Induction of the Viable-but-Nonculturable State in Salmonella Contaminating Dried Fruit.

Salmonella can become viable but nonculturable (VBNC) in response to environmental stressors, but the induction of the VBNC state in Salmonella contaminating ready-to-eat dried fruit is poorly characterized. Dried apples, strawberries, and raisins were mixed with a five-strain cocktail of Salmonella at 4% volume per weight of dried fruit at 109 CFU/g. The inoculated dried fruit were then dried in desiccators at 25°C until the water activity (aw) approximated that of the uninoculated dried fruit. However, Salmonella could not be recovered after drying, not even after enrichment, suggesting a population reduction of approximately 8 log CFU/g. To assess the potential impact of storage temperature on survival, dried apples were spot-inoculated with the Salmonella cocktail, dried under ambient atmosphere at 25°C, and stored at 4 and 25°C. Spot inoculation permitted recovery of Salmonella on dried apple after drying, with the population of Salmonella decreasing progressively on dried apples stored at 25°C until it was undetectable after about 46 days, even following enrichment. The population decline was noticeably slower at 4°C, with Salmonella being detected until 82 days. However, fluorescence microscopy and laser scanning confocal microscopy with the LIVE/DEAD BacLight bacterial viability system at time points at which no Salmonella could be recovered on growth media even following enrichment showed that a large proportion (56 to 85%) of the Salmonella cells on the dried fruit were viable. The data suggest that the unique combination of stressors in dried fruit can induce large numbers of VBNC cells of Salmonella. IMPORTANCE Salmonella is a leading foodborne pathogen globally causing numerous outbreaks of foodborne illnesses and remains the leading contributor to deaths attributed to foodborne disease in the United States and other industrialized nations. Therefore, efficient detection methods for Salmonella contaminating food are critical for public health and food safety. Culture-based microbiological methods are considered the gold standard for the detection and enumeration of Salmonella in food. Findings from this study suggest that unique stressors on dried fruit can induce the VBNC state in Salmonella, thus rendering it undetectable with culture-based methods even though the bacteria remain viable. Therefore, strong consideration should be given to using, in addition to culture-based methods, microscopic and molecular methods for the accurate detection of all viable and/or culturable cells of Salmonella contaminating dried fruit, as all of these cells have the potential to cause human illness.

The synergic interaction between environmental factors (pH and NaCl) and the physiological state (vegetative cells and spores) provides new possibilities for optimizing processes to manage risk of C. sporogenes spoilage.

Clostridium sporogenes has been widely used as a surrogate for proteolytic C. botulinum for validating thermal processes in low-acid cans. To limit the intensity of heat treatments, industrials must use other ways of control as an association of acidic and saline environment after a low heat treatment. The probability of growth of pH (7-4.4), sodium chloride concentration (0-11%) and heat treatment (80°C-10 min; 100°C-1.5 min and 5.2 min) were studied on C. sporogenes PA 3679 spores and vegetative cells. Vegetative cells or heat-treated spores were inoculated in PYGm broth at 30 °C for 48 days in anaerobic conditions. Vegetative cells growth (pH 4.6-pH 4.5; 7%-8% NaCl) range is larger than the spore one (pH 5.2-pH 5.0; 6%-7% NaCl). Spores germination and outgrowth rage is decreased if the spores are heat-treated at 100 °C for 1.5 min (pH 5.5-5.3; 4%-5% NaCl) and 5.2 min (pH 5.7-5.3; 4%-5% NaCl). The C. sporogenes PA 3679 spores germination and outgrowth is impacted by their physiological state. The synergic interaction between environmental factors (pH and NaCl) and the physiological state (vegetative cells and spores) opening new possibilities for optimizing food formulation processes to manage the risks of C. sporogenes spoilage.

Assessment of the sanitary quality of ready to eat sesame, a low moisture street food from Burkina Faso.

BACKGROUND: Microbial contamination of edible low moisture food poses a significant public health risk for human. In this study, the microbial quality of sweet dehulled sesame seed croquettes, salted dehulled sesame seed and the raw sesame seed, sold under ambient conditions were examined. The samples were collected in the cities of Burkina Faso. The first type is sweet dehulled sesame seed croquettes (n1 = 25); the second type is salted dehulled sesame seed (n2 = 25) and the third type is raw sesame seed (n3 = 25). Assessment of the microbial quality was based on the total aerobic mesophilic bacteria, the thermotolerant coliforms, the yeasts and moulds, the E. coli, and the Salmonella spp. using ISO methods. RESULTS: The results showed the presence of microorganisms varying from <1.0 to 1.72 × 105 CFU g- 1 for thermotolerant coliforms, from <1.0 to 6,12 × 106 CFU g- 1 for the total mesophilic aerobic flora and from <1.0 to 8.10 × 105 CFU g- 1 for yeasts and moulds. The higher contaminations rates were mostly observed in raw sesame seed samples. No E coli or Salmonella pathogens were detected. Based on international standards of dehydrated food, 50.67% of the ready to eat sesame are satisficing while 17.33% are acceptable and 32% are not satisficing. CONCLUSION: Attention should be emphasized on the processing practices, especially in crowded places where RTE sesames seeds are mostly sold. The high numbers of all microbial groups in these sesame seed samples suggested that the production of RTE sesame seed should be improved by better hygiene. This study highlights also that RTE sesame seed might harbor a wide range of microorganisms when processes are weak of hygiene.

WGS based analysis of acquired antimicrobial resistance in human and non-human Acinetobacter baumannii isolates from a German perspective.

BACKGROUND: Acinetobacter baumannii ability to develop and acquire resistance makes it one of the most critical nosocomial pathogens globally. Whole-genome sequencing (WGS) was applied to identify the acquired or mutational variants of antimicrobial resistance (AMR) genes in 85 German A. baumannii strains utilizing Illumina technology. Additionally, the whole genome of 104 German isolates deposited in the NCBI database was investigated. RESULTS: In-silico analysis of WGS data revealed wide varieties of acquired AMR genes mediating resistance mostly to aminoglycosides, cephalosporins, carbapenems, sulfonamides, tetracyclines and macrolides. In the 189 analyzed genomes, the ant (3″)-IIa conferring resistance to aminoglycosides was the most frequent (55%), followed by blaADC.25 (38.6%) conferring resistance to cephalosporin, blaOXA-23 (29%) and the blaOXA-66 variant of the intrinsic blaOXA-51-likes (26.5%) conferring resistance to carbapenems, the sul2 (26%) conferring resistance to sulfonamides, the tet. B (19.5%) conferring resistance to tetracycline, and mph. E and msr. E (19%) conferring resistance to macrolides. blaTEM variants conferring resistance to cephalosporins were found in 12% of genomes. Thirteen variants of the intrinsic blaOXA-51 carbapenemase gene, blaOXA-510 and blaADC-25 genes were found in isolates obtained from dried milk samples. CONCLUSION: The presence of strains harboring acquired AMR genes in dried milk raises safety concerns and highlights the need for changes in producing dried milk. Acquired resistance genes and chromosomal gene mutation are successful routes for disseminating AMR determinants among A. baumannii. Identification of chromosomal and plasmid-encoded AMR in the genome of A. baumannii may help understand the mechanism behind the genetic mobilization and spread of AMR genes.

A PCR method to identify ochratoxin A-producing Aspergillus westerdijkiae strains on dried and aged foods.

Ochratoxins are a group of mycotoxins that frequently occur as contaminants in agricultural commodities and foods, including dry-cured meats and cheeses. The fungus Aspergillus westerdijkiae is frequently isolated from aged foods and can produce ochratoxin A (OTA). However, individual strains of the fungus can have one of two OTA production phenotypes (chemotypes): OTA production and OTA nonproduction. Monitoring and early detection of OTA-producing fungi in food are the most effective strategies to manage OTA contamination. Therefore, we examined genome sequence data from five A. westerdijkiae strains isolated from the surface of cheese from southern Italy to identify genetic markers indicative of the twoOTA chemotypes. This analysis revealed a naturally occurring deletion of the OTA regulatory gene, otaR, in an OTA-nonproducing isolate.We used this information to design a polymerase chain reaction (PCR) method that could identify A. westerdijkiae and distinguish between the two OTA chemotypes. In this method, the PCR primers were complementary to conserved sequences flanking otaR and yielded different-sized amplicons from strains with the different chemotypes. The primers did not yield ota-region-specific amplicons from other OTA-producing species. Because the method is specific to A. westerdijkiae and can distinguish between the two OTA chemotypes, it has potential to significantly improve OTA monitoring programs.

Competitiveness of three biocontrol candidates against ochratoxigenic Penicillium nordicum under dry-cured meat environmental and nutritional conditions.

The environmental conditions during the ripening of dry-cured meats and their nutritional composition promote the colonisation of their surface by Penicillium spp., including P. nordicum producer of ochratoxin A (OTA). The objective of this work was to study the competitiveness of three potential biocontrol candidates (Debaryomyces hansenii FHSCC 253H, Enterococcus faecium SE920 and Penicillium chrysogenum CECT, 20922) against the ochratoxigenic P. nordicum FHSCC4 under environmental and nutritional conditions simulating the ripening of dry-cured meat products. For this, the nutritional utilisation pattern, niche overlap index (NOI), interactions by dual-culture assays and OTA production were determined. The number of carbon sources (CSs) metabolised depended on the microorganism and the interacting water activity (aw) x temperature conditions. The number of CSs utilised by both filamentous fungi was quite similar and higher than those utilised by D. hansenii and E. faecium. The yeast isolate metabolised a number of CSs much larger than the bacterium. The NOI values showed that, in general, P. nordicum nutritionally dominated E. faecium and D. hansenii regardless of the environmental conditions evaluated. The relationship between the toxigenic and non-toxigenic fungal isolates depended on the aw x temperature combinations, although in none of the conditions a dominance of P. nordicum was observed. According to the interaction assays, both D. hansenii and P. chrysogenum decreased the growth of P. nordicum. The effect of D. hansenii could be attributed to the production of some extra-cellular compounds, while the action of P. chrysogenum is likely related to nutritional competition. In addition, both P. chrysogenum and D. hansenii reduced the OTA levels produced by P. nordicum. The effect of the yeast was more pronounced decreasing the concentration of OTA at quantities lower than the limit established by the Italian legislation. Therefore, P. chrysogenum and D. hansenii can be suggested as biocontrol candidates in the manufacture of dry-cured meat products.

Survival and thermal resistance of Salmonella in dry and hydrated nonfat dry milk and whole milk powder during extended storage.

Foodborne pathogens such as Salmonella can endure dry environments of milk powders for extended periods due to the increased adaptability at a low water activity (aw) and proliferate when powders are hydrated. This study compared the survivability and the thermal resistance of a 5-serovar Salmonella cocktail in dry and hydrated nonfat dry milk (NFDM) and whole milk powder (WMP) stored for 180 days at ambient temperature (~20 °C). This study was designed as two factorial (storage days and milk powder type) randomized complete block design with three replications as blocks. The milk powders were spray inoculated with 5-serovar Salmonella cocktail and dried back to the original pre-inoculation aw. The D-values of Salmonella in inoculated NFDM and WMP were determined periodically (every 30 days, starting from day one). The milk powders were also individually hydrated on each analysis day to determine D- and z-values of Salmonella in hydrated powders. The D-values were determined using thermal-death-time disks and hot-water baths at 80, 85 and 90 °C for milk powders, and 59, 62 and 65 °C for hydrated powders. The D- and z-values of Salmonella at specific temperatures within dry or hydrated powders during the storage period were compared at P ≤ 0.05 using two-way ANOVA and Tukey's Test. The D-values of Salmonella in WMP on day 1 were 18.9, 9.9 and 4.4 min at 80, 85 and 90 °C, respectively, which increased to 29.4, 13.6 and 6.5 min at 80, 85 and 90 °C, respectively, on day 180. Whereas, D-values of Salmonella in NFDM on day 1 were 17.9, 9.2 and 4.4 min at 80, 85 and 90 °C, respectively, and stayed similar during the storage. The D-values of Salmonella in milk powder remained similar throughout the storage once hydrated. The overall z-value of Salmonella in NFDM and WMP was 16.3 °C, whereas in hydrated NFDM and WMP, the overall z-value was 6.4 °C.

Distribution of Burkholderia cepacia complex species isolated from industrial processes and contaminated products in Argentina.

Burkholderia cepacia complex (Bcc) members have clinical relevance as opportunistic pathogens in patients with cystic fibrosis and are responsible of numerous nosocomial infections. These closely related bacteria are also reported as frequent contaminants of industrial products. In this retrospective study, we use PCR and recA gene sequence analysis to identify at species level Bcc isolates recovered from massive consumption products and industrial processes in Argentina during the last 25 years. The sequences obtained were also compared with recA sequences from clinical Bcc isolates deposited in GenBank database. We detected Bcc in purified water and preserved products from pharmaceutics, cosmetics, household cleaning articles, and beverages industries. B. contaminans (which is prevalent among people with cystic fibrosis in Argentina) was the most frequent Bcc species identified (42% of the Bcc isolates studied). B. cepacia (10%), B. cenocepacia (5%), B. vietnamiensis (16%), B. arboris (3%), and the recently defined B. aenigmatica (24%) were also detected. Rec A sequences from all B. cepacia and most B. contaminans industrial isolates obtained in this study displayed 100% identity with recA sequences from isolates infecting Argentinean patients. This information brings evidence for considering industrial massive consumption products as a potential source of Bcc infections. In addition, identification at species level in industrial microbiological laboratories is necessary for a better epidemiological surveillance. Particularly in Argentina, more studies are required in order to reveal the role of these products in the acquisition of B. contaminans infections.

Survival of Listeria monocytogenes during storage on dried apples, strawberries, and raisins at 4 °C and 23 °C.

The survival of Listeria monocytogenes was assessed during long-term storage on three dried fruits: dried apples, raisins and dried strawberries. Using sand as a carrier, the dried fruits were dry-inoculated with a four-strain cocktail of L. monocytogenes to achieve numbers of 4.0 to 4.6 log CFU/g. The inoculated foods were stored at 4 °C, 25-81% relative humidity (RH) and 23 °C, 30-35% RH for 336 days. Colonies of L. monocytogenes could not be recovered from the dried apples after inoculation, i.e., day 0. Concentrations of L. monocytogenes decreased rapidly on the raisins and dried strawberries during storage at 23 °C, with enhanced survival observed at 4 °C. Linear rates of decline for populations of L. monocytogenes during storage at 4 °C on the raisins and dried strawberries were 0.1 and 0.2 log CFU/g/month, respectively. The relative distribution of the four L. monocytogenes strains making up the cocktail was determined by multiplex PCR at the beginning of storage and after 336 days on the dried fruits. At day 0, L. monocytogenes populations were predominantly composed of the serotype 1/2a and 3a strains on both the raisins and dried strawberries. After long-term storage at 4 °C, a relative decrease in serotype 1/2a was observed on both fruits, coupled with relative increases in the serotype 3a strain during storage on both fruits, in addition to the serotype 1/2b strain on the raisins. These results demonstrate that L. monocytogenes is rapidly inactivated during storage on raisins and dried strawberries at 23 °C, but it is capable of long-term survival at 4 °C. Improved knowledge on the survival of L. monocytogenes on these commodities is important for predictive modeling and can be used to better inform microbial health risk assessments.

Applications of nonthermal plasma technology on safety and quality of dried food ingredients.

Cold plasma technology is an efficient, environmental-friendly, economic and noninvasive technology; and in recent years these advantages placed this novel technology at the centre of diverse studies for food industry applications. Dried food ingredients including spices, herbs, powders and seeds are an important part of the human diet; and the growing demands of consumers for higher quality and safe food products have led to increased research into alternative decontamination methods. Numerous studies have investigated the effect of nonthermal plasma on dried food ingredients for food safety and quality purposes. This review provides critical review on potential of cold plasma for disinfection of dried food surfaces (spices, herbs and seeds), improvement of functional and rheological properties of dried ingredients (powders, proteins and starches). The review further highlights the benefits of plasma treatment for enhancement of seeds performance and germination yield which could be applied in agricultural sector in near future. Different studies applying plasma technology for control of pathogens and spoilage micro-organisms and modification of food quality and germination of dried food products followed by benefits and current challenges are presented. However, more systemic research needs to be addressed for successful adoption of this technology in food industry.

Occurrence of Listeria and Escherichia coli in frozen fruit and vegetables collected from retail and catering premises in England 2018-2019.

Frozen vegetables have previously been associated with outbreaks of listeriosis in both the USA and Europe. An outbreak of Listeria monocytogenes serogroup 4 caused 53 cases in five European countries between 2015 and 2018. Whole genome sequencing (WGS) indicated that frozen sweet corn from a producer in Hungary was the source of illness. However, limited data is available on the prevalence of Listeria in frozen produce. A study of frozen fruit and vegetables from catering and retail premises in England was therefore carried out to assess their microbiological quality with respect to Listeria and Escherichia coli. Between December 2018 and April 2019, 1050 frozen fruit and vegetable samples were collected. Of these, 99% were of a satisfactory or borderline microbiological quality. Eleven samples (1%) contained ≥100 cfu/g of Escherichia coli (considered unsatisfactory in products labelled as ready-to-eat). Listeria monocytogenes or other Listeria species were detected in six samples (2%) of fruit compared to 167 samples (24%) of vegetables and six samples (26%) of fruit and vegetable mixes, but none at a level of ≥100 cfu/g. Characterisation by WGS of 74 L. monocytogenes isolates identified ten genetic clusters indicating a common source. For 8 of the 10 clusters, the isolates came from homogenous food types: four were sweet corn, and there was one cluster each for beans, peas, peppers and broccoli. There were five genetic associations between isolates from frozen vegetables and from clinical cases of listeriosis, including two cultures from frozen beans that were indistinguishable from the 2015-2018 sweet corn outbreak strain. This study indicates that L. monocytogenes was present in 10% of frozen vegetables and even though products are generally not ready-to-eat and are intended to be cooked prior to consumption, these have the potential to cause illness. Clear cooking and handling instructions are therefore required on these products to ensure that the health of consumers is not put at risk, and appropriate Good Manufacturing Practice measures should be followed by all fruit and vegetable freezing plants in order to reduce contamination with Listeria during processing.

Microbial and quality improvement of boiled gansi dish using carbon dots combined with radio frequency treatment.

Use of carbon dots (CDs) combined with radio frequency (RF) was applied to pasteurize and reduce the microorganism population in order to improve the quality of boiled gansi dish. CDs were prepared from banana using hydrothermal method, and characterized by using TEM, XRD and FTIR. The minimal inhibitory concentration (MIC) test showed CDs can efficiently inactivate Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) and Bacillus subtilis (B. subtilis). This study also evaluated the effectiveness of five treatments, including CDs alone, CDs combined RF (CDRF) heating for different time (8 min, 12 min, and 16 min), and high pressure steam (HPS) sterilization of boiled gansi dish inoculated with B. subtilis. After CDRF treated for 8 min, 12 min, and 16 min, the center temperature of samples reached to 78.92, 87.77 and 93.82 °C, and the colony forming units (CFU) of B. subtilis reduced by 2.13, 3.62, and 4.63 log, respectively. Samples with CDRF12 treatment, exhibited better product quality as evidenced by reduced loss of texture, flavor, and sensory as compared with HPS sample. The results indicated that CDRF treatment has a great potential to produce packaged boiled gansi dish with high product quality.

Mycobiota in Chilean chilli Capsicum annuum L. used for production of Merkén.

This work aims to provide the first study on the mycobiota present in Chilean pepper Capsicum annuum L. cv. "Cacho de Cabra" throughout the early production stages. Two hundred and forty berry fruits were sampled: 1) at the ripe fruits harvest day; 2) during drying; and 3) smoking processes. A total of 192 strains, encompassing 11 genera and 44 species, were identified through analysis of ß-tubulin (benA) gene and internal transcribed spacer of ribosomal DNA (ITS) region. All collection points showed samples with high fungal contamination, but the mycobiota composition varied as a result of different environmental conditions. Alternaria spp. and Fusarium spp. were predominantly isolated from fresh fruits of C. annuum. Penicillium spp. was the most frequent genus in all analysed points. Penicillium brevicompactum and P. crustosum were the most abundant species. Among Aspergillus, A. niger and A. flavus were dominant after the drying phase. In our study, none of the analysed strains of Penicillium (113) and Aspergillus (35) produced Ochratoxin A at detectable levels. The broad characterization of the fungal community of C. annuum carried out in this study, could be a guideline for future mycotoxin analyses performed directly on the pod. Understanding the role and dynamics of mycobiota and its relationship with the toxins present in this substrate, will be useful to establish and improve control measures considering the specificities of each point in the C. annuum production chain.

Rehydration of dry corn preserves the desirable bacterial community during ensiling.

This study evaluated the rehydration approach of mature corn grains as an alternative for high-moisture corn grain silage production in distinct corn hybrids, storage period, cultivation locations and kernel maturity at plant harvest. High-moisture corn was used as a control. The dry matter content and pH of the silage were measured, and the bacterial community associated with corn grains pre- and post-ensiling was also assessed through 16S rRNA high-throughput sequencing. The decrease in pH value was directly linked to an ecological microbial succession of Enterobacteriales and Actinomycetales to Lactobacillales in the silage at 120 days after storage, either in rehydrated or high-moisture corn. These results were similar for both maize production locations and hybrids tested. Finally, the similarity between the ensiling processes including rehydrated corn and the high-moisture corn grain silages proves the reliability of the rehydration approach as an alternative for the maintenance of a successful bacterial community structure and composition capable of producing high-quality silages from dent and flint corn hybrids in tropical conditions.

High prevalence of Clostridium botulinum in vegetarian sausages.

Clostridium botulinum is a significant food safety concern due to its ability to produce highly potent neurotoxin and resistant endospores. Vegetarian sausages have become a popular source of plant protein and alternative for meat products. While vegetarian sausages have not been linked to botulism, numerous outbreaks due to preserved vegetables suggest a frequent occurrence of C. botulinum spores in the raw material. The product formulation of vegetarian sausages involves limited NaCl and preservatives, and shelf-lives may be several months. The safety of vegetarian sausages thus relies mainly on heat treatment and chilled storage. The main food safety concern is C. botulinum Group II that can grow and produce toxin at refrigeration temperatures. Here we show a high overall prevalence (32%) of C. botulinum in 74 samples of vegetarian sausages from seven producers. Both Groups I and II strains and genes for neurotoxin types A, B, E and F were detected in the products. The highest cell counts (1200 spores/kg) were observed for C. botulinum Group II in products with remaining shelf-lives of 6 months at the time of purchase. We conclude that vacuum-packaged vegetarian sausage products frequently contain C. botulinum spores and may possess a high risk of C. botulinum growth and toxin production. Chilled storage below 3°C and thorough reheating before consumption are warranted.

Multi Mycotoxin Determination in Dried Beef Using Liquid Chromatography Coupled with Triple Quadrupole Mass Spectrometry (LC-MS/MS).

Dried beef meat, a locally processed meat from the cow, is vulnerable to contamination by mycotoxins, due to its exposure to the environmental microbiota during processing, drying, and point of sale. In this study, 108 dried beef samples were examined for the occurrence of 17 mycotoxins. Samples were extracted for mycotoxins using solid-liquid phase extraction method, while liquid chromatography coupled with triple quadrupole mass spectrometry (LC-MS/MS) via the dilute and shoot method was used to analyze the mycotoxins. Aflatoxin was found in 66% of the samples (average value of 23.56 µg/kg). AFB1 had a mean value of 105.4 µg/kg, AFB2 mean value of 6.92 µg/kg, and AFG1 and AFG2 had an average mean value of 40.49 µg/kg and 2.60 µg/kg, respectively. The total aflatoxins exceed the EU (4 µg/kg) permissible level in food. The α-Zea average mean value was 113.82 µg/kg for the various selling locations. Also, cyclopiazonic acid had an average mean value of 51.99 µg/kg, while some of the beef samples were contaminated with more than nine different mycotoxins. The occurrence of these mycotoxins in dried beef is an indication of possible exposure of its consumers to the dangers of mycotoxins that are usually associated with severe health problems. This result shows that there are mycotoxin residues in the beef sold in Ekiti State markets.

Assessment of Microbiological Quality and Mycotoxin in Dried Chili by Morphological Identification, Molecular Detection, and Chromatography Analysis.

The growing interest in spicy foods leads to the global demand for spices, particularly dried chili. This study aimed to assay both aflatoxin (AFs) and ochratoxin A (OTA) contamination using an integrative method of morphological identification, molecular detection, and chromatography analysis on dried chili provided from traditional and modern markets in Indonesia. The results showed that total fungal infection ranged from 1-408 × 103 CFU/g. Eighty percent of the chili obtained from both the traditional and the modern markets were infected by Aspergillus spp., in which 50% of the infections were identified as A. parasiticus and A. flavus. A complete set of targeted genes involved in AF production and OTA were detected in two isolates of A. flavus and one isolate of A. carbonarius, respectively. The levels of AFs B1, B2, and OTA in the contaminated dried chilies were in the range of 39.3-139.5 µg/kg, 2.6-33.3 µg/kg, and 23.7-84.6 µg/kg, respectively. In contrast, no AFs G1 and G2 were detected. This study showed that the fungal infection of Indonesian dried chili occurs both in the field and during storage; thus, it is suggested to implement good agricultural and handling processes.

Aspergillus, Penicillium and Cladosporium species associated with dried date fruits collected in the Perugia (Umbria, Central Italy) market.

A total of 20 dried date samples, chosen as representative among those available on the Perugia (Umbria, Central Italy) market, were analyzed for the possible occurrence of fungal species and related contamination by fungal secondary metabolites. Twenty-six isolates, representative of the total mycobiota, were obtained and morphologically identified as belonging to the genera Aspergillus, Penicillium and Cladosporium. Inside each genus, molecular characterization (by partial sequencing of ITS region and/or ß-tubulin and calmodulin regions for Aspergillus and Penicillium isolates or actin region for Cladosporium isolates) and in vitro mycotoxigenic profile characterization (by LC-MS/MS analysis) showed the presence of the following species: A. flavus, A. tubingensis, P. brevicompactum, P. chrysogenum, P. crustosum, P. glabrum, P. solitum, P. venetum, C. cladosporioides, C. limoniforme and C. halotolerans, with A. tubingensis as the prevalent species and P. crustosum, P. solitum, P. venetum and C. limoniforme first reported here on dates. Date packaging and format showed an effect on the incidence of isolated fungi, with the lowest incidence recovered from whole dates and in hermetic bag packaging. These findings can be useful both for dried dates producers and consumers, guiding them towards choices of packaging and format with a lower risk of mycotoxigenic species presence. However, no fungal metabolites were detected in the dried date samples analyzed, which were therefore regarded as safe for human consumption, underlining the absence of correspondence between fungal isolation and mycotoxin contaminations.

Lactobacillus hegangensis sp. nov., Lactobacillus suibinensis sp. nov., Lactobacillus daqingensis sp. nov., Lactobacillus yichunensis sp. nov., Lactobacillus mulanensis sp. nov., Lactobacillus achengensis sp. nov., Lactobacillus wuchangensis sp. nov., Lactobacillus gannanensis sp. nov., Lactobacillus binensis sp. nov. and Lactobacillus angrenensis sp. nov., isolated from Chinese traditional pickle and yogurt.

Fourteen Gram-stain-positive bacterial strains were isolated from Chinese traditional pickle and yogurt. The strains were characterized using a polyphasic taxonomic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, fatty acid methyl ester analysis, determination of DNA G+C content, determination of average nucleotide identity (ANI), in silico DNA-DNA hybridization (isDDH) and an analysis of phenotypic features. The data demonstrated that the 14 strains represented ten novel species belonging to the genus Lactobacillus, strains 73-4T, 247-3T, 143-4(a)T, 33-1T, 143-6T, 247-4T, 17-4T, 143-1T, 735-2T and M1530-1T were designated as the type strains. Strains 73-4T and 247-3T were phylogenetically related to the type strains of Lactobacillus camelliae and Lactobacillus jixianensis, having 97.0-98.9 % 16S rRNA gene sequence similarities, 83.9-87.2 % pheS gene sequence similarities and 86.8-93.3 % rpoA gene sequence similarities. Strains 143-4(a)T and 33-1T were phylogenetically related to the type strains of Lactobacillus rhamnosus, Lactobacillus paracasei and Lactobacillus casei, having 93.6-96.5 % 16S rRNA gene sequence similarities, 73.9-77.2 % pheS gene sequence similarities and 76.1-77.6 % rpoA gene sequence similarities. Strains 143-6T, 247-4T, 17-4T and 143-1T were phylogenetically related to the type strains of Lactobacillus concavus, Lactobacillus dextrinicus and Lactobacillus bayanensis, exhibiting 95.5-99.9 % 16S rRNA gene sequence similarities, 76.5-83.1 % pheS gene sequence similarities and 83.6-98.3 % rpoA gene sequence similarities. Strain 735-2T was phylogenetically related to the type strains of Lactobacillus zhaoyuanensis, Lactobacillus jiayinensis and Lactobacillus coryniformis, having 98.2-99.1 % 16S rRNA gene sequence similarities, 82.8-84.1 % pheS gene sequence similarities and 93.0-93.9 % rpoA gene sequence similarities. Strain M1530-1T was phylogenetically related to the type strains of Lactobacillus suantsaiihabitans and Lactobacillus brevis, having 99.5 and 99.0 % 16S rRNA gene sequence similarities, 90.3 and 81.7 % pheS gene sequence similarities and 97.7 and 91.1 % rpoA gene sequence similarities. The ANI and isDDH values between strains 73-4T, 247-3T, 143-4(a)T, 33-1T, 143-6T, 247-4T, 17-4T, 143-1T, 735-2T, M1530-1T and type strains of phylogenetically related species were less than 86.8 % and 33.9 % respectively, confirming that they represent ten novel species within the genus Lactobacillus. Based upon the data of polyphasic characterization obtained in the present study, ten novel species, Lactobacillus hegangensis sp. nov., Lactobacillus suibinensis sp. nov., Lactobacillus daqingensis sp. nov., Lactobacillus yichunensis sp. nov., Lactobacillus mulanensis sp. nov., Lactobacillus achengensis sp. nov., Lactobacillus wuchangensis sp. nov., Lactobacillus gannanensis sp. nov., Lactobacillus binensis sp. nov. and Lactobacillus angrenensis sp. nov., are proposed and the type strains are 73-4T (=NCIMB 15177T=CCM 8912T=CCTCC AB 2018407T), 247-3T (=NCIMB 15176T=JCM 33275T), 143-4(a)T (=NCIMB 15173T=CCM 8948T=JCM 33273T=CCTCC AB 2018390T), 33-1T (=NCIMB 15169T=CCM 8947T=JCM 33272T=CCTCC AB 2018405T), 143-6T (=NCIMB 15162T=CCM 8951T=JCM 33274T=CCTCC AB 2018411T), 247-4T (=NCIMB 15155T=CCM 8897T=LMG 31059T=CCTCC AB 2018410T), 17-4T (=NCIMB 15161T=CCM 8946T=JCM 33271T=CCTCC AB 2018406T), 143-1T (=NCIMB 15157T=CCM 8937T=CCTCC AB 2018409T), 735-2T (=NCIMB 15190T=CCM 8925T=LMG 31186T) and M1530-1T (=NCIMB 15150T=CCM 8893T=LMG 31046T=CCTCC AB 2018402T), respectively.

Lethality of high-pressure carbon dioxide on Shiga toxin-producing Escherichia coli, Salmonella and surrogate organisms on beef jerky.

Low water activity (aW) foods permit the survival of low-infectious dose pathogens including Escherichia coli and Salmonella. Desiccation of non-heat resistant E. coli and Salmonella enterica increases their heat resistance; therefore, alternative methods are necessary to ensure the safety of low aW foods. High-pressure carbon dioxide (HPCD) reduced microbial contaminants in high aW foods. This study aimed to identify HPCD conditions that reduce pathogenic E. coli and Salmonella in low aW conditions. Four strains of Shiga toxin-producing E. coli (STEC) and one strain of enteropathogenic E. coli were treated as a cocktail, and five strains of Salmonella were treated individually. The suitability of E. coli AW1.7, Pediococcus acidilactici FUA 3072, Enterococcus faecium NRRL B-2354 and Staphylococcus carnosus R6 FUA 2133 as surrogate organisms was evaluated. Treatments were validated in beef jerky. Samples were equilibrated to aW 0.75 and treated with heat, HPCD or pressurized N2. Treatment of desiccated E. coli AW1.7 and the STEC cocktail with dry gaseous CO2 (5.7 MPa and 65 °C) did not reduce cell counts; however, treatment with gaseous CO2 saturated with water reduced cell counts of all strains of E. coli. Treatment of beef jerky inoculated with E. coli and Salmonella with saturated gaseous CO2 resulted in >5-log reductions for all strains. E. faecium NRRL B-2354 and S. carnosus R6 were suitable surrogates for Salmonella on beef jerky treated with HPCD. Treatment of beef jerky with water-saturated gaseous CO2 was more effective than treatment with supercritical CO2 or treatments with N2 at the same temperature and pressure. Overall, the treatment of low aW foods with water-saturated gaseous HPCD can meet industry standards by achieving a >5-log reductions of E. coli and Salmonella. Additionally, surrogate organisms representing pathogenic E. coli and Salmonella have been validated.