioxide mineral aggregate on days 3, 7, and 21. Osteocalcin activity showed a significant difference on days 3, 7, 14, and 21. There was a significant difference in DSPP levels on days 7 and 21. DPSCs exposed to NMT and to trioxide mineral aggregate showed extracellular matrix formation on day 7 and 14, respectively. Furthermore, NMT may effectively increase the proliferation and differentiation of DPSCs as well as their maturation toward odontoblasts.

The effect of coating chitosan on Porphyromonas gingivalis biofilm formation in the surface of orthodontic mini-implant.

Infection is the main problem for the failure of orthodontic mini-implant. Modern prevention of infection is now focused on local antibacterial coatings on implant devices. Chitosan is biocompatible and has antibacterial properties. Azithromycin is a synthetic antibiotic with immunomodulatory properties in which it has an advantage over the rest of antibiotics. This study aimed to evaluate the effect coating chitosan on the orthodontic mini-implant in Porphyromonas gingivalis biofilm formation. This is an experimental study using 25 orthodontic mini-implants. Five samples were coated with chitosan, 5 samples were coated with chitosan-azithromycin, 5 samples were coated with azithromycin, 5 samples were uncoated, and 5 samples were uncoated and were not exposed to P. gingivalis. P. gingivalis biofilms on the surface of the orthodontic mini-implant were observed after 24 h of incubation. P. gingivalis biofilm mass inhibition was highest in the azithromycin-treated group, followed by chitosan + azithromycin and chitosan only. The one-way ANOVA statistic test and post hoc Bonferroni statistic test of P. gingivalis biofilm mass show a significant difference between and within groups of experiments (P < 0.05). The Pearson correlation test with a value of R = +0.88, indicated that the bacterial viability count and the biofilm mass have a strong positive correlation. In conclusion, orthodontic mini-implant coated with chitosan, chitosan with azithromycin, or azithromycin only effectively suppressed P. gingivalis biofilm formation.

Identification of active compounds in ethyl acetate, chloroform, and N-hexane extracts from peels of Citrus aurantifolia from Maribaya, West Java, Indonesia.

Lime peel contains essential oils are used as anti-oxidants or anti-cancer compounds. As a traditional medicine, lime has been widely used as a substitute for antibiotics. This study aimed to identify active compounds in peel extracts from Citrus aurantifolia that grows in Indonesia. Extraction was carried out by maceration using three different solvents: ethyl acetate, chloroform, and n-hexane. The extracts were analyzed using gas chromatography-mass spectrometry. The results showed that lime peel contained many important compounds and that 28, 27, and 24 different chemical compounds, both minor and major constituents, were extracted by ethyl acetate, chloroform, and n-hexane, respectively. Four compounds were found in all three solvent extracts, namely, D-limonene, phytol, α-tocopherol, and 5, 7-dimethoxycoumarin. Forty-seven compounds were uniquely present in one solvent, including 17 in ethyl acetate, 17 in chloroform, and 13 in n-hexane. Among the active compounds extracted, several are of biological importance, for example, stigmasterol, D-limonene, Vitamin E, and α-tocopherol. It can be concluded that a variety of distinct compounds are extracted from the same lime peel sample when different solvents are used and that some of these are bioactive compounds with anti-oxidant, anti-microbial, or anti-cancer properties.

Expression of TNF, IL1B, and iNOS2 in the neural cell after induced by Porphyromonas gingivalis with and without coating antibody anti -Porphyromonas gingivalis.

Porphyromonas gingivalis has virulence factors such as gingipain and lipopolysaccharide, causing bacteremia to reach the brain and activate neuroinflammatory release cytokines. This study analyzed the effect of the co-culture of neuron cells with P. gingivalis coated with anti -P. gingivalis antibodies against cytokines produced by neuron cells. The gene expressions of the TNF, IL1B, NOS2 in neurons was evaluated using RT-qPCR. The results showed that P. gingivalis coated with anti -P. gingivalis antibody before co-culture with neuron cells could decrease the gene expression of TNF, IL1B, and NOS2 of neuron cells.

Effect of Implantation Ti-6Al-4V ELI in femoral bone defect regeneration of Sprague Dawley rat.

Ti-6Al-4V ELI is one of the most commonly used dental implant restore function. The solution treatment temperature variation can significantly increase the strength, but it is not yet known the effect of these temperature variations on the alloy's biocompatibility properties. Twelve female Sprague Dawley rats were divided into six groups as follows: the treated group, the control group, and the defect group without implant material. In the treated group, the femur bone defect was implanted with as-cast Ti-6Al-4V ELI, 850°C, 950°C, and 1050°C heat-treated Ti-6Al-4V ELI implant material. The rats were euthanized after 30 days postimplantation and evaluated histologically. The results show that the histological scoring of the specimen for femur defect without implant material is 2 (fibrous union and fibrocartilaginous), score with implant as-cast is 2.5, the sample with 850°C heat treatment material is 2.5, 950°C is 2.5, and the temperature at 1050°C is 2.5. The score of 2.5 is between score 2 and score 3: hemorrhage, fibrous union, fibrocartilaginous microhemorrhage, and mineralized cartilage union. In conclusion, there is no effect of different heat treatment temperatures for Ti-6Al-4V ELI implant material in rat bone regeneration's maturation level.

Enhanced bone regeneration capability of chitosan sponge coated with TiO2 nanoparticles.

Chitosan has been a popular option for tissue engineering, however exhibits limited function for bone regeneration due to its low mechanical robustness and non-osteogenic inductivity. Here we hybridized chitosan with TiO2 nanoparticles to improve its bone regeneration capability. Morphology and crystallographic analysis showed that TiO2 nanoparticles in anatase-type were distributed evenly on the surface of the chitosan sponges. Degradation test showed a significant effect of TiO2 nanoparticles addition in retaining its integrity. Biomineralization assay using simulated body fluid showed apatite formation in sponges surface as denoted by PO4- band observed in FTIR results. qPCR analysis supported chitosan - TiO2 sponges in bone regeneration capability as indicated by DMP1 and OCN gene upregulation in TiO2 treated group. Finally, cytotoxicity analysis supported the fact that TiO2 nanoparticles added sponges were proved to be biocompatible. Results suggest that chitosan-50% TiO2 nanoparticles sponges could be a potential novel scaffold for bone tissue engineering.

Nanochitosan antimicrobial activity against Streptococcus mutans and Candida albicans dual-species biofilms.

OBJECTIVE: Chitosan nanoparticle (nanochitosan) has a broad antimicrobial spectrum against diverse pathogenic microorganisms. However, its effect on dental caries-associated microorganisms, such as Streptococcus mutans and Candida albicans is yet to be explored. These microorganisms are known for causing early childhood caries. Therefore, this study was aimed at investigating nanochitosan inhibition capacity against dual-species biofilms of S. mutans and C. albicans. In this study, nanochitosan antimicrobial activity is reported against mono and dual biofilm species of S. mutans and/or C. albicans at 3 and 18 h incubation time. Nanochitosan inhibition capacity was observed through biofilm mass quantity and cell viability. RESULTS: The present study successfully synthesized nanochitosan with average diameter of approximately 20-30 nm, and also established dual-species biofilms of S. mutans and C. albicans in vitro. With nanochitosan treatment, the cell viability of both microorganisms significantly decreased with the increasing concentration of nanochitosan. There was no significant decrease in biofilm mass both in the dual and single-species biofilms after 3 h of incubation. However, greater inhibition of biofilm was observed at 18 h incubation.

Genotyping analysis of the Pax9 Gene in patients with maxillary canine impaction.

Background: Paired-box gene 9 ( PAX9) mutation is potentially associated with impaction in some patient populations. Here, we analyzed the relationship between PAX9 polymorphism and the occurrence of maxillary canine impaction. Methods: Patients with and without maxillary canine impaction were selected based on specific inclusion criteria, and samples of genomic DNA were obtained from a buccal mucosa swab. DNA was amplified by polymerase chain reaction and sequenced for further bioinformatics analysis to identify single nucleotide polymorphism (SNP) genotypes. Genotype and allele counting was performed in both case and control groups prior to conducting statistical analysis. Results: Four SNPs were identified in patients with maxillary canine impaction, with relative confidence determined based on chromatogram-peak assessment. All SNPs were located in exon 3 of PAX9 and in the region sequenced by the primer pair -197Fex3 and +28Rex3. Three of the SNPs (rs375436662, rs12881240, and rs4904210) were reported previously and are annotated in NCBI (dbSNP version 150), whereas another SNP mapped to chromosome 14 has not been reported. Patients with a CC genotype at SNP 3 [odds ratio (OR): 2.61 vs. TT; 1.28 vs. CT] and a CC genotype at SNP 4 [OR: 0.71 vs. GG; 0.79 vs. CG] were more likely to have maxillary canine impaction. Conclusions: These results demonstrated that the presence of SNPs 3 and 4 is associated with increased likelihood of suffering from maxillary canine impaction.

Salivary nitric oxide, Simplified Oral Hygiene Index, and salivary flow rate in smokers and non-smokers: a cross-sectional study.

Background: Salivary nitric oxide plays an important role as an antibacterial agent in the oral cavity. Here, we analyze salivary nitric oxide, Simplified Oral Hygiene Index (OHI-S) scores and the salivary flow rate in smokers and non-smokers which has not been done previously. Methods: A cross sectional study included 25 smokers and 25 non-smokers. Their OHI-S results were categorized as "good," "medium," or "bad." Unstimulated saliva samples were collected, and their nitric oxide concentration was measured using the Griess method. Results: The salivary flow rate in smokers was lower, at 0.30 ml/minute, compared to non-smokers who had a salivary flow rate of 0.33 ml/minute. This was statistically insignificant. There was a significant difference in the concentrations of nitric oxide between smokers and non-smokers (p < 0.05). Smokers had higher concentrations than non-smokers (185.4 µM Vs 114.60 µM). In addition, there was a moderate positive correlation (r = 0.305) between the concentration of salivary nitric oxide level and the OHI-S results. Conclusions: It was concluded that salivary nitric oxide concentration was higher in smokers, and the oral hygiene condition of smokers was poor.

The Anti-inflammatory Effects of Glycerol-supplemented Probiotic Lactobacillus reuteri on Infected Epithelial cells In vitro.

BACKGROUND: One of the most interesting effects of probiotics is their ability to modulate the immune system through the induction of cytokines and to enhance the host immune response. AIMS: The purpose of this study was to evaluate the anti-inflammatory effect of glycerol-supplemented Lactobacillus reuteri on the transcription level of interleukin (IL)-8 and human-beta-defensin (hBD)-2 expressed by epithelial cells after exposure to bacteria. MATERIALS AND METHODS: The confluent-cultured HaCat cell line (105 cells/mL) was exposed to Streptococcus mutans ATCC-25175 and Porphyromonas gingivalis ATCC-33277 (107 colony-forming units [CFU]/mL) for 24 h and challenged with probiotic L. reuteri ATCC-55730 (107 CFU/mL) supplemented with glycerol. Subsequently, the transcription levels of IL-8 and hBD-2 in HaCat cells were analyzed using reverse-transcription polymerase chain reaction (RT-PCR). In addition, cell viability was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. All the obtained data were statistically analyzed using the one-way analysis of variance test, with P < 0.05 set as the level of significance. RESULTS: The MTT assays confirmed no cytotoxic effects of glycerol-supplemented L. reuteri on HaCat cells (viability >90%). mRNA expression of IL-8 and hBD-2 increased after exposure to both bacteria. The presence of glycerol-supplemented L. reuteri significantly reduced the expression of IL-8 and hBD-2 on HaCat cells (P < 0.05). CONCLUSION: Glycerol-supplemented L. reuteri reduced the expression of IL-8 and hBD-2, and the results may be proof of principle for a probiotic approach to combating inflammation. However, further studies are needed to validate this probiotic effect.

Challenges in Implementing a Biorisk Management Program at Universitas Indonesia: A Checklist Tool for Biorisk Management.

Rapid development and advancement of bioresearch at a university's laboratories can have both positive and negative implications for public health and the environment. Many research activities in which biological materials have been created, modified, stored, and manipulated require safety procedures to keep the negative effects on humans and the environment as low as possible. The Occupational Health, Safety and Environmental (OHS&E) Department of the University of Indonesia (UI) is trying to increase the awareness and responsibility of its university members and laboratory staffs who work with biohazard materials by creating a biorisk checklist. The checklist was developed based on WHO guidelines and the National University of Singapore (NUS) Laboratory Manual, which contains 311 questions about the management, administration, and handling of various hazards, recombinant experiments, and animal and plant experiments. A gap analysis was run against the checklist in 14 laboratories at the University of Indonesia Salemba campus, which daily works with highly infectious pathogens and high-risk agents. Overall result showed that none of these laboratories had met all of the checklist items, and there were only 2 laboratories that had implemented more than half of the items. This checklist was proven to be a simple tool for assessing laboratories that handle and store biohazard materials, and it could be used as a monitoring tool for biorisk programs as well. It also could be further developed as a laboratory software application to increase its effectiveness and its accuracy.