Rapid and Sensitive Detection of Staphylococcus aureus in Food Using Selective Enrichment and Real-Time PCR Targeting a New Gene Marker

Staphylococcus aureus is a bacterial pathogen considered a principal etiological agent of food poisoning. The aim of this study was to develop and evaluate a rapid and sensitive method for the detection of S. aureus in food by using selective enrichment and a new species-specific real-time polymerase chain reaction (PCR). Specific primers and a TaqMan probe targeted to specific S. aureus gene encoding for acriflavine resistance protein were designed. The real-time PCR was highly specific for S. aureus with 100% inclusivity and 100% exclusivity determined using 83 S. aureus strains and 64 non-S.-aureus strains. PCR detection limit of 6.8 × 10¹ and 3.4 × 10¹ CFU ml⁻¹ were obtained with 100% and 70% detection probability, respectively. The single selective enrichment based on the study of different enrichment conditions was selected and a lysis by boiling was used to obtain bacterial DNA. Out of 112 food samples analyzed, 61 were positive by the PCR-based method and 53 by the standard method. Out of ten food matrices artificially contaminated at a level of 10° CFU g⁻¹, ten and six were positive by the respective methods. Moreover, 10° CFU 10 g⁻¹ was detected in all ten artificially contaminated samples after a large-scale enrichment using PCR-based detection, in contrast to seven false negative by standard detection. The developed method facilitated the detection of S. aureus on the next day after the sample reception. This method can be used for S. aureus detection as a faster, highly specific, and more sensitive alternative to microbiological method with the potential for providing of improved food-processing hygiene control.     

Tensile behavior of Basalt/Glass single and multilayer-woven fabrics

High strength fabrics are perfect materials for use in automotive and aerospace systems, where high performance and light weight structures are demanded. Mechanical characterization under constant loading is of great importance for these materials. The purpose of this study is to analyze the tensile properties of Basalt and Glass-woven structures at different pick density, weave design, stitch distance, and number of layers. For this purpose, single and double layerwoven fabric structures have been prepared using Basalt and Glass yarns on sample weaving loom. Their uniaxial testing has been performed to analyze their tensile curves. Analysis of variance showed the statistical significance of material, pick density, direction of applied load, weave, and stitching in strength of fabric, while the effect of stitch distance was insignificant.     

Endotoxin and biogenic amine levels in Atlantic mackerel (Scomber scombrus), sardine (Sardina pilchardus) and Mediterranean hake (Merluccius merluccius) stored at 22 °C

Whole Atlantic mackerel (Scomber scombrus), sardine (Sardina pilchardus) and Mediterranean hake (Merluccius merluccius) from the Croatian Adriatic were stored at 22 °C and changes in histamine, putrescine, tyramine and cadaverine levels were monitored in relation to bacterial endotoxin. After 12 h, histamine levels in sardine were above the legal limit of 50 mg kg^?1, set by the US Food and Drug Administration, and an increase in putrescine content preceded the increase in histamine. After 24 h, histamine contents in mackerel and sardine reached 1090 ± 101 and 577 ± 275 mg kg^?1, respectively, which exceeded the toxic threshold of 500 mg kg^?1. At the same time, the putrescine content was also high in both fish (353–420 mg kg^?1). The time-course of endotoxin production was similar in all fish species stored at 22 °C. A high correlation was found between endotoxin and histamine, and between endotoxin and putrescine in mackerel and sardine. On the other hand, high endotoxin levels in hake, after 24 h, were associated with the low histamine and putrescine content (40–60 mg kg^?1).     

Temperature-dependent kinetics of grape seed phenolic compounds extraction: Experiment and model

The kinetics of a batch solid–liquid extraction of total phenolic compounds (PC) from milled grape seed (Vitis vinifera L. cv. “Frankovka”) using 50% ethanol at different extraction temperatures (25–80 °C) was studied. The maximum yield of PC was 0.13 kg_GAE/kg_db after 200 min of extraction in agitated vessel at 80 °C. A new model based on the assumptions of a first order kinetics mechanism for the solid–liquid extraction and a linear equilibrium at the solid–liquid interface was developed. The model involves the concept of broken and intact cells in order to describe two successive extraction periods: a very fast surface washing process followed by slow diffusion of phenolic compounds from grape seeds to the solvent.The proposed model is suited to fit experimental data and to simulate the extraction of phenolic compounds, which was confirmed by the correlation coefficient (r ? 0.965), the root mean square error (RMSE ? 0.003 kg_GAE/kg_db) and the mean relative deviation modulus (E ? 2.149%). The temperature influenced both equilibrium partition coefficients of phenolic compounds and transport properties, which is manifested by a relatively high value of activation energy (23–24) kJ/mol and by values of effective diffusivity in seed particles.     

Suitability of some selected maize hybrids from Serbia for the production of bioethanol and dried distillers' grains with solubles

BACKGROUND: Bioethanol is mostly produced from starchy parts of the corn grain kernel leaving significant amounts of valuable by‐products such as dried distillers' grains with solubles (DDGS) which can be used as a substitute for traditional feedstuff. The suitability of six maize hybrids from Serbia was investigated for bioethanol and DDGS production. The correlation between physical and chemical characteristics of the grain, bioethanol yield and quality of the corresponding DDGS was assessed. RESULTS: All hybrids had very different chemical composition and physical characteristics which could allow various applications. The highest bioethanol yield (94.5% of theoretical) and volumetric productivity (2.01 g l^?1 h^?1) were obtained with hybrid ZP 434 and the lowest with ZP 611k. Regarding chemical composition, all DDGS samples manifested good properties as feed components. Their protein content was higher compared to the kernel. In addition, the samples showed high digestibility and high mineral content, especially of calcium and phosphorus. CONCLUSION: A hybrid ZP 434 was selected as the most promising bioethanol producer. This property is attributed to the highest level of soft endosperm which is more susceptible to starch‐hydrolysing enzymes. A high yield potential per hectare makes it the best candidate for commercial bioethanol production. © 2012 Society of Chemical Industry     

Validation of a Method for Analysis of Aroma Compounds in Red Wine using Liquid–Liquid Extraction and GC–MS

An analytical method for determination of volatile composition of wines using sample preparation by liquid–liquid extraction and gas chromatography coupled to mass spectrometry for separation and detection has been developed and validated. Extraction of volatile compounds was performed in dichloromethane, and 1-octanol was added as an internal standard. Kékfrankos red wine produced in Villány wine region in Hungary was used as a model wine for testing and validation of the method. The developed method allowed satisfactory determination of 33 volatile compounds in the wines. Compounds analyzed include alcohols, esters, lactones, fatty acids, furans, and nitrogen compounds. The calibration curves of the four reference compounds used (2-phenyl ethanol, ethyl nonanoate, butyrolactone, and tyrosol) were linear in all cases with correlation coefficients (R ²) ranging from 0.9951 to 0.9992. The accuracy of the method was checked with a standard addition method (recovery 92.2–103 %), showing good repeatability and reproducibility (RSD?<?10 %).