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.

Dry surface biofilms in the food processing industry: An overview on surface characteristics, adhesion and biofilm formation, detection of biofilms, and dry sanitization methods.

Bacterial biofilm formation in low moisture food processing (LMF) plants is related to matters of food safety, production efficiency, economic loss, and reduced consumer trust. Dry surfaces may appear dry to the naked eye, however, it is common to find a coverage of thin liquid films and microdroplets, known as microscopic surface wetness (MSW). The MSW may favor dry surface biofilm (DSB) formation. DSB formation is similar in other industries, it occurs through the processes of adhesion, production of extracellular polymeric substances, development of microcolonies and maturation, it is mediated by a quorum sensing (QS) system and is followed by dispersal, leading to disaggregation. Species that survive on dry surfaces develop tolerance to different stresses. DSB are recalcitrant and contribute to higher resistance to sanitation, becoming potential sources of contamination, related to the spoilage of processed products and foodborne disease outbreaks. In LMF industries, sanitization is performed using physical methods without the presence of water. Although alternative dry sanitizing methods can be efficiently used, additional studies are still required to develop and assess the effect of emerging technologies, and to propose possible combinations with traditional methods to enhance their effects on the sanitization process. Overall, more information about the different technologies can help to find the most appropriate method/s, contributing to the development of new sanitization protocols. Thus, this review aimed to identify the main characteristics and challenges of biofilm management in low moisture food industries, and summarizes the mechanisms of action of different dry sanitizing methods (alcohol, hot air, UV-C light, pulsed light, gaseous ozone, and cold plasma) and their effects on microbial metabolism.

Microbial resistance to sanitizers in the food industry: review.

Hygiene programs which comprise the cleaning and sanitization steps are part of the Good Hygiene Practices (GHP) and are considered essential to ensure food safety and quality. Inadequate hygiene practices may contribute to the occurrence of foodborne diseases, development of microbial resistance to sanitizers, and economic losses. In general, the sanitizer resistance is classified as intrinsic or acquired. The former is an inherent characteristic, naturally present in some microorganisms, whereas the latter is linked to genetic modifications that can occur at random or after continuous exposure to a nonnormal condition. The resistance mechanisms can involve changes in membrane permeability or in the efflux pump, and enzymatic activity. The efflux pump mechanism is the most elucidated in relation to the resistance caused by the use of different types of sanitizers. In addition, microbial resistance to sanitizers can also be favored in the presence of biofilms due to the protection given by the glycocalyx matrix and genetic changes. Therefore, this review aimed to show the main microbial resistance mechanisms to sanitizers, including genetic modifications, biofilm formation, and permeability barrier.

Salmonella spp. in low water activity food: Occurrence, survival mechanisms, and thermoresistance.

The occurrence of disease outbreaks involving low-water-activity (aw ) foods has gained increased prominence due in part to the fact that reducing free water in these foods is normally a measure that controls the growth and multiplication of pathogenic microorganisms. Salmonella, one of the main bacteria involved in these outbreaks, represents a major public health problem worldwide and in Brazil, which highlights the importance of good manufacturing and handling practices for food quality. The virulence of this pathogen, associated with its high ability to persist in the environment, makes Salmonella one of the main challenges for the food industry. The objectives of this article are to present the general characteristics, virulence, thermoresistance, control, and relevance of Salmonella in foodborne diseases, and describe the so-called low-water-activity foods and the salmonellosis outbreaks involving them.

Incidence of Bacillus cereus, Bacillus sporothermodurans and Geobacillus stearothermophilus in ultra-high temperature milk and biofilm formation capacity of isolates.

The presence of mesophilic and thermophilic spore-forming bacteria in UHT milk, as well as biofilm formation in dairy plants, are concerning. The current study explored the spore-forming bacilli diversity in 100 samples of UHT milk (skimmed and whole). Through this work, a total of 239 isolates from UHT milk samples were obtained. B. cereus s.s. was isolated from 7 samples, B. sporothermodurans from 19 and, G. stearothermophilus from 25 samples. Genes encoding hemolysin (HBL), and non-hemolytic (NHE) enterotoxins were detected in B. cereus s.s. isolates. All isolates of B. cereus s.s. (12) B. sporothermodurans (38), and G. stearothermophilus (47) were selected to verify the ability of biofilm formation in microtiter plates. The results showed all isolates could form biofilms. The OD595 values of biofilm formation varied between 0.14 and 1.04 for B. cereus, 0.20 to 1.87 for B. sporothermodurans, and 0.49 to 2.77 for G. stearothermophilus. The data highlights that the dairy industry needs to reinforce control in the initial quality of the raw material and in CIP cleaning procedures; avoiding biofilm formation and consequently a persistent microbiota in processing plants, which can shelter pathogenic species such as B. cereus s.s.

Competitive and/or Cooperative Interactions of Listeria monocytogenes With Bacillus cereus in Dual-Species Biofilm Formation.

Microorganisms in dairy industries can form monospecies, dual-species, or multispecies biofilms, showing cooperative or competitive behaviors, which might contribute to the reduction of efficiency of cleaning and sanitization processes and eventually turn into a potential source of contamination. This study proposes to evaluate the behavior of Listeria monocytogenes in monospecies biofilms, cocultured with Bacillus cereus. The isolates were of dairy origin, and the selection occurred after studies of competition among species. The biofilm formations on AISI 304 stainless steel at 25°C in a stationary culture were analyzed to observe the cooperative or competitive interactions among species, as well as the effect of pre-adhered cells. Biofilm formation assays were performed in four experiments: Experiment 1: in the presence of strains of antagonistic substance producer B. cereus (+); Experiment 2: extract of the antagonistic substance of B. cereus; Experiment 3: pre-adhered cells of B. cereus; and Experiment 4: pre-adhered cells of L. monocytogenes. Subsequently, cooperative behavior was observed by scanning electron microscopy. The L. monocytogenes monospecies biofilm counts of greater than 5 log colony-forming units (CFU)/cm2 were also observed in dual-species biofilms in the presence of B. cereus (non-producers of antagonist substance), showing cooperative behavior between species. However, in the presence of antagonistic substance produced by B. cereus, the counts were lower, 1.39 and 1.70 log CFU/cm2 (p > 0.05), indicating that the antagonistic substance contributes to competitive interactions. These data are relevant for the development of new studies to control L. monocytogenes in the dairy industry.

Avaliação microscópica de superfícies de aço inoxidável usados em ensaios de biofilmes / Microscopic evaluation on stainless steel surfaces used in biofilm assays

Superfícies de aço inoxidável com especificações determinadas pela American Society for Testing and Materials (ASTM) são usadas em testes in vitro para simular a formação e a remoção de biofilmes. Muitas vezes estas superfícies são reutilizadas nos ensaios de formação de biofilmes. O objetivo deste trabalho foi avaliar se cupons de aço inoxidável anteriormente utilizados para formação de biofilmes multiespécies podem ser reutilizados em novos ensaios. Assim, cupons submetidos a diferentes procedimentos de higienização foram analisados por microscopia eletrônica de varredura (MEV) e perfilometria . A reutilização das superfícies em novos experimentos deve ser realizada com cautela, aplicando procedimentos que removam as células bacterianas e a substância polimérica extracelular (EPS) aderidas na superfície. Além disso, observações da superfície (topografia e rugosidade) devem ser avaliadas, comprovando as especificações da ASTM.

Bacillus cereus e Geobacillus stearothermophilus em leite ultra alta temperatura (UAT) / Bacillus cereus and Geobacillus steartothermophilusin ultra high temperature (UHT) milk

O objetivo deste trabalho foi avaliar a presença de B. cereus e G. stearothermophilus em 100 amostras de leite UAT (integral e desnatado). O isolamento dos esporos e das células vegetativas seguiu metodologias oficiais, com pequenas modificações. B. cereus foi isolada de 7% amostras de leite UHT, de 6 diferentes marcas. As contagens máximas de células vegetativas e esporos de B. cer eus foram de 3,54 Log UFC/mL e 3,93 Log esporos/mL, respectivamente. A presença dos genes codificadores de enterotoxina não hemolítica (NHE) foi observada em 33% dos isolados e da hemolisina (HBL ) em 100% dos isolados. O gene hblA foi encontrado em 91,6 % dos isolados, porém nenhum isolado apresentou os 3 genes do complexo HBL. G. stearothermophilus foi identificada em 22,8% (34/149) dos isolados de esporo altamente resistente ao calor (HRRS), representando 18% das amostras de leite UAT e as contagens de esporos variaram de < 1Log a 3,40 Log esporos/mL.