Vegetative Cells Research Articles

Microbial preference for chlorate over perchlorate under simulated shallow subsurface Mars-like conditions

The Martian surface and shallow subsurface lacks stable liquid water, yet hygroscopic salts in the regolith may enable the transient formation of liquid brines. This study investigated the combined impact of water scarcity, UV exposure, and regolith depth on microbial survival under Mars-like environmental conditions. Both vegetative cells of Debaryomyces hansenii and Planococcus halocryophilus, alongside with spores of Aspergillus niger, were exposed to an experimental chamber simulating Martian environmental conditions (constant temperatures of about − 11 °C, low pressure of approximately 6 mbar, a CO2 atmosphere, and 2 h of daily UV irradiation). We evaluated colony-forming units (CFU) and water content at three different regolith depths before and after exposure periods of 3 and 7 days, respectively. Each organism was tested under three conditions: one without the addition of salts to the regolith, one containing sodium chlorate, and one with sodium perchlorate. Our results reveal that the residual water content after the exposure experiments increased with regolith depth, along with the organism survival rates in chlorate-containing and salt-free samples. The survival rates of the three organisms in perchlorate-containing regolith were consistently lower for all organisms and depths compared to chlorate, with the most significant difference being observed at a depth of 10–12 cm, which corresponds to the depth with the highest residual water content. The postulated reason for this is an increase in the salt concentration at this depth due to the freezing of water, showing that for these organisms, perchlorate brines are more toxic than chlorate brines under the experimental conditions. This underscores the significance of chlorate salts when considering the habitability of Martian environments.

The Effectiveness of LISTEXTM P100 in Reducing the Biofilm of Listeria spp. on Steel, Plastic, and Galvanised Surfaces.

Eliminating and managing L. monocytogenes, L. welshimeri, and L. ivanovii biofilms is a significant problem for food safety, as listeriosis is among the worst foodborne illnesses. The Listex P100 bacteriophage's bactericidal and inhibitory properties have been investigated in relation to varying strains of vegetative cells and biofilms of L. monocytogenes, L. welshimeri, and L. ivanovii. The phage concentrations of 109 and 1010 PFU/ml showed strong antibacterial activity against L. monocytogenes, L. welshimeri, and L. ivanovii at both 10°C and 30°C (P<0.05). In 96- well microplate experiments, bacteriophage treatment inhibited biofilm development and reduced biofilm by up to 57.6% (P ≤ 0.05). When compared to controls, Listex P100 bacteriophage significantly reduced the populations of L. monocytogenes, L. welshimeri, and L. ivanovii biofilms on the surfaces of galvanised, stainless steel, and plastic surfaces where holes were produced and the structure of Listeria spp. was disturbed. This study clearly demonstrated that L. monocytogenes, L. welshimeri, and L. ivanovii biofilms on galvanised, stainless steel, and plastic surfaces might be removed by using Listex P100 bacteriophage.

Inactivation mechanism of phenyllactic acid against Bacillus cereus spores and its application in milk beverage

Phenyllactic acid (PLA) as a natural phenolic acid exhibits antibacterial activity against non-spore-forming bacteria, while the inhibitory effect against bacterial spore remained unknown. Herein, this study investigated the inactivation effect of PLA against Bacillus cereus spores. The results revealed that the minimum inhibitory concentration of PLA was 1.25 mg/mL. PLA inhibited the outgrowth of germinated spores into vegetative cells rather than germination of spores. PLA disrupted the spore coat, and damaged the permeability and integrity of inner membrane. Moreover, PLA disturbed the establishment of membrane potential due to the inhibition of oxidative metabolism. SEM observations further visualized the morphological changes and structural disruption caused by PLA. Besides, PLA caused the degradation of DNA of germinated spores. Finally, PLA was applied in milk beverage, and showed promising inhibitory effect against B. cereus spores. This finding could provide scientific basis for the application of PLA against spore-forming bacteria in food industry.

Inhibition and Mechanism of Citral on Bacillus cereus Vegetative Cells, Spores, and Biofilms.

Bacillus cereus causes food poisoning by producing toxins that cause diarrhea and vomiting and, in severe cases, endocarditis, meningitis, and other diseases. It also tends to form biofilms and spores that lead to contamination of the food production environment. Citral is a potent natural antibacterial agent, but its antibacterial activity against B. cereus has not been extensively studied. In this study, we first determined the minimum inhibitory concentrations and minimum bactericidal concentrations, growth curves, killing effect in different media, membrane potential, intracellular adenosine triphosphate (ATP), reactive oxygen species levels, and morphology of vegetative cells, followed by germination rate, morphology, germination state of spores, and finally biofilm clearance effect. The results showed that the minimum inhibitory concentrations and minimum bactericidal concentrations of citral against bacteria ranged from 100 to 800 μg/mL. The lag phase of bacteria was effectively prolonged by citral, and the growth rate of bacteria was slowed down. Bacteria in Luria-Bertani broth were reduced to below the detection limit by citral at 800 μg/mL within 0.5 h. Bacteria in rice were reduced to 3 log CFU/g by citral at 4000 μg/mL within 0.5 h. After treatment with citral, intracellular ATP concentration was reduced, membrane potential was altered, intracellular reactive oxygen species concentration was increased, and normal cell morphology was altered. After treatment with citral at 400 μg/mL, spore germination rate was reduced to 16.71%, spore morphology was affected, and spore germination state was altered. It also had a good effect on biofilm removal. The present study showed that citral had good bacteriostatic activity against B. cereus vegetative cells and its spores and also had a good clearance effect on its biofilm. Citral has the potential to be used as a bacteriostatic substance for the control of B. cereus in food industry production.

Mechanism of Bacillus coagulans T242 in prevention of Salmonella infection in mice

Salmonella is recognized as an important foodborne pathogen, which seriously threatens human safety. Bacillus coagulans T242 (T242) could prevent the growth of Salmonella and alleviate Salmonella infection in Caco-2 cells in vitro. The main objective of this study was to investigate the mechanism of action of T242 in preventing Salmonella infection in mice. Both T242 vegetative cells and spore prevention groups could reduce the bacterial loads in the ileum, liver and spleen of Salmonella typhimurium (S. typhi) SL1344 infected mice, and reduce the levels of Interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) in serum and also increase the expression of Interleukin-1β (IL-1β) in serum, Monocyte chemoattractant protein-1 (MCP-1) and Inducible nitric oxide synthase (iNOS) in the liver, Interleukin-22 (IL-22) and Interleukin-23 (IL-23) in the ileum, and tight junction proteins (Claudin-1, Occludin and ZO-1) in the colon, and regulate intestinal flora. Pretreatment of T242 spores could downregulate the relative abundance of Desulfovibrionaceae in Salmonella infected mice. The relative abundance of Ruminococcus in Salmonella-infected mice was down-regulated by T242 vegetative cells preconditioning. The results showed that T242 prevented the systemic spread of Salmonella by regulating intestinal flora and enhancing intestinal barrier function against Salmonella invasion, inducing anti-inflammatory effects and regulating immune responses. T242 had great potential in controlling the epidemic of foodborne pathogens and was expected to be an effective alternative to antibiotic therapy, which is of great significance for the control and defense of Salmonella infection.

Synchronic distribution of the dinoflagellate Protoceratium reticulatum and yessotoxins in a high stratified fjord system: Tidal or light modulation?

Protoceratium reticulatum is the main yessotoxin-producer along the Chilean coast. Thus far, the yessotoxin levels recorded in this region have not posed a serious threat to human health. However, a bloom of P. reticulatum during the austral summer of 2022 caused the first ban of shellfish collection, due to the high toxin levels. A bloom of P. reticulatum during the austral summer of 2020 allowed an evaluation of the fine-scale distribution of the dinoflagellate during a tidal cycle. High-resolution measurements of biophysical properties were carried out in mid-summer (February 18–19) at a fixed sampling station in Puyuhuapi Fjord, Chilean Patagonia, as part of an intensive 24-h biophysical experiment to monitor the circadian distributions of P. reticulatum vegetative cells and yessotoxins. High P. reticulatum cell densities (>20 × 103 cells L–1) were found in association with a warmer (14.5–15 °C) and estuarine (23.5–24.5 g kg–1) sub-surface water layer (6–8 m). P. reticulatum cell numbers and yessotoxins followed a synchronic distribution pattern consistent with the excursions of the pycnocline. Nevertheless, the surface aggregation of the cells was modulated by the light cycle, suggesting daily vertical migration. The yessotoxin content per P. reticulatum cell ranged from 9.4 to 52.2 pg. This study demonstrates both the value of fine-scale resolution measurements of biophysical properties in a highly stratified system and the potential ecosystem impact of P. reticulatum strains producing high levels of yessotoxins.

Optimization of continuous astaxanthin production by Haematococcus pluvialis in nitrogen-limited photobioreactor

The industrial production of astaxanthin from Haematococcus pluvialis is mainly operated following a two-stage cultivation strategy in photobioreactors (PBRs) operating in batch mode. This study provides a strategy for optimizing continuous astaxanthin production in a one-stage approach, by optimizing both astaxanthin accumulation and biomass productivity of H. pluvialis under nitrogen-limited condition. To achieve the good balance in culture conditions to maintain growth under nitrogen-limited conditions, while triggering significant astaxanthin accumulation in reddish vegetative cells, the role of light absorption, as represented by the mean rate of photon absorption (MRPA), and nitrate concentration was especially investigated. For that purpose, H. pluvialis cultures were grown in a flat-panel PBR with constant dilution rate D of 0.015 h−1, different photon flux densities PFDs ranging from 75 to 750 μmolhν·m−2·s−1 and different nitrogen concentration levels in the feeding medium [NO3−] of 1, 3 and 8.8 mM. A parabolic relationship between MRPA and astaxanthin production rate was obtained. This indicated that astaxanthin synthesis was limited by the MRPA, opening further optimization by adjusting incident light intensity, nitrogen concentration in the feeding medium or culture dilution rate. After optimization of operating conditions, we demonstrated that a large quantity of astaxanthin can be produced in continuous mode, following then a one-stage strategy which may be advantageous for industrial use. A maximum astaxanthin productivity of 1.27 ± 0.03 g·m−2·d−1 with an astaxanthin accumulation of 3.9 ± 0.2 % DW was reached for the culture featuring MRPA of 9000 μmolhν·kgx−1·s−1. The process was also proved reversible, allowing to tailor the biomass composition and physiological state (from green to red cells enriched in astaxanthin) by adjusting operating conditions, opening perspectives from an optimized coupling with downstream processing steps.

Особливості онтогенезу бацил впродовж розвитку від спори до вегетативної клітини

In the review, on the example of aerobic spore-forming bacteria, the problems of the development and ontogenesis of the bacterial cell are considered along with the possibilities of influencing the course of these processes. The characteristics of the main concepts "growth", "differentiation", and "development" as independent processes with their dynamic interrelation are presented. Attention is focused on the analysis of literature data on the peculiarities of vegetative cell development, starting from a spore in a dormant state and finishing with vegetative form formation. In particular, the mechanisms that maintain the spore dormant state and subsequent processes of activation, initiation, outgrowth, and vegetative cell formation are described. There are emphasized certain problems with research on the ontogenesis of bacterial cells due to the deficiency of appropriate methods, as well as the lack of a single opinion regarding individual stages of the development and vegetative form formation. It was concluded that the study of individual stages of the development of prokaryotes, which differ in spore-forming and non-spore-forming microorganisms, is still relevant. Knowledge of these processes will help scientists to develop mechanisms of influence on the ontogenesis of microorganisms.

Visualization of Bacillus subtilis spore structure and germination using quick-freeze deep-etch electron microscopy.

Bacterial spores, known for their complex and resilient structures, have been the focus of visualization using various methodologies. In this study, we applied quick-freeze and replica electron microscopy techniques, allowing observation of Bacillus subtilis spores in high-contrast and three-dimensional detail. This method facilitated visualization of the spore structure with enhanced resolution and provided new insights into the spores and their germination processes. We identified and described five distinct structures: (i) hair-like structures on the spore surface, (ii) spike formation on the surface of lysozyme-treated spores, (iii) the fractured appearance of the spore cortex during germination, (iv) potential connections between small vesicles and the core membrane and (v) the evolving surface structure of nascent vegetative cells during germination.

The roles of yeast formins and their regulators Bud6 and Bil2 in the pheromone response.

In response to pheromone Saccharomyces cerevisiae extend a mating projection. This process depends on the formation of polarized actin cables which direct secretion to the mating tip and translocate the nucleus for karyogamy. Here, we demonstrate that proper mating projection formation requires the formin Bni1, as well as the actin nucleation promoting activities of Bud6, but not the formin Bnr1. Further, Bni1 is required for pheromone gradient tracking. Our work also reveals unexpected new functions for Bil2 in the pheromone response. Previously we identified Bil2 as a direct inhibitor of Bnr1 during vegetative cell growth. Here, we show that Bil2 has Bnr1-independent functions in spatially focusing Bni1-GFP at mating projection tips, and in vitro Bil2 and its binding partner Bud6 organize Bni1 into clusters that nucleate actin assembly. bil2∆ cells also display entangled Bni1-generated actin cable arrays and defects in secretory vesicle transport and nuclear positioning. At low pheromone concentrations, bil2∆ cells are delayed in establishing a polarity axis, and at high concentrations they prematurely form a second and a third mating projection. Together, these results suggest that Bil2 promotes the proper formation and timing of mating projections by organizing Bni1 and maintaining a persistent axis of polarized growth.

In Silico Design of Potential Small-Molecule Antibiotic Adjuvants against Salmonella typhimurium Ortho Acetyl Sulphydrylase Synthase to Address Antimicrobial Resistance.

The inhibition of O-acetyl sulphydrylase synthase isoforms has been reported to represent a promising approach for the development of antibiotic adjuvants. This occurs via the organism developing an unpaired oxidative stress response, causing a reduction in antibiotic resistance in vegetative and swarm cell populations. This consequently increases the effectiveness of conventional antibiotics at lower doses. This study aimed to predict potential inhibitors of Salmonella typhimurium ortho acetyl sulphydrylase synthase (StOASS), which has lower binding energy than the cocrystalized ligand pyridoxal 5 phosphate (PLP), using a computer-aided drug design approach including pharmacophore modeling, virtual screening, and in silico ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) evaluation. The screening and molecular docking of 4254 compounds obtained from the PubChem database were carried out using AutoDock vina, while a post-screening analysis was carried out using Discovery Studio. The best three hits were compounds with the PubChem IDs 118614633, 135715279, and 155773276, possessing binding affinities of -9.1, -8.9, and -8.8 kcal/mol, respectively. The in silico ADMET prediction showed that the pharmacokinetic properties of the best hits were relatively good. The optimization of the best three hits via scaffold hopping gave rise to 187 compounds, and they were docked against StOASS; this revealed that lead compound 1 had the lowest binding energy (-9.3 kcal/mol) and performed better than its parent compound 155773276. Lead compound 1, with the best binding affinity, has a hydroxyl group in its structure and a change in the core heterocycle of its parent compound to benzimidazole, and pyrimidine introduces a synergistic effect and consequently increases the binding energy. The stability of the best hit and optimized compound at the StOASS active site was determined using RMSD, RMSF, radius of gyration, and SASA plots generated from a molecular dynamics simulation. The MD simulation results were also used to monitor how the introduction of new functional groups of optimized compounds contributes to the stability of ligands at the target active site. The improved binding affinity of these compounds compared to PLP and their toxicity profile, which is predicted to be mild, highlights them as good inhibitors of StOASS, and hence, possible antimicrobial adjuvants.

Impact of species and strain variability on non-thermal plasma decontamination efficacy

Microwave non-thermal plasma (NTP) is a promising preservation technology for surface decontamination, alternative to conventional methods such as heat treatments and chemical additives. This study explores the impact of plasma processed air (PPA) exposure on 50 strains from five diverse microbial species relevant to food quality and safety. Bacterial vegetative cells were effectively inactivated in an agar plate model system. The average reduction after 60 s of treatment was 4.9 log10, 5.8 log10, and 5.9 log10 for Listeria monocytogenes, Escherichia coli, and Lactiplantibacillus plantarum respectively, with limited strain variability. Saccharomyces cerevisiae reached an average of 5.3 log10 reduction after 30 s of treatment, with one resistant strain (AD1890). Bacillus subtilis spores were inactivated by 2.0 log10 on average after 30 min, with two distinct strain groups. These results showed that PPA can be used as a mild decontamination treatment applicable to dry and heat-sensitive surfaces, suitable for various food applications.

Heat resistance of five spoilage microorganisms in a carbonated broth

Despite their acidic pH, carbonated beverages can be contaminated by spoilage microorganisms. Thermal treatments, before and/or after carbonation, are usually applied to prevent the growth of these microorganisms. However, the impact of CO2 on the heat resistance of spoilage microorganisms has never been studied. A better understanding of the combined impact of CO2 and pH on the heat resistance of spoilage microorganisms commonly found in carbonated beverages might allow to optimize thermal treatment.Five microorganisms were selected for this study: Alicyclobacillus acidoterrestris (spores), Aspergillus niger (spores), Byssochlamys fulva (spores), Saccharomyces cerevisiae (vegetative cells), and Zygosaccharomyces parabailii (vegetative cells). A method was developed to assess the impact of heat treatments in carbonated media on microbial resistance.The heat resistances of the five studied species are coherent with the literature, when data were available. However, neither the dissolved CO2 concentration (from 0 to 7 g/L), nor the pH (from 2.8 to 4.1) have an impact on the heat resistance of the selected microorganisms, except for As. niger, for which the presence of dissolved CO2 reduced the heat resistance. This study improved our knowledge about the heat resistance of some spoilage microorganisms in presence of CO2.

Mechanistic insights into the adaptive evolvability of spore heat resistance in Bacillus cereus sensu lato

Wet heat treatment is a commonly applied method in the food and medical industries for the inactivation of microorganisms, and bacterial spores in particular. While many studies have delved into the mechanisms underlying wet heat killing and spore resistance, little attention has so far been dedicated to the capacity of spore-forming bacteria to tune their resistance through adaptive evolution. Nevertheless, a recent study from our group revealed that a psychrotrophic strain of the Bacillus cereus sensu lato group (i.e. Bacillus weihenstephanensis LMG 18989) could readily and reproducibly evolve to acquire enhanced spore wet heat resistance without compromising its vegetative cell growth ability at low temperatures. In the current study, we demonstrate that another B. cereus strain (i.e. the mesophilic B. cereus sensu stricto ATCC 14579) can acquire significantly increased spore wet heat resistance as well, and we subjected both the previously and currently obtained mutants to whole genome sequencing. This revealed that five out of six mutants were affected in genes encoding regulators of the spore coat and exosporium pathway (i.e. spoIVFB, sigK and gerE), with three of them being affected in gerE. A synthetically constructed ATCC 14579 ΔgerE mutant likewise yielded spores with increased wet heat resistance, and incurred a compromised spore coat and exosporium. Further investigation revealed significantly increased spore DPA levels and core dehydration as the likely causes for the observed enhanced spore wet heat resistance. Interestingly, deletion of gerE in Bacillus subtilis 168 did not impose increased spore wet heat resistance, underscoring potentially different adaptive evolutionary paths in B. cereus and B. subtilis.

Retraction for Sarker et al., "Comparative Experiments To Examine the Effects of Heating on Vegetative Cells and Spores of Clostridium perfringens Isolates Carrying Plasmid Genes versus Chromosomal Enterotoxin Genes".

Retraction for Sarker et al., "Comparative Experiments To Examine the Effects of Heating on Vegetative Cells and Spores of Clostridium perfringens Isolates Carrying Plasmid Genes versus Chromosomal Enterotoxin Genes".

Pilot-scale production of Bacillus subtilis MSCL 897 spore biomass and antifungal secondary metabolites in a low-cost medium.

Bacillus subtilis is a plant growth promoting bacterium (PGPB) that acts as a microbial fertilizer and biocontrol agent, providing benefits such as boosting crop productivity and improving nutrient content. It is able to produce secondary metabolites and endospores simultaneously, enhancing its ability to survive in unfavorable conditions and eliminate competing microorganisms. Optimizing cultivation methods to produce B. subtilis MSCL 897 spores on an industrial scale, requires a suitable medium, typically made from food industry by-products, and optimal temperature and pH levels to achieve high vegetative cell and spore densities with maximum productivity. This research demonstrates successful pilot-scale (100L bioreactor) production of a biocontrol agent B. subtilis with good spore yields (1.5×109 spores mL-1) and a high degree of sporulation (>80%) using a low-cost cultivation medium. Culture samples showed excellent antifungal activity (1.6-2.3cm) against several phytopathogenic fungi. An improved methodology for inoculum preparation was investigated to ensure an optimal seed culture state prior to inoculation, promoting process batch-to-batch repeatability. Increasing the molasses concentration in the medium and operating the process in fed-batch mode with additional molasses feed, did not improve the overall spore yield, hence, process operation in batch mode with 10g molasses L-1 is preferred. Results also showed that the product quality was not significantly impacted for up to 12months of storage at room temperature. An economically-feasible process for B. subtilis-based biocontrol agent production was successfully developed at the pilot scale.

Microalgae for freshwater arsenic bioremediation: examining cellular toxicity, bioconcentration factor and eluding an alternative arsenic detoxification pathway

Microalgae are photoautotrophic organisms in freshwater systems known to uptake and bioremediate arsenic, a heavy metal. In this study, we compared the growth and arsenic uptake of two microalgae strains, Nostoc and Chlorella, to determine their suitability for arsenic bioremediation. As compared to the control, our results showed that treatment with As (III) enhanced the Nostoc growth by approximately 15% when grown in the absence of phosphate. The highest bioconcentration factor of Nostoc at this treatment was 1463.6, whereas 0.10 mg L−1 As (V) treatment improved the Chlorella growth by 25%, in the presence of phosphate. However, arsenic uptake reduced from 175.7 to 32.3 throughout the cultivation period for Chlorella. This suggests that Nostoc has an upper advantage in the bioremediation of arsenic as compared to the Chlorella strain. To gain insights into the potential of Nostoc in arsenic bioremediation, we further conducted SEM analysis on the vegetative cell surface. The SEM results showed that As (III) disrupted the Nostoc vegetative cell surface and structure. Further to this, pathway analysis and polymerase chain reaction (PCR) were conducted to identify the potential arsenic pathway regulated by Nostoc. The primary As (III)-related pathways elucidated include the arsA transporter and arsD complex that require ATP and As (III) methylation to S-adenosylmethionine. The phosphate deficiency condition resulting in the inability to generate ATP caused As (III) could not be excreted from the Nostoc cells, potentially contributing to the high arsenic concentration accumulated under phosphate-depleted conditions. These insights contribute to understanding the efficacy of microalgae strains in freshwater arsenic bioremediation.

Inactivation of Alicyclobacillus acidoterrestris in apple juice using pulsed light

We aimed to investigate the effect of pulsed light (PL) on inactivating Alicyclobacillus acidoterrestris in apple juice, the mechanism behind the inactivation, and the resultant effects on the quality of the juice. Our results showed that PL had a good inactivation effect on both spores and vegetative cells of A. acidoterrestris in apple juice, with a reduction of 3.5 ± 0.1 and 2.1 ± 0.1 log CFU/mL, respectively, following 3 s of treatment. This effect was better when lower concentrations of apple juice and lower liquid layer depths were used. Transmission electron microscopy (TEM) showed that the morphology and structure of A. acidoterrestris spores did not change significantly before and after treatment. When we compared the dipicolinic acid (DPA) release rate and death rate of the spores, we hypothesized that PL might have damaged the cortical proteins on A. acidoterrestris that are associated with the resistance of the spores, thus leading to their death. However, TEM revealed that the cell membranes of vegetative cells of A. acidoterrestris were intact and smooth before the treatment, but significantly wrinkled afterward. Raman spectroscopy revealed that the main targets of PL on A. acidoterrestris vegetative cells were: amide I on the proteins, nucleic acids [adenine(A) and guanine (G), in particular] and DNA (O–P–O‾ stretching) structures, and lipids (C–H2 deformation). The PL treatment increased the sugar content significantly, while having little to no effect on the other basic physicochemical parameters of apple juice.

A Dual-Mode Fluorescent Nanoprobe for the Detection and Visual Screening of Pathogenic Bacterial Spores.

Bacterial vegetative cells turn into metabolically dormant spores in certain environmental situations. Once suitable conditions trigger the germination of spores belonging to the pathogenic bacterial category, public safety and environmental hygiene will be threatened, and lives will even be endangered when encountering fatal ones. Instant identification of pathogenic bacterial spores remains a challenging task, since most current approaches belonging to complicated biological methods unsuitable for onsite sensing or emerging alternative chemical techniques are still inseparable from professional instruments. Here we developed a polychromatic fluorescent nanoprobe for ratiometric detection and visual inspection of the pathogenic bacterial spore biomarker, dipicolinic acid (DPA), realizing rapidly accurate screening of pathogenic bacterial spores such as Bacillus anthracis spores. The nanoprobe is made of aminoclay-coated silicon nanoparticles and functionalized with europium ions, exhibiting selective and sensitive response toward DPA and Bacillus subtilis spores (simulants for Bacillus anthracis spores) with excellent linearity. The proposed sensing strategy allowing spore determination of as few as 0.3 × 105 CFU/mL within 10 s was further applied to real environmental sample detection with good accuracy and reliability. Visual quantitative determination can be achieved by analyzing the RGB values of the corresponding test solution color via a color recognition APP on a smartphone. Different test samples can be photographed at the same time, hence the efficient accomplishment of examining bulk samples within minutes. Potentially employed in various on-site sensing occasions, this strategy may develop into a powerful means for distinguishing hazardous pathogens to facilitate timely and proper actions of dealing with multifarious security issues.

When the Weibull model helps in deciphering bacterial resistance variability related to survival behaviour

Survival curves of bacterial vegetative cells or spores subjected to an inactivation process are often log-linear and then described by the d-value parameter. However, non log-linear, convex, shapes might be also observed particularly when mild inactivation treatments are applied. Our objective was to investigate whether the 3-parameters Weibull model (logN0, δ, p) could be used to go beyond a simple fitting of convex curve by providing information related to bacterial variability. First, survival curves were simulated to mimic the behaviour of a cocktail containing bacterial vegetative cells or spores undergoing an inactivation treatment, on which the Weibull model was fitted. Second, a mathematical model was developed to describe the link between the Weibull parameters p and delta with the d-values of sub-populations of bacterial vegetative cells or spores (considering as well the percentage of each sub-population). Based on this model, it was shown that the Weibull model can be used to go beyond a simple description of a convex curve. For instance, if p is estimated around 0.8, that means the presence of a resistant sub-population, but with a limited resistant variability (ratio of resistance from 1.5 to 4). In contrast, if p is estimated to 0.3–04 that means the presence of a resistant sub-population in a small proportion (less than 10 %) combined with a large resistant variability (ratio of 10 or more). This study shows that the Weibull model can be used in combination with the new model developed here to decipher vegetative cell or spore resistance variability, with application in food industry processes such as thermal or physical inactivation treatment as well as cleaning and disinfection verification procedure.