Biofilm formation and biocides sensitivity of Pseudomonas marginalis isolated from a maple sap collection system

The susceptibility of planktonic and biofilm cells of Pseudomonas marginalis toward four commonly used biocides at different temperatures (15 and 30 degrees C) and biofilm growth times (24 and 48 h) was assessed. Using the MBEC biofilm device, biofilm production in maple sap was shown to be highly reproducible for each set of conditions tested. Biofilm formation was influenced by growth temperature and time. A temperature of 15 degrees C and incubation time of 24 h yielded fewer CFU per peg and showed fewer adhered cells and typical biofilm structures, based on scanning electron microscopy observations as compared with other conditions. Minimal biofilm eradication concentration values for P. marginalis were significantly greater (P. < 0.001) than were MBCs for planktonic cells and for every biocide tested, with the exception of minimal biofilm eradication concentration values for peracetic acid at 15 degrees C and 24 h. Sodium hypochlorite and peracetic acid sanitizers were able to eliminate P. marginalis biofilms at lower concentrations as compared with hydrogen peroxide- and quaternary ammonium-based sanitizers (P < 0.001). According to the results obtained, sodium hypochlorite and peracetic acid sanitizers would be more appropriate for maple sap collection system sanitation.     

Compositions of maple sap microflora and collection system biofilms evaluated by scanning electron microscopy and denaturing gradient gel electrophoresis

The bacterial microflora of maple sap and biofilms in collection system tubing were studied through the use of bacterial counts, scanning electron microscopy (SEM) of surfaces and the analysis of 16S rRNA gene by denaturing gradient gel electrophoresis (DGGE). Samples were taken at five times during the 2002 and 2003 seasons in order to follow the changes in the microflora of this complex ecosystem. Bacterial counts showed the growth of bacterial populations as the season advanced. These populations were mainly composed of psychrotrophic bacteria and Pseudomonas spp. SEM results confirmed the suspected presence of biofilms on the inner surfaces of tubing samples. Bacterial colonization and biofilm formation progressively increased during the season for both lateral and main line surfaces, and biofilms were mainly composed of rod shape bacteria. The bacterial microflora profiles obtained for sap and corresponding biofilm by DGGE showed up to 12 major bands. The Shannon-Weaver index of diversity (H) calculated from DGGE bands were statistically higher for sap samples compared to biofilm. The diversity index was relatively stable or increasing for lateral line sap and biofilm samples during the season while the diversity index for sap and biofilm samples of the main line showed a decreasing profile as the season progressed. Sequence analysis of major DGGE bands revealed the predominance of bacteria from the genera Pseudomonas, Rahnella and another, unidentified genus. The results describe the composition of sap collection system microflora as well as the formation of biofilms and will be useful for further studies on factors affecting maple product quality.     

The effect of drinking milk containing conjugated linoleic acid on fecal microbiological profile,enzymatic activity,and fecal characteristics in humans

Background  

The primary objective was to determine whether consumption of conjugated linoleic acids (CLAs) affected the fecal microbiota composition, fecal enzyme activity or fecal composition.     

Psychrotrophs in dairy products: Their effects and their control

Health concerns and technological effects of psychrotrophic bacteria in dairy products are reviewed, as well as methods to control their presence and development. The various Gram‐negative and Gram‐positive psychrotrophic species are listed and, with respect to pathogenic psychrotrophs, emphasis is given on Listeria monocytogenes, Yersinia enterocolytica, and Bacillus cereus. The influence of psychrotrophic bacteria on the quality of raw milk, pasteurized and UHT milks, butter, ice cream, cheese, and powders is examined. Public health considerations of Listeria monocytogenes, Yersinia enterocolytica, and Bacillus cereus of these various dairy products are also presented. Methods that can be used to eliminate or control the development of psychrotropic bacteria include low or high temperatures, chemicals, gases, the lactoperoxidase system, lactic acid bacteria, microfiltration, bactofugation, lactoferrin‐related proteins, sanitation, flavors, and naturally occurring spore germinants.     

Efficiency of sanitizing agents for destroying Listeria monocytogenes on contaminated surfaces

In an effort to control the potential hazard of dairy product contamination by contact with processing surfaces, the efficiency of four commercial sanitizing agents was evaluated using the AOAC use-dilution method for their bactericidal activity at 4 and 20 degrees C against Listeria monocytogenes strain Scott-A attached on four types of surfaces (stainless steel, glass, polypropylene, and rubber). Our results indicate that all sanitizers tested were more efficient against L. monocytogenes attached to nonporous surfaces than to porous surfaces. After 10 min of contact time, the limit concentration of disinfectants was at least 5 to 10 times higher for sanitizing rubber than stainless steel or glass surfaces. Concentrations of each sanitizer needed to be higher at sanitation at 4 degrees C than at 20 degrees C to destroy L. monocytogenes attached to stainless steel, glass and rubber when surface contamination was achieved at 4 or 20 degrees C.     

Evidence for Escherichia coli O157:H7 attachment to water distribution pipe materials by scanning electron microscopy

Scanning electron microscopy observation was used to investigate the adhesion of Escherichia coli O157:H7 on water distribution pipe surfaces such as copper and polyethylene plastic at different contact times and storage temperatures. Our results indicated that E. coli cells could easily attach to both surface types after exposures as short as 1 or 4 h at ambient (20 degrees C) and refrigeration temperatures (4 degrees C). Also, we found that copper surfaces have a higher number of attached E. coli cells than plastic surfaces. The number of cells attached to each type of material depended on the nature of the water distribution pipe surfaces and the length of contact time. In addition, the surface energy value of each surface estimated by contact angle measurements using water, alpha-bromonaphthalene, and dimethyl sulfoxide as wetting agents showed that both copper (41.2 megajoules [MJ] m(-2)) and plastic (45.8 MJ x m(-2)) have a low energy surface. In no cases could evidence of extracellular material be observed on surfaces with either exposure condition.     

Efficacy of egg cleaning compounds on eggshells contaminated with Salmonella enterica serovar Enteritidis

Salmonella Enteritidis infections of egg contents can be related to external contamination of the shell. In this study, the efficacy of three commercial cleaning and/or sanitizing compounds (sodium carbonate, sodium hypochlorite, and potassium hydroxide) was evaluated for bactericidal activity at pH values of 10, 11, and 12 against various concentrations (10(2), 10(4), or 10(6) CFU/ml) of Salmonella Enteritidis inoculated onto the eggshell surface. Efficacy of these chemical agents was also assessed against Salmonella Enteritidis in aqueous suspension. Our results indicated that none of the chemicals applied at the recommended manufacturer's concentrations (sodium carbonate, 36 ppm; other treatments, 200 ppm) could eliminate Salmonella Enteritidis from eggshells artificially contaminated with the highest bacterial concentrations (10(4) or 10(6) CFU/ml). Higher concentrations of each product, at least 5 to 20 times greater than recommended doses, were needed to destroy the bacteria on egg surfaces. However, at or slightly above the manufacturer's recommended use concentrations, all three formulations were effective against Salmonella Enteritidis in aqueous suspension (10(8) CFU/ml) or on eggshells contaminated with 10(2) CFU/ml. For both shell and suspension assays, inactivation of Salmonella Enteritidis occurred at lower concentrations at pH 12 than at pH 11 and 10. Contact time between chemicals and Salmonella apparently influenced bacterial inactivation. Extended contact times (2 to 10 min) reduced minimum chemical concentrations necessary to inactivate the bacteria. However, neither pH nor contact time influenced Salmonella Enteritidis inactivation when the initial bacterial numbers on eggshells were high.     

Substitution of Two Active‐Site Residues Alters C9‐Hydroxylation in a Class II Diterpene Synthase

Diterpenes form a vast and diverse class of natural products of both ecological and economic importance. Class II diterpene synthase (diTPS) enzymes control the committed biosynthetic reactions underlying diterpene chemical diversity. Homology modelling with site‐directed mutagenesis identified two active‐site residues in the horehound (Marrubium vulgare) class II diTPS peregrinol diphosphate synthase (MvCPS1); residue substitutions abolished the unique MvCPS1‐catalysed water‐capture reaction at C9 and redirected enzyme activity toward formation of an alternative product, halima‐5(10),13‐dienyl diphosphate. These findings contributed new insight into the steric interactions that govern diTPS‐catalysed regiospecific oxygenation reactions and highlight the feasibility of diTPS engineering to provide a broader spectrum of bioactive diterpene natural products.     

Attachment of Arcobacter butzleri,a new waterborne pathogen,to water distribution pipe surfaces

The capability of Arcobacter butzleri to attach to various water distribution pipe surfaces, such as stainless steel, copper, and plastic, was evaluated using scanning electron microscopy. Our results indicated that Arcobacter cells could easily attach to all surface types and the number of attached cells depended on the length of exposure and temperatures (4 and 20 degrees C). Extracellular fibrils were also observed on the stainless steel surface, especially after 72 h of contact times at both refrigeration and ambient temperatures. In addition, the surface energy value of each material was estimated by contact angle measurements using water, alpha-bromonaphthalene, and dimethylsulfoxide. The surface energy of A. butzleri was 58.6 mJ x m(-2) and the surface energy values of the three surfaces studied showed that plastic had a low energy surface (26.1 mJ x m(-2)) as did copper (45.8 mJ x m(-2)) and stainless steel (65.7 mJ x m(-2)).