Eurasian consumers' food safety beliefs and trust issues in the age of COVID-19: evidence from an online survey in 15 countries.

BACKGROUND: This investigation provides an important insight into Eurasian consumers' food safety beliefs and trust issues influenced by the COVID-19 pandemic. An online survey was conducted in 15 European and Asian countries involving more than 4000 consumers. RESULTS: It has confirmed that different socioeconomic characteristics, cultural aspects and education levels shape food safety perceptions within Eurasian countries. The COVID-19 pandemic influenced their beliefs and trust in food safety, which is relatively low on average. However, it is significantly higher for European consumers (especially European Union ones) compared to their Asian counterparts. Both Asian and European respondents agreed that food fraud and climate changes represent a food safety issue. However, European consumers were less concerned regarding the food safety of genetically modified foods and meat and dairy analogs/hybrids. Asian consumers were, to a greater extent, worried about the risk of getting COVID-19 from food, restaurants, food retail establishments and home food deliveries. CONCLUSION: Eurasian consumers have put their greatest extent of trust, when food safety assurance is concerned, into food scientists and food producers holding a food safety certificate. Broadly, they are uncertain to what extent their federal governments and food inspectors are competent, able and efficient in ensuring food safety. Higher education of Eurasian consumers was followed by increased food safety confidence in all parts of the food chain. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Environmental risk factors associated with the survival, persistence, and thermal tolerance of Cronobacter sakazakii during the manufacture of powdered infant formula.

Cronobacter sakazakii is an opportunistic foodborne pathogen of concern for foods having low water activity such as powdered infant formula (PIF). Its survival under desiccated stress can be attributed to its ability to adapt effectively to many different environmental stresses. Due to the high risk to neonates and its sporadic outbreaks in PIF, C. sakazakii received great attention among the scientific community, food industry and health care providers. There are many extrinsic and intrinsic factors that affect C. sakazakii survival in low-moisture foods. Moreover, short- or long-term pre-exposure to sub-lethal physiological stresses which are commonly encountered in food processing environments are reported to affect the thermal resistance of C. sakazakii. Additionally, acclimation to these stresses may render C. sakazakii resistance to antibiotics and other antimicrobial agents. This article reviews the factors and the strategies responsible for the survival and persistence of C. sakazakii in PIF. Particularly, studies focused on the influence of various factors on thermal resistance, antibiotic or antimicrobial resistance, virulence potential and stress-associated gene expression are reviewed.

"Multidimensional correlation analysis of temperature and contact time on eradication of biofilms of Cronobacter sakazakii on abiotic surfaces by combination of hypochlorite and malic acid".

AIM: In the present study, malic acid in combination with sodium hypochlorite is evaluated for eradication of biofilms formed by Cronobacter sakazakii strains individually and in a cocktail on different abiotic surfaces. METHOD AND RESULTS: The biofilm formation by five strains of C. sakazakii and their cocktail culture on different substrates was studied in Tryptone Soy Broth (TSB) and reconstituted Powdered Infant Formula (PIF). Further, the effect of temperature (4, 27, 37 and 50°C) and contact time (10, 20, 30, 40, 50 and 60 min) on antibiofilm potential of test solution (0.0625 mol l-1 malic acid and 0.00004 mol l-1 sodium hypochlorite) against biofilm formed by C. sakazakii cocktail culture was investigated on these surfaces. The effect was evaluated in terms of viable cell count and biofilm texture using scanning electron microscopy (SEM). Principal Component Analysis (PCA) revealed that the maximum biofilm reduction was observed for stainless steel at 4°C after 60 min of contact whereas at 25, 37 and 50°C, maximum biofilm reduction was observed for polycarbonate. For glass and polyurethane, maximum log reductions were observed at 50°C. The SEM images revealed cell surface deformation and disruption in biofilms after treatment with the test solution. CONCLUSIONS: The antibiofilm potential was observed to be greatly affected by contact time and temperature. These results indicated that the combination of malic acid NaOCl can effectively kill and remove C. sakazakii biofilms from food contact surfaces and enteral feeding tubes.

"Multidimensional correlation analysis of temperature and contact time on eradication of biofilms of Cronobacter sakazakii on abiotic surfaces by combination of hypochlorite and malic acid".

AIM: In the present study, malic acid in combination with sodium hypochlorite is evaluated for eradication of biofilms formed by Cronobacter sakazakii strains individually and in a cocktail on different abiotic surfaces. METHOD AND RESULTS: The biofilm formation by five strains of C. sakazakii and their cocktail culture on different substrates was studied in Tryptone Soy Broth (TSB) and reconstituted Powdered Infant Formula (PIF). Further, the effect of temperature (4, 27, 37 and 50°C) and contact time (10, 20, 30, 40, 50 and 60 min) on antibiofilm potential of test solution (0.0625 mol l-1 malic acid and 0.00004 mol l-1 sodium hypochlorite) against biofilm formed by C. sakazakii cocktail culture was investigated on these surfaces. The effect was evaluated in terms of viable cell count and biofilm texture using scanning electron microscopy (SEM). Principal Component Analysis (PCA) revealed that the maximum biofilm reduction was observed for stainless steel at 4°C after 60 min of contact whereas at 25, 37 and 50°C, maximum biofilm reduction was observed for polycarbonate. For glass and polyurethane, maximum log reductions were observed at 50°C. The SEM images revealed cell surface deformation and disruption in biofilms after treatment with the test solution. CONCLUSIONS: The antibiofilm potential was observed to be greatly affected by contact time and temperature. These results indicated that the combination of malic acid NaOCl can effectively kill and remove C. sakazakii biofilms from food contact surfaces and enteral feeding tubes.

Transcriptome Analysis of Podoscypha petalodes Strain GGF6 Reveals the Diversity of Proteins Involved in Lignocellulose Degradation and Ligninolytic Function.

The present study reports transcriptomic profiling of a Basidiomycota fungus, Podoscypha petalodes strain GGF6 belonging to the family Podoscyphaceae, isolated from the North-Western Himalayan ranges in Himachal Pradesh, India. Podoscypha petalodes strain GGF6 possesses significant biotechnological potential as it has been reported for endocellulase, laccase, and other lignocellulolytic enzymes under submerged fermentation conditions. The present study attempts to enhance our knowledge of its lignocellulolytic potential as no previous omics-based analysis is available for this white-rot fungus. The transcriptomic analysis of P. petalodes GGF6 reveals the presence of 280 CAZy proteins. Furthermore, bioprospecting transcriptome signatures in the fungi revealed a diverse array of proteins associated with cellulose, hemicellulose, pectin, and lignin degradation. Interestingly, two copper-dependent lytic polysaccharide monooxygenases (AA14) and one pyrroloquinolinequinone-dependent oxidoreductase (AA12) were also identified, which are known to help in the lignocellulosic plant biomass degradation. Overall, this transcriptome profiling-based study provides deeper molecular-level insights into this Basidiomycota fungi, P. petalodes, for its potential application in diverse biotechnological applications, not only in the biofuel industry but also in the environmental biodegradation of recalcitrant molecules. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-022-01037-6.

Sukshmjeevanu in Vedas: The Forgotten Past of Microbiology in Indian Vedic Knowledge.

No one questions the existence of presumptive knowledge of invisible organisms causing disease, decay and destruction mentioned before the discovery of the microbial world by Antonie Van Leeuwenhoek, who was the first to describe the invisible world as per literature available today. However, the knowledge about microbial world by Indian Rishis presented in Sanskrit shlokas or suktas of our traditional manuscripts such as Vedas remained unseen, where the Rishis had predicted the role of microorganisms known as Krimi or Jeevanu years before Leeuwenhoek. This note is an attempt to bring an emphasis to revisit our traditional Vedic knowledge and establish them through research based facts for wider acceptance globally.

Lactobacillus gastricus BTM 7 prevents intestinal colonization by biofilm forming Cronobacter sakazakii in Caenorhabditis elegans model host.

The study reports protective role of potential probiotic cultures against infection by biofilm forming Cronobacter sakazakii in Caenorhabditis elegans model system. Among the fifteen indigenous potential probiotics, the cell free supernatant of Lactobacillus gastricus BTM7 possessed highest antimicrobial action and biofilm inhibition against C. sakazakii. The competitive exclusion assays revealed that preconditioning with probiotics resulted in increased mean life span of the nematode to 12-13 days as compared to 5-6 days when the pathogen was administered alone. Enhanced expression of the marker genes (pmk-1, daf-16 and skn-1) was observed during the administration of probiotic cultures. The highest expression of pmk-1 (2.5 folds) was observed with administration of L. gastricus BTM7. The principal component analysis on selected variables revealed that L. gastricus BTM7 has the potential to limit the infection of C. sakazakii in C. elegans and enhance the expression of key genes involved in extending life span of the worm.

Probiotic mediated colonization resistance against E.coli infection in experimentally challenged Caenorhabditis elegans.

This study investigates the antimicrobial potential of sixteen indigenous probiotic bacteria in Caenorhabditis elegans infected with different serotypes of clinical Escherichia coli isolates. The probiotic cultures exhibited varying degree of antimicrobial activity against the pathogenic strains. The Cell Free Supernatant (CFS) of Lactobacillus plantarum K90 exhibited maximum antimicrobial activity against all indicator strains. Further, the pathogenic potential of the clinical strains was determined using liquid killing assay in C. elegans, where the pathogenic strains resulted in complete killing of the worm in 5 days as compared to 60% survival of worms fed with standard food of E. coli OP50. The clinical strains also resulted in impaired pharynx and internal hatching of the eggs in the worms. The protective effects of probiotics against the pathogenic strains was determined via competition, exclusion and displacement assays with different stages of intervention of probiotic culture. No significant increase in mean life span (MLS) of the worm was observed in competition and displacement assay. Among the tested strains in exclusion assay, a pretreatment with L. gastricus BTM 7 was found to result in better protection of the worm against infection with pathogenic E. coli strains by extending its life by three days and no other adverse effect on physiology and morphology of the worm. The results suggest that preconditioning with probiotic strains can be used as an effective way to reduce the invasion and colonization by the pathogens.

Statistical assessment of DNA extraction methodology for culture-independent analysis of microbial community associated with diverse environmental samples.

Cost-effectiveness, quality, time-effectiveness and ease of the methodology are the most crucial factors in isolating quality DNA from wide variety of samples. Thus, research efforts focusing on the development of an efficient DNA extraction protocol is the need of the hour. The present study therefore, focuses on development of an efficient, rapid and free of inhibitory substances based methodology for extracting metagenomic DNA from diverse environmental samples viz. anaerobic biogas digesta, ruminant stomach, human feces, soil, and microbial starter cultures used for preparation of fermented food. PCR-DGGE based analysis and quality metagenomic library preparation, using DNA extraction methodology, validates the developed protocol. The developed protocol is cost effective, capable of isolating DNA from small sample size (100-1000 µl), time efficient (1.5-2.0 h protocol) and results in significantly higher DNA yield (4-8 times increased yield) when compared to previously available DNA extraction method and a commercial DNA extraction kit. The DNA extracted from the samples using different protocols was evaluated based on its ability to identify diverse microbial species using PCR-DGGE profiles targeting variable region within the 16S rRNA gene. The results of microbial community analysis revealed comparability of the developed protocol to commercial kits, in effectively identifying dominant representatives of the microbial community in different samples. Using the DNA extracted from the presented methodology, metagenomic libraries were prepared, which were found suitable for sequencing on Illumina platform.

Profiling of Virulence Determinants in Cronobacter sakazakii Isolates from Different Plant and Environmental Commodities.

Cronobacter sakazakii is an emerging pathogen causing meningitis, sepsis and necrotizing enterocolitis in neonates and immune-compromised adults. The present study describes the profiling of different virulence factors associated with C. sakazakii isolates derived from plant-based materials and environmental samples (soil, water, and vacuum dust). All the isolates exhibited ß-hemolysis and chitinase activity, and were able to utilize inositol. Among the nine virulence-associated genes, hly gene coding for hemolysin was detected in all the isolates followed by ompA (outer membrane protein); however, plasmid-borne genes were detected at a level of 60% for both cpa (cronobacter plasminogen activator) and eitA (Ferric ion transporter protein) gene, respectively. Furthermore, the isolate C. sakazakii N81 showed cytotoxicity for Caco-2 cells. The presence of the virulence determinants investigated in this study indicates the pathogenic potential of C. sakazakii with their plausible connection with clinical manifestations.

Inhibition of quorum-sensing-mediated biofilm formation in Cronobacter sakazakii strains.

The present study investigated plant extracts for their anti-quorum-sensing (QS) potential to inhibit the biofilm formation in Cronobacter sakazakii strains. The bioassay based on loss of pigment production by Chromobacterium violaceum 026 and Agrobacterium tumefaciens NTL4(pZLR4) was used for initial screening of the extracts. Further, the effect of extracts on the inhibition of QS-mediated biofilm in C. sakazakii isolates was evaluated using standard crystal violet assay. The effect on biofilm texture was studied using SYTO9 staining and light and scanning electron microscopy. Among the tested extracts, Piper nigrum and Cinnamomum verum at 100 ppm resulted in 78 and 68 % reduction in the production of violacein as well as blue-green colour in both biosensor strains. A higher inhibitory activity (>50 %) on biofilm formation in C. sakazakii was observed for Pip. nigrum and Cin. verum, whereas the other extracts possessed moderate (25-50 %) and minimal (<25 %) inhibitory activities. Further, the fluorescent and scanning electron microscopic images indicated a major disruption in the architecture of biofilms of tested strains by Pip. nigrum. This study points to the possibility of using Pip. nigrum and Cin. verum as inhibitor of QS-mediated biofilm formation by C. sakazakii that could be further explored for novel bioactive molecules to limit the emerging infections of C. sakazakii.

Nano-LC-Q-TOF Analysis of Proteome Revealed Germination of Aspergillus flavus Conidia is Accompanied by MAPK Signalling and Cell Wall Modulation.

Aspergillus flavus is the second most leading cause of aspergillosis. The ability of A. flavus to adapt within the host environment is crtical for its colonization. Onset of germination of conidia is one of the crucial events; thus, in order to gain insight into A. flavus molecular adaptation while germination, protein profile of A. flavus was obtained. Approximately 82 % of conidia showed germination at 7 h; thus, samples were collected followed by protein extraction and subjected to nLC-Q-TOF mass spectrometer. Q-TOF data were analysed using Protein Lynx Global Services (PLGS 2.2.5) software. A total of 416 proteins were identified from UniProt Aspergillus species database. Orthologues of A. flavus was observed in A. fumigatus, A. niger, A. terreus, A. oryzae, etc. Proteins were further analysed in NCBI database, which showed that 27 proteins of A. flavus are not reported in UniProt and NCBI database. Functional characterization of proteins resulted majorly to cell wall synthesis and degradation, metabolisms (carbohydrate and amino acid metabolism), protein synthesis and degradation. Proteins/enzymes associated with aflatoxin biosynthesis were observed. We also observed Dicer-like proteins 1, 2 and autophagy-related proteins 2, 9, 18, 13, 11, 22. Expression of protein/enzymes associated with MAPK signalling pathway suggests their role during the germination process. Overall, the data present a catalogue of proteins/enzymes involved in the germination of A. flavus conidia and could also be applied to other Aspergillus species.

Retraction: Characterization of cellulolytic activities of newly isolated Thelephora sowerbyi from North-Western Himalayas on different lignocellulosic substrates.

Characterization of cellulolytic activities of newly isolated Thelephora sowerbyi from North-Western Himalayas on different lignocellulosic substrate J. Basic Microbiol. 2015, 55, 1-11 - DOI: 10.1002/jobm.201500107 The above article from the Journal of Basic Microbiology, published online on 08 June 2015 in Wiley Online Library as Early View (http://onlinelibrary.wiley.com/doi/10.1002/jobm.201500107/pdf), has been retracted by agreement between the authors, the Editor-in-Chief and Wiley-VCH GmbH & Co. KGaA. The retraction has been agreed because the microorganism studied in the described experiments has been identified as the fungus Cotylidia pannosa (Gene Accession No. KT008117) instead of Thelephora sowerbyi. The culture has been identified on the basis of the sequence of the amplified ITS region of the microorganism which was submitted by the authors to the NCBI database.

Autochthonous microbial community associated with pine needle forest litterfall influences its degradation under natural environmental conditions.

The slow natural degradation of chir pine (Pinus roxburghii) needle litterfall and its accumulation on forest floors have been attributed to its lignocellulosic complexities of the biomass. The present study offers a microbiological insight into the role of autochthonous microflora associated with pine needle litterfall in its natural degradation. The denaturing gradient gel electrophoresis (DGGE) fingerprinting indicated actinomycetes (Saccharomonospora sp., Glycomyces sp., Agrococcus sp., Leifsonia sp., Blastocatella sp., and Microbacterium sp.) as a dominant microbial community associated with pine needle litterfall with the absence of fungal decomposers. On exclusion of associated autochthonous microflora from pine litterfall resulted in colonization by decomposer fungi identified as Penicillium chrysogenum and Aspergillus sp., which otherwise failed to colonize the litterfall under natural conditions. The results, therefore, indicated that the autochthonous microbial community of pine needle litterfall (dominated by actinomycetes) obstructs the colonization of litter-degrading fungi and subsequently hinders the overall process of natural degradation of litterfall.

Prevalence and Characterization of Cronobacter spp. from Various Foods, Medicinal Plants, and Environmental Samples.

Dairy or non-dairy based products were explored to determine the prevalence, molecular characterization, and antibiotic susceptibility of Cronobacter spp. The isolation was done as per ISO 22964:2006 on chromogenic media followed by further confirmation by biochemical- and 16S rRNA-based identification. From 219 samples, the chromogenic agar assay and biochemical tests yielded presumptive 45 isolates. Among them, only 36 isolates showed 282 bp band amplified from ITS-G gene confirming as Cronobacter sakazakii. The Cronobacter spp. prevalence was highest in herbs and spices (34 %) while environmental samples had contamination rates of 23 % indicating plants as a possible reservoir of this pathogen. All the isolates were resistant to ß-lactam derivatives (68 %), macrolides (88.6 %), and aminoglycosides (79.9 %) but susceptible to phenicoles (31.6 %) and tetracyclines (15 %) derivatives. The results emphasize the screening of plant materials before their incorporation in food matrices.

Probiotic potential of gluten degrading Bacillus tequilensis AJG23 isolated from Indian traditional cereal-fermented foods as determined by Multiple Attribute Decision-Making analysis.

The present study reported the characterization of gluten hydrolyzing strains of Bacillus sp. from fermented cereal dough. The strains were characterized for probiotic as well as technological attributes. A total of 45 presumptive gluten degrading isolates were obtained on gliadin agar plate assay. Based on hemolytic and antibiotic susceptibility pattern, only six isolates were considered safe which also indicated gliadinase activity on zymography. All the six strains were able to resist the pH 2.0, 0.25% bile and also possessed ability to adhere to the organic solvents and mucin. The cell free supernatant of five strains exhibited antimicrobial activities against Gram-positive and Gram-negative pathogens. A more than 50% survival of the isolated strains was obtained at a salt concentration of 2%, phenol concentration of 0.1% and temperature upto 45 °C. All the strains exhibited antioxidant activities and biofilm forming ability. Furthermore, the ranking of strains based on probiotic as well as other functional attributes was determined using multidimensional Technique for Order of Preference by Similarity to Ideal Solution (TOPSIS). A matrix of multidimensional indicators was prepared using alternatives and criteria, the analysis indicated the strain Bacillus tequilensis AJG23 as the potential probiotic candidate based on all screening criteria. Further work still needs to be done about the protective role of the potential strain against gluten sensitivity using in vitro models.

Nano-Conjugated Food-Derived Antimicrobial Peptides As Natural Biopreservatives: A Review of Technology and Applications.

In recent years, microbial food safety has garnered a lot of attention due to worldwide expansion of the food industry and processed food products. This has driven the development of novel preservation methods over traditional ones. Food-derived antimicrobial peptides (F-AMPs), produced by the proteolytic degradation of food proteins, are emerging as pragmatic alternatives for extension of the shelf-life of food products. The main benefits of F-AMPs are their wide spectrum antimicrobial efficacy and low propensity for the development of antibiotic resistance. However, direct application of F-AMPs in food limits its efficacy during storage. Therefore, the development of nanocarriers for the conjugation and distribution of potential AMPs may hold great potential to increase their bioactivity. This review highlights the significance of F-AMPs as a feasible and sustainable alternative to conventional food preservatives. The most recent developments in production, characterization, and mode of action of these AMPs against planktonic and biofilm forming pathogens are thoroughly discussed in this work. Moreover, nano-conjugation of F-AMPs with different nano-carriers and potential future application in food packaging are emphasized. This review may aid in comprehending the nano-conjugation of F-AMPs and offer insightful recommendations for further exploration and potential uses in the food processing industry.

Consumer attitudes and perceptions towards chilled ready-to-eat foods: a multi-national study.

Understanding consumers' behavior and their handling of high-risk foods at home is essential for reducing the number of foodborne illnesses. This study shows the results of a cross-national analysis of consumers' perception from nine countries, and the identification of customers' clusters and its characteristics in order to understand customers' behavior, and to build safe chilled ready-to-eat (RTE) foods prevention strategies. The cluster analysis resulted in two clusters: (1) "Precautious consumers" characterized by the orientation towards pre-packed RTE foods, with consumers mainly coming from Bosnia and Herzegovina, India, Poland, Portugal, Spain, and Turkey. Their attitudes and self-reported practices may be categorized as less risky in terms of food-borne illnesses connected with the consumption of RTE foods; (2) "Unconcerned consumers" preferred cutting and slicing RTE foods freshly at the point of purchase, usually sold at the delicatessen department in a supermarket or at open markets. Those consumers mostly came from Croatia, Serbia and Slovenia and their attitudes and self-reported practices were riskier. These results allow a better understating of what characterizes consumers of RTE foods in different countries.

Trending biocontrol strategies against Cronobacter sakazakii: A recent updated review.

Cronobacter sakazakii is an emerging foodborne pathogen, causing life-threatening infections in newborns and premature infants. Cronobacter spp. can survive under difficult processing conditions thereby contaminate the Powdered Infant Formula (PIF) during the manufacturing process. Infantile infections are associated with the consumption of contaminated PIF that was either contaminated intrinsically or extrinsically. This necessitates the development of sustainable strategies to manage the risk of Cronobacter infections. Natural methods of preservation holds promise as a viable alternative strategy to address the critical problem of emerging antimicrobial resistance and also to limit the negative effects of commonly used physico-chemical methods in food processing. The present study reviews the efficacies, potentials and developmental trends of biological antagonists and a combinatorial therapy to eliminate C. sakazakii using in vitro and in vivo methods. The mode of action of each biocontrol method has been discussed comprehensively. Most of these biocontrol agents interfere with the cell membrane integrity and its functions. However, none of the individual methods are able to eliminate the pathogen completely from the model food system i.e. reconstituted PIF. Each of the biological control strategies (agent) has its limitations in terms of their dose and method of application. A synergistic effect has been observed between the biological agent and physico-chemical treatments that may have the potential to ensure pathogen-free foods. Future research studies should evaluate the synergistic activities of these methods for their implication in infant foods as well as to understand the mechanisms of inactivation.

Impairment of Cronobacter sakazakii and Listeria monocytogenes biofilms by cell-free preparations of lactobacilli of goat milk origin.

Biofilm-associated bacterial infections represent one of the major threats to modern medical treatments. Bacteria encased in biofilm matrix are more resistant towards antimicrobials and thus the capability of microbes to persist and nurture in a biofilm seems to be the foremost aspect of pathogenesis and therapeutic failure. Therefore, there is a pressing demand for new drugs active against microbial biofilms. In the current study, anti-biofilm potential of Lactobacillus spp. cell-free supernatants (CFSs) against Cronobacter sakazakii and Listeria monocytogenes was characterized using crystal violet staining and MTT assay. CFSs of goat milk origin lactobacilli not only prevented biofilm formation but also disrupted preformed biofilms. Neutralized and heat-treated preparations of Lactobacillus CFSs also inhibited biofilm formation by test pathogens. The results were quantitatively confirmed by light and fluorescent microscopy observations. Biofilms developed under static conditions displayed typical compact microcolonies with uniform distribution over the surface, while upon CFS challenge, biofilms were disrupted with presence of dead cells. These findings highlight the anti-biofilm potency of Lactobacillus spp. strains of goat milk origin and their potential application in food industries.