Search Results (1,115)

Background: Intensive agriculture in fragile, dry–hot valleys degrades coffee plantation soils. Combining leguminous cover crops with microbial inoculants is promising, yet their synergy remains unresolved. Methods: In a field trial, we established Medicago sativa L. (ZB1) and Vicia villosa Roth var. glabrescens (ZB2) cover crops following Bacillus cereus inoculation, then assessed soil chemistry, nitrate–nitrogen (NO₃⁻-N), key enzyme activities (catalase, CAT; sucrase, IA; urease, UA), and bacterial communities; redundancy analysis linked edaphic variables to community structure. Results: Co-application remodeled the soil microenvironment. ZB1 moderated pH from 7.92 (weakly alkaline) to 7.46 (near neutral) and increased total nitrogen (TN) and potassium (K). NO₃⁻-N rose 1.38-fold (ZB1) and 2.14-fold (ZB2), indicating improved N retention and reduced leaching risk. CAT, IA, and UA activities increased concurrently. The bacterial community shifted from Acidobacteria toward Bacteroidetes and was enriched in taxa including Flavobacterium. Redundancy analysis identified total nitrogen as the primary environmental driver of community change. Conclusions: Leguminous cover crops combined with B. cereus synergistically improved soil conditions and reorganized bacterial communities in dry–hot valley coffee systems, providing field-scale evidence and practical guidance for sustainable agroecosystem management. Full article

The growing popularity of plant-based meat alternatives (PBMAs) has increased interest in their microbiological safety, particularly during refrigerated storage. However, despite the rapid expansion of this product category, limited information is available on the microbiological status of industrially produced, heat-treated PBMAs. This study evaluated the microbiological safety of heat-treated PBMAs during refrigerated storage using a descriptive approach. A total of 100 formulations, including salami-type and frankfurter-type products, were produced under standardized industrial conditions and subjected to thermal processing at a core temperature of ≥92 °C. Microbiological analyses were carried out at four storage intervals (day 0, 15, 35, and 60 during storage at 0–4 °C), focusing on selected foodborne pathogens and hygiene indicator microorganisms, together with key physicochemical parameters, including pH, water activity, moisture content, and salt content. Salmonella spp., Listeria monocytogenes and Enterobacteriaceae were not detected in any sample throughout the storage period. Hygiene indicator microorganisms were not detected during early storage (day 0–15), while limited occurrence was recorded at extended storage (day 60), including Escherichia coli (3%), coagulase-positive Staphylococcus aureus (20%), and Bacillus cereus (15%). Detected Staphylococcus aureus levels ranged between 10³ and 10⁵ CFU/g. Overall, the results suggest good microbiological stability during early refrigerated storage, with some microbial presence appearing at extended storage. These findings provide a practical overview of the microbiological safety of heat-treated PBMAs under typical industrial processing and storage conditions. From an industrial perspective, these results may support the establishment of appropriate refrigerated storage limits, post-processing hygiene control, and routine microbiological monitoring strategies for heat-treated PBMAs. Full article

Cereulide is a heat-stable cyclic depsipeptide toxin produced by Bacillus cereus and is responsible for foodborne emetic syndrome. Recent reports of Bacillus cereus contamination and cereulide occurrence in infant formula have raised increasing food safety concerns. Due to the immature immune and metabolic systems of infants, exposure to cereulide through contaminated formula may lead to potential health risks. However, direct application of existing cereulide analytical methods to infant formula remains challenging because of the unique processing technologies, encapsulated nutrients, and variable matrix composition of this product category, which may hinder toxin release and cause significant matrix interference. In practical analysis, inter-laboratory comparisons revealed that existing methods exhibited relatively large deviations and insufficient sensitivity, making them not specifically optimized for infant formula matrices. The present study was motivated by the need for a matrix-specific, sensitive, and reliable analytical method for cereulide determination in infant formula. In this study, a method based on solid-phase extraction coupled with ultra-performance liquid chromatography–tandem mass spectrometry (SPE–UPLC–MS/MS) was developed and validated. To improve the applicability of cereulide analysis to infant formula, this method incorporates a hydration-assisted extraction step tailored to infant formula, which increased the detected cereulide response by approximately fourfold, together with optimized SPE clean-up and improved chromatographic conditions to reduce matrix effects and enhance quantitative reproducibility. The method showed good linearity (0.1–10 μg·L⁻¹, R² > 0.999), low values for limit of detection (LOD) (0.03 μg·kg⁻¹) and limit of quantification (LOQ) (0.1 μg·kg⁻¹), and acceptable recoveries (94.4–110.3%) with RSDs below 3.7%. The developed method was successfully applied to commercial infant formula samples, and cereulide-positive samples were identified. This method provides a reliable analytical tool for the monitoring of cereulide in infant formula and contributes to improved food safety surveillance and exposure risk assessment. Full article

Polluted mangroves are ecologically sensitive habitats that provide ecosystem services. In a selected polluted forest of Simaisma, viable aerobic, halophilic, and heterotrophic hydrocarbon-degrading bacterial strains were isolated from both rhizosphere and non-rhizosphere regions. The chemical composition of sediment showed a clear distinction between the rhizosphere and non-rhizosphere sites, as well as coastal and non-coastal sediments, as per Principal Component Analysis (PCA) clustering. Anthracene, an indicator of oil pollution, was present along with vanadium, another marker of oil pollution. Through selective enrichment cultures, a total of 25 hydrocarbon-degrading bacterial strains were isolated, including Lysinibacillus xylanilyticus, Bacillus cereus, Lysinibacillus sphaericus, Pseudomonas stutzeri, Acinetobacter calcoaceticus, and Staphylococcus warneri. To link the adaptation of bacteria to sediment chemistry, nine B. cereus strains were investigated using their MALDI-TOF MS protein profiles combined with their dendrogram. The relationship between protein profiles of B. cereus strains with their biosurfactant production capabilities was explained by a tanglogram. The tanglegram suggests that biosurfactant production is an important functional trait in B. cereus, but it is not consistently reflected in the overall protein profile. This suggests that bacterial adaptation in the polluted mangrove sediments may involve changes at multiple cellular levels, including metabolic activity and variation in protein expression profiles. These findings confirm the involvement of mangrove-associated bacteria in the sustainability of mangrove forests by promoting bioremediation of oil pollution, thereby protecting coastal ecosystems and their environmental and socio-economic aspects. Full article

Antimicrobial blue light (aBL) within the visible violet–blue spectrum has emerged as a promising non-chemical strategy for microbial control, yet its performance across environmentally realistic matrices and surfaces remains insufficiently characterised. Here, we evaluate a continuous-exposure aBL LED system operating within the visible 407–421 nm range for its antimicrobial efficacy against Escherichia coli K-12 MG1655 and Bacillus cereus NCTC 11143 across liquid cultures, agar surfaces, and representative built-environment materials (glass and steel bar). Bacterial inactivation was quantified using culture-based enumeration and flow cytometric viability profiling. The system delivered a controlled irradiance of 0.72 mW/cm² at 58 cm, corresponding to cumulative doses of 2.59–62.23 J cm⁻² over 1–24 h of exposure. Significant, time-dependent reductions in viability were observed across all matrices relative to fluorescent-light controls, with near-complete or complete loss of recoverable cells on solid surfaces following prolonged exposure. Flow cytometric analyses revealed progressive transitions from viable to injured and dead cell populations, consistent with photodynamic inactivation mediated by endogenous photosensitiser activation and reactive oxygen species generation. These findings demonstrate that continuous visible-light aBL illumination can achieve effective multisurface microbial inactivation under moderate irradiance conditions compatible with occupied environments, supporting its translational potential as a sustainable, non-chemical decontamination strategy for healthcare, food-processing, and built environments. Full article

Silage is a fundamental component of cattle feed, and its microbiological quality is critical for animal health and human safety. Improper ensiling conditions, such as oxygen exposure or inadequate acidification, can promote the growth of pathogens like Listeria monocytogenes, Clostridium botulinum, and Bacillus cereus. This study aimed to evaluate the microbial status of maize silages and identify pre-ensiling factors influencing its hygienic safety. Over a four-year period, 406 silage samples were collected from cattle farms across Poland. The research evaluated general hygiene indicators and screened for specific pathogens using standard culture methods, polymerase chain reaction toxotyping, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The impact of agricultural practices, including soil quality, organic fertilization, and microbial inoculation, was also analyzed. The analysis revealed that 32.1% of silages fell outside the reference pH range, indicating potential aerobic instability. While Salmonella and Campylobacter were not detected, Clostridium spp. were highly prevalent (81.0%), and C. perfringens was confirmed in 24.9% of samples. Listeria species occurred in 2.9% of silages, with L. innocua being the most frequent isolate. Statistical analysis showed that organic fertilization was significantly linked to specific C. perfringens toxotypes, though it did not increase the overall microbial burden. Conversely, microbial inoculation generally reduced the counts of several undesirable bacteria, although these differences were not statistically significant across all parameters. High pH values and significant contamination with Clostridium, B. cereus, and fungi remain critical challenges for silage safety. The results underscore the necessity for improved agricultural practices—specifically the minimization of soil and manure contamination during harvest—and the broader adoption of microbial inoculation to ensure the microbiological stability of fermented forage. Full article

Antimicrobial resistance (AMR) is increasingly becoming a major global public health concern, as antibiotics are losing their effectiveness at an alarming rate due to drug resistance. The ß-lactam group of antibiotics are widely used in dairy farms to treat animal infections, and their presence in the food chain is a significant concern. Addressing this issue requires the development of effective analytical tools for the rapid detection of antibiotics. In this work, a miniaturized Biosensor-on-Pillar platform was developed for detecting ß-lactam antibiotics in milk, which operates in a rapid, cost-effective, and user-friendly format, making it particularly suitable for resource-limited settings. The platform employs an enzyme induction-based approach, wherein Bacillus cereus spores germinate in the presence of β-lactam antibiotics, leading to the production of β-lactamase enzyme, which is then recognized using a chromogenic substrate functionalized on paper associated with the pillar platform. The developed biosensor can detect 12 β-lactam antibiotics with limits of detection (LODs) ranging from 1 to 1000 ppb, achieving sensitivity at or below the maximum residue limits (MRLs) set by regulatory bodies (FSSAI/CODEX) for the majority of the tested antibiotics. The performance of the platform, including the design, fabrication, and working principle, was further evaluated by analyzing six blind milk samples, yielding significant results compared to the commercially available AOAC-approved gold-standard method. Hence, the developed biosensor demonstrates promising potential for the rapid, cost-effective and high-throughput screening of milk samples for β-lactam antibiotics, benefiting the dairy industry and ensuring food safety. Full article

Recently, polyhydroxyalkanoates (PHAs) have gained significant attention as a bioactive material for replacing petrochemical plastics. PHAs can be produced by microorganisms growing on sludge substrates. In this study, fish-processing wastewater was investigated as an alternative substrate for PHA production using Bacillus cereus. Wastewater dilution, carbon-to-nitrogen ratio modification, and the addition of fish oil as a lipidic substrate were examined, and bacterial growth and biopolymer production were optimized. First, wastewater was diluted (25–100%) and examined. The 50% dilution treatment was selected, yielding a CDM of 0.426 g/L and a PHA content of 6.69%. In subsequent steps, the effects of wastewater fermentation and bacterial adaptation prior to the main production processes were investigated. According to the results, the 50% and 100% fermented treatments exhibited higher CDM values (0.970–1.022 g/L) compared to the non-fermented treatments. Cultures inoculated with adapted bacteria showed superior performance (CDM: 1.455 g/L, PHA: 0.499 g/L, PHA content: 34.63%) relative to non-adapted treatments. The effect of the carbon-to-nitrogen (C/N) ratio was also optimized by supplementing two carbon sources: glucose and crude fish oil. The optimal treatment T1 (effluent + 0.6 g/L glucose) had a CDM of 1.32 g/L and a PHA content of 0.215 g/L. Treatment 1, which consisted solely of effluent and fish oil, exhibited higher values (CDM: 1.12 g/L, PHA: 0.65 g/L) and was therefore considered the cost-effective treatment. Subsequently, a scale-up process was conducted in a 4 L bioreactor over 300 h under semi-continuous, long-term cultivation. The optimal harvesting time for the biopolymer was achieved during the fourth cycle (180–240 h). The produced biopolymer was characterized using FTIR, NMR, TGA, DSC, SEM, and XRD analyses, confirming the production of a copolymer, specifically poly(3-hydroxybutyrate-co-3-hydroxyvalerate). This study used wastewater from the fish industry for the production of biodegradable polyhydroxyalkanoates. Full article

Despite its rich bioactive composition, orange peel dust (OPD), a fine industrial by-product generated during citrus processing in the filter tea industry, has not received much attention as a valuable matrix. Using antioxidant activity (2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and reducing power (RP)), α-amylase inhibitory activity, antimicrobial potential, and sugar composition as function-oriented indicators, this study aimed to compare four green extraction technologies: subcritical water extraction (SWE), pressurized ethanol extraction (PEE), ultrasound-assisted extraction (UAE), and sequential supercritical CO₂–UAE (Sc-CO₂–UAE) applied to OPD derived from Citrus sinensis L. Among thermally driven techniques, PEE at 220 °C had the highest radical-scavenging activity, while UAE showed the broadest antifungal activity against Fusarium spp. and Alternaria alternata, along with selective antibacterial activity against Bacillus cereus. Sequential Sc-CO₂ pretreatment at 300 bar followed by UAE resulted in the highest α-amylase inhibitory activity. Sugar analysis indicated that thermal conditions enhanced carbohydrate hydrolysis, while UAE and Sc-CO₂-UAE maintained structural sugars under mild conditions. All green extraction approaches outperformed conventional maceration. These findings validate OPD as a valuable industrial by-product suitable for sustainable valorization, supporting circular economy concepts in the citrus processing sector. Full article

The high mortality rate from microbial infections underscores the need to discover new antimicrobials. This work produced antibacterial Chitosan biofilms with and without the incorporation of the ethanolic extract of Libidibia ferrea stem bark and its ethyl acetate and aqueous fractions. The extract and fractions were subjected to FTIR and ¹H NMR analysis. The biofilms were characterized by FTIR, scanning electron microscopy, thermogravimetry, and differential scanning calorimetry analysis. The ¹H NMR and FTIR data, as well as the colorimetric quantification of total phenolics, demonstrated the presence of phenolic compounds. Staphylococcus aureus and Bacillus cereus were the most susceptible bacteria for Chitosan/L. ferrea biofilms and fractions (growth inhibition zones values in the range of 10.8 ± 0.1 to 14.0 ± 0.1 mm, and minimum inhibitory or bactericidal concentration, MIC or MBC values of the fractions were in the range of 125 to 250 µg mL⁻¹. Only the fractions inhibited Pseudomonas aeruginosa (MIC = 250 µg mL⁻¹). Chitosan/L. ferrea biofilms exhibited efficient interactions between chitosan functional groups and secondary metabolites, good thermal stability, and increased rigidity in mechanical tests. This study reinforces the pharmacological potential of biodegradable Chitosan/L. ferrea biofilms as antibacterial agents biofilms. Full article

Bacterial β-barrel pore-forming toxins, including Staphylococcus aureus α-toxin (Hla) and Bacillus cereus toxins hemolysin II (HlyII) and cytolytic toxin K2 (CytK-2), are secreted by bacterial cells as water-soluble monomers. These monomers assemble within lipid bilayers to form cylindrical pores, leading to lysis of target eukaryotic cells. We created mutant forms of these toxins that, based on the results of X-ray structural analysis of Hla and the prediction of the 3D structure of HlyII and CytK2, can form intramolecular disulfide bonds in monomers. The substitutions were made in the region responsible for toxin insertion into the target membrane. The mutant forms reversibly altered their hemolytic activity depending on the presence of reducing reagents and were non-toxic when injected into experimental animals. The immune response to injection of the mutant forms of Hla and CytK-2 toxins resulted in higher antibody titers against the wild-type toxins and a higher level of immunological memory than with injection of the HlyII mutant. The mutant form of CytK-2 demonstrates the properties of a prototype vaccine, as immunization with this protein protects animals against the effects of the wild-type toxin. Full article

This study aimed to evaluate the physicochemical characterization and antibacterial activity of the essential oil (EO) extracted from the leaves of Mentha rotundifolia (L.) Huds. Molecular interactions between bioactive ligand compounds, target bacterial proteins and DNA gyrase subunit B (GyrB), as well as an in silico ADMET prediction study, were also conducted. The EO was obtained by hydrodistillation of the plant leaves. The Gas Chromatography–Tandem Mass Spectrometry (GC-MS/MS) analysis revealed Rotundifolone (27.95%) and carvacrol (19.48%) as the major constituents. Other components identified included Piperitenone (6.09%), Cinerolon (4.73%), and Pulegone (4.47%). Antibacterial activity was assessed against six bacterial strains: Enterococcus faecalis CIP 103214, Salmonella Typhi CIP 5535, Staphylococcus aureus ATCC 9144, Bacillus cereus ATCC 33019, Streptococcus agalactiae IPM 24842, and Providencia alcalifaciens CIP 82.90T. The disk diffusion assay showed a strong inhibitory effect against E. faecalis (inhibition zone: 19.66 ± 0.3 mm), while the lowest minimum inhibitory concentration (MIC) was observed for B. cereus (0.58 ± 0.01 µL/mL). The time-kill kinetics assay showed a progressive inactivation of all tested bacterial strains after their exposure to EO for 8 h at MICs. Furthermore, Molecular docking showed remarkable affinities between EO components, target proteins and DNA gyrase subunit B (GyrB). Moreover, the in silico ADMET predictions provided preliminary insights into the safety-related properties of the major EO components. In addition, EO compounds have the potential to interact with bacterial structures. These findings highlight the in vitro antibacterial potential of the M. rotundifolia EO and suggest its promise as a natural source of bioactive compounds. Full article

This study investigated the effects of dietary Bacillus cereus, administered alone or in combination with biofloc technology, on the growth performance, immune response, disease resistance, and intestinal microbiota of Litopenaeus vannamei. Shrimp fed diets supplemented with B. cereus, either directly or via biofloc systems, exhibited significantly increased final body weight and specific growth rate, together with a reduced feed conversion ratio compared with the control group. The expression levels of key hepatopancreatic immune-related genes, including lysozyme, prophenoloxidase, superoxide dismutase, Toll, immune deficiency, and Relish, were significantly upregulated in probiotic-associated treatments. Following challenge with Vibrio parahaemolyticus, cumulative mortality was markedly lower in all treatments involving B. cereus or biofloc compared with the control. Although alpha diversity indices were not significantly affected, beta diversity analysis demonstrated that supplementation frequency and delivery mode altered intestinal microbial community structure. The phyla Bacteroidota, Firmicutes, and Proteobacteria predominated across treatments, while members of Marinilabiliaceae and Shewanellaceae were enriched under probiotic-associated conditions, suggesting enhanced nutrient transformation potential. Co-occurrence network analysis further revealed increased microbial network complexity and positive interactions in probiotic and biofloc treatments, indicating improved community stability. These findings demonstrate that the synergistic application of B. cereus and biofloc technology enhances growth performance, immune capacity, and intestinal microbial resilience in intensive shrimp culture, and that supplementation strategy plays a critical role in optimizing probiotic efficacy. Full article

Synthetic dyes, such as methylene blue (MB), constitute a major category of environmental pollutants due to their toxicity, persistence, and resistance to standard treatment methods. In this study, Bacillus cereus BC WW Saida was isolated from the heavily polluted Saida dumpsite in Lebanon and evaluated for its MB degradation efficiency. The isolate was identified through whole-genome sequencing, which revealed the presence of key enzymatic systems involved in azo dye degradation. Under optimized conditions, the strain achieved 82% decolorization, as determined by optical density measurements using a microplate reader. The process was further examined using High-Performance Liquid Chromatography (HPLC), which revealed a significant reduction in the original dye peak and the emergence of new intermediate products. These findings suggest the strong biodegradation capability of B. cereus BC WW Saida isolated from contaminated environments and highlight its potential application in the eco-friendly treatment of azo dye-contaminated wastewater. Full article

The presence of pharmaceutical residues, such as ibuprofen, in aquatic environments poses a growing environmental challenge due to their persistence and potential ecotoxicological effects. In this study, a novel biohybrid composite based on pyrolysed rice husk (biochar) modified with Bacillus cereus cells was developed for the efficient removal of ibuprofen from aqueous solutions. The material was comprehensively characterised using SEM, BET, TGA, CHN analysis, and FTIR spectroscopy. Pyrolysis significantly increased the surface area (up to 300 m² g⁻¹) and porosity compared to raw rice husk, while bacterial immobilisation introduced additional functional groups, enhancing surface heterogeneity. Batch adsorption experiments demonstrated a clear improvement in adsorption capacity in the order of rice husk < biochar < composite. The maximum Langmuir adsorption capacities were 4.86, 11.68, and 13.73 mg g⁻¹ for rice husk, biochar, and the composite, respectively. Isotherm modelling indicated that ibuprofen adsorption was best described by the Langmuir and the Freundlich models, suggesting a combination of monolayer adsorption and heterogeneous surface interactions. Isotherm analyses (D–R energy values < 9 kJ mol⁻¹) indicate that ibuprofen removal occurs predominantly through physisorption, governed by π–π interactions, hydrogen bonding, and surface heterogeneity rather than chemisorption. Kinetic studies revealed rapid adsorption behaviour, with pseudo-first-order and pseudo-second-order models providing the best fit (R² up to 0.997). The Weber–Morris model confirmed that intraparticle diffusion contributed to the process but was not the sole rate-limiting step. The enhanced performance of the composite is attributed to synergistic effects between physicochemical adsorption on the porous carbon matrix and interactions with bacterial cell wall functional groups. The developed composite represents a low-cost, sustainable, and highly effective material for ibuprofen removal from contaminated water. Full article