A Paper-Based Wearable Moist-Electric Generator for Sustained High-Efficiency Power Output and Enhanced Moisture Capture.

Disposable wearable electronics are valuable for diagnostic and healthcare purposes, reducing maintenance needs and enabling broad accessibility. However, integrating a reliable power supply is crucial for their advancement, but conventional power sources present significant challenges. To address that issue, a novel paper-based moist-electric generator is developed that harnesses ambient moisture for power generation. The device features gradients for functional groups and moisture adsorption and architecture of nanostructures within a disposable paper substrate. The nanoporous, gradient-formed spore-based biofilm and asymmetric electrode deposition enable sustained high-efficiency power output. A Janus hydrophobic-hydrophilic paper layer enhances moisture harvesting, ensuring effective operation even in low-humidity environments. This research reveals that the water adsorption gradient is crucial for performance under high humidity, whereas the functional group gradient is dominant under low humidity. The device delivers consistent performance across diverse conditions and flexibly conforms to various surfaces, making it ideal for wearable applications. Its eco-friendly, cost-effective, and disposable nature makes it a viable solution for widespread use with minimal environmental effects. This innovative approach overcomes the limitations of traditional power sources for wearable electronics, offering a sustainable solution for future disposable wearables. It significantly enhances personalized medicine through improved health monitoring and diagnostics.

Unveiling the matrix effect on Bacillus licheniformis and Bacillus subtilis spores heat inactivation between plant-based milk alternatives, bovine milk and culture medium.

This study examined the inactivation of spores of Bacillus licheniformis and Bacillus subtilis in four pea-based milk alternatives, semi-skimmed bovine milk and Brain Heart Infusion (BHI) broth to assess the matrix impact on the thermal inactivation of bacterial spores. Heat inactivation was performed with the method of capillary tubes in temperature range 97-110 °C. A four-parameter non-linear model, including initial level, shoulder duration, inactivation rate and tailing, was fitted to the data obtained. D-values were estimated and secondary ZT-value models were developed for both species. A secondary model for the shoulder length of B. licheniformis in a plant-based milk alternative formulation was built too. Models were validated at a higher temperature, 113.5 °C. D-values in the different matrices ranged between 2.3 and 8.2 min at 97 °C and 0.1-0.3 min at 110 °C for B. licheniformis. D-values for B. subtilis ranged between 3.9 and 6.3 min at 97 °C and 0.2-0.3 min at 110 °C. ZT-values in the different matrices ranged between 7.3 and 8.9 °C and 8.9-10.0 °C for B. licheniformis and B. subtilis, respectively. Significant differences in inactivation parameters were found within the pea-based formulations as well as when compared to bovine milk. Heat resistance was higher in pea-based matrices. Shoulders observed were temperature- and matrix-dependent, while no such trend was found for the tailings. These results provide insights, useful on designing safe thermal processing, limiting spoilage in plant-based milk alternatives and thus, reducing global food waste.

[Genome Stability of Bacillus velezensis after Two-Year Exposure in Open Space].

Spore-forming bacteria have a unique resistance to negative environmental conditions, including aggressive space factors, and are an excellent model for studying adaptation mechanisms and survival strategies at the molecular level. The study analyzed the genome of Bacillus velezensis, which remained viable after a 2-year exposure in outer space on the outer surface of the ISS as part of the Test space experiment. A comparative analysis of the draft genomes of the exhibit strain and the ground control did not reveal significant changes; the average nucleotide identity was 99.98%, which indicates the ability of microorganisms to maintain genome stability in space conditions, due to both increased stress resistance of bacterial spores and efficient operation of the system of repair of accumulated changes. The study of a single nucleotide polymorphism in the genome of B. velezensis revealed nine point substitutions, three of which are in intergenic regions, six in protein-coding genes, three of them are missense mutations, two nucleotide deletions leading to a shift in the reading frame, and one synonymous substitution. The profiles of the housekeeping genes were determined during MLST typing and it was found that the allelic profiles obtained for B. velezensis T15.2 and 924 strains do not correspond to any of the previously described sequence types. The presented results indicate the ability of B. velezensis bacteria to maintain the viability of spores and the integrity of the genome for a long time under extreme conditions of outer space, which is important for the problem of planetary protection, as well as the potential possibility of performing biotechnological processes based on B. velezensis during space exploration.

Counts of mesophilic aerobic, mesophilic anaerobic, thermophilic aerobic sporeforming bacteria and persistence of Bacillus cereus spores throughout cocoa powder processing chain.

Sporeforming bacteria are a concern in some food raw materials, such as cocoa powder. Samples (n = 618) were collected on two farms and at several stages during cocoa powder manufacture in three commercial processing lines to determine the impact of each stage on bacterial spore populations. Mesophilic aerobic, mesophilic anaerobic, thermophilic aerobic, and Bacillus cereus spore populations were enumerated in all the samples. Genetic diversity in B. cereus strains (n = 110) isolated from the samples was examined by M13 sequence-based PCR typing, partial sequencing of the panC gene, and the presence/absence of ces and cspA genes. The counts of different groups of sporeforming bacteria varied amongst farms and processing lines. For example, the counts of mesophilic aerobic spore-forming (MAS) populations of cocoa bean fermentation were lower than 1 log spore/g in Farm 1 but higher than 4 log spore/g in Farm 2. B. cereus isolated from cocoa powder was also recovered from cocoa beans, nibs, and samples after roasting, refining, and pressing, which indicated that B. cereus spores persist throughout cocoa processing. Phylogenetic group IV was the most frequent (73%), along with processing. Strains from phylogenetic group III (14 %) did not show the ces gene's presence.

Impact of sporulation temperature on germination of Bacillus subtilis spores under optimal and adverse environmental conditions.

Bacillus subtilis spores are important food spoilage agents and are occasionally involved in food poisoning. In foods that are not processed with intense heat, such bacterial spores are controlled by a combination of different hurdles, such as refrigeration, acidification, and low water activity (aw), which inhibit or delay germination and/or growth. Sporulation temperature has long been regarded as a relevant factor for the assessment of germination in chemically defined media, but little is known about its impact on food preservation environments. In this study, we compared germination dynamics of B. subtilis spores produced at optimal temperature (37 °C) with others incubated at suboptimal (20 °C) and supraoptimal (43 °C) temperatures in a variety of nutrients (rich-growth medium, L-alanine, L-valine, and AGFK) under optimal conditions as well as under food-related stresses (low aw, pH, and temperature). Spores produced at 20 °C had a lower germination rate and efficiency than those incubated at 37 °C in all the nutrients, while those sporulated at 43 °C displayed a higher germination rate and/or efficiency in response to rich-growth medium and mostly to L-alanine and AGFK under optimal environmental conditions. However, differences in germination induced by changes in sporulation temperature decreased when spores were activated by heat, mainly due to the greater benefit of heat for spores produced at 20 °C and 37 °C than at 43 °C, especially in AGFK. Non-heat-activated spores produced at 43 °C still displayed superior germination fitness under certain stresses that had considerably impaired the germination of the other two populations, such as reduced temperature and aw. Moreover, they presented lower temperature and pH boundaries for the inhibition of germination in rich-growth medium, while requiring a higher NaCl concentration threshold compared to spores obtained at optimal and suboptimal temperature. Sporulation temperature is therefore a relevant source of variability in spore germination that should be taken into account for the accurate prediction of spore behaviour under variable food preservation conditions with the aim of improving food safety and stability.

Bacterial spores respond to humidity similarly to hydrogels.

Bacterial spores have outstanding properties from the materials science perspective, which allow them to survive extreme environmental conditions. Recent work by [S. G. Harrellson et al., Nature 619, 500-505 (2023)] studied the mechanical properties of Bacillus subtilis spores and the evolution of these properties with the change of humidity. The experimental measurements were interpreted assuming that the spores behave as water-filled porous solids, subjected to hydration forces. Here, we revisit their experimental data using literature data on vapor sorption on spores and ideas from polymer physics. We demonstrate that upon the change of humidity, the spores behave like rubber with respect to their swelling, elasticity, and relaxation times. This picture is consistent with the knowledge of the materials comprising the bacterial cell walls-cross-linked peptidoglycan. Our results provide an interpretation of the mechanics of bacterial spores and can help in developing synthetic materials mimicking the mechanical properties of the spores.

Enhancement of microbicidal efficacy of chemical disinfectants when combined with ultrasound technology.

AIMS: This study investigated the antimicrobial efficacy of ultrasound technology (US) in combination with two different disinfectants (Disinfectant A and Disinfectant B), containing peracetic acid (PAA) and quaternary ammonium compounds (QACs), respectively, against two sporigenic pathogens, Aspergillus brasiliensis and Bacillus subtilis. METHODS AND RESULTS: The microbicidal activity of the coupled treatment was compared with the use of the disinfectants alone, and the efficacy of the disinfection strategies was evaluated by the log reduction of the population of the microorganism inoculated onto stainless-steel surface. The combination treatment resulted in a log reduction of 5.40 and 3.88 (Disinfectant A + US) against A. brasiliensis and B. subtilis, at 850 and 500 ppm PAA, compared to 265 and 122 (Disinfectant A only). For Disinfectant B, in combination with US, showed a logarithmic reduction of 5.04 and 4.79 against A. brasiliensis and B. subtilis at 078% v v-1 and 392% v v-1 QACs, respectively, vs. 1.58 and 1.64 (Disinfectant B only). Moreover, no colonies or not statistically significant growth was observed within the US bath containing the disinfectant. CONCLUSIONS: The antimicrobial efficacy of the two disinfectants was greatly enhanced when used in combination with US, and this also makes it possible to avoid the overuse of chemicals for disinfection.

Construction of Graphene@Ag-MLF composite structure SERS platform and its differentiating performance for different foodborne bacterial spores.

A new surface-enhanced Raman spectroscopy (SERS) biosensor of Graphene@Ag-MLF composite structure has been fabricated by loading AgNPs on graphene films. The response of the biosensor is  based on plasmonic sensing. The results showed that the enhancement factor of three different spores reached 107 based on the Graphene@Ag-MLF substrate. In addition, the SERS performance was stable, with good reproducibility (RSD<3%). Multivariate statistical analysis and chemometrics were used to distinguish different spores. The accumulated variance contribution rate was up to 96.35% for the top three PCs, while HCA results revealed that the spectra were differentiated completely. Based on optimal principal components, chemometrics of KNN and LS-SVM were applied to construct a model for rapid qualitative identification of different spores, of which the prediction set and training set of LS-SVM achieved 100%. Finally, based on the Graphene@Ag-MLF substrate, the LOD of three different spores was lower than 102 CFU/mL. Hence, this novel Graphene@Ag-MLF SERS substrate sensor was rapid, sensitive, and stable in detecting spores, providing strong technical support for the application of SERS technology in food safety.

Inactivating and damaging properties of the disinfectant "MultiDez" when exposed to bacteria and spores.

Disinfectants play a crucial role in controlling the spread of infectious diseases caused by bacteria and spore-forming organisms. Bacteria and spores can persist on surfaces and in the environment for extended periods, posing a significant risk to public health. Disinfectants are designed to inactivate or kill these microorganisms by disrupting their cellular structures and functions. Effective disinfectants are essential for preventing the spread of infectious diseases in hospitals, laboratories, food processing facilities, and other settings where the risk of contamination is high. This study evaluated the effectiveness of a disinfectant called "MultiDez" on Y.pestis bacteria and Bacillus anthracis spores using microbiological and electron microscopic methods. Results showed that after exposure to a 0.5 % solution of the disinfectant, the death of all Y.pestis bacteria was achieved after 90 min, while the death of Bacillus anthracis spores was achieved after 240 min. Electron microscopy revealed that the disinfectant caused complete destruction of both bacterial cells and spores by enveloping their outer surfaces with polymer molecules, disrupting the structure and function of their membranes, and destroying their cytoplasm and nucleode. The mechanism of action of the disinfectant on bacteria and spores involved different processes, with the disinfectant causing rapid hydration of dehydrated spores and blocking the functions of spore membranes in the case of bacterial spores.

Fluorescent silica nanoparticles as nano-chemosensors for the sequential detection of Pb2+ ions and bacterial-spore biomarker dipicolinic acid (DPA) in aqueous solution.

Herein, we report fluorescein-labelled silica nanoparticles (FSNP) which serve as fluorescent nano-chemosensors for sequential detection of Pb2+ (which is a toxic heavy metal) and dipicolinic acid (DPA) (which is a distinctive indicator biomarker of bacterial spores) with high sensitivity and selectivity. The fluorescence of FSNP is quenched because of the complex formation between Pb2+ ions and surface amide groups, however, the fluorescence is recovered in contact with DPA, resulting from the association of DPA with surface bound Pb2+ ions. FSNP-Pb2+ complexes show high sensitivity towards DPA with a low detection limit of 850 nM which is approximately seventy times lower than the infectious dosage of bacterial spores (60 µM). Lateral flow test platform was further developed to show the applicability and practicability of our system.

Bacterial Spore-Based Delivery System: 20 Years of a Versatile Approach for Innovative Vaccines.

Mucosal vaccines offer several advantages over injectable conventional vaccines, such as the induction of adaptive immunity, with secretory IgA production at the entry site of most pathogens, and needle-less vaccinations. Despite their potential, only a few mucosal vaccines are currently used. Developing new effective mucosal vaccines strongly relies on identifying innovative antigens, efficient adjuvants, and delivery systems. Several approaches based on phages, bacteria, or nanoparticles have been proposed to deliver antigens to mucosal surfaces. Bacterial spores have also been considered antigen vehicles, and various antigens have been successfully exposed on their surface. Due to their peculiar structure, spores conjugate the advantages of live microorganisms with synthetic nanoparticles. When mucosally administered, spores expressing antigens have been shown to induce antigen-specific, protective immune responses. This review accounts for recent progress in the formulation of spore-based mucosal vaccines, describing a spore's structure, specifically the spore surface, and the diverse approaches developed to improve its efficiency as a vehicle for heterologous antigen presentation.

The chronicle of the ups and downs that made Bacillus thuringiensis a natural insecticide

Bacillus thuringiensis is a worldwide known bacterium for its capacity to control insect pests thanks to the action of its parasporal crystal. The objective of this paper deals with the history, in some cases unknown, of the study of Bacillus thuringiensis that led it to be a crucial biological alternative in controlling pest insects. How the mode of action for killing insects was understood, as well as the field tests that were carried out to evaluate its effectiveness and to develop the first commercial products, are reflected in this review that presents and discusses the scientific successes and failures that marked the course of B. thuringiensis.

Bacillus thuringiensis es una bacteria conocida mundialmente por su capacidad para controlar insectos plaga, gracias a la acción de su cristal parasporal. El objetivo de esta revisión trata de la historia, en algunos casos desconocida, del estudio de Bacillus thuringiensis que la llevó a ser una importante alternativa biológica en el control de insectos plaga. Cómo se llegó a comprender el modo de acción para matar insectos, así como las pruebas de campo que se realizaron para evaluar su efectividad y lograr desarrollar los primeros productos comerciales están plasmados en esta revisión que presenta y discute los aciertos y desaciertos científicos que marcaron el rumbo de B. thuringiensis.

The underlying mechanism of bacterial spore germination: An update review.

Bacterial spores are highly resilient and universally present on earth and can irreversibly enter the food chain to cause food spoilage or foodborne illness once revived to resume vegetative growth. Traditionally, extensive thermal processing has been employed to efficiently kill spores; however, the relatively high thermal load adversely affects food quality attributes. In recent years, the germination-inactivation strategy has been developed to mildly kill spores based on the circumstance that germination can decrease spore-resilient properties. However, the failure to induce all spores to geminate, mainly owing to the heterogeneous germination behavior of spores, hampers the success of applying this strategy in the food industry. Undoubtedly, elucidating the detailed germination pathway and underlying mechanism can fill the gap in our understanding of germination heterogeneity, thereby facilitating the development of full-scale germination regimes to mildly kill spores. In this review, we comprehensively discuss the mechanisms of spore germination of Bacillus and Clostridium species, and update the molecular basis of the early germination events, for example, the activation of germination receptors, ion release, Ca-DPA release, and molecular events, combined with the latest research evidence. Moreover, high hydrostatic pressure (HHP), an advanced non-thermal food processing technology, can also trigger spore germination, providing a basis for the application of a germination-inactivation strategy in HHP processing. Here, we also summarize the diverse germination behaviors and mechanisms of spores of Bacillus and Clostridium species under HHP, with the aim of facilitating HHP as a mild processing technology with possible applications in food sterilization. Practical Application: This work provides fundamental basis for developing efficient killing strategies of bacterial spores in food industry.

Safety assessment of surfactin-producing Bacillus strains and their lipopeptides extracts in vitro and in vivo.

Beneficial Bacillus strains can be administered to livestock as probiotics to improve animal health. Cyclic lipopeptides produced by Bacillus such as surfactins may be responsible for some of the beneficial effects due to their anti-inflammatory and immunomodulatory activity. The aim of the present study was to isolate and evaluate the biocompatibility of native Bacillus spp. strains and their surfactin-like lipopeptides in vitro and in vivo to determine their potential to be used on animals. Biocompatibility of endospore suspensions (108 UFC/mL), and different dilutions (1:10; 1:50; 1:100; 1:500, and 1:1000) of Bacillus lipopeptide extracts containing surfactin was tested on Caco-2 cells by microculture tetrazolium-based colorimetric assay. Genotoxicity was tested on BALB/c mice (n = 6) administered 0.2 mL of endospore suspensions by the bone marrow erythrocyte micronuclei assay. All the isolates tested produced between 26.96 and 239.97 µg mL- 1 of surfactin. The lipopeptide extract (LPE) from isolate MFF1.11 demonstrated significant cytotoxicity in vitro. In contrast, LPE from MFF 2.2; MFF 2.7, TL1.11, TL 2.5, and TC12 had no cytotoxic effect (V% > 70%) on Caco-2 cells, not affecting cell viability signifficantly in most treatments. Similarly, none of the endospore suspensions affected cell viability (V% > 80%). Likewise, endospores did not cause genotoxicity on BALB/c mice. This study was elementary as a first step for a new line of research, since it allowed us to choose the safest isolates to keep working on the search of new potentially probiotic strains destined to production animals to improve their performance and health.

Novel Sources of Biodiversity and Biomolecules from Bacteria Isolated from a High Middle Ages Soil Sample in Palermo (Sicily, Italy).

The urban plan of Palermo (Sicily, Italy) has evolved throughout Punic, Roman, Byzantine, Arab, and Norman ages until it stabilized within the borders that correspond to the current historic center. During the 2012 to 2013 excavation campaign, new remains of the Arab settlement, directly implanted above the structures of the Roman age, were found. The materials investigated in this study derived from the so-called Survey No 3, which consists of a rock cavity of subcylindrical shape covered with calcarenite blocks: it was probably used to dispose of garbage during the Arabic age and its content, derived from daily activities, included grape seeds, scales and bones of fish, small animal bones, and charcoals. Radiocarbon dating confirmed the medieval origin of this site. The composition of the bacterial community was characterized through a culture-dependent and a culture-independent approach. Culturable bacteria were isolated under aerobic and anaerobic conditions and the total bacterial community was characterized through metagenomic sequencing. Bacterial isolates were tested for the production of compounds with antibiotic activity: a Streptomyces strain, whose genome was sequenced, was of particular interest because of its inhibitory activity, which was due to the Type I polyketide aureothin. Moreover, all strains were tested for the production of secreted proteases, with those belonging to the genus Nocardioides having the most active enzymes. Finally, protocols commonly used for ancient DNA studies were applied to evaluate the antiquity of isolated bacterial strains. Altogether these results show how paleomicrobiology might represent an innovative and unexplored source of novel biodiversity and new biotechnological tools. IMPORTANCE One of the goals of paleomicrobiology is the characterization of the microbial community present in archaeological sites. These analyses can usually provide valuable information about past events, such as occurrence of human and animal infectious diseases, ancient human activities, and environmental changes. However, in this work, investigations about the composition of the bacterial community of an ancient soil sample (harvested in Palermo, Italy) were carried out aiming to screen ancient culturable strains with biotechnological potential, such as the ability to produce bioactive molecules and secreted hydrolytic enzymes. Besides showing the biotechnological relevance of paleomicrobiology, this work reports a case of germination of putatively ancient bacterial spores recovered from soil rather than extreme environments. Moreover, in the case of spore-forming species, these results raise questions about the accuracy of techniques usually applied to estimate antiquity of DNA, as they could lead to its underestimation.

Hypervirulent R20291 Clostridioides difficile spores show disinfection resilience to sodium hypochlorite despite structural changes.

BACKGROUND: Clostridioides difficile is a spore forming bacterial species and the major causative agent of nosocomial gastrointestinal infections. C. difficile spores are highly resilient to disinfection methods and to prevent infection, common cleaning protocols use sodium hypochlorite solutions to decontaminate hospital surfaces and equipment. However, there is a balance between minimising the use of harmful chemicals to the environment and patients as well as the need to eliminate spores, which can have varying resistance properties between strains. In this work, we employ TEM imaging and Raman spectroscopy to analyse changes in spore physiology in response to sodium hypochlorite. We characterize different C. difficile clinical isolates and assess the chemical's impact on spores' biochemical composition. Changes in the biochemical composition can, in turn, change spores' vibrational spectroscopic fingerprints, which can impact the possibility of detecting spores in a hospital using Raman based methods. RESULTS: We found that the isolates show significantly different susceptibility to hypochlorite, with the R20291 strain, in particular, showing less than 1 log reduction in viability for a 0.5% hypochlorite treatment, far below typically reported values for C. difficile. While TEM and Raman spectra analysis of hypochlorite-treated spores revealed that some hypochlorite-exposed spores remained intact and not distinguishable from controls, most spores showed structural changes. These changes were prominent in B. thuringiensis spores than C. difficile spores. CONCLUSION: This study highlights the ability of certain C. difficile spores to survive practical disinfection exposure and the related changes in spore Raman spectra that can be seen after exposure. These findings are important to consider when designing practical disinfection protocols and vibrational-based detection methods to avoid a false-positive response when screening decontaminated areas.

New Thoughts on an Old Topic: Secrets of Bacterial Spore Resistance Slowly Being Revealed.

The quest for bacterial survival is exemplified by spores formed by some Firmicutes members. They turn up everywhere one looks, and their ubiquity reflects adaptations to the stresses bacteria face. Spores are impactful in public health, food safety, and biowarfare. Heat resistance is the hallmark of spores and is countered principally by a mineralized gel-like protoplast, termed the spore core, with reduced water which minimizes macromolecular movement/denaturation/aggregation. Dry heat, however, introduces mutations into spore DNA. Spores have countermeasures to extreme conditions that are multifactorial, but the fact that spore DNA is in a crystalline-like nucleoid in the spore core, likely due to DNA saturation with small acid-soluble spore proteins (SASPs), suggests that reduced macromolecular motion is also critical in spore dry heat resistance. SASPs are also central in the radiation resistance characteristic of spores, where the contributions of four spore features-SASP; Ca2+, with pyridine-2,6-dicarboxylic acid (CaDPA); photoproduct lyase; and low water content-minimize DNA damage. Notably, the spore environment steers UV photochemistry toward a product that germinated spores can repair without significant mutagenesis. This resistance extends to chemicals and macromolecules that could damage spores. Macromolecules are excluded by the spore coat which impedes the passage of moieties of ≥10 kDa. Additionally, damaging chemicals may be degraded or neutralized by coat enzymes/proteins. However, the principal protective mechanism here is the inner membrane, a compressed structure lacking lipid fluidity and presenting a barrier to the diffusion of chemicals into the spore core; SASP saturation of DNA also protects against genotoxic chemicals. Spores are also resistant to other stresses, including high pressure and abrasion. Regardless, overarching mechanisms associated with resistance seem to revolve around reduced molecular motion, a fine balance between rigidity and flexibility, and perhaps efficient repair.

Expression of the 2Duf protein in wild-type Bacillus subtilis spores stabilizes inner membrane proteins and increases spore resistance to wet heat and hydrogen peroxide.

AIMS: This work aimed to characterize spore inner membrane (IM) properties and the mechanism of spore killing by wet heat and H2O2 with spores overexpressing the 2Duf protein, which is naturally encoded from a transposon found only in some Bacillus strains with much higher spore resistance than wild-type spores. METHODS AND RESULTS: Killing of Bacillus subtilis spores by wet heat or hydrogen peroxide (H2O2) was slower when 2Duf was present, and Ca-dipicolinic acid release was slower than killing. Viabilities on rich plates of wet heat- or H2O2 -treated spores +/- 2Duf were lower when NaCl was added, but higher with glucose. Addition of glucose but not Casamino acids addition increased treated spores' viability on minimal medium plates. Spores with 2Duf required higher heat activation for germination, and their germination was more wet-heat resistant than that of wild-type spores, processes that involve IM proteins. IM permeability and lipid mobility were lower in spores with 2Duf, although IM phospholipid composition was similar in spores +/- 2Duf. CONCLUSIONS: These results and previous work suggests that wet heat and H2O2 kill spores by damaging an IM enzyme or enzymes involved in oxidative phosphorylation.

The persistence of time: the lifespan of Bacillus anthracis spores in environmental reservoirs.

Anthrax is a lethal bacterial zoonosis primarily affecting herbivorous wildlife and livestock. Upon host death Bacillus anthracis vegetative cells form spores capable of surviving for years in soil. Anthrax transmission requires host exposure to large spore doses. Thus, conditions that facilitate higher spore concentrations or promote spore survival will increase the probability that a pathogen reservoir infects future hosts. We investigated abiotic and pathogen genomic variation in relation to spore concentrations in surface soils (0-1 cm depth) at 40 plains zebra (Equus quagga) anthrax carcass sites in Namibia. Specifically, how initial spore concentrations and spore survival were affected by seasonality associated with the timing of host mortality, local soil characteristics, and pathogen genomic variation. Zebras dying of anthrax in wet seasons-the peak season for anthrax in Etosha National Park-had soil spore concentrations 1.36 orders of magnitude higher than those that died in dry seasons. No other variables considered affected spore concentrations, and spore survival rates did not differ among sites. Surface soils at these pathogen reservoirs remained culture positive for a range of 3.8-10.4 years after host death. Future research could evaluate if seasonal patterns in spore concentrations are driven by differences in sporulation success or levels of terminal bacteremia.

Effect of High Hydrostatic Pressure Treatment Combined with Alkaline Electrolyzed Water on the Injury and Growth Characteristics of Bacterial Spores.

The characteristics of growth and injury of Bacillus subtilis spores whose heat resistance was reduced by the treatment of high hydrostatic pressure processing (HPP) combined with/without alkaline electrolyzed water (AlEW) were investigated. The delay in the lag phase of growth curve observed when used in combination with AlEW, especially at 50 MPa or less, and was prolonged by about 4 to 6 hours. However, the effects of temperature and time during treatment on the lag phase were not significantly different between solo-HPP and in combination with AlEW. The injury characteristics of the treated bacterial spores were evaluated by using antibiotics (penicillin G, rifampicin and chloramphenicol) supplemented trypticase soy broth. In the chloramphenicol supplemented TSB, although the lag phase of bacterial spores treated by HPP with AlEW was not prolonged as compared with the normal-TSB, the decrease in growth rate during logarithmic phase and increase in maximum growth amount were observed. This result could suggest due to a damage by combined treatment of HPP with AlEW such as the inhibition of protein synthesis. Furthermore, the combined treatment with AlEW increased the catalase activity by 1.45, 1.63 and 4.25 times at 30, 80 and 100 MPa, respectively, as compared with solo-HPP, therefore the combined treatment could cause high oxidative stress on bacterial spores.