Utilization of Piper betle L. Extract for Inactivating Foodborne Bacterial Biofilms on Pitted and Smooth Stainless Steel Surfaces.

Biofilms are a significant concern in the food industry. The utilization of plant-derived compounds to inactivate biofilms on food contact surfaces has not been widely reported. Also, the increasing negative perception of consumers against synthetic sanitizers has encouraged the hunt for natural compounds as alternatives. Therefore, in this study we evaluated the antimicrobial activities of ethanol extracts, acetone extracts, and essential oils (EOs) of seven culinary herbs against Salmonella enterica serotype Typhimurium and Listeria innocua using the broth microdilution assay. Among all tested extracts and EOs, the ethanol extract of Piper betle L. exhibited the most efficient antimicrobial activities. To evaluate the biofilm inactivation effect, S. Typhimurium and L. innocua biofilms on pitted and smooth stainless steel (SS) coupons were exposed to P. betle ethanol extract (12.5 mg/ml), sodium hypochlorite (NaClO; 200 ppm), hydrogen peroxide (HP; 1100 ppm), and benzalkonium chloride (BKC; 400 ppm) for 15 min. Results showed that, for the untreated controls, higher sessile cell counts were observed on pitted SS versus smooth SS coupons. Overall, biofilm inactivation efficacies of the tested sanitizers followed the trend of P. betle extract ≥ BKC > NaClO > HP. The surface condition of SS did not affect the biofilm inactivation effect of each tested sanitizer. The contact angle results revealed P. betle ethanol extract could increase the surface wettability of SS coupons. This research suggests P. betle extract might be utilized as an alternative sanitizer in food processing facilities.

Transmission electron microscopy of precipitation in fine-grained heat-affected zone of Grade91 steel weld during creep exposure.

Precipitation during creep exposure at 550 °C in the fine grain heat-affected zone of a Grade91 steel weld produced by gas tungsten arc welding with no filler and post-weld heat treatment at 760 °C for 30 min. has been investigated. The applied tension load during creep testing was 165 MPa, the creep rupture time was 1138 hrs. (100% creep aging), and the creep exposure times were 911, 569, 341, 223 and 114 hrs. for 80%, 50%, 30%, 20% and 10% creep aging, respectively. Light microscopy and scanning electron microscopy were used to examine the heat-affected zone in the steel weld. Grain size in the fine grain heat-affected zone was measured by a circular intercept method on secondary electron images from scanning electron microscopy. Precipitates were detached from the fine grain heat-affected zone by a carbon-film extraction replica technique and investigated by transmission electron microscopy. During creep, a continual increase in grain size was observed in the fine grain heat-affected zone, starting from about 1.70 µm after post-weld heat treatment to about 2.30 µm at 100% creep aging. Precipitates found during creep were M23C6, M7C3, M6C, MX, M2X and high-Si Laves phase. M23C6 was found in all creep conditions until rupture, mainly at prior austenite grain boundaries and sub-boundaries. M7C3, M6C and high-Si Laves phase were observed during creep aging, mainly nearby grain boundaries and sub-boundaries. Relatively fine MX and M2X were found after 20% creep aging until rupture, mainly within grain interiors. Detailed criteria for identification of these precipitate phases, based mainly on energy dispersive X-ray spectrometry and selected area electron diffraction using transmission electron microscopy, is given and a major cause of premature loss of creep strength in Grade91 steel weld is discussed. This is important in understanding creep behavior of steel parts used for boiler tubing, heat exchangers, and steam piping systems in fossil-fuel power plants.

In-Situ Monitoring of Real-Time Loop-Mediated Isothermal Amplification with QCM: Detecting Listeria monocytogenes.

Functionalized DNA sequences are promising sensing elements to combine with transducers for bio-sensing specific target microbes. As an application example, this paper demonstrates in situ detection of loop-mediated isothermal amplification products by hybridizing them with thiolated-ssDNA covalently anchored on the electrodes of a quartz crystal microbalance (QCM). Such hybridization leads to a frequency signal, which is suitable for monitoring real-time LAMP amplification based on mass-sensing: it detects interactions between the complementary nucleobases of LAMP products in solution and the thiolated-ssDNA probe sequence on the gold surface. Target DNA LAMP products cause irreversible frequency shifts on the QCM surfaces during hybridization in the kHz range, which result from both changes in mass and charge on the electrode surface. In order to confirm the LAMP assay working in the QCM sensing system at elevated temperature, the sky blue of positive LAMP products solution was achieved by using the Hydroxy Naphthol Blue (HNB) and agarose gel electrophoresis. Since on-QCM sensing of DNA hybridization leads to irreversible sensor responses, this work shows characterization by X-ray photoelectron spectroscopy (XPS) core spectra of S2p, N1s, Mg1s, P2p and C1s. XPS results confirmed that indeed both DNA and by-products of LAMP attached to the surface. Listeria monocytogenes DNA served to study in-situ detection of amplified LAMP products on DNA-functionalized surfaces.

Removal of Salmonella Typhimurium Biofilm from Food Contact Surfaces Using Quercus infectoria Gall Extract in Combination with a Surfactant.

Quercus infectoria (nutgall) has been reported to possess antimicrobial activities against a wide range of pathogens. Nevertheless, the biofilm removal effect of nutgall extract has not been widely investigated. In this study, we therefore evaluated the effect of nutgall extract in combination with cetrimonium bromide (CTAB) against preformed biofilm of Salmonella Typhimurium on polypropylene (PP) and stainless steel (SS) coupons in comparison with other sanitizers. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of nutgall extract and surfactants (CTAB and sodium dodecyl sulfate; SDS) were assessed. CTAB showed a more efficient antimicrobial activity than SDS and was selected to use in combination with nutgall extract for removing biofilm. To determine the biofilm removal efficacy, the PP and SS coupons were individually submerged in 2x MBC of nutgall extract (256 mg/ml) + 2x MBC of CTAB (2.5 mg/ml), nutgall extract alone (256 mg/ml), CTAB alone (2.5 mg/ml), distilled water, and 100 ppm sodium hypochlorite for 5, 15, and 30 min. The remaining sessile cells in biofilm were determined. Overall, the greatest biofilm removal efficacy was observed with nutgall extract + CTAB; the biofilm removal efficacy of sanitizers tended to increase with the exposure time. The SEM analysis demonstrated that S. Typhimurium biofilm on PP and SS coupons after exposure to nutgall extract + CTAB for 30 min displayed morphological alterations with wrinkles. This study suggests nutgall extract + CTAB may be an alternative to commonly used sanitizers to remove biofilm from food contact surfaces in the food industry and household.