Heat Resistance Of Spores Research Articles

Evidence of defined temporal expression patterns that lead agram-negative cell out of dormancy.

Many bacterial species are capable of forming long-lived dormant cells. The best characterized are heat and desiccation resistant spores produced by many Gram-positive species. Less characterized are dormant cysts produced by several Gram-negative species that are somewhat tolerant to increased temperature and very resistant to desiccation. While there is progress in understanding regulatory circuits that control spore germination, there is scarce information on how Gram-negative organisms emerges from dormancy. In this study, we show that R. centenum cysts germinate by emerging a pair of motile vegetative cells from a thick cyst cell wall coat ~ 6 hrs post induction of germination. Time-lapse transcriptomic analysis reveals that there is a defined temporal pattern of gene expression changes during R. centenum cyst germination. The first observable changes are increases in expression of genes for protein synthesis, an increase in expression of genes involved in the generation of a membrane potential and the use of this potential for ATP synthesis via ATPase expression. These early events are followed by expression changes that affect the cell wall and membrane composition, followed by expression changes that promote chromosome replication. Midway through germination, expression changes occur that promote the flow of carbon through the TCA cycle to generate reducing power and parallel synthesis of electron transfer components involved in oxidative phosphorylation. Finally, late expression changes promote the synthesis of a photosystem as well as flagellar and chemotaxis components for motility.

Role of Bacillus species in biofilm persistence and emerging antibiofilm strategies in the dairy industry.

Biofilm-forming Bacillus species are often involved in persistent contamination and spoilage of dairy products. They therefore present a major microbiological challenge in the field of dairy food quality and safety. Due to their substantial physiological versatility, Bacillus species can survive in various parts of dairy manufacturing plants, leading to a high risk of product spoilage and potential dissemination of foodborne diseases. Furthermore, biofilm and heat-resistant spore formation make these bacteria challenging to eliminate. Thus, some strategies have been employed to remove, prevent, or delay the formation of Bacillus biofilms in the dairy industry, but with limited success. Lack of understanding of the Bacillus biofilm structure and behavior in conditions relevant to dairy-associated environments could partially account for this situation. The current paper reviews dairy-associated biofilm formation by Bacillus species, with particular attention to the role of biofilm in Bacillus species adaptation and survival in a dairy processing environment. Relevant model systems are discussed for the development of novel antimicrobial approaches to improve the quality of dairy food. © 2020 Society of Chemical Industry.

Application of (super)cavitation for the recycling of process waters in paper producing industry.

In paper production industry, microbial contaminations of process waters are common and can cause damage to paper products and equipment as well as the occurrence of pathogens in the end products. Chlorine omission has led to the usage of costly reagents and products of lower mechanical quality. In this study, we have tested a rotation generator equipped with two sets of rotor and stator assemblies to generate developed cavitation (unsteady cloud shedding with pressure pulsations) or supercavitation (a steady cavity in chocked cavitation conditions) for the destruction of a persistent bacteria Bacillus subtilis. Our results showed that only supercavitation was effective and was further employed for the treatment of waters isolated from an enclosed water recycle system in a paper producing plant. The water quality was monitored and assessed according to the chemical (COD, redox potential and dissolved oxygen), physical (settleable solids, insolubles and colour intensity) and biological methods (yeasts, aerobic and anaerobic bacteria, bacterial spores and moulds). After one hour of treatment, a strong 4 logs reduction was achieved for the anaerobic sulphate reducing bacteria and for the yeasts; a 3 logs reduction for the aerobic bacteria; and a 1.3 logs reduction for the heat resistant bacterial spores. A 22% reduction in COD and an increase in the redox potential (37%) were observed. Sediments were reduced by 50% and the insoluble particles by 67%. For bacterial destruction in real industrial process waters, the rotation generator of supercavitation spent 4 times less electrical energy in comparison to the previously published cavitation treatments inside the Venturi constriction design.

High pressure CO2 reduces the wet heat resistance of Bacillus subtilis spores by perturbing the inner membrane

Spores of wild-type Bacillus subtilis PS533 were treated by wet heat at 75 °C for 30 min, and high pressure CO2 (HPCD) at 6.5 MPa and 30 °C or 75 °C for 30 min. The spores were analyzed for wet heat resistance (85 °C, 90 °C, 95 °C) and typical germination events including DPA release and cortex hydrolysis, inner membrane permeability, and germination triggered by nutrient (L-valine and AGFK) or non-nutrient (dodecylamine and high pressure at 150 MPa or 550 MPa) germinants. The results showed that (i) HPCD-treated spores exhibited reduced wet heat resistance compared to the untreated or wet heat-treated spores; (ii) HPCD-treated spores did not undergo typical germination events such as DPA release or cortex hydrolysis compared to normally germinated spores; (iii) HPCD-treated spores released more metal ions and exhibited decreased ability to maintain DPA, indicating that the permeability of inner membrane of HPCD-treated spores was increased; (iv) HPCD-treated spores exhibited reduced germination rate when triggered by L-valine or 150 MPa, but increased germination rate when triggered by dodecylamine or 550 MPa, suggesting that the fluidity of the inner membrane of HPCD-treated spores might be increased. These results indicated that HPCD could reduce the wet heat resistance of spores, and this resistance decrease was probably due to the modification of the inner membrane caused by HPCD. Industrial relevanceThe extremely high wet heat resistance of spores makes them a significant problem in the thermal processing of foods. Thus, it of great interest to develop a process to reduce the wet heat resistance of spores. In this work, we found that HPCD can significantly reduce the wet heat resistance of B. subtilis spores, and this was achieved by perturbing the inner membrane of spores. These results can improve our understanding of the inactivation mechanism of spores by HPCD, and also provide an alternative approach for spore inactivation in foods.

Biofilm-Forming Ability and Effect of Sanitation Agents on Biofilm-Control of Thermophile Geobacillus sp. D413 and Geobacillus toebii E134.

Geobacillus sp. D413 and Geobacillus toebii E134 are aerobic, non-pathogenic, endospore-forming, obligately thermophilic bacilli. Gram-positive thermophilic bacilli can produce heat-resistant spores. The bacteria are indicator organisms for assessing the manufacturing process’s hygiene and are capable of forming biofilms on surfaces used in industrial sectors. The present study aimed to determine the biofilm-forming properties of Geobacillus isolates and how to eliminate this formation with sanitation agents. According to the results, extracellular DNA (eDNA) was interestingly not affected by the DNase I, RNase A, and proteinase K. However, the genomic DNA (gDNA) was degraded by only DNase I. It seemed that the eDNA had resistance to DNase I when purified. It is considered that the enzymes could not reach the target eDNA. Moreover, the eDNA resistance may result from the conserved folded structure of eDNA after purification. Another assumption is that the eDNA might be protected by other extracellular polymeric substances (EPS) and/or extracellular membrane vesicles (EVs) structures. On the contrary, DNase I reduced unpurified eDNA (mature biofilms). Biofilm formation on surfaces used in industrial areas was investigated in this work: the D413 and E134 isolates adhered to all surfaces. Various sanitation agents could control biofilms of Geobacillus isolates. The best results were provided by nisin for D413 (80%) and α-amylase for E134 (98%). This paper suggests that sanitation agents could be a solution to control biofilm structures of thermophilic bacilli.

The effect of phosphate on the heat resistance of spores of dairy isolates of Geobacillus stearothermophilus

In this study, we show that phosphate decreases the spore heat resistance by accelerating the rate of loss of cations from spores. Heat resistance of spores of Geobacillus stearothermophilus A1, D1, P3 and ATCC 12980 were determined in distilled water containing varying concentrations (0.1, 1 and 2% w/v) of di‑sodium phosphate. The average decimal reduction times (D value) for strains A1, D1, P3 and ATCC 12980 in distilled water were 5.8, 6.8, 5.7 and 9 min at 110 °C respectively. On the addition of 0.1, 1 and 2% w/v of di‑sodium phosphate, the average D110 values of all the strains in distilled water were lowered by 50, 61 and 70% respectively. Addition of 0.05% w/v of Na-EDTA to distilled water resulted in lowering of the average D110 value of all the strains by 55%. Heat resistance of spores of A1, D1, P3 and ATCC 12980 was found to be associated with the Dipicolinic Acid (DPA) content whose concentrations were 0.25, 0.30, 0.27 and 1.6 pg per spore respectively. Analysis by atomic absorption spectroscopy revealed that the phosphate present in the heating medium causes excess release of calcium from spores with 2% w/v phosphate being highly effective, thus confirming the chelating effect of phosphate. This study provides insight into the heat resistance and the increased heat sensitivity of spores of G. stearothermophilus A1, D1 and P3 in the presence of phosphate, which can be used in the design of Cleaning in Place (CIP) systems involving phosphate based cleaning agents to combat biofilms and spores in the dairy industry.

SpoIVA-SipL Complex Formation Is Essential for Clostridioides difficile Spore Assembly.

Spores are the major infectious particle of the Gram-positive nosocomial pathogen Clostridioides difficile (formerly Clostridium difficile), but the molecular details of how this organism forms these metabolically dormant cells remain poorly characterized. The composition of the spore coat in C. difficile differs markedly from that defined in the well-studied organism Bacillus subtilis, with only 25% of the ∼70 spore coat proteins being conserved between the two organisms and with only 2 of 9 coat assembly (morphogenetic) proteins defined in B. subtilis having homologs in C. difficile We previously identified SipL as a clostridium-specific coat protein essential for functional spore formation. Heterologous expression analyses in Escherichia coli revealed that SipL directly interacts with C. difficile SpoIVA, a coat-morphogenetic protein conserved in all spore-forming organisms, through SipL's C-terminal LysM domain. In this study, we show that SpoIVA-SipL binding is essential for C. difficile spore formation and identify specific residues within the LysM domain that stabilize this interaction. Fluorescence microscopy analyses indicate that binding of SipL's LysM domain to SpoIVA is required for SipL to localize to the forespore while SpoIVA requires SipL to promote encasement of SpoIVA around the forespore. Since we also show that clostridial LysM domains are functionally interchangeable at least in C. difficile, the basic mechanism for SipL-dependent assembly of clostridial spore coats may be conserved.IMPORTANCE The metabolically dormant spore form of the major nosocomial pathogen Clostridioides difficile is its major infectious particle. However, the mechanisms controlling the formation of this resistant cell type are not well understood, particularly with respect to its outermost layer, the spore coat. We previously identified two spore-morphogenetic proteins in C. difficile: SpoIVA, which is conserved in all spore-forming organisms, and SipL, which is conserved only in the clostridia. Both SpoIVA and SipL are essential for heat-resistant spore formation and directly interact through SipL's C-terminal LysM domain. In this study, we demonstrate that the LysM domain is critical for SipL and SpoIVA function, likely by helping recruit SipL to the forespore during spore morphogenesis. We further identified residues within the LysM domain that are important for binding SpoIVA and, thus, functional spore formation. These findings provide important insight into the molecular mechanisms controlling the assembly of infectious C. difficile spores.

Spores in dairy – new insights in detection, enumeration and risk assessment

The performance of the ISO method for the ‘Enumeration of the Specially Thermoresistant Spores of Thermophilic Bacteria in Dried Milk’ (ISO/TS27265; 2009) was compared with a more practical method. Both were tested for predictability of spoilage of UHT treated reconstituted milk. The data show that heating for 30 min at 100 °C has the same predictive value as heating for 30 min at 106 °C, provided that specifications are increased 1 log10 and the use of TSA as a cultivation medium is recommended over PCMA. Predictability of spoilage using classical plating methods is furthermore discussed in relation to variation in spore heat resistance of spores commonly present in the dairy industry.

Investigating the influence of pH and selected heating media on thermal destruction kinetics of Geobacillus stearothermophilus (ATCC10149)

Geobacillus stearothermophilus is a thermophilic spore-forming bacterium which is used for intense thermal process validations (10 min–15 min). Several factors including the pH and heating medium influence the heat resistance of such microbial spores. In this study, the influence of using different buffer types including McIlvaine and phosphate was compared to distilled water at pH 7. Also, the effect of pH variations was studied using McIlvaine buffer at pH 3, 4, 5, 6, 7, and 8. All kinetics studies were conducted at 110 to 120 (°C). Results showed higher heat sensitivity (higher Z value) in distilled water (8.27 °C) than in phosphate (9.21 °C) and McIlvaine buffers (9.59 °C); likely being influenced by the high percentage of inorganic salts present in these buffers. The spores had the highest heat resistance at neutral pH, and the associated D values decreased both below and above pH 7.0. The pH effect on enhancing the heat sensitivity was higher at 110 °C, but the overall temperature influence was not significantly different (p > 0.05). Further, both first-order and Weibull models were compared for thermal destruction kinetics of G. stearothermophilus. Results demonstrated some concavity in the log-linear survival curves suggesting a slightly better fit for the Weibull model in some cases, but the model did not show any better prediction than the first-order model for a 3D lethality (Fo) value of 15 min. The combined effect of pH and temperature also showed cubic trends, and the Ea ranged between 240 and 398 (kJ) at pH 4–7.

Clostridia in Commercial Fish of the Azov and Black Seas and in Aquaculture Facilities in the Southern Region of Russia

The paper presents studies on the infection with clostridia of fish with skin lesions and ulcers on the surface of the body. The objects of study were syrman goby from the eastern part of the Taganrog Bay of the Sea of Azov, turbot from the shelf zone of the north-eastern Black Sea and carps reared under aquaculture conditions. Using an Autoflex speed III Bruker Daltonics (Germany) mass spectrometer, by the MALDI-TOF mass spectrometry was showed that sulfite-reducing clostridia (Clostridium perfringens, C. sporogenes) have been shown to infect the organs and tissues of syrman goby with vibriosis and turbot with ulcerative skin lesions of unknown etiology. Species such as Clostridium difficile, Clostridium novyi are of clinical importance and were found in the parenchymal organs of carp suffering from chronic aeromonas infection on pond fish farms in the southern Russia. These bacteria are its cosmopolitan distribution ability them to generate heat-resistant spores and cause food poisoning, which makes control and prevention measures needed in the food chain.

Convergence of Bigelow and Arrhenius models over a wide range of heating temperatures

The heat resistance of the bacterial spores of Moorella thermoacetica, Clostridium sporogenes, Geobacillus stearothermophilus and Bacillus coagulans was determined over a wide range of temperatures using the capillary method and thermoresistometer Mastia. The results showed that the two experimental methods gave similar heat resistance values excepted for Geobacillus stearothermophilus.The effect of temperature on thermal resistance was evaluated using the Arrhenius and Bigelow models. The fit of the heat sensitivity parameters of the Arrhenius and Bigelow models on the heat resistance parameter values obtained over a wide temperature range was equally good. Despite the apparent mathematical incompatibility of the two equations, it is recognized that they yield the same goodness of fit. This paper finds a mathematical reason for this convergence and explains why inside a temperature range of at least 100 °C, no significant difference in the quality of fit between these two models can be found.

Heat resistance of spores of 18 strains of Geobacillus stearothermophilus and impact of culturing conditions

In this study, different methods were evaluated for enumeration of spores of G. stearothermophilus, different sporulation methods were assessed for yields and wet heat resistances of obtained spores, and subsequently, the variation in heat resistances of spores was determined. Overall, tryptone soya agar (TSA) was the most suitable medium for enumeration of spores of this thermophilic bacterium. Sporulation on different media both at 55 and at 61 °C led to considerable variation in spore heat resistance. The heat resistance of spores was highest upon sporulation on medium supplemented with free ions of calcium, potassium, magnesium and manganese (CaKMgMn). For 18 different G. stearothermophilus strains that were isolated from various sources, spores were subsequently produced on nutrient agar supplemented with CaKMgMn at 55 °C. Strain ATCC 12980T, also known as 9A20, which is commonly used in steam sterilization tests was included. The survival of spores of all strains was assessed at 125 °C and 130 °C using two independent spore batches per strain. The mean D125°C for spores of the 18 strains was 1.1 min (95% PI 0.48–2.3 min) and the mean D130°C was 0.37 min (95% PI 0.17–0.82 min). For spore inactivation of these 18 strains, a z-value of 11.1 °C was estimated, resulting in an estimated D-value of 2.4 min (95% PI 1.1–5.2) at the reference temperature 121.1 °C. Based on the data sets obtained in this study, it was found that the variability in spore heat resistance could largely be attributed to strain variability and conditions used during sporulation (especially the sporulation medium); reproduction and experimental variabilities were much smaller. The established variabilities were compared with the overall variability in spore heat resistance of G. stearothermophilus based on a meta-analysis of reported D-values. The data presented indicate that strain variability and history of sporulation each account for approximately half of the overall variability observed with respect to the heat resistance of spores of G. stearothermophilus. The findings presented in this study allow for optimal recovery of G. stearothermophilus spores from foods and a better understanding of factors that determine the heat resistance properties of spores of G. stearothermophilus. Moreover, this study once more underlines the limited effects of heat treatments used in the food industry on inactivation of spores of this bacterium.

Differential requirements for conserved peptidoglycan remodeling enzymes during Clostridioides difficile spore formation.

Spore formation is essential for the bacterial pathogen and obligate anaerobe, Clostridioides (Clostridium) difficile, to transmit disease. Completion of this process depends on the mother cell engulfing the developing forespore, but little is known about how engulfment occurs in C. difficile. In Bacillus subtilis, engulfment is mediated by a peptidoglycan degradation complex consisting of SpoIID, SpoIIP and SpoIIM, which are all individually required for spore formation. Using genetic analyses, we determined the functions of these engulfment-related proteins along with the putative endopeptidase, SpoIIQ, during C. difficile sporulation. While SpoIID, SpoIIP and SpoIIQ were critical for engulfment, loss of SpoIIM minimally impacted C. difficile spore formation. Interestingly, a small percentage of ∆spoIID and ∆spoIIQ cells generated heat-resistant spores through the actions of SpoIIQ and SpoIID, respectively. Loss of SpoIID and SpoIIQ also led to unique morphological phenotypes: asymmetric engulfment and forespore distortions, respectively. Catalytic mutant complementation analyses revealed that these phenotypes depend on the enzymatic activities of SpoIIP and SpoIID, respectively. Lastly, engulfment mutants mislocalized polymerized coat even though the basement layer coat proteins, SpoIVA and SipL, remained associated with the forespore. Collectively, these findings advance our understanding of several stages during infectious C. difficile spore assembly.

Synergistic inactivation and mechanism of thermal and ultrasound treatments against Bacillus subtilis spores

Some Bacillus species are causative agents of food spoilage and a wide array of diseases. Due to their ability to form highly heat-resistant spores, it is of great interest to develop more effective inactivation strategies whereby these spores could be inactivated. Therefore, this work assessed inactivation of thermal and ultrasound treatments against Bacillus subtilis spores. The study further investigated the thermosonication (thermal and ultrasound, TS) -induced inactivation to the spores through a combination of morphology observation and internal factor analyses. The results of TS indicated that the TS combination synergistically inactivated spores by the maximum log reduction of 2.43 ± 0.08 at 80 °C and 20 W/ml and caused severe cell damage. The visual images revealed that the destructive mode of action of TS had multitarget sites, including coat, cortex, and inner membrane. Three distinct sub-populations were detected by Flow cytometry (FCM), and an unknown step with some physical compromise of the spore's inner membrane and partially hydrolyzed cortex involving the three steps model of inactivation was suggested. The combination of DPA (pyridine-2,6 dicarboxylic acid) content and the relative viabilities of the fractions suggested that during the TS treatment DPA release took place largely after spore death. The dead spores that retained DPA germinated relatively normally, but outgrow poorly, indicating that some key enzymes of intermediary metabolism has been damaged by TS treatment. Such understanding of the lethal action of TS may lead to the development of novel strategies involving spore destruction.

Paenibacillus xerothermodurans sp. nov., an extremely dry heat resistant spore forming bacterium isolated from the soil of Cape Canaveral, Florida.

A Gram-stain-positive, motile, endospore-producing, facultative anaerobic bacterial strain, designated ATCC 27380T, was isolated from heat-stressed soil of Cape Canaveral, Florida, USA. Growth was observed at 20-42 °C (optimum, 37 °C), at pH 6.0-10.0 (optimum pH 7.0) and in the presence of 0.5-3 % NaCl (optimum 0.5 %). The cell wall contained meso-diaminopimelic acid as the diagnostic amino acid and the isoprenoid quinone was MK-7. The polar lipids present were phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol and one unknown phospholipid. The main fatty acids were iso-C15 : 0 and anteiso-C15 : 0. Phylogenetic analysis based on 16S rRNA gene sequencing affiliated strain ATCC 27380T to the genus Paenibacillus, and showed the highest sequence similarity to Paenibacillus rigui JCM 16352T (97.0 %). The other closely related type strains exhibited 16S rRNA gene sequence similarity values below 95.9 %. The draft genome of ATCC 27380T had a size of 4,361,187 bases, with a G+C content of 51.0 %. The average nucleotide identity and in silico DNA-DNA hybridization values between strain ATCC 27380T and P. rigui JCM 16352T were 72.5% and 18.5 %, respectively, which were below the threshold suggested for species differentiation (96% and 70 %, respectively). The average amino acid identity between strain ATCC 27380T and P. rigui JCM 16352T was 68.72 %, which was above the suggested genus level demarcation of 65 %. Based on phenotypic, genotypic and chemotaxonomic data, strain ATCC 27380T represents a novel species in the genus Paenibacillus, for which the name Paenibacillusxerothermodurans sp. nov. (=DSM 520T=NRRL NRS-1629T=ATCC 27380T) is proposed.

Genetic and Functional Analyses of the DKxanthene Biosynthetic Gene Cluster from Myxococcus stipitatus DSM 14675.

DKxanthenes are a class of yellow secondary metabolites produced by myxobacterial genera Myxococcus and Stigmatella. We identified a putative 49.5 kb DKxanthene biosynthetic gene cluster from Myxococcus stipitatus DSM 14675 by genomic sequence and mutational analysis. The cluster was comprisedof 15 genes (MYSTI_06004-MYSTI_06018) encoding polyketide synthases, non-ribosomal peptide synthases, and proteins with unknown functions. Disruption of the genes by plasmid insertion resulted in defects in the production of yellow pigments. High-performance liquid chromatography and liquid chromatography-tandem mass spectrometry analysis indicated that the yellow pigments produced by M. stipitatus DSM 14675 might be noble DKxanthene derivatives. M. stipitatus did not require DKxanthenes for the formation of heat-resistant viable spores, unlike Myxococcus xanthus. Furthermore, DKxanthenes showed growth inhibitory activity against the fungi Aspergillus niger, Candida albicans, and Rhizopus stolonifer.

A quantitative microbiological exposure assessment of Bacillus cereus group IV in couscous semolina, Algeria

Couscous is widely consumed in Algeria, but it has been associated with several outbreaks. Couscous semolina may contain heat-resistant spores of Bacillus cereus group IV. These bacteria are mesophilic and potential producers of diarrheal toxin. The present study aimed at predicting (i) the prevalence and concentration of B. cereus group IV during couscous preparation at the time of consumption, and (ii) the number of consumers exposed to 105 cfu/g or more B. cereus in Algeria, due to couscous consumed at home or during collective events. In a probabilistic exposure assessment model, couscous semolina preparation was divided into three modules (i) contamination at retail, (ii) inactivation of B. cereus during couscous steaming and (iii) growth during storage before serving. Each module was built using a combination of newly gathered data, literature studies, predictive models and expert opinions. Algeria was split into four regions: North, South, High Plateau and Kabylia. The main differences between the regions were the seasonal temperature and the consumption pattern. Annually, a total of 3000 people (0.007% total population) and 170 people (0.0004%) were estimated to be exposed to a concentration equal to or above 105 cfu/g, due to home and collective consumption, respectively. The highest exposure came from couscous semolina kept at ambient temperature, in the south of Algeria during summer. Cold storage and consumption within the first 12 h after steaming are factors which limit the exposure to B. cereus group IV in couscous.

Forespore Targeting of SpoVD in Bacillus subtilis Is Mediated by the N-Terminal Part of the Protein.

SpoVD and PBP4b are structurally very similar high-molecular-weight, class B penicillin-binding proteins produced early during sporulation in Bacillus subtilis SpoVD is known to be essential for endospore cortex synthesis and thereby the production of heat-resistant spores. The role of PBP4b is still enigmatic. Both proteins are synthesized in the cytoplasm of the mother cell. PBP4b remains in the cytoplasmic membrane of the mother cell, whereas SpoVD accumulates in the forespore outer membrane. By the use of SpoVD/PBP4b chimeras with swapped protein domains, we show that the N-terminal part of SpoVD, containing the single transmembrane region, determines the forespore targeting of the protein.IMPORTANCE Beta-lactam-type antibiotics target penicillin-binding proteins (PBPs), which function in cell wall peptidoglycan synthesis. Bacteria of a subset of genera, including Bacillus and Clostridium species, can form endospores. The extreme resistance of endospores against harsh physicochemical conditions is of concern in clinical microbiology and the food industry. Endospore cortex layer biogenesis constitutes an experimental model system for research on peptidoglycan synthesis. The differentiation of a vegetative bacterial cell into an endospore involves the formation of a forespore within the cytoplasm of the sporulating cell. A number of proteins, including some PBPs, accumulate in the forespore. An understanding of the molecular mechanisms behind such subcellular targeting of proteins in bacterial cells can, for example, lead to a means of blocking the process of sporulation.

Perspectives of high power ultrasound in food preservation

High Power ultrasound can be used to alter physicochemical properties and improve the quality of foods during processing due to a number of mechanical, chemical, and biochemical effects arising from acoustic cavitation. Cavitation creates pressure waves that inactivate microbes and de-agglomerate bacterial clusters or release ascospores from fungal asci. Bacterial and heat resistant fungal spores’ inactivation is a great challenge in food preservation due to their ability to survive after conventional food processing, causing food-borne diseases or spoilage. In this work, a showcase of application of high power ultrasound combined with heat or thermosonication, to inactivate bacterial spores i.e. Bacillus cereus spores in beef slurry and fungal spores i.e. Neosartorya fischeri ascospores in apple juice was presented and compared with thermal processing. Faster inactivation was achieved at higher TS (24 KHz, 0.33 W/g or W/mL) temperatures. Around 2 log inactivation was obtained for B. cereus spores after1 min (70 °C) and N. fischeri ascospores after 30 min (75 °C). Thermal treatments caused <1 log in B. Cereus after 2 min (70 °C) and no inactivation in N. Fischeri ascospores after 30 min (80 °C). In conclusion, temperature plays a significant role for TS spore inactivation and TS was more effective than thermal treatment alone. The mould spores were more resistant than the bacterial spores.

Clostridium perfringens in foods and fish

Foodborne diseases are considered an important public health problem at a global level due to their levels of incidence and mortality as well as their negative consequences in economic and social aspects. Foodborne diseases are defined as those that are generated by the ingestion of food and water contaminated by chemical or biological agents commonly affecting health at the level of the gastrointestinal system. Among the risks and dangers to health from food are the diseases caused by Clostridium perfringens, a common bacterium which inhabits water, soil, vegetables, fish, the gastrointestinal system of human and animals and of course foods. The importance of this bacterium in health and food lies both in its cosmopolitan distribution, ability to generate heat-resistant spores and food poisoning, which makes control and prevention actions indispensable along the food chain. This article presents a general description of foodborne diseases, including those caused by consumption of food, such as fish, derived from contamination by C. perfringens; likewise, the actions and recommendations undertaken around the world for the prevention and control of these diseases are shown, including aspects related to the antimicrobial resistance phenomenon and its impact on public health.