1186 Journal of Food Protection, Vol. 78, No. 6, 2015, Pages 1186–1190 doi:10.4315/0362-028X.JFP-14-440 Copyright G, International Association for Food Protection Research Note Adhesion of Asaia bogorensis to Glass and Polystyrene in the Presence of Cranberry Juice HUBERT ANTOLAK,* DOROTA KREGIEL, AND AGATA CZYZOWSKA Institute of Fermentation Technology and Microbiology, Lodz University of Technology, Wolczanska 171/173, 90-924 Lodz, Poland MS 14-440: Received 10 September 2014/Accepted 29 January 2015 Downloaded from http://meridian.allenpress.com/jfp/article-pdf/78/6/1186/1686447/0362-028x_jfp-14-440.pdf by guest on 02 October 2021 ABSTRACT The aim of the study was to evaluate the adhesion abilities of the acetic acid bacterium Asaia bogorensis to glass and polystyrene in the presence of American cranberry (Vaccinium macrocarpon) juice. The strain of A. bogorensis used was isolated from spoiled commercial fruit-flavored drinking water. The cranberry juice was analyzed for polyphenols, organic acids, and carbohydrates using high-performance liquid chromatography and liquid chromatography–mass spectrometry techniques. The adhesive abilities of bacterial cells in culture medium supplemented with cranberry juice were determined using luminometry and microscopy. The viability of adhered and planktonic bacterial cells was determined by the plate count method, and the relative adhesion coefficient was calculated. This strain of A. bogorensis was characterized by strong adhesion properties that were dependent upon the type of surface. The highest level of cell adhesion was found on the polystyrene. However, in the presence of 10% cranberry juice, attachment of bacterial cells was three times lower. Chemical analysis of juice revealed the presence of sugars, organic acids, and anthocyanins, which were identified as galactosides, glucosides, and arabinosides of cyanidin and peonidin. A-type proanthocyanidins responsible for the antiadhesion properties of V. macrocarpon also were detected. Red cranberries (Vaccinium macrocarpon Ait., family Helicobacter pylori to human erythrocytes and human Ericaceae) are native to North America. Cranberries were gastric mucous (3) and can inhibit coaggregation and reduce first used by Native Americans, who discovered the wild salivary counts of oral bacteria (24). The antiadhesive berry’s versatility as a food, fabric dye, and healing agent. properties of cranberry can affect various microorganisms The name ‘‘cranberry’’ derives from ‘‘craneberry,’’ the (20), but less attention has been paid to cranberry juice as an name used by early European settlers in America because antiadhesive and antibacterial agent against acidophilic or the small pink or red blossoms resemble the head and bill of acid-tolerant food spoilage bacteria. a crane. Today, cranberries are used for juices, cocktails, The aims of this study were to investigate the effects of jellies, and sauces. cranberry juice on the food-spoiling acetic acid bacterium Scientific studies have revealed that cranberry com- Asaia bogorensis. Many Asaia species share the ability to pounds have potential health effects against cancer, aging coaggregate, forming both flocks in beverages and biofilms and neurological diseases, inflammation, diabetes, and on production surfaces (11–13). These biofilms are very bacterial infections (18). The cranberry also has been stable and difficult to remove using standard cleaning and recognized for its contribution to maintenance of a healthy disinfection methods (25). The type of food packaging used urinary tract. Although the earliest studies suggested that can affect the development of spoilage microflora (8). acidification of urine was responsible for the cranberry’s Polystyrene and glass are packaging materials commonly effect on urinary tract health, research since the 1980s has used in the beverage industry (1, 22). We studied the ability focused on the specific antiadhesive properties of cranberry of A. bogorensis cells to adhere to two carriers, glass and juice. Zafriri and coworkers (29) were the first to postulate polystyrene, in the presence of 10% (vol/vol) cranberry that various compounds in cranberries could affect P and juice. This work is the first to describe the use of cranberry type 1 fimbriae of Escherichia coli, and they found that juice as a natural antiadhesive agent against the acidophilic fructose inhibited the adhesion of bacteria with type 1 food spoilage bacteria Asaia spp. fimbriae. In 1998, Howell et al. (10) identified specific proanthocyanidin compounds in cranberries that caused P- MATERIALS AND METHODS fimbriated E. coli to exhibit antiadhesion properties, and the Bacterial cultures. A. bogorensis strain FFMW (GenBank chemical structure of these compounds was later described KC756841.1) was isolated from flavored mineral water in Poland, (6). Cranberry juice inhibits the adhesion of three strains of and polyphasic identification was based on both physiological and genetic testing (12, 13). For aerobic bacterial cultures, commercial * Author for correspondence. Tel: z48426313475; Fax: z48426365976; fruit-flavored mineral water was used as the liquid culture medium: E-mail: [email protected]. 8.1% saccharose (wt/vol), 0.05% strawberry flavor (wt/vol), J. Food Prot., Vol. 78, No. 6 CRANBERRY JUICE INHIBITS A. BOGORENSIS ADHESION 1187 Downloaded from http://meridian.allenpress.com/jfp/article-pdf/78/6/1186/1686447/0362-028x_jfp-14-440.pdf by guest on 02 October 2021 FIGURE 1. Chromatographic profiles of cranberry juice for acids and saccharides (A) and polyphenols (B). (A) Peak 1, citric acid; peak 2, glucose; peak 3, malic acid; peak 4, fructose and quinic acid. (B) Peak 1, cyanidin-3-galactoside; peak 5, cyanidin-3-arabinoside; peak 7, peonidin-3-galactoside; peak 8, peonidin-3-glucoside; peak 11, peonidin-3-arabinoside. 0.16% citric acid (wt/vol), 0.02% sodium benzoate (wt/vol), and Carriers. Polystyrene slides (76 by 26 mm; Packor 0.02% velcorin (wt/vol), pH 3.5. The fruit-flavored mineral water Packaging, Skierniewice, Poland) were used as sample carriers. was sterilized by membrane filtration (0.45-mm-pore-size filter; This material is approved by the Polish National Institute of Public Millipore, Billerica, MA). The culture medium (20 ml) was poured Health for contact with food. White glass slides (76 by 26 mm; Star into a 25-ml Erlenmeyer flask, and sterile sample carriers were Frost, Knittel Glass, Braunschweig, Germany) were used as the placed vertically so that half of the carrier was immersed in reference material. medium, leaving the other part suspended over the medium. Culture media with and without cranberry juice (10%, vol/vol) Adhesion analysis. The amount of inoculum was standard- were used. Two types of medium contained cranberry juice: (i) ized to obtain levels in culture medium of approximately 104 CFU/ without pH correction and (ii) with pH correction using 0.1 M ml at the beginning of the experiment. The samples were incubated NaOH. at 25uC on a laboratory shaker (130 rpm) for 6 days. Bacterial growth and adhesion in the presence of cranberry juice were Cranberry juice. Cranberries (V. macrocarpon) were analyzed by comparing results obtained after 0, 3, and 6 days of purchased from a local retail market (Lodz, Poland). The fresh incubation. The results of experiments without cranberry supple- fruits were washed with sterile water, lightly dried with a sterile mentation were used as the control. The analysis of adhesion to the paper towel, frozen at 220uC, and stored frozen for 3 months. carriers was done by luminometry and the plate count method. For After defrosting, the fruits were pressed with a juice extractor luminometric tests, the carrier plate was removed from the culture (MES3000, Bosch, Warsaw, Poland). The cloudy juice was filtered medium, washed with sterile distilled water, and swabbed with with a qualitative paper filter (Whatman, Clifton, NJ) and then with pens for sampling free ATP (Merck, Darmstadt, Germany). a sterile 0.45-mm-pore-size membrane filter (Millipore). Measurement was reported in relative light units (RLU) from a 1188 ANTOLAK ET AL. J. Food Prot., Vol. 78, No. 6 FIGURE 2. Adhesion of Asaia bogorensis to glass and polysty- rene in flavored mineral water (light shading), flavored mineral Downloaded from http://meridian.allenpress.com/jfp/article-pdf/78/6/1186/1686447/0362-028x_jfp-14-440.pdf by guest on 02 October 2021 water with cranberry juice (medium shading), and flavored mineral water with cranberry juice after pH correction (dark shading). Data were analyzed with a one-way repeated measures ANOVA. * P , 0.05; ** P , 0.01; *** P , 0.001 compared with the control medium (flavored mineral water). HY-LiTE2 luminometer (Merck). For the determination of viable bacterial cells, both on the tested surface and in the culture medium, the colony method was used. For the determination of the number of attached bacterial cells, previously removed biofilm was vortexed in 0.85% saline with 0.1% Tween 80, and dilutions were transferred onto GC agar (2% D-glucose, 0.3% peptone [wt/vol], 0.3% yeast extract [wt/vol], and 0.7% CaCO3 [wt/vol]). After incubation (25uC for 96 h), the Asaia colonies were counted, and FIGURE 3. Relative adhesion coefficient A of Asaia bogorensis the number of attached bacterial cells per square centimeter of the calculated for glass and polystyrene in flavored mineral water carrier was determined. Corresponding medium and conditions (light shading), flavored mineral water with cranberry juice were used for