Enterotoxigenic Staphylococcus aureus in Brazilian artisanal cheeses: Occurrence, counts, phenotypic and genotypic profiles.

The present study aimed to assess the occurrence and counts of Staphylococcus aureus in Brazilian artisanal cheeses (BAC) produced in five regions of Brazil: Coalho and Manteiga (Northeast region); Colonial and Serrano (South); Caipira (Central-West); Marajó (North); and Minas Artisanal cheeses, from Araxá, Campos das Vertentes, Cerrado, Serro and Canastra microregions (Southeast). The resistance to chlorine-based sanitizers, ability to attach to stainless steel surfaces, and antibiogram profile of a large set of S. aureus strains (n = 585) were assessed. Further, a total of 42 isolates were evaluated for the presence of enterotoxigenic genes (sea, seb, sec, sed, see, seg, sei, sej, and ser) and submitted to typing using pulsed-field gel electrophoresis (PFGE). BAC presented high counts of S. aureus (3.4-6.4 log CFU/g), varying from 25 to 62.5%. From the S. aureus strains (n = 585) assessed, 16% could resist 200 ppm of sodium hypochlorite, whereas 87.6% produced strong ability to attach to stainless steel surfaces, corroborating with S. aureus ability to persist and spread in the environment. Furthermore, the relatively high frequency (80.5%) of multidrug-resistant S. aureus and the presence of enterotoxin genes in 92.6% of the strains is of utmost attention. It reveals the lurking threat of SFP that can survive when conditions are favorable. The presence of enterotoxigenic and antimicrobial-resistant strains of S. aureus in cheese constitutes a potential risk to public health. This result calls for better control of cheese contamination sources, and taking hygienic measures is necessary for food safety. More attention should be paid to animal welfare and hygiene practices in some dairy farms during manufacturing to enhance the microbiological quality of traditional cheese products.

Molecular characterization and toxigenic profiles of Bacillus cereus isolates from foodstuff and food poisoning outbreaks in Brazil.

Bacillus cereus sensu stricto (s.s.) is a well-known foodborne pathogen that produces a range of enterotoxins and is able to cause two different types of foodborne illnesses-the emetic and the diarrheal syndromes. In this study, 54 B. cereus s.s. strains isolated from foodstuff and foods involved in food poisoning outbreaks were characterized according to the presence of toxin-encoding genes, virulence-encoding genes, and panC typing. Most isolates were assigned to panC groups IV (61.1%) and III (25.9%), but members of groups II and V could also be found. Investigation of specific alleles revealed high numbers of isolates carrying toxin and other virulence genes including nheA (100%), nheB (100%), hblA (79.6%), hblC (79.6%), hblD (74.1%), cytK-2 (61.1%), clo (100%), pc-plc (75.9%), sph (68.5%), pi-plc (66.6%), hlyIII (62.9%), and hlyII (24.1%). All isolates were negative for ces and cytK-1. In summary, we detected various enterotoxin and other virulence factor genes associated with diarrheal syndrome in strains analyzed, implicated or not with food poisoning. Furthermore, the most isolates analyzed belong to high-risk phylogenetic groups' panC types III and IV. Our study provides a convenient molecular scheme for characterization of B. cereus s.s. strains responsible for food poisoning outbreaks in order to improve the monitoring and investigation and assess emerging clusters and diversity of strains.

Chilled Pacu (Piaractus mesopotamicus) fillets: Modeling Pseudomonas spp. and psychrotrophic bacteria growth and monitoring spoilage indicators by 1H NMR and GC-MS during storage.

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 (µmax, 1/h), lag time (tLag, h), and (Nmax, Log10 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 R2 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). 1H 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.

Predicting the impact of temperature and relative humidity on Salmonella growth and survival in sliced chard, broccoli and red cabbage.

This study assessed the fate of a Salmonella enterica cocktail (S. Typhimurium, S. Enteritidis, S. Newport, S. Agona and S. Anatum; initial counts 3.5 log CFU/g) in minimally processed sliced chard, broccoli and red cabbage at 16 conditions of different temperature (7, 14, 21 and 37 °C) and relative humidity (RH; 15, 35, 65 and 95%) over six days (144 h). Linear regression was used to estimate the rate change of Salmonella in cut vegetables as a function of temperature and relative humidity (RH). R2 value of 0.85, 0.87, and 0.78 were observed for the rates of change in chard, broccoli, and red cabbage, respectively. The interaction between temperature and RH was significant in all sliced vegetables. Higher temperatures and RH values favored Salmonella growth. As temperature or RH decreased, the rate of S. enterica change varied by vegetable. The models developed here can improve risk management of Salmonella in fresh cut vegetables.

Investigating desiccation resistance, post-rehydration growth, and heat tolerance in desiccation-injured cells of Salmonella enterica isolated from the soybean production chain.

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-aw 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.

Quantitative risk assessment of the presence of fumonisin in corn produced in different regions of Brazil: Influence of climatic conditions.

In this study, the probability of occurrence of fumonisins in corn in the states of greatest production in Brazil was determined. The data were analyzed through quantitative risk analysis using the Monte Carlo simulation. The results indicated that there is a strong correlation between fumonisins contamination levels and the geographical region due to the influence of climatic characteristics, with temperature having the main influence. The Southern states presented higher risks of occurrence and concentration levels of fumonisins in corn due to the temperate climate with lower average temperature and higher relative humidity and precipitation indices. Cultivation in the best season indicates a significant reduction in the production of fumonisins when this period was evaluated, with average concentration levels up to 42% lower. The generated data are important for regulatory agencies and the agricultural sector, which needs to be aware that the chance of success in grain production depends on efficient planning of the growing season, mainly concerning the climatic conditions to which it is subject to minimize the risks.

Salmonella enterica in soybean production chain: Occurrence, characterization, and survival during soybean storage.

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.

Survival kinetics, membrane integrity and metabolic activity of Salmonella enterica in conventionally and osmotically dehydrated coconut flakes.

Many outbreaks involving Salmonella enterica in dehydrated coconut have been reported. Little is known about the survival of S. enterica in dehydrated coconut flakes at common retail or domestic storage conditions. This study evaluated the behavior of a S. enterica cocktail (S. Enteritidis PT4, S. Typhimurium PT4, S. Bredeney, S. Muenster and S. Agona) in conventionally and osmotically dehydrated coconut flakes under four storage regimes: 25 °C for 120 days, 25 °C for 30 days then 7 °C for 90 days, 7 °C for 30 days then 25 °C for 90 days, and 7 °C for 120 days. S. enterica membrane integrity (using with propidium iodide and bis-1,3-dibutylbarbutyric acid) and metabolic activity (using 5-cyano-2,3-ditolyl tetrazolium chloride) were assessed by flow cytometry analysis after dehydration and storage at 7 °C or 25 °C for 120 days. Lower S. enterica inactivation rates (kmax 0.02 to 0.04 1/days) were observed in conventionally dehydrated coconut flakes compared to osmotically dehydrated coconut flakes (kmax 0.16 to 0.20 1/days). Changes in storage temperature did not affect the behavior of S. enterica in conventionally or osmotically dehydrated coconut flakes. Results show that S. enterica inactivation in conventionally dehydrated coconut flakes could be described by log-linear with tail models. S. enterica inactivation in osmotically dehydrated coconut flakes could be described by log-linear with shoulder and tail models. Subpopulations of S. enterica cells with damaged membranes and without metabolic activity were larger in conventionally (32.1% and 90.9%, respectively) than osmotically dehydrated coconut (18.5% and 82.2%, respectively) flakes at the beginning of the storage. Subpopulations of S. enterica cells with damaged membrane decreased by 9.4-14.4%, while cells with membrane potential and intact membrane increased by 23.7 and 24.2% in conventionally dehydrated coconut flakes after 120 days of storage at 7 °C or 25 °C, respectively. Subpopulations of S. enterica with damaged membranes did not change significantly in osmotically dehydrated coconut flakes. Our findings suggest that S. enterica populations decline during storage occurs due in part to membrane integrity losses. These data can contribute to the development of risk management strategies for S. enterica in dehydrated coconut flakes.

Growth behavior of low populations of Listeria monocytogenes on fresh-cut mango, melon and papaya under different storage temperatures.

The growth behavior of Listeria monocytogenes low population (1-4 cells/sample) on fresh-cut mango, melon, papaya and fruit mix stored at 4, 8, 12 and 16 °C was evaluated over 10 days. Mango showed the lowest counts for L. monocytogenes during 10 days regardless of storage temperature (<1.7 log cfu.g-1). Melon supported high bacterial growth over 10 days, reaching 5 log cfu.g-1 at 16 °C. Both the fruit and storage temperature influenced the Listeria low population growth potential (δ). Cumulative frequency distribution of L. monocytogenes showed that after 10 days, 100% of fresh-cut fruits and fruit mix stored at 4 °C remained ≤2 log cfu.g-1, while at 12 and 16 °C 100% of melon, papaya and fruit mix samples exceeded this limit. At 8 °C, 100% of mango and fruit mix samples remained below this limit after 10 days, whereas 100% of melon and papaya reached it after 7 days. Results indicate 4 °C as the ideal to store safely fresh-cut mango, melon, papaya and fruit mix for 10 days. Besides, 8 °C can also be an option, but not for melon and papaya. Findings highlight the ability of L. monocytogenes to survive and grow in fresh-cut fruits even at a very low initial population levels.

A worldwide systematic review, meta-analysis, and health risk assessment study of mycotoxins in beers.

Mycotoxins, including aflatoxins (AFs), ochratoxin A (OTA), deoxynivalenol (DON), fumonisins (FBs), and zearalenone (ZEN), have been reported as beer contaminants. This systematic review and meta-analysis provide the prevalence and concentration of mycotoxins in beers and their worldwide distribution. Mycotoxin's exposure and cancer risk through beer consumption were determined. The overall pooled prevalence of mycotoxins in beers was 31% (95% confidence interval [CI] = 28%-35%; I2  = 90%, p = .00). The most prevalent mycotoxins in beers were DON and its derivatives (53%), OTA (52%), FBs (47%), followed by AFs (12%). Iran (99%), Hungary (95%), Denmark (92%), Armenia (83%), and Cyprus (83%) had the highest mycotoxin prevalence in beers. The global mycotoxins average concentration in beers was 12.52 µg/L (95% CI = 10.70-14.75 µg/L; I2  = 100%, p = .00). DON and its derivatives showed the highest concentration (26.91 µg/L), followed by FBs (23.19 µg/L), ZEN and its derivatives (20.25 µg/L), and AFs (15.65 µg/L). African region had the highest mycotoxins concentration (73.95 µg/L) mostly due to the high levels reported in beers from Cameroon (293.02 µg/L), Malawi (132.34 µg/L), and Eastern Cape province (126.12 µg/L). The meta-regression indicated stability (p ≥ .05) of the global pooled concentration of mycotoxins in beers over the years, whereas FBs concentration increased. The intake of DON and its derivatives, FBs, ZEN and its derivatives, and OTA through beers is of concern in African countries. OTA is also of concern in Brazil and Belgium. Results show high mycotoxins concentration in beers worldwide and highlight the health risks through contaminated beer consumption.

High throughput screening of technological and biopreservation traits of a large set of wild lactic acid bacteria from Brazilian artisanal cheeses.

This study aimed to evaluate technological (acidification, proteolysis, lipolysis, resistance to low pH, NaCl, and bile salts) and biopreservation (antimicrobial activity against foodborne pathogens) features of 1002 LAB by high throughput screening (HTS) methods. The LAB was isolated from 11 types of Brazilian artisanal cheeses (BAC) marketed in the main 5 producing regions. Remarkable intra-species variability in acidification rates have been found, which was most pronounced between isolates from Mina's artisanal cheeses, Caipira and Coalho cheeses. Lacticaseibacillus paracasei and Levilactobacillus brevis showed the fastest acidification rate; however, all isolates showed slower acidification rates than a lactococcal control strain (4.3 × lower). When testing inhibitory effects, > 75% of LAB isolates could inhibit the growth of Staphylococcus aureus ATCC 19095 and Listeria monocytogenes ATCC 7644. Two of these isolates, identified as Lactiplantibacillus plantarum and Lentilactobacillus buchneri, the sterile and neutral supernatants alone, were sufficient to inhibit L. monocytogenes growth. Principal component analysis (PCA) allowed the identification of functional groups based on proteolytic and lipolytic activity, osmotic stress resistance, and inhibition of L. monocytogenes. The type of cheese the isolates were recovered from influenced properties such as anti-listerial compounds and lipolytic enzyme production. The use of HTS and multivariate statistics allowed insights into a diverse set of LAB technological and biopreservation properties. These findings allow a profound knowledge of the heterogeneity of a large set of isolates, which can be further used to design starter cultures with varied and combined properties, such as biopreservation and technological features. Besides that, HTS makes it possible to analyze a vast panel of LAB strains, reducing costs and time within laboratory analysis, while avoiding the loss of information once all LAB are tested at the same time (differently from the traditional labor-intensive approach, in which a few numbers of strains is tested per time).

Using extended Bigelow meta-regressions for modelling the effects of temperature, pH, °Brix on the inactivation of heat resistant moulds.

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 zpH = 7.56; 95% CI: 5.13-13.5) and Neosartorya (overall zpH = 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 zpH = 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.

Evaluation of fruta-do-lobo (Solanum lycocarpum St. Hill) starch on the growth of probiotic strains.

Fruta-do-lobo (Solanum lycocarpum St. Hill) is a native fruit commonly used in Brazilian folk medicine as a hypoglycemic agent. These properties are attributed to their starch, mainly its resistant fraction. Resistant starch has shown to increases the growth of Bifidobacterium and Lactobacillus in the gut, even though not being selective for these strains. In this scenario, this study aimed to investigate the potential prebiotic activity of fruta-do-lobo starch (FLS). FLS showed around 30% of resistant starch and their prebiotic potential was evaluated with five probiotic strains L. acidophilus (LA3 and LA5), L. casei (LC01) and B. animalis (BB12) and B. lactis (BLC1) in a concentration range of 1.0-2.0% of starch. In a preliminary screening, we evaluated, during 48 h, the viability of the starch with promoting growth agent. An increase in the growth of the probiotic strains tested was observed. We also evaluated the microorganism's metabolic activity by assessing the short-chain fatty acid (SCFA) production, using the best starch growth promotion conditions (2% of FLS and strains BLC1, LA5, and LC01). As expected, MRS and lactose were preferentially metabolized by BLC1, with the highest growth rates: 0.231 and 0.224 h-1, respectively. However, for this strain, the FLS growth rate (0.222 h-1) was 65% higher than FOS (0.144 h-1). Also, for LA5 FLS promoted higher growth (0.150 h-1) than FOS (0.135 h-1). Additionally, FLS promoted acetate production. These data are promising and indicate that FLS may have prebiotic potential and more studies need to be done with pathogenic microorganisms.

An ordinal logistic regression approach to predict the variability on biofilm formation stages by five Salmonella enterica strains on polypropylene and glass surfaces as affected by pH, temperature and NaCl.

This study assessed the adhesion and formation of biofilm by five Salmonella enterica strains (S. Enteritidis 132, S. Infantis 176, S. Typhimurium 177, S. Heidelberg 281 and S. Corvallis 297) on polypropylene (PP) and glass (G) surfaces as affected by pH (4-7), NaCl concentration (0-10% w/v) and temperature (8-35 °C). Sessile counts <3 log CFU/cm2 were considered lack of adhesion (category 1), while counts ≥ 3 and < 5 log CFU/cm2 corresponded to adhesion (category 2) and counts ≥ 5 log CFU/cm2 corresponded biofilm formation (category 3). The obtained results categorized in these three responses were used to develop ordinal regression models to predict the probability of biofilm stages on PP- and G-surfaces. The experimental outcomes for lack of adhesion were >90% on PP- and G-surfaces. Generally, adhesion outcomes corresponded to approximately 36% of the total, whereas biofilm outcomes were close to 65% in both PP- and G-surfaces. The biofilm stages varied among the strains studied and with the material surface under the same experimental conditions. According to the generated ordinal models, the probability of adhesion and biofilm formation on PP-surface by the five S. enterica strains tested decreased at pH 4 or 5 in NaCl concentrations >4% and at a temperature <20 °C. On G-surface, the probability of adhesion increased pH 6 or 7, in the absence of NaCl and temperatures <20 °C, while, the probability of biofilm formation increased in the same pH, NaCl concentration up to 4% and temperatures ≥20 °C. This is the first study assessing the biofilm formation through categorical, ordinal responses and it shows that ordinal regression models can be useful to predict biofilm stages of S. enterica as a function of pH, NaCl, and temperature or their interactions.

The fate of Bacillus cereus and Geobacillus stearothermophilus during alkalization of cocoa as affected by alkali concentration and use of pre-roasted nibs.

Alkalization is a step of cocoa processing and consists of the use of alkali and high temperature to improve the sensorial and technological qualities of cocoa. Intense food processing can select spores, which can compromise safety and quality of the final product. Thus, the aim of this study was to evaluate the fate of B. cereus and G. stearothermophilus spores during the alkalization of pre-roasted (Pr) nibs (held at 120 °C) and unroasted (Ur) nibs (held at 90 °C) using potassium carbonate (0, 2, 4 and 6% w/w). In all conditions, log-linear inactivation kinetics with a tail was observed. The inactivation rate (kmax) for B. cereus varied from 0.065 to 1.67 min-1, whereas the kmax for G. stearothermophilus varied from 0.012 to 0.063 min-1. For both microorganisms, the lowest kmax values were observed during Ur nibs alkalization. The carbonate concentration increase promoted kmax values reduction. The highest tail values were observed for G. stearothermophilus in Ur nibs alkalization, reaching 3.04 log spores/g. Tail formation and low kmax values indicated that cocoa alkalization does not cause significant reductions on bacterial spore population. Therefore, the microbiological control should be primarily ensured by the raw material quality and by avoiding recontamination in the cocoa chain.

Brazilian Artisanal Cheeses: An Overview of their Characteristics, Main Types and Regulatory Aspects.

A large variety of cheeses exist in Brazil, reflecting historical and cultural aspects. Brazilian artisanal cheeses present differences in the processing, ripening time (when applied), type of milk used, texture, size, shape, color, moisture content, flavor, the use or not of starter cultures, among others. This review describes the main artisanal cheeses produced in Brazil, focusing on general and particular characteristics associated with their making process and geographical identity. Overall, the high variability of the physicochemical data and deficiency of information on sensorial properties of Brazilian artisanal cheeses were noticed. On the other hand, culture-dependent methods were mostly used to expand the knowledge into the microbiology of these cheeses, whereas their microbial diversity has been recently discovered through the use of 16S rRNA gene sequencing-based methods. The certification of a geographical indication for Brazilian artisanal cheeses may encompass an essential milestone for adding value to these products. Regardless of their significance in the diet, culture, and economy of producing regions, taken together, the reviewed literature discloses the need of insightful studies to generate scientific data to support the expansion of the market, while ensuring the protection of historic aspects related to the production of Brazilian artisanal cheeses.

Corrigendum: Quantifying the Responses of Three Bacillus cereus Strains in Isothermal Conditions and During Spray Drying of Different Carrier Agents.

[This corrects the article on p. 1113 in vol. 9, PMID: 29904375.].

Survival variability of 12 strains of Bacillus cereus yielded to spray drying of whole milk.

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 ±â€¯0.2 log10 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 (Tg), 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 Tg: 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 Tg 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.

Quantifying the Responses of Three Bacillus cereus Strains in Isothermal Conditions and During Spray Drying of Different Carrier Agents.

Spray drying is a widely used method for producing milk powder. This process is not aimed to cause microbial inactivation, thus sporeforming bacteria may be abundant in the microbiota of milk powder. The first aim of this study was to determine the inactivation kinetics parameters in capillary tubes of three Bacillus cereus strains (436, B63, 540) in three menstrua (whole milk, phosphate buffer, and talc suspension) at 90, 100, and 110°C. D-values for B. cereus in the three menstrua were not significantly different at the highest tested temperature (p > 0.05). Thus, talc was chosen as a carrier agent to allow the recovery of B. cereus from spray dried materials given its low interference on inactivation kinetics. B. cereus spores were also inoculated in whole milk and skim milk following spray drying at 95, 105, and 110°C (outlet temperature). After the spray drying runs, B. cereus spores were counted and the number of decimal reductions (γ) calculated. A correlation between the small diameter of the particles with the survival of spores of three B. cereus strains was found, and B. cereus 436 presented consistently the lowest γ no matter temperature and a carrier agent. The highest γ was found when talc powder was used, which suggest that this carrier agent does not protect B. cereus spores during spray drying. Spray drying of milk can lead to up to 4 γ (strain 540) of B. cereus spores but depending on the strain less than one γ (strain 436) could be observed. This study contributes to the knowledge on the microbiology of low water activity foods by providing novel findings regarding the fate of three B. cereus strains to different spray drying conditions. Acknowledging the variability of inactivation of B. cereus during spray drying is key in the current context of food safety in which the quantification of effects of unit operations must be known for the validation of processes and development of more robust formulations.