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This study presents comprehensive insights into the microbiological profile across all concentrated chicken broth processing stages, utilizing a combination of amplicon sequencing based on metataxonomic and culturing techniques. Samples were systematically collected throughout the production chain, with each batch yielding 10 samples per day across eight different dates. These samples underwent thorough analysis, including 16S rRNA and ITS sequencing (n = 30), culture-dependent microbiological tests (n = 40), and physical-chemical characterization (n = 10). Culturing analysis revealed the absence of Listeria monocytogenes and Salmonella spp. at any stage of processing, counts of various microorganisms such as molds, yeasts, Enterobacteria, and others remained below detection limits. Notably, spore counts of selected bacterial groups were observed post-processing, indicating the persistence of certain species, including Bacillus cereus and Clostridium perfringens, albeit in low counts. Furthermore, the study identified a diverse array of bacterial and fungal species throughout the processing chain, with notable occurrence of spore-forming bacteria. The presence of spore-forming bacteria in the final product, despite thermal processing, suggests the need for enhanced strategies to mitigate their introduction and persistence in the processing premises. Thus, this study offers valuable insights into microbial dynamics and diversity through processing concentrated chicken broth., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Gelatin, a versatile protein derived from collagen, is widely used in the food, pharmaceutical and medical sectors. However, bacterial contamination by spore-forming bacteria during gelatin processing represents a significant concern for product safety and quality. In this study, an investigation was carried out to explore the heat and chemical resistance, as well as the identification and characterization of spore-forming bacteria isolated from gelatin processing. The methodologies involved chemical resistance tests with drastic pH in microplates and thermal resistance tests in capillary tubes of various isolates obtained at different processing stages. In addition, phenotypic and genotypic analyses were carried out to characterize the most resistant isolates of spore-forming bacteria. The findings of this study revealed the presence of several species, including Bacillus cereus, Bacillus licheniformis, Bacillus sonorensis, Bacillus subtilis, Geobacillus stearothermophilus, and Clostridium sporogenes, with some isolates exhibiting remarkable chemical and heat resistances. In addition, a significant proportion of the most resistant isolates showed gelatinase activity (n = 19/21; 90.5 %) and the presence of heat resistance (n = 5/21; 23.8 %), and virulence genes (n = 11/21; 52.4 %). The results of this study suggest that interventions should be done in quality control practices and that process parameter adjustments and effective contamination reduction strategies should be implemented through gelatin processing., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Cronobacter sakazakii is a potentially pathogenic bacterium that is resistant to osmotic stress and low a w , and capable of persisting in a desiccated state in powdered infant milks. It is widespread in the environment and present in various products. Despite the low incidence of cases, its high mortality rates of 40 to 80 % amongst neonates make it a microorganism of public health interest. This current study performed a comparative assessment between current reduction methods applied for C. sakazakii in various food matrices, indicating tendencies and relevant parameters for process optimization. A systematic review and meta-analysis were conducted, qualitatively identifying the main methods of inactivation and control, and quantitatively evaluating the effect of treatment factors on the reduction response. Hierarchical clustering dendrograms led to conclusions on the efficiency of each treatment. Review of recent research trend identified a focus on the potential use of alternative treatments, with most studies related to non-thermal methods and dairy products. Using random-effects meta-analysis, a summary effect-size of 4-log was estimated; however, thermal methods and treatments on dairy matrices displayed wider dispersions - of τ 2  = 8.1, compared with τ 2  = 4.5 for vegetal matrices and τ 2  = 4.0 for biofilms. Meta-analytical models indicated that factors such as chemical concentration, energy applied, and treatment time had a more significant impact on reduction than the increase in temperature. Non-thermal treatments, synergically associated with heat, and treatments on dairy matrices were found to be the most efficient., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Despite the wide variety of native and exotic fruits in Brazil, there is limited understanding of their ability to support pathogens during storage. This study aimed to evaluate the behavior of Salmonella enterica and Listeria monocytogenes inoculated into the pulp of eight fruits native and exotic to Brazil: Jenipapo (Genipa americana L.), Umbu (Spondias tuberosa Arruda), Maná (Solanum sessiliflorum), Cajá-manga (Spondias dulcis), Physalis (Physalis angulata L.), Feijoa (Acca sellowiana), Cupuaçu (Theobroma grandiflorum) (average pH < 3.3) and in a low acidy fruit: Abiu (Pouteria caimito) (pH 6.11). The pathogens were inoculated into the different fruits and stored at 10, 20, 30 and 37 °C for up to 12 h and 6 days, respectively. Among the fruits evaluated, Abiu was the only one that allowed Salmonella growth, showing higher δ-values at 20 and 30 °C (5.6 log CFU/g for both temperatures). For Physalis and Feijoa, there was a small reduction in the pathogen concentration (<1 log-cycle), mainly at 10 and 20 °C, indicating its ability to remain in the matrices. For the other fruits, notable negative δ-values were obtained, indicating a tendency towards microbial inactivation. The survival potential was significantly affected by temperature in Abiu, Maná, Cupuaçu, and Cajá-manga (p < 0.05). The same phenomena regarding δ-value were observed for L. monocytogenes population, with the greatest survival potential observed at 20 °C in Abiu (3.3 log CFU/g). Regarding the exponential growth rates in Abiu, the highest values were observed at 30 and 37 °C, both for Salmonella (4.6 and 4.9 log (CFU/g)/day, respectively) and for L. monocytogenes (2.8 and 2.7 log (CFU/g)/day, respectively), with no significant difference between both temperatures. Regarding microbial inactivation, L. monocytogenes showed greater resistance than Salmonella in practically all matrices. Jenipapo and Umbu were the pulps that, in general, had the greatest effect on reducing the population of pathogens. Furthermore, the increase in storage temperature seems to favor the increase on inactivation rates. In conclusion, Salmonella and L. monocytogenes can grow only in Abiu pulp, although they can survive in some acidic tropical fruits kept at refrigeration and abusive temperatures., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Alicyclobacillus spp. is the cause of great concern for the food industry due to their spores' resistance (thermal and chemical) and the spoilage potential of some species. Despite this, not all Alicyclobacillus strains can spoil fruit juices. Thus, this study aimed to identify Alicyclobacillus spp. strains isolated from fruit-based products produced in Argentina, Brazil, and Italy by DNA sequencing. All Alicyclobacillus isolates were tested for guaiacol production by the peroxidase method. Positive strains for guaiacol production were individually inoculated at concentration of 10 3  CFU/mL in 10 mL of orange (pH 3.90) and apple (pH 3.50) juices adjusted to 11°Brix, following incubation at 45 °C for at least 5 days to induce the production of the following spoilage compounds: Guaiacol, 2,6-dichlorophenol (2,6-DCP) and 2,6-dibromophenol (2,6-DBP). The techniques of micro-solid phase extraction by headspace (HS-SPME) and gas-chromatography with mass spectrometry (GC-MS) were used to identify and quantify the spoilage compounds. All GC-MS data was analyzed by principal component analysis (PCA). The effects of different thermal shock conditions on the recovery of Alicyclobacillus spores inoculated in orange and apple juice (11°Brix) were also tested. A total of 484 strains were isolated from 48 brands, and the species A. acidocaldarius and A. acidoterrestris were the most found among all samples analyzed. In some samples from Argentina, the species A. vulcanalis and A. mali were also identified. The incidence of these two main species of Alicyclobacillus in this study was mainly in products from pear (n = 108; 22.3 %), peach (n = 99; 20.5 %), apple (n = 86; 17.8 %), and tomato (n = 63; 13 %). The results indicated that from the total isolates from Argentina (n = 414), Brazil (n = 54) and Italy (n = 16) were able to produce guaiacol: 107 (25.8 %), 33 (61.1 %) and 13 (81.2 %) isolates from each country, respectively. The PCA score plot indicated that the Argentina and Brazil isolates correlate with higher production of guaiacol and 2,6-DCP/2,6-DBP, respectively. Heatmaps of cell survival after heat shock demonstrated that strains with different levels of guaiacol production present different resistances according to spoilage ability. None of the Alicyclobacillus isolates survived heat shocks at 120 °C for 3 min. This work provides insights into the incidence, spoilage potential, and thermal shock resistance of Alicyclobacillus strains isolated from fruit-based products., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study aimed to assess the growth of Pseudomonas spp. and psychrotrophic bacteria in chilled Pacu (Piaractus mesopotamicus), a native South American fish, stored under chilling conditions (0 to 10 °C) through the use of predictive models under isothermal and non-isothermal conditions. Growth kinetic parameters, maximum growth rate (μ max, 1/h), lag time (t Lag, h), and (N max, Log 10 CFU/g) were estimated using the Baranyi and Roberts microbial growth model. Both kinetic parameters, growth rate and lag time, were significantly influenced by temperature (P < 0.05). The square root secondary model was used to describe the bacteria growth as a function of temperature. Secondary models, √μ = 0.016 (T + 10.13) and √μ =0.017 (T + 9.91) presented a linear correlation with R 2 values >0.97 and were further validated under non-isothermal conditions. The model's performance was considered acceptable to predict the growth of Pseudomonas spp. and psychrotrophic bacteria in refrigerated Pacu fillets with bias and accuracy factors between 1.24 and 1.49 (fail-safe) and 1.45-1.49, respectively. Fish biomarkers and spoilage indicators were assessed during storage at 0, 4, and 10 °C. Volatile organic compounds, VOCs (1-hexanol, nonanal, octenol, and indicators 2-ethyl-1-hexanol) showed different behavior with storage time (P > 0.05). 1 H NMR analysis confirmed increased enzymatic and microbial activity in Pacu fillets stored at 10 °C compared to 0 °C. The developed and validated models obtained in this study can be used as a tool for decision-making on the shelf-life and quality of refrigerated Pacu fillets stored under dynamic conditions from 0 to 10 °C., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Fungi are the main microorganisms responsible for the spoilage of bakery products, and their control and subsequent reduction of food waste are significant concerns in the agri-food industry. Synthetic preservatives are still the most used compounds to reduce bakery product spoilage. On the other hand, studies have shown that biopreservation can be an attractive approach to overcoming food and feed spoilage and increasing their shelf-life. However, limited studies show the preservation effects on real food matrices. Therefore, this study aimed to investigate the influence of microorganisms such as lactic acid bacteria (LAB) and yeasts on the growth of spoilage filamentous fungi (molds) on bread and panettones. In general, on conventional and multigrain bread, treatments containing Limosilactobacillus fermentum IAL 4541 and Wickerhamomyces anomalus IAL 4533 showed similar results when compared to the negative control (calcium propionate) in delaying the fungal growth of the tested species (Aspergillus chevalieri, Aspergillus montevidensis, and Penicillium roqueforti). Different from bread, treatments with W. anomallus in panettones delayed the A. chevalieri growth up to 30 days, 13 days longer than observed on negative control (without preservatives). This study showed that biopreservation is a promising method that can extend bakery products' shelf-life and be used as an alternative to synthetic preservatives., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study aimed to estimate the risk of listeriosis from the consumption of pasteurized milk in Brazil, comparing conventional treatment with the technology of thermosonication. The Quantitative Microbiological Risk Assessment (QMRA) model was developed, covering the entire milk production chain, from milking to the moment of consumption. In general, higher risks were observed in association with higher initial concentrations of the pathogen and the vulnerable population. The highest risk predicted (3.67 × 10 -5 ) was related to the scenario considering the initial concentration range of L. monocytogenes between 4 and 6 log CFU/mL, with conventional treatment and considering the vulnerable population, resulting in one case of listeriosis every 27,248 servings. When considering thermosonication treatment, lower risks have been predicted. The scenario analysis indicated that the steps related to storage conditions in retail and at the consumer's home (post-processing steps) are the most influential in the associated risk, in all scenarios. The predictive parameters of inactivation related to the applied treatment also have a considerable influence on the risk. The results point to the influence of the stages of the dairy production chain and the thermosonication treatment applied in the food safety of milk, subsidizing information for industrial application and for regulatory agencies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

This study compared the resistance to different desiccation conditions of 190 Salmonella enterica strains previously isolated from the soybean meal production chain and belonging to 23 serovars. Additionally, the post-rehydration growth and heat tolerance of the strains previously exposed to desiccation were determined. Variability in desiccation resistance was observed both within and between serovars. Strains belonging to S. Havana and S. Schwarzengrund serovars were the most resistant, regardless of storage condition. The drying temperature (20 °C and 30 °C) did not influence the desiccation resistance of the Salmonella strains. On the other hand, increasing drying time from 1 to 7 days reduced Salmonella counts. The origin (isolation sources) also influenced the desiccation resistance of the Salmonella strains. The growth of the Salmonella strains after rehydration varied considerably depending on the drying conditions and incubation temperature during cultivation. An increase in the time and temperature of drying led to a reduction in population of most Salmonella strains after rehydration. Salmonella strains previously desiccated also showed differences in the heat tolerance in all temperature-time binomials tested. Some strains were highly resistant to heat tolerance conditions, presenting <1 log CFU/mL reduction from the initial population. The results obtained in this study suggest that the strategies to mitigate Salmonella in low-a w foods must consider the existence of high-stress resistant strains and their multiple-stress adaptability profiles, including effects of processing, food composition, and storage conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

This study aimed to determine Salmonella enterica occurrence along the soybean meal production chain (raw material, in-processing samples, final products, and in the environment of five processing plants), characterize the isolates, and assess the survival of Salmonella Senftenberg 775W in soybeans stored under different temperature conditions. Among 713 samples analyzed, 12.9% (n = 92) were positive for Salmonella enterica. Dust collected inside and outside processing plants (n = 148) comprised the samples with the highest positivity for Salmonella enterica, 47.3%. The occurrence of Salmonella enterica varied among the different processing plants. Twenty-nine (n = 29) Salmonella serotypes were isolated, with S. Mbandaka as the most frequent serotype, whereas S. Typhimurium was mainly linked to final product samples (soybean meal). S. Senftenberg 775W did not survive for a long time in soybean stored at 20-37 °C, but at 20 °C, cells were viable for more than 60 days. This study suggests that soybean meal may harbor Salmonella serotypes related to foodborne disease outbreaks in humans and can be responsible for Salmonella introduction into livestock and, consequently, in foods of animal origin. This study provides crucial data on contamination pathways of Salmonella in the soybean production chain, contributing to the understanding of Salmonella epidemiology which is strategic for the development of preventive and control measures to reduce the burden of salmonellosis linked to products of animal origin., (Copyright © 2022 Elsevier B.V. All rights reserved.)

This study aimed to assess the counts and biodiversity characterization of aerobic sporeforming bacteria (ASB) in powdered infant formula (PIF). Fifty-four (n = 54) samples of PIF of three brands were analyzed for mesophilic aerobic bacteria, and ASB counts. ASB isolated from PIF were assessed for their ability to produce spoilage enzymes and hemolytic activity and further identified by MALDI-TOF mass spectrometry. Then, the isolates were subjected to rpoB gene typing and assessment of bceT, entFM, nhe (A, B, C), and hbl (A, B, C) toxin genes. The main species isolated were B. licheniformis (54%), followed by B. cereus (33%) and B. subtilis (5%). The ASB counts ranged from 1 to 4 log CFU/g, and the mean was 2.9 log CFU/g for mesophilic aerobic sporeforming bacteria (MSC) and 2.5 log CFU/g for thermophilic aerobic sporeforming bacteria (TSC). Most PIF samples presented MSC and TSC counts between 2 and 3 log CFU/g. A total of 13%, 50%, and 37% of the samples presented MSC counts from <2 log CFU/g, between 2 and 3 log CFU/g and between 3 and 4 log CFU/g, respectively. Among the ASB isolates, 97% had protease, 84% hydrolyzed starch, 66% had hemolytic activity, and 61% had lecithinase activity. A total of 44 out of 120 isolates harbored at least one toxin gene; 56% for B. cereus, 34% for B licheniformis, and less than 5% for B. subtilis, B pumilus, and L. sphaericus. All B. cereus isolates harbored the nhe gene, 60% entFM, 44% cytK, 32% bceT, and 28% hbl genes. Besides, 17% of B. licheniformis harbored nhe. A small proportion of B. subtilis, B. pumilus, and L. sphaericus carried toxin genes. The rpoB based phylogenetic tree provided high resolution among Bacillus species. The findings of this study provide insights into the phenotypic and genotypic biodiversity of Bacillus present in PIF., (Copyright © 2022 Elsevier B.V. All rights reserved.)

This study aimed to evaluate the occurrence and diversity of yeasts in frozen concentrated orange juice (FCOJ) and assess the resistance of yeasts to peracetic acid. One thousand five hundred (n = 1500) samples of frozen concentrated orange juice (FCOJ) were analyzed, and 280 yeast strains were isolated and identified. Candida represented 37% of all isolates, and the main species identified were Candida pseudointermedia and C. orthopsilosis. Other yeasts identified were Starmerella, Wickerhamiella, Wickerhamiella, Clavispora, Kodamaea, Meyerozyma, Rhodotorula, Trichosporon, Wickerhamomyces, Kluyveromyces, Hanseniaspora, Saccharomyces, Torulaspora, and Zygosaccharomyces. The exogenous origin of the contamination in FCOJ samples analyzed was shown by the high diversity, corroborated by the Simpson (D) and Shannon (H') indices. From a total of 227 yeasts strains tested, more than 20% were able to withstand peracetic acid concentrations >200 ppm, with emphasis on W. anomalus (300 ppm), W. sergipensis (350 ppm), C. rugopelliculosa (350 ppm), K. marxianus (450 ppm), C. parapsilosis (500 ppm), C. pseudointermedia (500 ppm), W. sorbosivorans (500 ppm), C. boleticola (600 ppm), S. cerevisiae (700 ppm) and C. orthopsilosis (750 ppm). This study adds novel data regarding the occurrence and diversity of yeasts present in FCOJ and their resistance to a chemical compound commonly employed in the sanitization of processing and distribution premises and vehicles. These findings are essential to support the development of measures for proper mitigation of contamination of orange juice towards reducing the risks of spoilage by yeasts during FCOJ transportation/storage or when FCOJ is used as an ingredient., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The survival kinetics of Listeria monocytogenes (9 log CFU/mL) as a post-fermentation contaminant in probiotic fermented milk (Lactobacillus acidophilus La-5, 8-9 log CFU/mL) processed with milk subjected to ohmic heating (0, 4, 6, and 8 V/cm; CONV, OH 4 , OH 6 , OH 8 , 90-95 °C/5 min) was investigated using Weibull predictive model. Additionally, the presence of bioactive compounds (antioxidant activity, inhibition of the enzymes α-glucosidase, α-amylase, and angiotensin-converting) and sensory analysis (consumer test) of probiotic fermented milks were evaluated. Overall, OH provided a decrease in the viability of Listeria monocytogenes, suitable Lactobacillus acidophilus counts, and satisfactory results in the gastrointestinal tract survival. The Weibull model presented an excellent fit to the data of all conditions. Furthermore, lower δ values (217-298 against 665 h, CONV), and increased R 2 values (0.99 against 0.98, CONV) were obtained for the OH-treated samples, emphasizing the best performance of OH data. In addition, OH improved the generation of bioactive compounds as well as the sensory acceptance. Indeed, considering functional and safety purposes, OH presented as an interesting technology to be used in milk for manufacturing probiotic fermented milk., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Meat and meat products are perishable products that require the use additives to prevent the spoilage by foodborne microorganisms and pathogenic bacteria. Current trends for products without synthetic preservatives have led to the search for new sources of antimicrobial compounds. Essential oils (EOs), which has been used since ancient times, meet these goals since their effectiveness as antimicrobial agents in meat and meat products have been demonstrated. Cinnamon, clove, coriander, oregano, rosemary, sage, thyme, among others, have shown a greater potential to control and inhibit the growth of microorganisms. Although EOs are natural products, their quality must be evaluated before being used, allowing to grant the Generally Recognized as Safe (GRAS) classification. The bioactive compounds (BAC) present in their composition are linked to their activity, being the concentration and the quality of these compounds very important characteristics. Therefore, a single mechanism of action cannot be attributed to them. Extraction technique plays an important role, which has led to improve conventional techniques in favour of green emerging technologies that allow to preserve better target bioactive components, operating at lower temperatures and avoiding as much as possible the use of solvents, with more sustainable processing and reduced energy use and environmental pollution. Once extracted, these compounds display greater inhibition of gram-positive than gram-negative bacteria. Membrane disruption is the main mechanism of action involved. Their intense characteristics and the possible interaction with meat components make that their application combined with other EOs, encapsulated and being part of active film, increase their bioactivity without modifying the quality of the final product., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The occurrence of black aspergilli in onions has been reported as frequent, and this group of fungi harbors potentially toxigenic species. In addition, Aspergillus niger has been reported as the causative agent of black mold rot, an important postharvest disease that causes damage throughout the world. Brazil stands out as one of the world's largest onion producers. However, few studies have been conducted to investigate the mycobiota in Brazilian onions. For this reason, we investigated the mycobiota of 48 market (n = 25) and field (n = 23) onion bulb samples. Nineteen soil samples were collected from the same fields and evaluated. In field onions and soil samples, Penicillium spp. was the prevalent fungal group, whereas in market samples A. section Nigri was the most frequent group. Due to the taxonomic complexity of this group, species identification was supported by phylogenetic data (CaM gene). A. welwitschiae was the most prevalent species in market samples. Black aspergillus strains were evaluated for fumonisin B 2 (FB 2 ) and ochratoxin A (OTA) production. Overall, 53% and 2.2% of the strains produced FB 2 and OTA, respectively. The occurrence of FB 2 and OTA was also investigated in onion bulb samples but none showed contamination with these mycotoxins., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The management of Heat Resistant Moulds (HRMs) is considered a great challenge for the juice fruit industry. Neosartorya, Byssochlamys and Talaromyces are three out of the main genera isolated from fruit juices that show great resistance to heat treatments. Several inactivation parameters can be found in the literature, however all of them were carried out in specific food matrices and using diverse inactivation methods. Thus, this meta-analysis study synthesizes the thermal resistance parameters of the three HRMs by adjusting extended Bigelow-based meta-regression models to data on inactivation experiments conducted in different liquid media. The meta-analytical data, extracted from publications between 1969 and 2017, was composed of decimal reduction time (D), inactivation method, temperature of inactivation, pH, °Brix, age of spores, and type of medium (model, juice, concentrates). Pooled D* values (D at 90 °C, pH 3.5 and 12° Brix) were estimated for B. fulva (1.95 min; 95% CI: 1.21-3.11 min), Talaromyces (4.03 min; 95% CI: 3.43-4.74 min), Neosartorya (0.5.35 min; 95% CI: 4.10-7.08 min), and B. nivea (10.32 min; 95% CI: 5.81-18.4 min). It was found that increasing the soluble solids in concentrates tends to cause a lower decrease in the heat resistance of Neosartorya and Talaromyces than increasing the soluble solids in model liquid or juices (p = 0.001; 0.012). In general, the screw-capped tubes and three neck round inactivation methods render higher D* values (p < 0.05) than the thermal death tubes, the polyethylene bag and the capillary methods. Spores of Talaromyces (overall z pH  = 7.56; 95% CI: 5.13-13.5) and Neosartorya (overall z pH  = 7.07; 95% CI: 5.04-10.8) appear to be more thermal sensitive to a decrease in medium pH than spores of Byssochlamys (overall z pH  = 4.34; 95% CI: 3.20-6.73). The meta-regression models presented in this study can be valuable for estimating pooled inactivation kinetic parameters to be used by the fruit juice industry in the management of thermal processes and in the determination of shelf-life., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Alicyclobacillus acidoterrestris is a cause of major concern for the orange juice industry due to its thermal and chemical resistance, as well as its spoilage potential. A. acidoterrestris spoilage of orange juice is due to off-flavor taints from guaiacol production and some halophenols. The present study aimed to evaluate the effectiveness of antimicrobial Photodynamic Treatment (aPDT) as an emerging technology to inactivate the spores of A. acidoterrestris. The aPDT efficiency towards A. acidoterrestris was evaluated using as photosensitizers the tetracationic porphyrin (Tetra-Py + -Me) and the phenothiazinium dye new methylene blue (NMB) in combination with white light-emitting diode (LED; 400-740 nm; 65-140 mW/cm 2 ). The spores of A. acidoterrestris were cultured on YSG agar plates (pH 3.7 ± 0.1) at 45 °C for 28 days and submitted to the aPDT with Tetra-Py + -Me and NMB at 10 μM in phosphate-buffered saline (PBS) in combination with white light (140 mW/cm 2 ). The use of Tetra-Py + -Me at 10 μM resulted in a 7.3 ± 0.04 log reduction of the viability of A. acidoterrestris spores. No reductions in the viability of this bacterium were observed with NMB at 10 μM. Then, the aPDT with Tetra-Py + -Me and NMB at 10 μM in orange juice (UHT; pH 3.9; 11°Brix) alone and combined with potassium iodide (KI) was evaluated. The presence of KI was able to potentiate the aPDT process in orange juice, promoting the inactivation of 5 log CFU/mL of A. acidoterrestris spores after 10 h of white light exposition (140 mW/cm 2 ). However, in the absence of KI, both photosensitizers did not promote a significant reduction in the spore viability. The inactivation of A. acidoterrestris spores artificially inoculated in orange peels (10 5 spores/mL) was also assessed using Tetra-Py + -Me at 10 and 50 μM in the presence and absence of KI in combination with white light (65 mW/cm 2 ). No significant reductions were observed (p < .05) when Tetra-Py + -Me was used at 10 μM, however at the highest concentration (50 μM) a significant spore reduction (≈ 2.8 log CFU/mL reductions) in orange peels was observed after 6 h of sunlight exposition (65 mW/cm 2 ). Although the color, total phenolic content (TPC), and antioxidant capacity of orange juice and peel (only color evaluation) seem to have been affected by light exposition, the impact on the visual and nutritional characteristics of the products remains inconclusive so far. Besides that, the results found suggest that aPDT can be a potential method for the reduction of A. acidoterrestris spores on orange groves., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Aspergillus fischeri ascospores are known as potential spoilage microorganisms of pasteurized fruit products due to their high incidence in fruits, the ability to survive pasteurization and to grow in acidic conditions. This study aimed to develop a quantitative microbial spoilage risk assessment (QMSRA) model approach to estimate the spoilage risk of packaged strawberry purees due to A. fischeri under various scenarios regarding product formulation, processing and storage conditions. The development of the risk assessment comprised three steps: (1) initial contamination level of raw material by ascospores (N 0 ), (2) inactivation of ascospores during thermal processing (N p ) and (3) determination of the number of ascospores which are able to survive thermal processing and develop visible mycelia (D = 2 mm) during storage (N f ). Data of visible growth (t v , days) comprised distributions previously obtained as function of water activity (a w ) (0.860-0.985), oxygen (0-21%), temperature (8-30 °C) and pasteurization (95-105 °C/15 s). The simulations were performed in triplicate with 100,000 iterations using the software R. The outcome "spoilage risk" was defined as the probability of having at least one ascospore (N f ) capable of forming visible colonies in 100 g-pack strawberry puree within the typical use-by dates. Overall, high probabilities of spoilage were estimated for purees pasteurized at milder treatments at 85 °C/15-60 s (67%) and 90 °C/15-60 s (≥40%) stored at ambient temperature (22 °C). The spoilage risk was only effectively reduced (0.02%) by increasing pasteurization conditions to 95 °C for at least 45 s. Moreover, the microbial stability of such purees, i.e., spoilage risk <0.001% (=less than 1 spoilage pack out of 10 5 produced units) was predicted to occur for purees treated at 100 °C/15 s or stored at chilled conditions (≤8 °C) or at strict anaerobic conditions or produced as concentrates (a w  ≤ 0.860). Based on the outcomes obtained, a set of specifications for Heat-Resistant Moulds (HRMs) in raw material and pasteurized purees aimed to be used as an ingredient was suggested. Furthermore, the results can be used to support risk management decisions in identifying and quantifying the impact of possible interventions during formulation, processing and storage conditions of fruit purees to effectively reduce this risk., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Even though spore-forming bacteria have been mainly linked to spoilage or foodborne pathogens vehiculated through foods, some strains of Bacillus can potentially present probiotic properties. The advantage of incorporating probiotic Bacillus strains in foods relies mainly on the fact that these microorganisms present high resistance to harsh processing conditions. "Requeijão cremoso" is a type of processed cheese highly appreciated in Brazil. During processing, this product is submitted to several harsh conditions (heating at 90 °C, for instance), leading to the inactivation of probiotic bacteria belonging to Lactobacillus and Bifidobacterium genera. That fact has precluded the development of probiotic "requeijão cremoso" products; however, probiotic Bacillus strains may comprise a promising alternative to overcome the low resistance of traditional probiotics to food processing. The objective of this study was to evaluate the behavior of different Bacillus strains with claimed probiotic properties throughout processed cheese ("requeijão cremoso") manufacturing. A total of five different Bacillus strains with claimed probiotic properties (B. coagulans MTCC 5856, B. coagulans GBI-30 6086, B. subtilis PXN 21, B. subtilis PB6, and B. flexus HK1) were individually inoculated at different stages of manufacture - curd pasteurization, coagulation, and fusion - of "requeijão cremoso" and their survival in each of these stages was determined. The survival of B. coagulans GBI-30 6086 was further assessed throughout "requeijão cremoso" production and shelf life (45 days at 6 °C). Besides, the chemical composition, level of proteolysis, and fatty acid profile of the treatments during shelf life were evaluated. The fusion stage was found as the most appropriate for the addition of B. coagulans GBI-30 6086, which allows the production of probiotic "requeijão cremoso" and facilitates the technological process while preventing the occurrence of final product recontamination., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The variability in spore survival during spray drying of 12 Bacillus cereus strains was evaluated. B. cereus spores were inoculated on whole milk (7.2 ± 0.2 log 10  spores/g dry weight) and processed in a spray-dryer. Twelve independent experiments were carried out in triplicate. The spore count was determined before and after each drying process, based on the dry weight of whole milk and powdered milk. Then, the number of decimal reductions (γ) caused by the spray drying process was calculated. B. cereus strains presented γ values ranging from 1.0 to 4.7 log10 spores/g dry weight, with a high coefficient of variation (CV) of 46.1%. Cluster analysis allowed to group B. cereus as sensitive (strains 511, 512, 540, 432 and ATCC 14579), intermediate (strains B18, B63, and B86) and resistant strains (strains B3, B94, B51 and 436). Three strains (one of each group) were selected for further investigation and characterization of their physicochemical and molecular (proteomics) differences. Fourier transform infrared spectroscopy (FTIR), and differential scanning calorimetry (DSC) were used to determine physicochemical characteristics and glass transition temperature (T g ), respectively. No differences in signs among the three strains were found in spectra ranging from 900 to 4000 cm -1 . The endothermic peak ranged between 54 and 130 °C for strain 540; between 81 and 163 °C for strain B63; and between 110 and 171 °C for strain 436. However, they showed different T g : 88.82 °C for strain 540; 114.32 °C for strain B63; and 122.70 °C for strain 436. A total of eleven spots were identified by mass spectrometry, with the spore coat protein GerQ, sporulation protein YtfJ (GerW), and peptidyl-prolyl cis-trans isomerase being found in at least two strains. Altogether, the results suggested that the high survival variability of B. cereus spores to the spray drying process seems to be mainly associated with different T g and protein content. The study highlights the importance of quantifying the effects of this unit operation over the target microorganisms. These data may be relevant for the development of effective measures aiming to control the occurrence of B. cereus in milk powder as well as to reduce spoilage or safety issues associated with the presence of this bacterium in foods, particularly those formulated with milk powder., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Heat-resistant moulds (HRMs) are well known for their ability to survive pasteurization and spoil high-acid food products, which is of great concern for processors of fruit-based products worldwide. Whilst the majority of the studies on HRMs over the last decades have addressed their inactivation, few data are currently available regarding their contamination levels in fruit and fruit-based products. Thus, this study aimed to quantify and identify heat-resistant fungal ascospores from samples collected throughout the processing of pasteurized high-acid fruit products. In addition, an assessment on the effect of processing on the contamination levels of HRMs in these products was carried out. A total of 332 samples from 111 batches were analyzed from three processing plants (=three processing lines): strawberry puree (n = 88, Belgium), concentrated orange juice (n = 90, Brazil) and apple puree (n = 154, the Netherlands). HRMs were detected in 96.4% (107/111) of the batches and 59.3% (197/332) of the analyzed samples. HRMs were present in 90.9% of the samples from the strawberry puree processing line (1-215 ascospores/100 g), 46.7% of the samples from the orange juice processing line (1-200 ascospores/100 g) and 48.7% of samples from the apple puree processing line (1-84 ascospores/100 g). Despite the high occurrence, the majority (76.8%, 255/332) of the samples were either not contaminated or presented low levels of HRMs (<10 ascospores/100 g). For both strawberry puree and concentrated orange juice, processing had no statistically significant effect on the levels of HRMs (p > 0.05). On the contrary, a significant reduction (p < 0.05) in HRMs levels was observed during the processing of apple puree. Twelve species were identified belonging to four genera - Byssochlamys, Aspergillus with Neosartorya-type ascospores, Talaromyces and Rasamsonia. N. fumigata (23.6%), N. fischeri (19.1%) and B. nivea (5.5%) were the predominant species in pasteurized products. The quantitative data (contamination levels of HRMs) were fitted to exponential distributions and will ultimately be included as input to spoilage risk assessment models which would allow better control of the spoilage of heat treated fruit products caused by heat-resistant moulds., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This study aimed to assess the capability of 97 epidemic S. enterica strains belonging to 18 serovars to form biofilm. Five strains characterized as strong biofilm-producers, belonging to distinct serovars (S. Enteritidis 132, S. Infantis 176, S. Typhimurium 177, S. Heidelberg 281 and S. Corvallis 297) were assayed for adhesion/biofilm formation on stainless steel surfaces. The experiments were conducted in different combinations of NaCl (0, 2, 4, 5, 6, 8 and 10% w/v), pH (4, 5, 6 and 7) and temperatures (8 °C, 12 °C, 20 °C and 35 °C). Only adhesion was assumed to occur when S. enterica counts were ≥3 and <5 log CFU/cm 2 , whereas biofilm formation was defined as when the counts were ≥5 log CFU/cm 2 . The binary responses were used to develop models to predict the probability of adhesion/biofilm formation on stainless steel surfaces by five strains belonging to different S. enterica serovars. A total of 99% (96/97) of the tested S. enterica strains were characterized as biofilm-producers in the microtiter plate assays. The ability to form biofilm varied (P < 0.05) within and among the different serovars. Among the biofilm-producers, 21% (20/96), 45% (43/96), and 35% (34/96) were weak, moderate and strong biofilm-producers, respectively. The capability for adhesion/biofilm formation on stainless steel surfaces under the experimental conditions studied varied among the strains studied, and distinct secondary models were obtained to describe the behavior of the five S. enterica tested. All strains showed adhesion at pH 4 up to 4% of NaCl and at 20 °C and 35 °C. The probability of adhesion decreased when NaCl concentrations were >8% and at 8 °C, as well as in pH values ≤ 5 and NaCl concentrations > 6%, for all tested strains. At pH 7 and 6, biofilm formation for S. Enteritidis, S. Infantis, S. Typhimurium, S. Heidelberg was observed up to 6% of NaCl at 35 °C and 20 °C. The predicted boundaries for adhesion were pH values < 5 and NaCl ≥ 4% and at temperatures <20 °C. For biofilm formation, the predicted boundaries were pH values < 5 and NaCl concentrations ≥ 2% and at temperatures <20 °C for all strains. The secondary models obtained describe the variability in boundaries of adhesion and biofilm formation on stainless steel by five strains belonging to different S. enterica serovars. The boundary models can be used to predict adhesion and biofilm formation ability on stainless steel by S. enterica as affected by pH, NaCl and temperature., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The major aims of this study were to assess inter- and intra-species variability of heat resistant moulds (HRMs), Byssochlamys fulva and Byssochlamys nivea, with regards to (i) heat resistance and (ii) effect of heat treatment intensity on subsequent outgrowth. Four-week-old ascospores were suspended in buffered glucose solution (13° Brix, pH 3.5) and heat treated in a thermal cycler adjusted at 85 °C, 90 °C and 93 °C. Two variants of the Weibull model were fitted to the survival data and the following inactivation parameters estimated: b (inactivation rate, min -1 ), n (curve shape) and δ (the time taken for first decimal reduction, min). In addition to the assessment of heat resistance, outgrowth of Byssochlamys sp. from ascospores heated at 70 °C, 75 °C, 80 °C, 85 °C and 90 °C for 10 min and at 93 °C for 30 and 70 s was determined at 22 °C for up to 30 days. The Baranyi and Roberts model was fitted to the growth data to estimate the radial growth rates (μ max , mm.day -1 ) and lag times (λ, days). Inter-species variability and significant differences (p < 0.05) were observed for both inactivation and growth estimated parameters among B. fulva and B. nivea strains. The effect of heat treatment intensity on outgrowth of B. fulva strains was more apparent at the most intense heat treatment evaluated (90 °C/10 min), which was also the condition in which greater dispersion of the estimated kinetic parameters was observed. On the other hand, B. nivea strains were more affected by heating, resulting in greater variability of growth parameters estimated at different heating intensities and in very long lag phases (up to 25 days). The results show that inter- and intra-species variability in the kinetic parameters of Byssochlamys sp. needs to be taken into account for more accurate spoilage prediction. Furthermore, the effect of thermal treatments on subsequent outgrowth from ascospores should be explored in combination with other relevant factors such as °Brix and oxygen to develop thermal processes and storage conditions which can prevent the growth of HRMs and spoilage of heat treated food products., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The aim of this study was to assess the incidence and to estimate the growth kinetic parameters (maximum growth rate, μ; lag time, λ; and maximum population, κ) of Salmonella on the peel and pulp of avocado (Perseaamericana var. americana) and custard apple (Annona squamosa L.) as affected by temperature (10-30°C). The incidence of Salmonella was assessed on the peel and pulp of the fruits (n=200 of each fruit), separately, totalizing 800 analyses. Only three samples of custard apple pulp were positive for Salmonella enterica and the three isolates recovered belonged to serotype S. Typhimurium. Salmonella was not recovered from avocado and custard apple peels and from avocado pulp. Generally, the substrate (pulp or peel) of growth did not affect μ values of S. enterica (p>0.05). Very similar μ values were found for S. enterica inoculated in custard apple and avocado. S. enterica presented the highest λ in the peel of the fruits. The growth of S. enterica resulted in larger λ in custard apple in comparison to avocado. For example, the λ of S. enterica in the pulp of custard apple and avocado were 47.0±0.78h and 10.0±3.78h, respectively. The lowest values of κ were obtained at the lower storage temperature conditions (10°C). For instance, κ values of 3.7±0.06log CFU/g and 2.9±0.03log CFU/g were obtained from the growth of S. enterica in avocado and custard apple pulps at 10°C (p<0.05), respectively. On the other hand, at 30°C, κ values were 6.5±0.25log CFU/g and 6.5±0.05log CFU/g, respectively. Significantly higher κ were obtained from the growth of S. enterica in the pulp than in the peel of the fruits (p<0.05). For instance, the growth of S. enterica in the pulp of avocado led to a κ value of 6.5±0.25log CFU/g, while in the peel led to a κ value of 4.6±0.23log CFU/g (p<0.05). In general, growth kinetic parameters indicated that avocado comprises a better substrate than custard apple for the growth of S. enterica. The square root model fitted to the data obtained in this study and to the growth data available in the literature for other tropical low acid fruits indicated high variability in μ and λ of Salmonella. The results obtained in this study show that whole low acid tropical fruits can harbor Salmonella, and that this foodborne pathogen can not only survive but also grow both on the peel and pulp of low acid tropical fruits, such as avocado and custard apple., (Copyright © 2016 Elsevier B.V. All rights reserved.)

In this study, the fate of Alicyclobacillus acidoterrestris spores in different types of juice concentrates stored under different conditions was investigated. In addition, the impact of dilution procedures during the enrichment step for the detection of Alicyclobacillus in lemon juice concentrates was studied. Pear, red grape, mango, tangerine, carrot and lemon juice concentrates (50-69.4°Brix, pH1.7-4.3) were inoculated with A. acidoterrestris spores (10(3) spore/mL) and stored at 4 °C and 20 °C, after which the spores were counted at 0, 2, 5, 9, 17, 21, 28, 36, 43, and 50 days. No significant differences in the number of Alicyclobacillus spores were observed at storage temperatures of 4 °C and 20 °C (p>0.05). The results also indicated that the number of spores of A. acidoterrestris remained stable in all types of juice concentrates during the storage period, except in lemon juice concentrate. In lemon juice concentrate, a decline in A. acidoterrestris spore populations of 0.3-0.8 log CFU/mL was observed within 5-10 days of storage. The decline in A. acidoterrestris spore populations was more pronounced in cloudy lemon juice concentrate, which contained higher concentrations of flavonoids (mainly eriocitrin and hesperidin) than clarified lemon juice concentrate. It was also found that dilution of lemon juice concentrate samples in the proportion of 1:19 allowed the germination of A. acidoterrestris spores and the growth of populations of up to 10(7) CFU/mL. In contrast, the proportion (1:9) recommended in internationally recognized methods led to a reduction in the population of this microorganism that would yield false negative results. Data presented in this study demonstrated that Alicyclobacillus spores remain stable in most juice concentrates during storage, but that natural antimicrobial compounds present in some of them may decrease spore counts and inhibit their recovery by detection procedures., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This study aimed to report the incidence of Alicyclobacillus and Alicyclobacillus acidoterrestris in apple and pear flavorings (n=42) and to assess the effect of guaiacol-producing A. acidoterrestris strains on apple flavorings stored at 4, 20 and 45 °C. A real-time polymerase chain reaction (RT-PCR) method was used for simultaneous confirmation of alicyclobacilli. A total of six isolates were identified as A. acidoterrestris, and only one strain was not able to produce guaiacol. The storage of apple flavoring at the optimum growth temperature of A. acidoterrestris (45 °C) resulted in the reduction in the spores' counts within 30 days of storage. On the other hand, during chilling (4 °C) and ambient storage conditions (20 °C) the counts of spores that remained stable for up to 60 days. An A. acidoterrestris strain inoculated in flavoring and further added to apple juice was able to grow and produce guaiacol in high amounts between 1-7 days of storage at 30 °C. In the current study it was shown that flavorings may be contaminated by deteriogenic A. acidoterrestris strains that are able to survive during storage in a wide range of temperature for long periods and further contaminate and spoil formulated fruit juices and beverages. A novel potential source of fruit juices and beverages contamination by deteriogenic Alicyclobacillus was shown. To the best of the author's knowledge, this is the first report on the incidence and fate of Alicyclobacillus and A. acidoterrestris in flavorings., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Growth potential (δ) is defined as the difference between the population of a microorganism at the end of shelf-life of specific food and its initial population. The determination of δ of Salmonella and Listeria monocytogenes in RTE vegetables can be very useful to determine likely threats to food safety. However, little is known on the behavior of these microorganisms in several RTE vegetables. Therefore, the aim of this study was to determine the δ of both pathogens in nine different types of RTE vegetables (escarole, collard green, spinach, watercress, arugula, grated carrot, green salad, and mix for yakisoba) stored at refrigeration (7°C) and abuse temperature (15°C). The population of aerobic microorganisms and lactic acid bacteria, including those showing antimicrobial activity has been also determined. Results indicated that L. monocytogenes was able to grow (δ≥0.5 log(10)) in more storage conditions and vegetables than Salmonella. Both microorganisms were inhibited in carrots, although a more pronounced effect has been observed against L. monocytogenes. The highest δ values were obtained when the RTE vegetables were stored 15°C/6days in collard greens (δ=3.3) and arugula (δ=3.2) (L. monocytogenes) and arugula (δ=4.1) and escarole (δ=2.8) (Salmonella). In most vegetables and storage conditions studied, the counts of total aerobic microorganisms raised significantly independent of the temperature of storage (p<0.05). Counts of lactic acid bacteria were higher in vegetables partially or fully stored at abuse temperature with recovery of isolates showing antimicrobial activity. In conclusion, the results of this study show that Salmonella and L. monocytogenes may grow and reach high populations in RTE vegetables depending on storage conditions and the definition of effective intervention strategies are needed to control their growth in these products., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Listeria monocytogenes is a foodborne pathogen of great concern due to the high fatality rates of listeriosis. The consumption of RTE vegetables has increased in Brazil over the last two decades, but little is known about the risks associated to the consumption of these products. This study evaluated the prevalence and counts of L. monocytogenes in 512 packages of ready-to-eat vegetables marketed in São Paulo. The isolates were characterized for their serotypes, ribotypes, positivity for virulence genes inlA, inlC and inlJ, resistance to chlorine, growth rate variability and capability to form biofilm on stainless steel (AISI 304, #4) coupons. L. monocytogenes was detected in 3.1% of the samples. Only five samples presented countable levels, with counts between 1.0×10(1) and 2.6×10(2)CFU/g. Isolates belonged to serotypes 1/2b or 4b and most were positive for genes inlC and inlJ. Ribotypable isolates were grouped into four groups: 1038 (69.4%), 19175 (11.3%), 19191 (17.7%) and 18604 (one isolate). Most isolates survived to exposure to 125 ppm of a chlorine-based disinfectant for 3 min. All isolates were capable to attach to the coupons, reaching counts above 4 log(10) CFU/cm(2) and the growth rate (μ) at 25°C of the majority of the isolates varied between 0.1 and 0.2 log OD/h, but for few strains the μ was as high as 0.26 log OD/h. Results of this survey indicate that RTE vegetables may be vehicles of L. monocytogenes strains with limited variation in serotype, ribotype and virulence factors but varying significantly in resistance to chlorine disinfectants, capability of forming biofilm and growth rate. Data obtained is of foremost importance to serve as baseline for the development of scientific-based policies to control the incidence of L. monocytogenes in RTE vegetables in Brazil., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The purpose of this study was to determine the incidence of Alicyclobacillus spp. in fruit/vegetable juices (concentrated pulps and clarified and non-clarified juices) marketed in Argentina between 1996 and 2009. The presence of Alicyclobacillus was determined in a total of 8556 samples of fruit and vegetable juices (apple, pear, grape, peach, blend of juices, tangerine, pineapple, orange, mango, plum, guava, apricot, lemon, banana, kiwi, carrot, strawberry, grapefruit, and beetroot) collected in seven Argentinean provinces. Multiple factor analysis (MFA) was carried out on a data matrix that contained the percentage of positive samples, type of juice, raw material and production year. Except for kiwi and orange, Alicyclobacillus was found in juices from all the evaluated raw materials. The highest percentage of positive samples was found for beetroot, strawberry, banana, peach, mango, carrot and plum juices. The percentage of positive samples for these juices ranged from 100% to 24%. Furthermore, the application of multivariate techniques provided an insight on the relationship between the incidence of Alicyclobacillus and production variables. This approach enabled the identification of the most relevant variables that increased the percentage of positive samples among the juices, which could help in developing strategies to avoid the incidence of this bacterium. By means of hierarchical cluster analysis seven groups (clusters) of juices which showed different percentages of positive samples for Alicyclobacillus spp. were identified. This analysis showed that pineapple, peach, strawberry, mango and beetroot juices had higher rates of positivity for Alicyclobacillus than the rest of the evaluated juices. MFA analysis also showed that some clear relationships could be highlighted between the percentage of samples positive for Alicyclobacillus and five types of fruit juices (strawberry, beetroot, grapefruit, pineapple and mango). It was observed that a large proportion of juices produced in 2000, 2005 and 2008 were located in clusters with higher incidence of Alicyclobacillus spp., whereas a larger proportion of clarified concentrate juice and concentrate pulp samples showed higher probability of incidence of Alicyclobacillus in these products. Data presented in this study brings a contribution to the ecology of Alicyclobacillus in fruit/vegetable juices marketed in Argentina. This information would be useful to enhance the microbiological stability of fruit juices regarding the presence of Alicyclobacillus spp., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The objectives of this study were to isolate psychrotrophic clostridia from Brazilian vacuum-packed beef cuts (spoiled or not) and to identify the isolates by using 16S rRNA gene sequencing. Anaerobic psychrotrophic microorganisms were also enumerated and samples were collected to verify the incidence of psychrotrophic clostridia in the abattoir environment. Vacuum-packed beef cuts (n=8 grossly distended and n=5 non-spoiled) and environmental samples were obtained from a beef packing plant located in the state of São Paulo, Brazil. Each sample was divided in three subsamples (exudate, beef surface and beef core) that were analyzed for vegetative forms, total spore-forming, and sulfide reducing spore-forming, both activated by alcohol and heat. Biochemical profiles of the isolates were obtained using API20A, with further identification using 16S rRNA gene sequencing. The growth temperature and the pH range were also assessed. Populations of psychrotrophic anaerobic vegetative microorganisms of up to 10(10)CFU/(g, mL or 100 cm(2)) were found in 'blown pack' samples, while in non-spoiled samples populations of 10(5)CFU/(g, CFU/mL or CFU/100 cm(2)) was found. Overall, a higher population of total spores and sulfide reducing spores activated by heat in spoiled samples was found. Clostridium gasigenes (n=10) and C. algidicarnis (n=2) were identified using 16S rRNA gene sequencing. Among the ten C. gasigenes isolates, six were from spoiled samples (C1, C2 and C9), two were isolated from non-spoiled samples (C4 and C5) and two were isolated from the hide and the abattoir corridor/beef cut conveyor belt. C. algidicarnis was recovered from spoiled beef packs (C2). Although some samples (C3, C7, C10 and C14) presented signs of 'blown pack' spoilage, Clostridium was not recovered. C. algidicarnis (n=1) and C. gasigenes (n=9) isolates have shown a psychrotrophic behavior, grew in the range 6.2-8.2. This is the first report on the isolation of psychrotrophic Clostridium (C. gasigenes and C. algidicarnis) in Brazil. This study shows that psychrotrophic Clostridium may pose a risk for the stability of vacuum-packed beef produced in tropical countries during shelf-life and highlights the need of adopting control measures to reduce their incidence in abattoir and the occurrence of 'blown pack' spoilage., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Although the production of patulin in apple fruits is mainly by Penicillium expansum, there is no information on the ability of heat resistant moulds that may survive pasteurization to produce this mycotoxin in juice packages during storage and distribution. In this study, the production of patulin by Byssochlamys spp (Byssochlamys nivea FRR 4421, B. nivea ATCC 24008 and Byssochlamys fulva IOC 4518) in cloudy and clarified apple juices packaged in laminated paperboard packages or in polyethylene terephthalate bottles (PET) and stored at both 21 degrees C and 30 degrees C, was investigated. The three Byssochlamys strains were able to produce patulin in both cloudy and clarified apple juices. Overall, the lower the storage temperature, the lower the patulin levels and mycelium dry weight in the apple juices (p<0.05). The greatest variations in pH and degrees Brix were observed in the juices from which the greatest mycelium dry weights were recovered. The maximum levels of patulin recovered from the juices were ca. 150 microg/kg at 21 degrees C and 220 microg/kg at 30 degrees C. HPLC-UV, HPCL-DAD and mass spectrometry analyses confirmed the ability of B. fulva IOC 4518 to produce patulin. Due to the heat resistance of B. nivea and B. fulva and their ability to produce patulin either in PET bottles or in laminated paperboard packages, the control of contamination and the incidence of these fungi should be a matter of concern for food safety. Control measures taken by juice industries must also focus on controlling the ascospores of heat resistant moulds., (Copyright 2010 Elsevier B.V. All rights reserved.)

In this study, the influence of the hot-fill water-spray-cooling process after continuous pasteurization on the number of decimal reductions (gamma) and growth parameters (lag time; lambda, ratio N(f)/N(o); kappa, maximum growth rate; mu) of Alicyclobacillus acidoterrestris CRA 7152 in orange juice stored at 35 degrees C were investigated. Two different inoculum levels of A. acidoterrestris CRA 7152 (10(2) and 10(3) spores/mL) in orange juice (11(0)Brix, pH 3.7) and a Microthermics UHT-HTST pilot plant were used to simulate industrial conditions. Results have shown that regardless of the inoculum level (10(2) or 10(3) spores/mL), the pasteurization processes were unable to cause even 1 gamma. Predictive modeling using the Baranyi model showed that only kappa and time to reach 10(4)spores/mL (t10(4) - time to juice spoilage) were affected by the spore inoculum used (p<0.05). It has been concluded that A. acidoterrestris was able to survive the hot-fill process and to grow and spoil orange juice in 5-6 days when the final storage temperature was 35 degrees C., ((c) 2009 Elsevier B.V. All rights reserved.)