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Food spoilage bacteria are responsible for food spoilage and often fabricate biofilms on various surfaces. Naturally, biofilms are constructed by multiple bacterial species. These biofilms are more difficult to control compared to single species biofilms. Effective multispecies biofilm treatments are necessary in the food industry. This research evaluated antibiofilm activities of Streptomyces sp. KP110 and Pseudomonas fluorescens JB 3B metabolites. Prior to this research, the two antibiofilm crude extracts had shown significant results against various single species biofilms, including Bacillus , Shewanella , Vibrio , etc. The crude extracts also showed quorum-quenching activities, further asserting the antibiofilm potential. This research further characterized these antibiofilm crude extracts against multispecies biofilms, as well as assessing the duration of biofilm formation inhibition. The food spoilage bacteria used in this study are Bacillus cereus , Bacillus subtilis , and Shewanella putrefaciens . Both crude extracts showed antibiofilm activities with better results against Gram-negative bacteria. The metabolites are also considered nontoxic, meaning they are potential candidates for industrial purposes. The dominant active compounds of Streptomyces sp. KP110 are ticlopidine and hexadecanoid acid, while Pseudomonas fluorescens JB 3B are phenol and indole.

Background Foodborne disease and food spoilage are the prime public health issue and food security round the globe. Significant disease outbreaks mostly linked to the existence of pathogenic bacteria that extremely challenging due to the persistence of biofilm-forming. Proteins and bacterial metabolites have been shown to have good antibacterial activity and effectively removal bacterial biofilm. Recently, bacteriophage and their encoded lytic proteins such as lysin have attracted attention as potential alternative agent to control undesirable pathogens in human body infection, increasing food safety as advance preservations and medical treatment such as phage therapy. For these reasons, the efficacy of bacteriophage and their potential in combination with bacterial metabolites from Phyllosphere and Actinomycetes bacteria ( Pseudomonas fluorescens JB3B and Streptomyces thermocarboxydus 18PM crude extracts) was the aim of this present study. Results In this study, bacteriophage BC-VP (1.28  ±  0.29 × 10 11 PFU/ml) and ETEC-phage-TG (8.9  ±  2.19 × 10 8 PFU/ml) isolated from artificial lake water from previous study showed potential activity to control Bacillus cereus (BC) and Enterotoxigenic Escherichia coli (ETEC) population. The combination of BC-VP with metabolite ( P. fluorescens JB3B and S. thermocarboxydus 18PM) which were known from previous study had antibiofilm activities were able to inhibit (86.1%; 83.3%) and destruct (41%; 45.5%) biofilm formation of B. cereus respectively. Likewise, the synergy of bacteriophage ETEC-phage-TG with the same crude extract also showed promising activity against biofilm of ETEC with percentage of inhibition (81.9%; 76.4%) and percentage of destruction (54.1%; 44.4%). Application in various food, combination of BC-VP and bacterial metabolite extract ( P. fluorescens JB3B; S. thermocarboxydus 18PM) were able to reduce Bacillus cereus population in mashed potato (99.6%; 99.4%) at cold temperature (4 °C) and (68.9%; 56.6%) at room temperature (28 °C), boiled pasta (99.5%; 99.4%) and (84.7%; 75.7%), also soymilk (96.9%; 96.7%) and (42.4%; 39.4%) respectively. Likewise, combination of ETEC-phage-TG and bacterial metabolite ( P. fluorescens JB3B; S. thermocarboxydus 18PM) potentially reduced ETEC population after two different temperatures (4 °C and 28 °C) incubation in bean sprouts (TFTC; TFTC) and (47.5%; 49.1%), chicken meat (TFTC; TFTC) and (58.1%; 54%), also minced beef (99.5%; 99.4%) and (41.1%; 28%). GC-MS determination performed, oxalic acid, phenol, phenylethyl alcohol, N-hexadecanoic acid, and pyrolol[1,2-a]pyrazine-1,4-dione, hexadro-3-92-methylpropyl was the most active compound in P. fluorescens JB3B. 2,4-Di-tert-butylphenol, phenyl acetic acid, N-Hexadecanoic acid, pyrolol[1,2-a]pyrazine-1,4-dione, hexadro-3-92-methylpropyl, and Bis(2-ethylhexyl) phthalate was most active compound in the S. thermocarboxydus 18PM isolates. Conclusions The combination of isolated bacteriophages and bacterial metabolite showed promising results to be used as biocontrol candidate to overcome biofilm formed by foodborne and food spoilage bacteria using their ability to produce antibiofilm compounds and lytic activity. In addition, this combination also potentially reduces the use or replace the drawbacks of common application such as antibiotic treatment.

Background Plastic waste accumulation is one of the main ecological concerns in the past decades. A new generation of plastics that are easier to degrade in the environment compared to conventional plastics, such as starch-based bioplastics and oxo-biodegradable plastics, is perceived as a solution to this issue. However, the fate of these materials in the environment are unclear, and less is known about how their presence affect the microorganisms that may play a role in their biodegradation. In this study, we monitored the dynamics of bacterial community in soil upon introduction of commercial carrier bags claimed as biodegradable: cassava starch-based bioplastic and oxo-low-density polyethylene (oxo-LDPE). Each type of plastic bag was buried separately in compost soil and incubated for 30, 60, 90, and 120 days. Following incubation, soil pH and temperature as well as the weight of remaining plastics were measured. Bacterial diversity in soil attached to the surface of remaining plastics was analyzed using Illumina high-throughput sequencing of the V3-V4 region of 16SrRNA gene. Results After 120 days, the starch-based bioplastic weight has decreased by 74%, while the oxo-LDPE remained intact with only 3% weight reduction. The bacterial composition in soil fluctuated over time with or without the introduction of either type of plastic. While major bacterial phyla remained similar for all treatment in this study, different types of plastics led to different soil bacterial community structure. None of these bacteria were abundant continuously, but rather they emerged at specific time points. The introduction of plastics into soil increased not only the population of bacteria known for their ability to directly utilize plastic component for their growth, but also the abundance of those that may interact with direct degraders. Bacterial groups that are involved in nitrogen cycling also arose throughout burial. Conclusions The introduction of starch-based bioplastic and oxo-LDPE led to contrasting shift in soil bacterial population overtime, which may determine their fate in the environment.

Background Tempeh, a famous traditional Indonesian fermented soybean product, reportedly contains vitamin B 12 . Although Enterobacteriaceae have been previously implicated in vitamin B 12 production in tempeh, the function of Sphingomonadaceae , which is abundant in some tempeh samples, remains unknown. This study aimed to identify and characterize vitamin B 12 -producing bacteria from Empang (EMP) tempeh, with a focus on the understudied Sphingomonad . Results Metagenomic analysis focusing on vitamin B 12 biosynthesis genes revealed that Sphingomonad genes had a complete set gene that required for producing the vitamin. A total of 44 yellow-pigmented isolates, characteristic of Sphingomonas , were screened via a vitamin B 12 assay, three of which demonstrated potential for de novo biosynthesis. On the basis of 16S rRNA gene analysis, all three isolates were identified as Sphingomonas paucimobilis . Whole-genome sequencing and annotation of the EMP5-4 isolate revealed a complete gene set for the aerobic vitamin B 12 biosynthesis pathway, including hem genes, cob genes and cobalamin riboswitches. Vitamin B 12 production was confirmed through fermentation in a TSB medium with cobalt and DMBI supplementation and quantified at 0.949 µg/mL via HPLC. A genome-based safety assessment identified only low confidence antibiotic resistance and virulence genes, and a hemolysis assay revealed no red blood cell lysis, suggesting minimal pathogenicity. Conclusion These findings demonstrate that S. paucimobilis from tempeh may serve as a novel microbial source of vitamin B 12 , supporting its potential application in functional foods for individuals who avoid animal-derived products.

Enterotoxigenic Escherichia coli (ETEC) contamination remains a major global concern, including in Indonesia, where efforts to develop an ETEC vaccine to prevent foodborne diseases are currently underway. This study aimed to perform genomic characterization of bacteriophage φETEC-S3 and evaluate the efficacy of its application in food samples targeting Enterotoxigenic Escherichia coli . The phage exhibited a high lytic activity, reaching a titer of 3.8 ± 0.50 × 10 10 PFU/mL and remained stable at both 4 °C and 37 °C based on the results of the thermal stability assays. Whole-genome bioinformatics analysis identified φETEC-S3 as a virulent double-stranded DNA phage belonging to the genus Phapecoctavirus within the class Caudoviricites . This genome contained structural and lytic genes, including those encoding the major capsid protein, tail fiber, and Rz-like spanin. Application studies demonstrated that phage φETEC-S3 significantly reduced the enterotoxigenic Escherichia coli population in all food samples from the initial day until the seventh day of incubation. The highest reduction percentages were for φETEC-S3 in soy milk on the initial day (93%) and in mashed potatoes on the seventh day (94%). Thus, φ ETEC-S3 is shown to be a promising candidate for a biocontrol agent against bacteria responsible for food spoilage.

Objectives Oxo-polyethylene (oxo-PE) is marketed as a biodegradable plastic, yet its environmental degradation remains poorly understood, particularly in marine contexts. This study aimed to screen for bacterial isolates with enzymatic potential relevant to oxo-PE breakdown using a simulated marine environment. Microbial communities were enriched in Winogradsky columns containing oxo-PE sheets, and isolates were screened for enzymes known to contribute to polymer breakdown. Results After 60 days of incubation, no significant weight loss was detected in oxo-PE sheets. Surface-level alterations, including cracks and roughness, were observed using field emission scanning electron microscopy. Biofilms formed on the polymer surface over time yielded numerous bacterial isolates, of which 25 exhibited one or more enzymatic activities associated with polymer degradation, including alkane monooxygenase, laccase, lipase, and manganese peroxidase. Among them, isolate D30065 ( Priestia megaterium ) demonstrated the most diverse enzyme profile, suggesting potential for oxidative enzyme activity relevant to polyethylene breakdown. However, further studies are needed to directly evaluate the ability of this isolate to degrade oxo-PE. These findings highlight early microbial responses to oxo-PE exposure and provide a foundation for future studies on biodegradable plastic-microbe interactions in marine systems.

Background Indonesia is a country that uses half or more aquatic foods as protein intake. The increased production in aquaculture industries might cause several problems, such as bacterial disease resulting in mass mortality and economic losses. Antibiotics are no longer effective because aquaculture pathogens can form biofilm. Biofilm is a microbial community that aggregates and firmly attaches to living or non-living surfaces. Biofilm formation can be caused by environmental stress, the presence of antibiotics, and limited nutrients. Therefore, it is important to explore antibiofilm to inhibit biofilm formation and/or eradicate mature biofilm. Phyllosphere bacteria can produce bioactive compounds for antimicrobial, antibiofilm, and anti-quorum sensing. Three aquaculture pathogens were used in this study, such as Aeromonas hydrophila , Streptococcus agalactiae , and Vibrio harveyi . Results Pseudomonas fluorescens JB3B and Morganella morganii JB8F extracts could disrupt single and multi-species biofilms. Both extracts could inhibit single biofilm formation from one to seven days of incubation time. We confirmed the destruction activity on multi-species biofilm using light microscope and scanning electron microscope. Using GC-MS analysis, indole was the most active fraction of the P. fluorescens JB3B extracts and octacosane from the M. morganii JB8F extract. We also conducted a toxicity test using brine shrimp lethality assay on P. fluorescens JB3B and M. morganii JB8F extracts. P. fluorescens JB3B, M. morganii JB8F, and a mixture of both extracts were confirmed non-toxic according to the LC 50 value of the brine shrimp lethality test. Conclusions P. fluorescens JB3B and M. morganii JB8F phyllosphere extracts had antibiofilm activity to inhibit single biofilm and disrupt single and multi-species biofilm of aquaculture pathogens. Both extracts could inhibit single species biofilm until seven days of incubation. Bioactive compounds that might contribute to antibiofilm properties were found in both extracts, such as indole and phenol. P. fluorescens JB3B, M. morganii JB8F extracts, and mixture of both extracts were non-toxic against Artemia salina .

Objectives Bacteriophages are viruses that specifically target and kill bacteria. Bacteriophages are considered safe for humans, making them suitable for use in food applications Therefore, this study was conducted to isolate, characterize, and apply bacteriophages from the environment to control food spoilage bacteria. Results Bacteriophages were isolated by using Bacillus cereus ( B. cereus ) as reference host. We found bacteriophage BC-VP titers (1.16 ± 0.18 × 10 9 PFU/mL) and BC-AJ (1.72 ± 0.19 × 10 8 PFU/mL). Both bacteriophages were polyvalent, targeting not only B. cereus , but also lysing enteropathogenic Escherichia coli (EPEC) and enterotoxigenic Escherichia coli (ETEC) with different efficiencies. Bacteriophage BC-VP and BC-AJ miMOI were determined to be 0.01 and 1, respectively. Bacteriophage BC-VP morphology was analyzed using Transmission Electron Microscope (TEM) and categorized as Myoviridae family from the order of Caudovirales . Both bacteriophages showed significant bacterial reduction when applied to artificially contaminated cooked rice and pasteurized milk. The reduction number was higher in pasteurized milk at 28℃ for BC-VP (93%) and BC-AJ (90%). Molecular characterizations were performed and showed that BC-VP was 96.88% a virulent bacteriophage. Also, no sign of antibiotic resistance gene in BC-VP and genes related to lytic cycle such as putative tail lysin and tail fiber, were annotated.

Background Child growth during the first 1000 days is crucial for developing the immune system, particularly through the digestive system. However, in the early childhood phase, most children facing feeding issues, which leading to insufficient nutrient uptake. Thereby, the external supplementation may be essential to support their nutrition needs. Therefore, this study evaluated the potential of a baby food supplement (BFS) to promote the growth of beneficial lactic acid bacteria (LAB) and their metabolite production. Results The food supplement did not significantly interfere with LAB growth. Instead, it significantly enhanced the antimicrobial activity of Lactobacillus acidophilus and Limosilactobacillus fermentum against Enterohemorrhagic Escherichia coli (EHEC) and Staphylococcus aureus , respectively. The addition of BFS also improved the adhesion capacity of the LAB used in this study to yeast cells. Moreover, gas chromatography‒mass spectrometry (GC-MS) analysis revealed greater amounts of bioactive compounds related to antimicrobial and immunomodulatory properties, such as lactic acid and trilinolein, in the supernatants with the addition of food supplements. Liquid chromatography-high resolution mass spectrometry (LC-HRMS) analysis also revealed that there were more compounds in M3 than in M1 or only, which are associated with the immune system and neurocognitive development, such as choline, rapastinel, octopamine, aceglutamide, indoleacrylic acid, 1-aminocyclopropanecarboxylic acid (ACPC), and GABA. Conclusions Baby food supplements are promising for improving the production of beneficial health metabolites of lactic acid bacteria used in this in vitro study, as they inhibit pathogens, promote competition, and produce many beneficial bioactive compounds. The limitation of this study is that it was performed in vitro, and further studies are needed.

Objectives This study aimed to isolate lytic bacteriophage to control EHEC and ETEC, characterize, and apply their ability to control these two bacteria in food. Results One of the bacteria that contaminate the food is Diarrheagenic Escherichia coli (DEC) which was consist of Enterohaemorrhagic E. coli and Enteropathogenic E. coli . Preservation methods were used to improve shelf-life to reduce microbial growth but it might have side effect to the nutrition and human health. Therefore, it is required to explore an alternative method that can be used to be a biocontrol agent for food, including the bacteriophage approach. Our study showed that these two phages are promising to be used as biocontrol agent in food. Genomic DNA sequencing performed that this phage being exclusively lytic.