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.     

Effects of Fish Gelatin and Tea Polyphenol Coating on the Spoilage and Degradation of Myofibril in Fish Fillet During Cold Storage

Fish fillet is easily spoiled during storage. Antimicrobial edible coating of gelatin extracted from fish skins and bones and tea polyphenol (TP) was developed to inhibit the spoilage of fish fillet during cold storage. For coating containing 0.4 % TP and 1.2 % gelatin, the pH only slightly increased from 6.17 at day 0 to 6.32 at day 17 of cold storage, while the pH of control coating increased to 6.87 at day 17. Atomic force spectrometry was used to analyse the nanostructure of myofibril, which is the major component of fish muscle. The results showed that the length of myofibril from 0.4 % TP and 1.2 % gelatin group was greater than 15 μm, while the diameter and height were 3.38 and 0.59 μm, respectively, which exhibited the most intact nanostructure after 17 days of cold storage. Meanwhile, matrix-assisted laser desorption–ionisation–time-of-flight mass spectrometry result showed that TP delayed the degradation of myosin light chain 3 and troponin T in myofibril. Gas chromatography–mass spectrometry of volatile organic compounds (VOCs) also showed that 0.4 % TP and 1.2 % gelatin coating group had minimal production of spoilage markers, such as 1-octen-3-ol, 2-methyl propanoic acid and dimethyl sulfide. The microbial analysis showed that the aerobic mesophilic/psychrotrophic count, yeasts and moulds of 0.4 % TP and 1.2 % gelatin group were significantly lower than the control group. Therefore, 0.4 % TP and 1.2 % gelatin coating showed the best antimicrobial effect and can be used to maintain the nanostructure of fish fillet and prevent the spoilage during cold storage.     

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).     

Bending performance of 3-layer beech (Fagus sylvativa L.) and Norway spruce (Picea abies (L.) Karst.) VTC composites bonded with phenol–formaldehyde adhesive and liquefied wood

Low quality beech (Fagus sylvatica L.) and Norway spruce (Picea abies (L.) Karst.) were densified with viscoelastic thermal compression (VTC) process to two different degrees of densification, and lamellas were used to manufacture different types of 3-layer VTC composites. Bending properties of 3-layer VTC composites bonded with phenol formaldehyde (PF) adhesive and liquefied wood (LW) were determined and compared to 3-layer composites produced with undensified beech or spruce wood lamellas. Morphology of VTC spruce wood of higher density was analysed with fluorescent microscopic technique. All composites produced with densified beech lamellas and bonded with PF adhesive had significantly higher values of modulus of rupture (MOR) and modulus of elasticity (MOE) than composites produced with undensified lamellas. Densified spruce lamellas contributed to better bending performance of 3-layer VTC composites bonded with PF adhesive to some extent. Furthermore, composites bonded with LW had significantly lower MOR and MOE values compared to composites bonded with PF adhesive. Study of VTC spruce wood microstructure showed that densification caused non-uniform deformation of cell wall structure, in which cell wall fractures were observed.     

Application of Matrix Solid-Phase Dispersion and Liquid Chromatography–Ion Trap Mass Spectrometry for the Analysis of Pesticide Residues in Fruits

This study presents an application of rapid and sensitive multiresidue method for the analysis of acephate, acetamipride, atrazine, carbendazim, carbaryl, carbofuran, dimethoate, imidacloprid, linuron, malathion, monocrotophos, monuron, propazine, simazine, and tebufenozide in fruits. The method involves an extraction procedure based on matrix solid-phase dispersion using diatomaceous earth as a dispersant and dichloromethane as the eluent. The target pesticides were determined using liquid chromatography–ion trap mass spectrometry. Quantification of the analytes was carried out using the most sensitive ion transition. Ion trap parameters, like activation q and time, were found to have a prominent influence on method sensitivity for some pesticides and they were optimized accordingly. The confirmation of residues detected in real samples was performed by repeated injection and acquiring additional ion transitions besides the ones used for quantification. The method was validated for accuracy, linearity, reproducibility, and sensitivity. Mean values for recoveries were in the range of 70–120 % for all tested matrices. Repeatability of the method, expressed as the relative standard deviation, was in general lower than 20 %. The applicability of the method to routine analysis was tested in real fruit samples with good performance.     

Determination of sterigmatocystin in cheese by high-performance liquid chromatography-tandem mass spectrometry

Different cheese samples produced in Latvia (eight) and Belgium (13) were analysed for their sterigmatocystin (STC) content. Only two (9.5%) of the samples were positive for STC with concentration levels of 1.23 and 0.52 µg kg^?1, respectively. Five (24%) samples contained STC above the limit of detection (0.03 µg kg^?1) but below the limit of quantification (0.1 µg kg^?1), A sensitive liquid chromatography-electrospray positive ionisation-tandem mass spectrometry (LC-MS/MS) method, which was previously developed for the analysis of STC in grains, was modified and applied to the analysis of STC in cheese. This method involved sample extraction with acetonitrile–water (90 : 10, v/v), defatting with n-hexane, solid-phase extraction, separation on a reversed-phase C₁₈ column, and STC detection by LC-MS/MS.     

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     

Health effects of olive oil polyphenols: Recent advances and possibilities for the use of health claims

The Mediterranean diet and consumption of olive oil have been connected in several studies with longevity and a reduced risk of morbidity and mortality. Lifestyle, such as regular physical activity, a healthy diet, and the existing social cohesion in Southern European countries have been recognised as candidate protective factors that may explain the Mediterranean Paradox. Along with some other characteristics of the Mediterranean diet, the use of olive oil as the main source of fat is common in Southern European countries. The benefits of consuming olive oil have been known since antiquity and were traditionally attributed to its high content in oleic acid. However, it is now well established that these effects must also be attributed to the phenolic fraction of olive oil with its anti‐oxidant, anti‐inflammatory and anti‐microbial activities. The mechanisms of these activities are varied and probably interconnected. For some activities of olive oil phenolic compounds, the evidence is already strong enough to enable the legal use of health claims on foods. This review discusses the health effects of olive oil phenols along with the possibilities of communicating these effects on food labels.