Yeast biofilms on abiotic surfaces: Adhesion factors and control methods.

Biofilms are highly resistant to antimicrobials and are a common problem in many industries, including pharmaceutical, food and beverage. Yeast biofilms can be formed by various yeast species, including Candida albicans, Saccharomyces cerevisiae, and Cryptococcus neoformans. Yeast biofilm formation is a complex process that involves several stages, including reversible adhesion, followed by irreversible adhesion, colonization, exopolysaccharide matrix formation, maturation and dispersion. Intercellular communication in yeast biofilms (quorum-sensing mechanism), environmental factors (pH, temperature, composition of the culture medium), and physicochemical factors (hydrophobicity, Lifshitz-van der Waals and Lewis acid-base properties, and electrostatic interactions) are essential to the adhesion process. Studies on the adhesion of yeast to abiotic surfaces such as stainless steel, wood, plastic polymers, and glass are still scarce, representing a gap in the field. The biofilm control formation can be a challenging task for food industry. However, some strategies can help to reduce biofilm formation, such as good hygiene practices, including regular cleaning and disinfection of surfaces. The use of antimicrobials and alternative methods to remove the yeast biofilms may also be helpful to ensure food safety. Furthermore, physical control measures such as biosensors and advanced identification techniques are promising for yeast biofilms control. However, there is a gap in understanding why some yeast strains are more tolerant or resistant to sanitization methods. A better understanding of tolerance and resistance mechanisms can help researchers and industry professionals to develop more effective and targeted sanitization strategies to prevent bacterial contamination and ensure product quality. This review aimed to identify the most important information about yeast biofilms in the food industry, followed by the removal of these biofilms by antimicrobial agents. In addition, the review summarizes the alternative sanitizing methods and future perspectives for controlling yeast biofilm formation by biosensors.

Potential of electrolyzed water to inactivate bread and cheese spoilage fungi.

Controlling food spoilage fungi remains a challenge for food industries, and regulations on the usage of chemical disinfectants are becoming restrictive. Then, this study aimed to evaluate electrolyzed water (EW) as a sustainable alternative for food spoilage fungi inactivation. The experiment was carried out according to the protocol for testing the antifungal effects of chemical sanitizers by the European Committee for Standardization (CEN), using acidic electrolyzed water (AEW-AAC: 85 ppm; pH: 2.65; ORP: 1120 mV) and a basic electrolyzed water (BEW- pH: 11.12; ORP: -209 mV) to inactivate spoilage fungi strains from bread (Hyphopichia burtonii and Penicillium roqueforti) and cheese (P. roqueforti and Penicillium commune), besides the standard fungi for this type of essay (Candida albicans and Aspergillus brasiliensis). AEW presented a higher antifungal effect, inactivating an average of 89 % of the exposed population when compared to its respective BEW, which inactivates about 81.5 %. In general, the standard strains A. brasiliensis (ATCC 16404) and Candida albicans (ATCC 24433) were more sensitive to both AEW and BEW than the food-spoilage strains. Among those, P. roqueforti strains were the most sensitive, followed by P. commune strains, while H. burtonii strains were the most tolerant. EW can be a sustainable alternative for product surface and facility cleaning with further antifungal action when a sanitization step is not mandatory or needed. Future studies searching for conditions to improve the antifungal action of EW could make their industrial usage more viable.

Efficacy of weak acid preservatives on spoilage fungi of bakery products.

Organic acids and their salts are usually the first choice in the bread industry to restrict fungal spoilage, but their efficacy is pH-dependent and spoilage by fungi remains as a common threat. Therefore, this study aimed to evaluate the susceptibility of spoilage fungi of bakery products to acetic, sorbic, and propionic acids at different pH. Penicillium roqueforti, Penicilium paneum, Aspergillus pseudoglaucus, Aspergillus montevidensis and Hyphopichia burtonii strains isolated from spoiled products had their minimum inhibitory concentration (MIC) defined by macrodilution. The concentrations tested were: (i) sorbic acid up to 32 mM; (ii) propionic acid up to 1024 mM and (iii) acetic acid up to 800 mM with pH adjusted in 4.5, 5.0, 5.0 and 6.0 after setting the agent concentration. The lowest MICs for all agents were obtained at pH 4.5, usually doubling with every 0.5 pH increase. P. roqueforti strains isolated from spoiled products were the most resistant to all tested preservatives; while strains of the related species P. paneum, showed similar tolerance to acetic and propionic acids but was double more susceptible to sorbic acid. Strains of A. pseudoglaucus and A. montevidensis were indistinctly susceptible to the preservatives and were the most susceptible species to propionic and acetic acids. H. burtonii strains demonstrated the most variable behaviour in comparison to the other strains being the most susceptible to sorbic acid, were like Aspergillus strains regarding propionic acid, but tolerate well acetic acid. Propionic acid concentrations usually allowed in baked goods are lower than the concentrations required to inhibit the most tolerant isolates tested in this study. The same is true for sorbic acid at higher pH levels. Spoilage species of bakery ware presents a distinct susceptibility profile to the preservatives commonly used in this sector, but the high tolerance observed is a cause of concern.

Influence of type, concentration, exposure time, temperature, and presence of organic load on the antifungal efficacy of industrial sanitizers against Aspergillus brasiliensis (ATCC 16404).

Parameters such as type and concentration of the active compound, exposure time, application temperature, and organic load presence influence the antimicrobial action of sanitizers, although there is little data in the literature. Thus, this study aimed to evaluate the antifungal efficacy of different chemical sanitizers under different conditions according to the European Committee for Standardization (CEN). Aspergillus brasiliensis (ATCC 16404) was exposed to four compounds (benzalkonium chloride, iodine, peracetic acid, and sodium hypochlorite) at two different concentrations (minimum and maximum described on the product label), different exposure times (5, 10, and 15 min), temperatures (10, 20, 30, and 40 °C), and the presence or absence of an organic load. All parameters, including the type of sanitizer, influenced the antifungal efficacy of the tested compounds. Peracetic acid and benzalkonium chloride were the best antifungal sanitizers. The efficacy of peracetic acid increased as temperatures rose, although the opposite effect was observed for benzalkonium chloride. Sodium hypochlorite was ineffective under all tested conditions. In general, 5 min of sanitizer exposure is not enough and >10 min are necessary for effective fungal inactivation. The presence of organic load reduced sanitizer efficacy in most of the tested situations, and when comparing the efficacy of each compound in the presence and absence of an organic load, a difference of up to 1.5 log CFU was observed. The lowest concentration recommended on the sanitizer label is ineffective for 99.9% fungal inactivation, even at the highest exposure time (15 min) or under the best conditions of temperature and organic load absence. Knowledge of the influence exerted by these parameters contributes to successful hygiene since the person responsible for the sanitization process in the food facility can select and apply a certain compound in the most favorable conditions for maximum antifungal efficacy.

Comparison of electrolized water and multiple chemical sanitizer action against heat-resistant molds (HRM).

This study aimed to evaluate the sensitivity of heat-resistant molds isolated from spoiled thermally processed foods to antimicrobial compounds used for food industry sanitation. An ortho-phenylphenol-based smoke generator sanitizer, liquid chemical sanitizers (benzalkonium chloride, biguanide, iodine, peracetic acid, and sodium hypochlorite), and acidic and alkaline electrolyzed water were used against Aspergillus australensis (MB 2579; NFF 02), Aspergillus aureoluteus (NFC1), Paecilomyces fulvus (PFF 01), Paecilomyces niveus (PNT 01; PNDC 01; PNB1 01), and Paecilomyces variotii (PV 01; PV 01; PVCH 03). The fungal strains were exposed separately to liquid sanitizers and electrolyzed water in stainless steel discs for 15 min following the European Committee for Standardization (CEN) recommendations. Moreover, the fungal strains were exposed to the smoke generator sanitizer for 7 h following French protocol NF-T-72281. The best results of fungal inactivation were achieved when the highest concentration specified in the label of these sanitizers was tested. On the opposite, the lowest concentration specified in the label should be avoided since it was ineffective in most cases (94%). The ortho-phenyphenol-based smoke generator sanitizer and peracetic acid (1%) showed the best results of spore inactivation, while iodine and benzalkonium chloride achieved satisfactory results against the strains evaluated. Sodium hypochlorite and biguanide were ineffective against most of the fungi studied at all concentrations tested. Acidic and basic electrolyzed water was also ineffective to achieve the 3-log CFU reduction required in the concentrations tested. In general, Paecilomyces spp. was more sensitive than Aspergillus spp. against all sanitizers evaluated, whereas A. aureoluteus NFC1 was resistant to all agents and concentrations tested. The heat-resistant fungal strains showed varied sensitivity against the different agents. Notably, the two most effective commercial sanitizers against the heat-resistant strains were ineffective against the filamentous fungi recommended for sanitizer testing (A. brasiliensis ATCC 16404), which demonstrates the relevance of testing fungal isolates that cause spoilage to choose the most effective compound and obtain the best results of fungal control.

Spoilage fungi in a bread factory in Brazil: Diversity and incidence through the bread-making process.

This study aimed to verify the main fungal species involved in the deterioration of different types of bread and to identify the possible sources of contamination of these products. Samples of raw materials (n = 127), environmental air (n = 50) and moldy bread (n = 90) were analyzed. Aspergillus candidus, Wallemia sebi, and Penicillium roqueforti were the predominant species in the raw materials and were isolated in samples of wheat flour, in two-thirds of the samples of rye and 62.5% of the wheat flour. Penicillium roqueforti was isolated from all types of moldy bread analyzed and Hyphopichia burtoni was also present in samples of moldy wheat and rye bread. These two species were also recovered during air sampling from baking industry facilities (cooling and slice and package areas), which may be crucial for product contamination after baking. Hygienic measures to reduce airborne contamination during the cooling and packaging of food should be taken to prevent the early deterioration of bread.

Ochratoxin A production by Aspergillus westerdijkiae in Italian-type salami.

The occurrence of ochratoxin A (OTA) in matured meat products can be attributed to mycotoxin coming from raw materials and/or molds that develop on the product surface during ripening. This work aimed to evaluate OTA production by Aspergillus westerdijkiae inoculated on the surface of sausages and its diffusion into the product throughout ripening, study the effect of relative humidity (RH) on the production of this mycotoxin, and investigate the presence of OTA in dry fermented sausages that naturally present intense contamination by A. westerdijkiae. Italian-type sausages were surface inoculated with A. westerdijkiae and two distinct experiments were performed. In the first, the sausages were matured in a chamber with decreasing RH (from 95 to 75%) for 35 days. In the other, the sausages were incubated under different RH (79, 85 or 95%) for 21 days. Samples were taken at the beginning of the experiments and every 7 days, subdividing into casing, outer border, and core for analyses. Sausage samples naturally spoiled by ochratoxigenic fungi were collected during sanitary inspection. Even in the presence of A. westerdijkiae mycelia, no OTA was detected for up to 7 days of sausage maturation. On the other hand, this study demonstrated that the growth of A. westerdijkiae on salami surface produces high amounts of OTA on the casing and allows its diffusion through the casing with contamination to the outer border of sausages. In the same way, it shows that under similar water activity values of substrate, RH influences the amount of OTA produced. Conversely, OTA was restricted to the casing in the naturally contaminated sausages.

Antifungal activity of commercial sanitizers against strains of Penicillium roqueforti, Penicillium paneum, Hyphopichia burtonii, and Aspergillus pseudoglaucus: Bakery spoilage fungi.

Information on the sensitivity of spoilage fungi of bakery products to sanitizing agents is scarce in the literature. Thus, the aim of this study was to evaluate the antifungal activity of different classes of commercial sanitizers, which have permitted use in the food industry, on the main fungi involved in spoiling bakery products. The tests were carried out according to the protocol for testing the antifungal effect of chemical sanitizers of the European Committee for Standardization (CEN), with adaptations. Different strains of six isolated fungal species responsible for spoiling bakery products (Penicillium roqueforti, Penicillium paneum, Hyphopichia burtonii, and Aspergillus pseudoglaucus) were tested against five sanitizers at three concentrations: benzalkonium chloride (0.3%, 2.5%, 5%), biguanide (2%, 3.5%, 5%), peracetic acid (0.15%, 1.5%, 3%), quaternary ammonium (0.3%, 2.5%, 5%), and sodium hypochlorite (0.01%, 0.1%, 0.2%). Peracetic acid was the most effective sanitizes considering the genera, species, and concentrations evaluated, generally being capable of reductions between 2 and 4 logs of initial control tested. Biguanide should not be the compound of choice when the main goal of the bakery industry is fungal control.

Evaluation of mediators of oxidative stress and inflammation in patients with acute appendicitis.

CONTEXT: This study aims to explore the potential of new inflammatory markers for improving the challenging diagnosis of acute appendicitis (AA). METHODS: Levels of IL-1, IL-6, IL-8, IL-10, CRP, INF-γ, and TNF-α in serum were measured in 73 patients with AA. Oxidative stress and antioxidant enzymes were analyzed. RESULTS: Serum levels of interleukins, TNF-α, and INF-γ were significantly elevated in patients with appendicitis (p < 0.0001), except for IL-10, which presented decreased levels. There were no significant differences in SOD (p = 0.29), CAT (p = 0.19), or TBARS levels (p = 0.18), whereas protein carbonyls presented significant increase (p < 0.0001). CONCLUSION: Evaluating these biomarkers could aid in diagnosing AA.