Detection of Salmonella spp., Yersinia enterocolitica,Listeria monocytogenes and Campylobacter spp. by real‐time multiplex PCR using amplicon DNA melting analysis and probe‐based assay

Syto9 and probe‐based multiplex real‐time PCR assays for simultaneous detection of a group of foodborne pathogens (named SYLC group), targeting Salmonella spp. (invA gene), Yersinia enterocolitica (ystA gene), Listeria monocytogenes (hly gene) and Campylobacter spp. (rrna gene), have been developed. The Syto9 assay generates amplicon DNA melting curve with four peaks of 86.5 ± 0.5, 84 ± 0.5, 81.5 ± 0.5 and 90.5 ± 0.5 °C corresponding Salmonella spp., Y. enterocolitica, L. monocytogenes and Campylobacter spp. targets, respectively. The sensitivities of the Syto9 and TaqMan assays in artificially inoculated chicken wing rinses were in a range of 3.2 × 10² to 3.1 × 10⁴ and 9.8 × 10² to 1.9 × 10⁴ colony‐forming units per millilitre, respectively, depending on the pathogen. All tested target strains (n = 100) were correctly detected by the both assays, whereas nontarget strains (n = 100) demonstrated no cross‐reactivity representing 100% specificity. The assays are suitable for application in qualitative and quantitative detection of SYLC group pathogens in food matrices.     

‘Lipase and protease production of dairy Penicillium sp. on milk‐protein‐based solid substrates’

The lipolytic and proteolytic activity of Penicillium camemberti PC TT033 and Penicillium roqueforti PR G3, cultured on the whey solids or simulated cheese media, were compared under several pH reaction conditions. Lipolytic activity was higher when both strains had been cultured on the whey medium than on the simulated cheese medium, whereas proteolytic activity was less influenced by the culture medium. The relationship between the reaction pH and these enzyme activities was dependent on the culture medium, which suggested that the expression level and balance of isozyme rely on the culture substrate.     

Evaluation of the antioxidant properties and oxidative stability of Pecorino cheese made from the raw milk of ewes fed Rosmarinus officinalis L. leaves

The aim of this work was to study the effect of rosemary leaf dietary supplementation on the antioxidant activities and total phenolic content of Pecorino cheese. Three hundred and twenty‐four sheep were randomly assigned to two dietary groups, which received a standard diet based on lucerne hay and concentrate (400 g per day). The concentrate of the rosemary supplemented group contained 2.50% dried rosemary leaves. The trial lasted 7 weeks. Cheesemaking was performed 3, 5 and 7 weeks from the start of the trial. The Pecorino cheese antioxidant activity was modified by the diet. Rosemary supplementation increased the total phenolic content, enhanced the antioxidant properties and decreased the lipid oxidation of cheese. A slight decrease in flavour was detected in cheeses after 7 weeks of rosemary administration.     

Effects of Mentha longifolia L. essential oil and Lactobacillus casei on the organoleptic properties and on the growth of Staphylococcus aureus and Listeria monocytogenes during manufacturing,ripening and storage of Iranian white‐brined cheese

This research evaluated the effects of Mentha longifolia L. essential oil (EO) in concentrations 0, 50, 150 and 300 ppm and Lactobacillus casei (10⁸–10⁹ CFU/mL) on the growth of Staphylococcus aureus and Listeria monocytogenes during the manufacturing, ripening and storage of Iranian white‐brined cheese. The growth of the two pathogens was significantly reduced (P < 0.01) by both EO concentrations ≥50 ppm and probiotic and their combination in the standard manufacturing and storage process conditions of the cheese. Furthermore, the treatment containing 150 ppm of this EO combined with probiotic had a favourable inhibitory effect on the growth of two pathogenic micro‐organisms and also was the most appropriate treatment in sensory assessment. The synergistic effects of the above‐mentioned concentration level between the essential oil and probiotic were significant compared to other treatments, including essential oil and probiotic only. Thus, a lower concentration of this EO can be used when it is combined with this probiotic.     

Antibacterial effects of chitosan coating containing Mentha aquatica L. essence against Escherichia coli,Staphylococcus aureus and Listeria monocytogenes in Iranian white cheese

The effect of a chitosan coating and Mentha aquatica L. essence on Iranian white cheese was investigated. Results showed 100% inhibition of Escherichia coli growth using 1.5% essence after 10 days. After 15 days of incubation, the Staphylococcus aureus population was reduced by 44.2%, 70.0%, and 88.5% using 0.5, 1.0 and 1.5% essence, respectively. After 15 days, Listeria monocytogenes growth was inhibited by 63.84%, 70.12%, and 85.9% using 0.5, 1.0, and 1.5% essence, respectively. Inhibition zone diameter studies also confirmed the antibacterial effects of applied coating against all the above‐mentioned bacteria in Iranian white cheese.     

Evaluation of antimicrobial effects of lauric arginate on reduction of Salmonella spp. in ground chicken

The efficacy of lauric arginate (LAE) on Salmonella survivability in ground chicken containing 9.8% fat was determined under refrigerated storage. The effect of LAE treatments on total aerobes, pH and objective colour were also assessed in ground chicken. LAE was highly effective against Salmonella in 0.1% peptone with no detectable survivors following treatment with 200 ppm and 400 ppm of LAE. No difference existed (P > 0.05) in the growth of Salmonella and total aerobes after treatment with 200 and 400 ppm of LAE in ground chicken inoculated with a four strain mixture of Salmonella. At these concentrations, LAE did not exhibit any treatment effect on the pH and colour of ground chicken (P > 0.05). In conclusion, although LAE possesses strong inhibitory (P < 0.05) effect against Salmonella in suspension in 0.1% peptone water, no inhibitory (P > 0.05) effect on growth of Salmonella was observed in ground chicken at the currently approved levels of 200 ppm of LAE.     

Mineral composition,antioxidant and cytotoxic biopotentials of wild‐growing Ganoderma species (Serbia): G. lucidum (Curtis) P. Karst vs. G. applanatum (Pers.) Pat.

The aim of this work was to analyse mineral composition and chemical profile of two nonedible fungal species: Ganoderma lucidum and Ganoderma applanatum (Fru?ka Gora, Serbia) vs. their antioxidant (ABTS and A.E.A.C. assay) and cytotoxic biopotentials (MTT assay on MCF‐7). Both species were analysed for their content of macro‐ and microelements by atomic absorption spectrophotometry, while phenolic profile of EtOH and H₂O extracts was examined by LC‐MS/MS technique. Both species mostly contained the following ions: K⁺ > Ca²⁺ > Mg²⁺ > Mn²⁺ > Zn²⁺ > Cu²⁺ > Cr³⁺. Among nine phenolic compounds, the highest content of vanillic acid was detected in G. applanatum extracts while protocatechuic acid in EtOH extract and quinic acid in H₂O extract were mostly contained in G. lucidum. Ganoderma applanatum EtOH extract showed the best antioxidant activities related to its phenolic and flavonoid content. Further, the best cytotoxic effect after 72 h was observed in this extract as well.     

Effects of Mentha longifolia L. essential oil on viability and cellular ultrastructure of Lactobacillus casei during ripening of probiotic Feta cheese

The effects of Mentha longifolia L. essential oil during ripening and storage probiotic Feta cheese were studied, in relation to viability and cellular ultrastructure of Lactobacillus casei. The addition of the essential oil in the concentrations from 0.0 to 0.03% was trialled: the 0.03% treatment resulted in the highest viability of L. casei and the lowest pH value compared with other treatments (P < 0.05). Electron microscopy showed that essential oil caused no harm to L. casei. This study demonstrated the favourable effects of M. longifolia on optimal maintenance of L. casei at the end of cheese storage period.     

Optimising detection of acidification kinetics diversity in Lactococcus lactis subsp. lactis using SDS‐PAGE protein fingerprinting as screening method

Cell‐free extracts from 157 Lactococcus lactis strains isolated from Artisan cheese were screened by protein fingerprinting previous to their technological characterisation. The strains were classified according to their electrophoretic patterns into five groups. A set of strains representing the different clusters were selected to study their acidifying activity in milk. Time and rate feature points, as well as the shape of the acidification curves, resulted in six different fermentation kinetics, mostly consistent with the electrophoretic groups. Thus, selection of native strains as starter cultures based on their acidifying activity could be optimised by protein fingerprinting.     

Evaluation of Pseudomonas spp. through O2 and CO2 headspace analysis

This article proposes an approach to determine the level of Pseudomonas spp. in milk, based on the evaluation of the content of oxygen and carbon dioxide produced in the headspace of sealed vials; the research was divided into two phases: model building and preliminary validation. Three different strains of Pseudomonas spp., Ps. putida (wild strain) and Ps. fluorescens (wild and collection isolates), were used as targets. Data of CO₂ and O₂ were modelled through a modified positive (CO₂) or a negative Gompertz equation (O₂) to estimate the Minimum Detection Time (MDT), defined as the time to attain 3% of CO₂ (MDT₁) or a decrease in the content of O₂ by 3% (MDT₂). Then, MDT₁ and MDT₂ were submitted to a linear regression procedure, using cell concentration as independent variable; the correlations ‘MDT₁/cell concentration’ and ‘MDT₂/cell concentration’ showed high determination coefficients (>0.983). Moreover, the regression procedure pointed out that both MDT₁ and MDT₂ decreased by ca. 3 h for an increase in cell count of 1 log cfu mL^?1. Preliminary validation in milk pointed out that the error associated with the regression line ‘MDT₂/cell concentration’ was below 5%.     

Tolerance evaluation in Salmonella enterica serovar Typhimurium challenged with sublethal amounts of Rosmarinus officinalis L. essential oil or 1,8‐cineole in meat model

The induction of direct bacterial tolerance and cross‐tolerance (NaCl, acid pH, high temperature) in Salmonella Typhimurium ATTC 14028 following the exposure to sublethal amounts of the essential oil from Rosmarinus officinalis L. (ROEO), and its major component 1,8‐cineole (CIN) was evaluated in this study. Direct protection was not induced when cells were exposed to 1/2 MIC and 1/4 MIC of ROEO or CIN in meat broth and in previously irradiated meat ground‐beef. Cells exposed to ROEO or CIN at sublethal amounts did not present cross‐protection to high temperature, lactic acid and NaCl. Likewise, cells progressively subcultured in meat broth containing increasing amounts of ROEO or CIN were able to survive only up to 1/4 MIC for both tested substances. From these results, S. Typhimurium ATCC 14028 was not capable to develop direct or cross‐tolerance when exposed to ROEO or CIN in a meat‐based growth media and was not able to develop direct tolerance in a meat‐based model.     

Metabolic analysis of the cutaneous fungi Malassezia globosa and M. restricta for insights on scalp condition and dandruff

Dandruff is a global consumer problem, characterized by flaking and scaling of the scalp, accompanied by itch and irritancy. However, the aetiology of the condition remains poorly understood, although there is a strong consensus that the cutaneous fungi Malassezia globosa and M. restricta are a major contributory factor. Although there is a paucity of understanding on how these commensal microorganisms adopt a pathogenic phenotype, a rich source of potential insights now exists in the shape of the recently published whole‐genome sequence of M. globosa, a functional annotation and metabolic reconstruction of which is freely accessible via the integrated microbial genomes (IMG) online community resource ( http://www.hmpdacc-resources.org/cgi-bin/imgm_hmp/main.cgi ). In these studies, we have taken a combined in‐silico and in‐vitro approach to investigate aspects of lipid and amino acid metabolism by M. globosa and M. restricta that have the potential to impact on scalp condition and dandruff. The IMG platform was employed to analyse the metabolism of triacylglycerols and fatty acids, as well as the aromatic amino acid tryptophan, by M. globosa, to investigate pro‐inflammatory pathways linked in the literature to dandruff and pityriasis versicolour, respectively. Results were equivocal, leaving question marks over the ability of M. globosa to fully degrade unsaturated fatty acids and metabolize tryptophan to indole‐3‐pyruvic acid. In‐vitro assay systems were then developed to study the biotransformation of these metabolites by both M. globosa and M. restricta, as well as their effect on human keratinocytes, and the results here indicated that neither unsaturated fatty acids nor indole derivatives are likely to be major aetiological factors in dandruff.     

The revised COLIPA in vitro UVA method

A multicentred study derived from the COLIPA in vitro UVA method was performed to assess the influence of test conditions on UVA protection factor (UVAPF) values in terms of amplitude, reproducibility between laboratories and correlation with in vivo UVA results. Eight products with a range of in vivo UVAPF from three to 29 were used. Two different types of plates, namely high‐roughness (5 μm) and low‐roughness (2 μm) plates, were used with a different application rate for each (1.3 mg cm^?2 and 0.75 mg cm^?2 respectively). The UVR dose applied to both plate types followed the same principle as the original test (1.2 J. cm^?2 × UVAPF0). Strong, significant correlations between in vitro and in vivo UVAPF values were observed for both plate types (Pearson correlation > 0.9, P ≤ 0.01). The correlation and slope obtained with the low‐roughness plates confirmed the previous results obtained by COLIPA. Across all laboratories, higher UVAPF values were obtained on the high‐roughness plates (P < 0.01). Reproducibility of UVAPF values between laboratories was comparable between the two plate roughness values (low roughness, COV = 8%; high roughness, COV = 12%). Considering the in vitro/in vivo comparisons, a regression slope of 0.83 was observed for the low‐roughness plates, in comparison with a value of 1.05 for the high‐roughness plates. The accuracy of the method was improved, therefore, with the use of the high‐roughness plates. With a constraint to recommend the use of only one plate type in the COLIPA UVA in vitro Test, the high‐roughness plate was selected on an on‐going basis to limit variability of results and to provide better accuracy with in vivo data.     

Nutritional composition and sensory attributes of Alaskan flatfishes compared to plaice (Pleuronectes platessa)

Proximate composition, fatty acids profiles and other nutritional values were evaluated for fillets of Limanda aspera (yellowfin sole), Lepidopsetta bilineata (southern rock sole) and Lepidopsetta polyxystra (northern rock sole) and compared to North Sea plaice (Pleuronectes platessa). Additional information is given on the composition of fillets from arrowtooth flounder (Atheresthes stomias). Plaice (0.8% lipid) and Alaska soles (1.0–1.2% lipid) can be classified as lean species, resulting in low 0.3–0.5 g ∑EPA+DHA/100 g muscle, although the fatty acid profiles of the extracted lipids were characterised by high amount of n‐3 fatty acids (33.2–47.3%). Arrowtooth flounder belong to the medium‐fat species (4.3%). Taurine was the most prevalent free amino acid; mean values ranging between 221 mg100 g^?1 and 247 mg100 g^?1 wet weight. The selenium content varied between 130 and 310 μg kg^?1 ww. Sensory attributes of Southern and Northern rock sole were comparable to plaice.     

First isolation of Brevibacterium sp. pigments in the rind of an industrial red‐smear‐ripened soft cheese

The smear‐ripened soft cheeses are characterised by a surface orange‐red‐brown colour, which has a microbial origin. For a long time, this colouration was mainly imputed to Brevibacterium linens. However, the latest published works, based on molecular biology, have shown a minor role for this bacterium. This study shows the results obtained with an industrial cheese named Vieux‐Pané, which is characterised by the presence of carotenoids from Brevibacterium linens group at its surface. This demonstrates that, under certain conditions, the Brevibacterium linens group (Brevibacterium linens and Brevibacterium aurantiacum sp. nov.) is able to produce pigments and to colour cheeses effectively.