Target Microorganisms Research Articles

The Bactericidal Effect of MA-T for Factitiously Contaminated and Used Masks.

Matching transformation system (MA-T), an on-demand aqueous chlorine dioxide solution, is an excellent safety disinfectant, because chlorine dioxide is not detected during storage or before use. The production of chlorine dioxide in MA-T is induced by a catalytic reaction in the presence of target microorganisms. In this study, we investigated MA-T disinfection of masks as a reuse method to eliminate mask shortages. After spraying Escherichia coli on sterilized surgical mask, samples (factitiously contaminated masks) were treated with MA-T spraying or immersion, and the bactericidal efficacy was assessed by culturing. Used surgical masks were also sprayed with MA-T or were immersed in MA-T, and then were cultured to verify the bactericidal effect. The performance of N95 masks was assessed before and after application of MA-T. After spraying with MA-T, the numbers of bacteria of factitiously contaminated masks and used masks were drastically reduced compared with control samples (not applicable and p = 0.002, respectively). After MA-T immersion, the bacterial counts of both masks (factitiously contaminated masks and used masks) were significantly reduced (both p = 0.002). Taken together, the disinfection test on factitiously contaminated with E. coli and used surgical masks showed that masks can be disinfected by MA-T spray and sterilized by immersion, respectively. The N95 mask performance test after 30 min of immersion in MA-T showed that MA-T disinfected the mask without degrading the performance of the mask. In conclusion, MA-T is useful for the reuse of masks because of its decontamination effect and safety while maintaining the function of the mask.

Characterization of Essential Oil and Wastewater from Thymus nummularius M. Bieb. and Micromorphological Examination of Glandular Trichomes

This study aimed to determine the volatile profile by GC-MS and antimicrobial activity of the essential oil isolated from Thymus nummularius using the microwave-assisted hydrodistillation method and morphologically characterize the glandular trichomes in leaf samples. Obtained essential oil, as well as wastewater obtained during essential oil isolation, were chemically (polyphenolic compounds, organic acids, total phenolic content, total flavonoid content and antioxidant activity) characterized. As a result of GC-MS analysis of the essential oil, different ratios of essential oil components were identified, predominantly thymol (38.91%), linalool (13.12%) and geraniol (6.51%). The antimicrobial activity results showed that T. nummularius essential oil has a broad-spectrum inhibitory effect against all target microorganisms (bacteria and yeast-like fungi). Screening of 32 polyphenolic compounds and 52 organic acids in the wastewater was performed by LC-MS/MS and the major polyphenolic compound and organic acid were determined as caffeic acid (97316.85 µg/kg) and malic acid (6898.88 mg/kg), respectively. While the total phenolic and flavonoid content of the wastewater were 35.91 mg GA/g and 34.91 mg QE/g, respectively, the antioxidant activity results were obtained using the CUPRAC, CERAC and ABTS tests were 86.28 mg TE/g, 72.85 mg TE/g and 47.88 mg TE/g. These results support that the T. nummularius, an important production area in Turkey, can be used in different industrial areas due to its rich bioactive content. In addition, it can be said that wastewater is also considered a valuable source of bioactive substances.

The Importance of Heating Unit Operations in the Food Industry to Obtain Safe and High-Quality Products.

Civilization has begun around 3,500 BCE in Mesopotamia and the realization by people that they could manipulate food to preserve it, through sun drying, fermentation, freezing in the snow, or cooking by fire, was an important factor for the nomadic humans to start settling. Food by nature is subject to spoilage and the application of any kind of preservation method enables storage and weighted consumption. Throughout human history, many techniques have been developed and improved such as heat treatment, drying, freezing, extraction, mixing and the use of preservatives, among others. In the food industry of the modern world, each technique is implemented through sequential steps, known as unit operations. This opinion paper presents an overview of the main heating unit operations used in the food industry, highlighting their benefits to converting raw materials into palatable products with high quality and safe for consumption. Examples are presented to illustrate how several food products available in the market were submitted only to physical transformations based on scientific knowledge. However, there is a range of intensity in physical processing and the applied energy level depends on the nature of the food, target microorganism, storage conditions, type of packaging, and desired shelf-life. The importance of food safety is stressed since processed foods have been criticized for confusion between nutritious values and processing steps. There are still many challenges to the food industry to design the process in optimal conditions for food quality and with less environmental impacts and novel thermal and non-thermal technologies have been studied and implemented.

Fate of Escherichia coli artificially inoculated in Tenebrio molitor L. larvae rearing chain for human consumption

The edible insect food chain represents a relatively novel food-producing system; hence, associated biological risks still need to be exhaustively evaluated. In the present study, the dynamics of Escherichia coli during the whole living period of Tenebrio molitor larvae (from eggs to pupae) were studied. To this end, a rearing substrate consisting of organic wheat middlings was spiked with E. coli cells at two initial contamination levels: 1 log cfu g−1 (low) and 6 log cfu g−1 (high). Microbial viability counting coupled with metataxonomic analyses was used to assess i) the persistence and growth of E. coli in the rearing substrate (wheat middlings); ii) the colonization and growth of E. coli in the insect larvae; and iii) the occurrence and load of E. coli in the frass (excrement from larvae mixed with substrate residues). The results highlighted a very limited persistence of the pathogen in all analyzed samples. In more detail, the results suggested that when E. coli was present at very low levels in the eggs of the insect, the pathogen was not able to reach concerning levels in the larvae. Moreover, when E. coli was present in the wheat middlings used for rearing, the environmental conditions of the substrate (low aw values) were not favorable for its survival and multiplication, irrespective of the presence of the larvae and their frass. Surprisingly, under the conditions applied in the present study, the larvae fed wheat middlings contaminated with E. coli seemed to be inhospitable or even hostile environments for microbial survival or multiplication. To explain the low levels of E. coli cells in the larvae reared in the present study, many factors can be considered, including the immune response of the host, microbial composition and interactions established in the gut of larvae, and insect species. Of note, part of the major fraction of the microbiota of larvae at the end of rearing was represented by Lactococcus, thus suggesting a possible effect of this lactic acid bacterium on E. coli decay. Further research is needed to better clarify the interactions between E. coli and the insect gut, as well as the interactions established among the target microorganism and those naturally harbored by the insect gut.

Essential oil in vapor phase in combination with Enterococcus mundtii STw38 to improve refrigerated hake fillets shelf-life

Essential oils (EOs) in vapor phase were applied together with the bacteriocinogenic strain E. mundtii STw38 to improve the shelf life of Argentinean hake -Merluccius hubbsi-. Evaluated EOs were oregano, thyme, rosemary, and lemongrass. In vitro results demonstrated their effectiveness against foodborne pathogen surrogate Listeria innocua, as well as against Lactobacillus plantarum, Shewanella putrefaciens, and fish indigenous biota. The most effective were oregano and lemongrass, showing the lowest minimal inhibitory concentration, and synergic activity when combined against L. innocua and S. putrefaciens. Furthermore, synergic mixtures promoted changes in target microorganisms cell wall. The composition of the volatile phase of a synergic EOs mixture showed a higher presence of oregano components. When applying the EOs alone in hake fillets, total bacteria count increased 3.3 log cycles whereas in the control system the increase was higher than 5 log cycles, and consumers evaluated similarly in taste and smell both the control and the EOs synergic mixtures. When adding the most concentrated synergic mixture (0.147 μL lemongrass/mL headspace and 0.195 μL oregano/mL headspace) together with the bioprotective culture, in 7 days total bacteria count only increased 3 log cycles, showing the effectiveness of the use of EOs together with E. mundtii STw38.

Enteric Pathogens Risk Factors Associated with Household Drinking Water: A Case Study in Ugu District Kwa-Zulu Natal Province, South Africa.

The occurrence of diarrheal infections depends on the level of water and sanitation services available to households of immunocompromised individuals and children of less than five years old. It is therefore of paramount importance for immunocompromised individuals to be supplied with safe drinking water for better health outcomes. The current study aimed at ascertaining the probability of infection that Escherichia coli, Salmonella typhimurium, Shigella dysenteriae, Vibrio cholerae, and rotavirus might cause to rural dwellers as compared to urban dwellers. Both culture-based and molecular-based methods were used to confirm the presence of target microorganisms in drinking water samples, while Beta-Poisson and exponential models were used to determine the health risk assessment. Results revealed the presence of all targeted organisms in drinking water. The estimated health risks for single ingestion of water for the test pathogens were as follows: 1.6 × 10−7 for S. typhimurium, 1.79 × 10−4 for S. dysenteriae, 1.03 × 10−3 for V. cholerae, 2.2 × 10−4 for E. coli O157:H7, and 3.73 × 10−2 for rotavirus. The general quantitative risk assessment undertaken in this study suggests that constant monitoring of household container-stored water supplies is vital as it would assist in early detection of microbial pathogens. Moreover, it will also allow the prompt action to be taken for the protection of public health, particularly for immunocompromised individuals and children who are prone to higher risk of infections.

Influence of Processing Parameters and Natural Antimicrobial on Alicyclobacillus acidoterrestris and Clostridium pasteurianum Using Response Surface Methodology.

The food industry must ensure the stability of the products, and this is often achieved by exposing foods to heat treatments that are able to ensure the absence of pathogenic or spoilage microorganisms. These treatments are different in terms of temperature and duration and could lead to a loss in nutritional and sensory value. Moreover, some types of microorganisms manage to survive these treatments thanks to the sporification process. The addition of antimicrobials can become necessary, but at present, consumers are more inclined toward natural products, avoiding synthetic and chemical additives. Antimicrobials from plants could be a valuable option and, in this context, a patent concerning an antimicrobial extract from fermented plant substrate was recently tested against foodborne pathogens revealing high antimicrobial activity. The objective of this study was the creation of a model for the evaluation and subsequent prediction of the combined effect of different process and product variables, including antimicrobial addition, on the inhibition and reduction of spore germination of target microorganisms, Alicyclobacillus acidoterrestris and Clostridium pasteurianum, responsible for spoilage of tomato-based products.

Use of static overpressure to assess the role of acoustic cavitation in ultrasound-mediated gene transfection

Ultrasound-mediated DNA delivery (UDD) is a technique that utilizes ultrasound to aid the transfer of genetic materials into bacterial cells of interest (either in their planktonic or biofilm modes), resulting in new functionalities being displayed by the target microorganisms in-situ. The physical mechanism of UDD is poorly understood, which hampers its widespread adoption and scaling. To investigate the role of cavitation in UDD, we utilized a static pressure vessel designed to suppress cavitation while leaving other acoustic effects unaltered. Pseudomonas putida UWC1, a soil bacterium that is not naturally competent, was suspended in water and exposed to 40 kHz burst-mode ultrasound (400 cycle burst, 10 Hz repetition frequency, 60 s total) at an ambient pressure of either 1 bar or 9.5 bars. Noise diagnostics yielded inertial cavitation levels and transformed cells were quantified using a doubly selective reporter plasmid DNA. Samples exposed at 9.5 bars overpressure exhibited a drastic reduction in both cavitation activity and the number of transformed cells compared to those exposed at 1 bar. (Cell viability was not affected by pressure change.) The fact that transformation rates are reduced significantly when cavitation is suppressed suggests that acoustic cavitation is the major contributing factor in UDD.

On the Efficacy of H2O2 or S2O82- at Promoting the Inactivation of a Consortium of Cyanobacteria and Bacteria in Algae-Laden Water.

Harmful algal blooms in coastal areas can significantly impact a water source. Microorganisms such as cyanobacteria and associated pathogenic bacteria may endanger an ecosystem and human health by causing significant eco-hazards. This study assesses the efficacy of two different reagents, H2O2 and S2O82−, as (pre-)treatment options for algae-laden waters. Anabaena sp. and Vibrio alginolyticus have been selected as target microorganisms. With the objective of activating H2O2 or S2O82−, additional experiments have been performed with the presence of small amounts of iron (18 µmol/L). For the cyanobacterial case, H2O2-based processes demonstrate greater efficiency over that of S2O82−, as Anabaena sp. is particularly affected by H2O2, for which >90% of growth inhibition has been achieved with 0.088 mmol/L of H2O2 (at 72 h of exposure). The response of Anabaena sp. as a co-culture with V. alginolyticus implies the use of major H2O2 amounts for its inactivation (0.29 mmol/L of H2O2), while the effects of H2O2/Fe(II) suggests an improvement of ~60% compared to single H2O2. These H2O2 doses are not sufficient for preventing the regrowth of V. alginolyticus after 24 h. The effects of S2O82− (+ Fe(II)) are moderate, reaching maximum inhibition growth of ~50% for Anabaena sp. at seven days of exposure. Nevertheless, doses of 3 mmol/L of S2O82− can prevent the regrowth of V. alginolyticus. These findings have implications for the mitigation of HABs but also for the associated bacteria that threaten many coastal ecosystems.

Decreased efficacy of antimicrobial agents in a polymicrobial environment

The airways of people with cystic fibrosis (CF) often harbour diverse polymicrobial communities. These airway infections can be impossible to resolve through antibiotic intervention, even though isolates of the individual species present are susceptible to the treatment when tested in vitro. In this work, we investigate how polymicrobial cultures comprised of key CF-associated pathogens respond to challenge with species-specific antimicrobial agents; colistin (targets Pseudomonas aeruginosa), fusidic acid (targets Staphylococcus aureus), and fluconazole (targets Candida albicans). We found that growth in a polymicrobial environment protects the target microorganism (sometimes by several orders of magnitude) from the effect(s) of the antimicrobial agent. This decreased antimicrobial efficacy was found to have both non-heritable (physiological) and heritable (genetic) components. Whole-genome sequencing of the colistin-resistant P. aeruginosa isolates revealed single nucleotide polymorphisms and indels in genes encoding lipopolysaccharide (LPS) biosynthesis and/or pilus biogenesis, indicating that a previously undescribed colistin resistance mechanism was in operation. This was subsequently confirmed through further genetic analyses. Our findings indicate that the polymicrobial nature of the CF airways is likely to have a significant impact on the clinical response to antimicrobial therapy.

Antimicrobial activity of oregano essential oil and resveratrol emulsions co-encapsulated by sodium caseinate with polysaccharides

Plant essential oils and polyphenol are promising antimicrobial agents, though their undesirable physical stability, volatilization, and hydrophobic property followed with unfavorable antimicrobial activity limited application in foods. In this study, oregano essential oil (OEO)-resveratrol (Res) emulsions were prepared using 0.25% sodium caseinate (SC) with and without 2.0% inulin/gum Arabic (GA). Antimicrobial activity of these emulsions against Staphylococcus aureus, Saccharomyces cerevisiae, and naturally occurring microorganisms in growth media and artificially inoculated on fresh-cut cabbage were assessed. Encapsulated OEO-Res with SC-GA had better antimicrobial effect than the encapsulated OEO-Res with SC-inulin followed by OEO-Res with SC alone. Incorporation of OEO-Res encapsulated by SC-GA resulted in a maximum reduction of 3.6-log and 5.77-log CFU/mL of S. aureus and S. cerevisiae in growth media and more than 90% reduction (1.53–1.93-log CFU/mL) of target microorganisms and naturally occurring microorganisms (reduced 1.36–2.19-log CFU/mL) on fresh cut cabbage. The antimicrobial ability of OEO-Res emulsions was highly related to retention of both OEO and Res. Low storage temperature (i.e., 4 °C) facilitated the retention of both bioactive compounds and reserved antimicrobial ability of them. The present study showed a potential antimicrobial ability of encapsulated OEO-Res emulsions on fresh food products.

Role is in the eye of the beholder-the multiple functions of the antibacterial compound tropodithietic acid produced by marine Rhodobacteraceae.

Many microbial secondary metabolites have been studied for decades primarily because of their antimicrobial properties. However, several of these metabolites also possess nonantimicrobial functions, both influencing the physiology of the producer and their ecological neighbors. An example of a versatile bacterial secondary metabolite with multiple functions is the tropone derivative tropodithietic acid (TDA). TDA is a broad-spectrum antimicrobial compound produced by several members of the Rhodobacteraceae family, a major marine bacterial lineage, within the genera Phaeobacter, Tritonibacter, and Pseudovibrio. The production of TDA is governed by the mode of growth and influenced by the availability of nutrient sources. The antibacterial effect of TDA is caused by disruption of the proton motive force of target microorganisms and, potentially, by its iron-chelating properties. TDA also acts as a signaling molecule, affecting gene expression in other bacteria, and altering phenotypic traits such as motility, biofilm formation, and antibiotic production in the producer. In microbial communities, TDA-producing bacteria cause a reduction of the relative abundance of closely related species and some fast-growing heterotrophic bacteria. Here, we summarize the current understanding of the chemical ecology of TDA, including the environmental niches of TDA-producing bacteria, and the molecular mechanisms governing the function and regulation of TDA.

LC-ESI-QTOF/MS characterization of antimicrobial compounds with their action mode extracted from vine tea (Ampelopsis grossedentata) leaves.

Vine tea (Ampelopsis grossedentata) is a tea plant cultivated south of the Chinese Yangtze River. It has anti‐inflammatory properties and is used to normalize blood circulation and detoxification. The leaves of vine tea are the most abundant source of flavonoids, such as dihydromyricetin and myricetin. However, as the main bioactive flavonoid in vine tea, dihydromyricetin was the main focus of previous research. This study aimed to explore the antibacterial activities of vine tea against selected foodborne pathogens. The antimicrobial activity of vine tea extract was evaluated by the agar well diffusion method. Cell membrane integrity and bactericidal kinetics, along with physical damage to the cell membrane, were also observed. The extract was analyzed using a high‐performance liquid chromatography‐diode array detector (HPLC‐DAD), and the results were confirmed using a modified version of a previously published method that combined liquid chromatography and electrospray‐ionized quadrupole time‐of‐flight mass spectrometry (LC‐ESI‐QTOF/MS). Cell membrane integrity and bactericidal kinetics were determined by releasing intracellular material in suspension and monitoring it at 260 nm using an ultraviolet (UV) spectrophotometer. A scanning electron microscope (SEM) was used to detect morphological alterations and physical damage to the cell membrane. Six compounds were isolated successfully: (1) myricetin (C15H10O8), (2) myricetin 3‐O‐rhamnoside (C21H20O12), (3) 5,7,8,3,4‐pentahydroxyisoflavone (C15H10O7), (4) dihydroquercetin (C15H12O7), (5) 6,8‐dihydroxykaempferol (C15H10O8), and (6) ellagic acid glucoside (C20H16O13). Among these bioactive compounds, C15H10O7 was found to have vigorous antimicrobial activity against Bacillus cereus (AS11846) and Staphylococcus aureus (CMCCB26003). A dose‐dependent bactericidal kinetics with a higher degree of absorbance at optical density 260 (OD260) was observed when the bacterial suspension was incubated with C15H10O7 for 8 h. Furthermore, a scanning electron microscope study revealed physical damage to the cell membrane. In addition, the action mode of C15H10O7 was on the cell wall of the target microorganism. Together, these results suggest that C15H10O7 has vigorous antimicrobial activity and can be used as a potent antimicrobial agent in the food processing industry.

Adaptation of O157:H7 and non-O157 Escherichia coli strains in orange juice and subsequent resistance to UV-C radiation

This study assessed the acid-adaptation of pathogenic and non-pathogenic strains of Escherichia coli in orange juice and the microbial resistance to the subsequent UV-C radiation treatment. Nine Shiga toxin-producing E. coli (STEC) and one strain of a non-pathogenic surrogate E. coli were used in this study. Each E. coli strain was inoculated in orange juice, following pre-exposure during 0, 1, 2, and 3 h at 10 °C. Then, the inoculated juices with the ten different strains separately were exposed to 0 and 2 J/cm2 of UV-C radiation. The D value (i.e., the UV-C dose in J/cm2 required to cause a one-log reduction in the target microorganism) was calculated. Further, the resistance coefficient [RC; i.e., the ratio between the D-values for the control condition (D0h) and each pre-exposure tested time (D1h,D2h, D3h)] were determined. The results indicated that the resistance of E. coli was influenced by the pre-exposure period in the orange juice, with increased resistance to UV-C observed for periods >2 h. Furthermore, the sensitivity of cells to subsequent UV-C treatment was found to be strain-dependent. The results may allow the development of more reliable UV-C radiation processes for orange juice processing aiming the inactivation of pathogenic E. coli.

UVA and solar driven photocatalysis with rGO/TiO2/polysiloxane for inactivation of pathogens in recirculation aquaculture systems (RAS) streams

In this study TiO2/polysiloxane(SiBi) thin films modified with different concentrations of graphene oxide (GO) were prepared by ink-jet printing on flexible polyethylene terephthalate (PET) substrates. Prepared coatings were characterized by means of scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), ultraviolet photoelectron spectroscopy (UPS), Raman and water contact angle measurements. During photocatalytic tests strong change of color of prepared coatings modified with GO was observed. XPS analysis of thin films after photocatalytic tests suggests that reduction of GO took place. Prepared coatings were studied for inactivation of microorganisms naturally occurring in aquaculture water under UVA and natural solar irradiation. Effect of rGO concentration in prepared coating on inactivation of target bacteria Aeromonas hydrophila and Citrobacter gillenii was evaluated under UVA irradiation. Aeromonas hydrophila was more sensitive to photocatalytic inactivation in comparison with Citrobacter gillenii. Higher photocatalytic inactivation of target microorganisms was attributed to thin films with concentration of rGO 1 and 5%. The rGO/TiO2/SiBi 5% were further tested for inactivation of Aeromonas salmocida, Serratia fonticola strain and Lactococcus lactis strain under natural solar light. Solar photocatalysis slightly enhanced inactivation of Aeromonas salmocida and Serratia fonticola strain, while opposite was observed for Lactococcus lactis strain.

Microbiological quality analysis of inoculants based on Bradyrhizobium spp. and Azospirillum brasilense produced \u201con farm\u201d reveals high contamination with non-target microorganisms

The use of inoculants carrying diazotrophic and other plant growth–promoting bacteria plays an essential role in the Brazilian agriculture, with a growing use of microorganism-based bioproducts. However, in the last few years, some farmers have multiplied microorganisms in the farm, known as “on farm” production, including inoculants of Bradyrhizobium spp. for soybean (Glycine max L. Merrill.) and Azospirillum brasilense for corn (Zea mays L.) or co-inoculation in soybean. The objective was to assess the microbiological quality of such inoculants concerning the target microorganisms and contaminants. In the laboratory, 18 samples taken in five states were serial diluted and spread on culture media for obtaining pure and morphologically distinct colonies of bacteria, totaling 85 isolates. Molecular analysis based on partial sequencing of the 16S rRNA gene revealed 25 genera of which 44% harbor species potentially pathogenic to humans; only one of the isolates was identified as Azospirillum brasilense, whereas no isolate was identified as Bradyrhizobium. Among 34 isolates belonging to genera harboring species potentially pathogenic to humans, 12 had no resistance to antibiotics, six presented intrinsic resistance, and 18 presented non-intrinsic resistance to at least one antibiotic. One of the samples analyzed with a shotgun-based metagenomics approach to check for the microbial diversity showed several genera of microorganisms, mainly Acetobacter (~ 32% of sequences) but not the target microorganism. The samples of inoculants produced on farm were highly contaminated with non-target microorganisms, some of them carrying multiple resistances to antibiotics.

ATIVIDADE ANTIMICROBIANA DE QUITOSANAS E SEUS DERIVADOS: INFLUÊNCIA DAS CARACTERÍSTICAS ESTRUTURAIS

ANTIMICROBIAL ACTIVITY OF CHITOSAN AND ITS DERIVATIVES: INFLUENCE OF ITS STRUCTURAL CHARACTERISTICS. In recent years, there has been an increasing interest in the development of antimicrobial materials for different applications, especially in the biomedical field. In this context, chitosan is one of the most employed materials due to its highly attractive properties, such as biocompatibility, biodegradability, non-toxicity, and antimicrobial activity against gram-negative and gram-positive bacteria and fungi. In this review, we discuss the different methods used to obtain chitosan and point out how they affect the structural and chemical properties of this material, followed by a detailed discussion on how these properties influence its antimicrobial activities. Additionally, we summarize the most accepted antimicrobial mechanisms of action for chitosans. Specifically, we aim to provide a comprehensive overview on the last findings and progress reported in the literature on the factors that influence the antimicrobial activity of chitosan, e.g., polymeric average molecular weight (MV ), average degree of acetylation (DA), water-soluble derivatization and sample degree of purity. The data available in the literature indicate that in addition to the structural properties, the effectiveness of such antimicrobial activity of chitosan is also dependent on the target microorganisms. This review will help researchers to better understand the mechanism of action of chitosan against microorganisms based on its structural characteristics.

Development of a predictive model of the microbial inactivation of L. monocytogenes during low thermal treatment of fruit juices in combination with carvacrol as aroma compound

Mild heat treatment of fruit juices in combination with natural aroma compounds has been reported as an alternative to conventional pasteurization to better preserve their nutritional value. However, its antimicrobial efficiency varies from one juice to another. This study aims at developing a secondary predictive model of microbial inactivation scale during such combined process. Carvacrol was used as aroma compound and acid-adapted L. monocytogenes as target microorganism. The inactivation kinetics of this bacteria were followed in simulated fruit juices using a Central Composite Design with pH (2-6), °Brix (0–24), temperature (55–65 °C), and carvacrol concentration (0–60 μL/L) as independent variables. Curves were fitted to the Weibull inactivation model, and data collected used to generate two predictive models of the inactivation scale parameter through multiple regression analysis following an empirical and a mechanistic (based on Gamma concept) approach. The best of the two models was then validated using real fruit (orange, pineapple, and watermelon) juices. The empirical model where only the four variables tested were considered showed a lower statistical performance compared to the mechanistic model where octanol-water partition coefficient (Ko/w) and vapour pressure (Vp) of carvacrol at the treatment temperature were integrated (R2 0.6 and 0.9; Accuracy factor 1.5 and 1.3; Sum of Squared Error 3.6 and 1.1, respectively). No significant difference was observed between inactivation scale values obtained with real juices and the predicted values calculated using this mechanistic model. The Ko/w and Vp of the aroma compound used are key parameters that determine the efficiency of the above-described combined treatment.

Inactivation of Escherichia coli K12 on raw almonds using supercritical carbon dioxide and thyme oil

Raw almonds could be contaminated by pathogens, but the current pasteurization practice using propylene oxide in the U.S. has flammability and carcinogenicity concerns. Supercritical carbon dioxide (scCO2) is a water-free technology and is a solvent of essential oils that are effective antimicrobial preservatives. The objective of this study was to investigate the possibility of combining scCO2 and thyme oil (TO) to reduce Escherichia coli K12 inoculated on raw almonds. Raw almonds inoculated with ∼6 log CFU/g E. coli K12 were batch-treated with scCO2 alone or the combination of presoaking in pure TO followed by scCO2 treatments at different combinations of temperature, pressure, and duration. Compared to scCO2 alone treatments, the combination of TO and scCO2 treatments significantly improved the disinfection effectiveness. Temperature had the most significant effect on the log reduction. At 70 °C, the log reduction by the combination treatment was over 4-log CFU/g and the maximum reduction was 5.16 log CFU/g. The findings suggest that the combination of TO and scCO2 may be a potential water-free technology to meet the requirement of over 4-log reduction of target microorganism for almond and other tree nut products.

Targeted Delivery of Miconazole Employing LL37 Fragment Mutant Peptide CKR12-Poly (Lactic-Co-Glycolic) Acid Polymeric Micelles.

We previously reported that conjugates of antimicrobial peptide fragment analogues and poly (lactic-co-glycolic) acid (PLGA) enhance antimicrobial activity and that the conjugated micelle structure is an effective tool for antimicrobial drug delivery. In recent years, the delivery of antimicrobial peptides to targets for antimicrobial activity has attracted attention. In this study, we targeted Candida albicans, a causative organism of catheter-related bloodstream infections, which is refractory to antimicrobial agents and is currently a problem in medical practice. We evaluated the antifungal activity of CKR12 (a mutant fragment of the human cathelicidin peptide, LL-37)-PLGA-miconazole (MCZ) micelles using nanotechnology with MCZ delivery. The prepared CKR12-PLGA-MCZ micelles were characterised by measuring dynamic light scattering, zeta potential, dilution stability, and drug release. CKR12-PLGA-MCZ micelles showed higher antifungal activity than CKR12-PLGA micelles and MCZ solution. Furthermore, scanning and transmission electron microscopy suggested that CKR12-PLGA-MCZ micelles disrupted both cell wall and cell membrane of C. albicans. Our results revealed a synergistic effect of antifungal activity using a combination of antimicrobial peptide fragment analogues and MCZ, and that MCZ is a promising tool for the delivery to target microorganisms.