Lack of growth of Listeria monocytogenes and Staphylococcus aureus in temperature abuse of E-beam treated ready-to-eat (RTE) cooked ham

The behaviour of Listeria monocytogenes and Staphylococcus aureus in vacuum-packed cooked ham slices treated with an electron beam and stored at 4, 7 and 10 °C was investigated. Cooked ham slices were inoculated with L. monocytogenes and S. aureus and electron beam treated at 2 and 3 kGy. After treatment, a long temperature-dependent death phase was observed, followed by growth at a slower rate than in untreated samples. Assuming a hypothetical load of 10 cells/g or cm² of L. monocytogenes and S. aureus as an original contamination in an industrial situation, an E-beam treatment of vacuum-packed cooked ham slices at 2 kGy guarantees the microbiological safety of the product along its shelf life, even if a noticeable temperature (10 °C) abuse occur during its storage period. Likewise, the E-beam treatment gave rise to a substantial increase of the RTE cooked ham shelf life off-sensory features associated to the spoilage only were detected in non-treated samples (controls) after 8 and 18 days of storage at 10 °C and 7 °C, respectively.     

Time course of peptide fingerprints in semimembranosus and biceps femoris muscles during Bayonne ham processing

The aim of this work was to define reliable markers of muscle and processing time in dry-cured ham using a rapid, precise semi quantitative method for the protein fraction soluble in low ionic strength buffer. For this purpose protein labchip Agilent was used to separate proteins and peptides and accurately determine their molecular weights and concentrations electrophoretically. In this way the protein fingerprinting of dry-cured ham at different process times was characterised, together with targets and products of proteolysis. In addition, the comparison of all the electrophoregrams indicated muscle and dry-curing process markers.     

Technology-induced selection towards the spoilage microbiota of artisan-type cooked ham packed under modified atmosphere

The microbiota associated with a highly-perishable Belgian artisan-type cooked ham was analyzed through plating and (GTG)₅-fingerprinting of isolates throughout its processing chain. The raw tumbled meat was characterized by the presence of a versatile microbiota around 4.8 log(cfu g⁻¹), consisting of lactic acid bacteria, staphylococci, Brochothrix thermosphacta, Gram-negative bacteria, and yeasts. Pasteurisation of the ham logs reduced bacterial counts below 2 log(cfu g⁻¹) and subsequent manipulations selected for leuconostocs and carnobacteria. Also, B. thermosphacta and several Enterobacteriaceae were found at this stage. During storage in an intermediate high-care area for 2 days, a selection towards certain Enterobacteriaceae (Hafnia alvei, Enterobacter spp., and Pantoea agglomerans) and lactic acid bacteria (mainly vagococci and Streptococcus parauberis) was observed. B. thermosphacta, Leuconostoc carnosum and carnobacteria were also detected, but only after allowing bacterial outgrowth by incubating the meat logs at 7 °C for four weeks. After a mild post-pasteurisation process and subsequent handling, incubation of the meat logs at 7 °C for four weeks led to outgrowth of Enterobacteriaceae (mainly Enterobacter spp. and Serratia spp.). B. thermosphacta, and lactic acid bacteria (Enterococcus faecalis, Leuc. carnosum, and Carnobacterium maltaromaticum) were also found. After slicing and packaging under modified atmosphere, the microbiota of the refrigerated end-product consisted of leuconostocs, carnobacteria, and B. thermosphacta.     

Effects of rapid chilling of carcasses and time of deboning on weight loss and technological quality of pork semimembranosus muscle

The effect of rapid air chilling of carcasses in the first 3 h of chilling at −31 °C (then at 2–4 °C, till 24 h post-mortem) and the possibility of earlier deboning (8 h post-mortem) after rapid air chilling, compared to conventional air chilling (at 2–4 °C, till 24 h post-mortem) on weight loss and technological quality (pH value, tenderness, drip loss, cooking loss and colour - L^*a^*b^* values) of pork M. semimembranosus was investigated. Under the rapid chilling conditions, weight loss was 0.8% at 8 h post-mortem and increased to 1.4% at 24 h post-mortem when weight loss was 2.0% under conventional chilling. Carcasses that were rapid chilled had significantly lower (P < 0.001) internal temperature in the deep leg at 4 (25.7 °C), 6 (13.0 °C), 8 (6.2 °C) and 24 h (3.8 °C) post-mortem compared to conventional chill treatment (32.7, 24.2, 19.1 and 5.1 °C, respectively). Rapid chilling reduced significantly (P < 0.05) the rate of pH value decline at 8 h (6.02) post-mortem in M. semimembranosus compared to conventional chill treatment (5.88). Compared to conventional chilling, in M. semimembranosus deboned in different time post-mortem, rapid chilling had a positive significant effect on drip loss (P < 0.05, muscles deboned 8 h post-mortem), cooking loss (P < 0.001) and incidence of pale colour (L^* value). Rapid chilling i.e. rapid chilling and earlier deboning had neither positive nor negative significant effects (P > 0.05) on other investigated technological quality parameters of M. semimembranosus (tenderness, a^* value and b^* value) compared to conventional chilling.     

Effect of high pressure treatment on microbial populations of sliced vacuum-packed cooked ham

In this study, culture-dependent and culture-independent approaches were used to reveal the microbial diversity and dynamic changes occurring in sliced vacuum-packed cooked ham after high pressure processing (HPP, 400MPa or 600MPa for 10min at 22°C) during refrigerated storage over 90days. Direct extraction of genome DNA and total RNA from meat samples, followed by PCR-denaturing gradient gel electrophoresis (DGGE) and RT-PCR-DGGE on 16S rDNA V3 region, was performed to define the structure of the bacterial populations and active species in pressurized cooked ham. Results showed that HPP affected differently the various species detected. The predominant spoilage organisms of cooked ham, such as Lactobacillus sakei and Lactobacillus curvatus, were found to be very sensitive to pressure as they were unable to be detected in HPP samples at any time during refrigerated storage. Weissella viridescens and Leuconostoc mesenteroides survived HPP at 600MPa for 10min at 22°C and were responsible for the final spoilage. An RNA-based DGGE approach clearly has potential for the analysis of active species that have survived in pressurized cooked ham. High pressure processing at 400 or 600MPa for 10min at room temperature (22°C) has a powerful inhibitory effect on the major spoilage bacteria of sliced vacuum-packed cooked ham. High pressure treatment may lead to reduced microbial diversity and improve the products' safety.     

In-home consumer and shear force evaluation of steaks from the M. serratus ventralis thoracis

The M. serratus ventralis thoracis was obtained from US Select arm chucks (n = 87) to investigate if this underutilized muscle can be used as a steak alternative. Muscles were assigned randomly into three treatment groups: (1) control; (2) blade tenderization; and (3) injection, containing salt, phosphate, and papain. Steaks were cut from each muscle for in-home consumer evaluation (n = 136) and Warner–Bratzler shear (WBS) force determination. The WBS values for injected steaks (13.1 N) were lower (P < 0.05) than for blade-tenderized (18.4 N) and control (19.9 N) steaks. Tenderness ratings for the injected steaks were higher (P < 0.05) compared to the other treatments when steaks were grilled, oven prepared or were cooked in a skillet; however, this improvement did not significantly influence overall like scores for steaks that were oven prepared or cooked in a skillet. For the most part, degree of doneness did not significantly impact consumer evaluations of steaks prepared by the various cooking methods. However, there was a treatment × degree of doneness interaction for grilled-cooked steaks where increased doneness for blade-tenderized and injected steaks resulted in increased palatability ratings, whereas increased doneness for control steaks generally resulted in lowered palatability ratings. Consumer ratings and WBS values for the M. serratus ventralis thoracis indicate that merchandising steaks from this muscle may be a viable option in the marketplace, especially if blade tenderization or injection processes are used for further enhancement.     

Effect of supplementing ewes’ diet with thyme (Thymus zygis ssp. gracilis) leaves on the lipid oxidation of cooked lamb meat

The aim of this study was to ascertain whether the inclusion of thyme leaves in the diet of pregnant sheep affects the sensorial characteristics and oxidative stability of cooked lamb meat. Thirty-six Segureña sheeps were randomly assigned into three homogeneous groups. One group was fed a basal diet as control, while the diet of the other two groups was modified by substituting 3.75% and 7.5% of the control diet with thyme leaves. TBARS, volatile compounds, and the sensory characteristics of cooked lamb meat were analysed on days 0, 2 and 4. The incorporation of thyme into the animal diet favoured the antioxidant capacity of these cooked lamb meat samples. The cooked meat of lambs from ewes fed thyme leaves showed lower TBARS, hexanal values, rancid odour and rancid flavour scores. No statistically significant differences were detected between the results obtained for the two leaf levels.     

Determination of mineral contents of semimembranosus muscle and liver from pure and crossbred pigs in Vojvodina (northern Serbia)

The content of phosphorous (P), potassium (K), sodium (Na), magnesium (Mg), calcium (Ca), zinc (Zn), iron (Fe), copper (Cu) and manganese (Mn) was investigated in the M. semimembranosus and liver of 69 pigs from 10 different genetic lines, produced in Vojvodina (northern Serbia). Phosphorous was determined by the standard spectrophotometric method. Metals were determined by the flame atomic absorption spectrometry after mineralisation by dry ashing. The difference in the mineral content among the different genetic lines of pigs, reared under the same conditions, was not significant (P > 0.05) in the analysed muscle and liver tissues. The order of the minerals in the meat samples and their content ranges in mg/100 g was K (214–328, on average 280) > P (200–263, on average 225) > Na (39.0–82.7, on average 59.8) > Mg (24.4–29.4, on average 26.6) > Ca (9.8–15.5, on average 11.8) > Zn (2.30–3.29, on average 2.70) > Fe (1.00–2.79, on average 1.42) > Cu (0.18–0.49, on average 0.32) > Mn (0.018–0.038, on average 0.025). Likewise, the order of the minerals in the liver samples and their content ranges in mg/100 g was P (336–448, on average 383) > K (156–306, on average 217) > Na (56.1–126.3, on average 82.2) > Mg (20.8–36.6, on average 25.7) > Ca (15.9–31.1, on average 20.4) > Fe (16.65–30.93, on average 21.78) > Zn (6.32–15.99, on average 9.82) > Cu (0.72–3.06, on average 1.61) > Mn (0.23–0.51, on average 0.35). A higher content of K and Mg was found in the muscle than in the liver tissue, with a highly significant difference for K (P < 0.001). The contents of P, Na, Ca, Zn, Fe, Cu and Mn were significantly higher (P < 0.001) in the liver tissue than those in the muscle tissue. The Vojvodian pork meat showed higher Cu and Mn contents, while the Vojvodian pork liver showed a higher Ca content compared with the values found in other countries.     

Essential oils of the aerial parts of three Salvia species from Jordan: Salvia lanigera, S. spinosa and S. syriaca

The compositions of the essential oils of three Jordanian Salvia species, S. lanigera and S. spinosa from a desert climate, and S. syriaca from a Mediterranean habitat, were studied. All three oils are rich in monoterpene derivatives (68–73%).     

Evaluation of antioxidant,antibacterial and anti-tyrosinase activities of four Macaranga species

The methanolic fresh leaf extracts of Macaranga gigantea, Macaranga pruinosa, Macaranga tanarius and Macaranga triloba were screened for their antioxidant properties (AOP), tyrosinase inhibition and antibacterial activities. Total phenolic content (TPC), 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging, ferric-ion reducing power (FRAP), ferrous-ion chelating (FIC) and lipid peroxidation inhibition (LPI) activities were used to evaluate the AOP. Modified 3,4-dihydroxy-L-phenylalanine (L-DOPA) method was used to determine tyrosinase inhibition activity, whereas antibacterial activity was determined using the disc-diffusion technique. TPC screening of the same species from different collection sites showed no significant difference between sites. M.triloba showed the highest ascorbic acid equivalent antioxidant activity (AEAC), FRAP and LPI values. M. tanarius, which showed the lowest TPC, AEAC, FRAP and LPI activities, exhibited the best FIC activity. M. pruinosa showed the best tyrosinase inhibition activity, whereas M. triloba showed the best antibacterial activity against Gram-positive bacteria species, with minimal inhibition dosage (MID) values as low as 10 μg/disc.     

Chemical and biological characteristics of Cuminum cyminum and Rosmarinus officinalis essential oils

Essential oils extracted by hydrodistillation from Cuminum cyminum and Rosmarinus officinalis were characterized by means of GC and GC–MS. C. cyminum and R. officinalis contained α-pinene (29.1%, 14.9%), 1,8-cineole (17.9%, 7.43%) and linalool (10.4%, 14.9%), respectively, as the major compounds. C. cyminum oil exhibited stronger antimicrobial activity than did R. officinalis oil against E. coli, S. aureus and L. monocytogenes. Complete death time on exposure to Cuminum cyminum L. and Rosmarinus officinalis L. oils were 20 and 25 min 180 and 240 min and 90 and 120 min for E. coli, S. aureus and L. monocytogenes, respectively. Radical-scavenging and antioxidant properties were tested by means of 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and the β-carotene bleaching test. These properties were compared to those of Thymus x-porlock essential oil, used as a reference ingredient. The radical scavenging performance of the rosemary oil was better than that of C. cyminum. Results from the antioxidant test were better than those provided by the radical-scavenging activity. C. cyminum and R. officinalis essential oils may be considered as potent agents in food preservation.     

Survival and death of Listeria monocytogenes on cooked bacon at three storage temperatures

Survival of Listeria monocytogenes on cooked bacon cubes (a_w 0.910 ± 0.080), strips (a_w 0.726 ± 0.054), and bits (a_w 0.620 ± 0.038) was determined during a 25 week storage period at −20, 4.4, and 22 °C. Selective enrichment and subsequent enzyme-linked fluorescent antibody (ELFA) detection were used to asses survival on samples inoculated at ca. 1-log₁₀ CFU/g (LI). Samples inoculated at ca. 5.5-log₁₀ CFU/g (HI) were analyzed over time by direct plating on modified Oxford medium (MOX). The Baranyi model was fitted to the inactivation curves of HI samples using the DMFit program. At −20 °C, a decline of about 1-log₁₀ CFU/g occurred on all HI cooked bacon types by 14 weeks, although most LI samples remained positive by the ELFA detection method for 25 weeks. At 4.4 and 22 °C, some strips and bits LI samples were negative for the pathogen within 3 weeks, and >1.5 log₁₀ CFU/g reductions occurred on HI strips and bits by 8 weeks. Reductions on cubes at refrigeration and ambient temperature were ca. 0.5 log₁₀ CFU/g, and cubes remained positive on LI samples for 25 weeks. Rate parameter estimates indicated that the population declined fastest on strips and bits at 22 °C compared to all other product and temperature combinations. This study demonstrates that cooked bacon does not support the growth of L. monocytogenes and that the pathogen gradually dies off during storage.     

Sensory,physical, chemical and microbiological changes in aquacultured meagre (Argyrosomus regius) fillets during ice storage

Commercial-sized meagre fillets were stored on ice at 4 °C for 18 days, in order to evaluate the loss of quality and freshness that occurs over this period of time. Physicochemical (pH, thiobarbituric acid (TBA), total volatile basic nitrogen (TVBN), trimethylamine (TMA), water activity, water-holding capacity, colour, texture and fatty acid profile), sensory and microbiological analyses were carried out at 0, 4, 7, 11, 14 and 18 days of storage. As part of the sensory analysis, attributes associated with fillet appearance, odour and texture were examined. Variations in pH, TBA, TVBN and TMA were observed throughout the storage period, although only TBA displayed a significant correlation with time (r = 0.96). L^∗ and b^∗ values increased, and the chroma and hue values decreased, reflecting the colour changes experienced by the fillets over time. With regards to the texture profile, hardness was significantly correlated with time (−0.68). All the sensory analysis attributes exhibited significant variations and correlations close to 1.00 with storage time, which is a reflection of the fillets’ loss of freshness. The correlation coefficients between aerobic mesophilic and psychrophilic bacteria, enterobacteria and coliform counts on the one hand and storage time on the other were also very high (0.99–1.00). A regression analysis using the acceptability limit set by the ICMSF standard (1986) for total aerobic mesophilic counts (7 log cfu/g) yielded a shelf-life for meagre fillets of 9 days. The TBA, sensory and microbiological analyses displayed very strong correlations with storage time, and they may be considered suitable indicators for evaluating meagre fillet spoilage during refrigerated storage.     

Characterisation of esterolytic activity from two wild mushroom species, Amanita vaginata var. vaginata and Tricholoma terreum

Characterisation of esterase activities from the edible mushroom species, Amanita vaginata var. vaginata and Tricholoma terreum, were investigated. Native electrophoresis of the crude extracts prepared from both mushroom samples showed the presence of esterolytic activities. The extracts had the greatest activity in the presence of p-nitrophenyl butyrate (pNPB) as a substrate. pH and temperature optima were found to be 8.0 and 30 °C for both enzymes, respectively. V_max and K_m values were determined as 14.2 U/l and 71 μM for A. vaginata var. vaginata and 34.6 U/l and 9.6 μM for T. terreum, respectively. The pH-stability profile showed a stationary line between 3.0 and 10.0 for both enzymes. The esterolytic activities from the extracts were maintained between 10 and 40 °C for 4 h and started to decrease at 50 °C. The effects of EDTA, NaN₃, DTT and PMSF on the enzyme activity were also investigated.     

Inactivation of Serratia liquefaciens on dry-cured ham by high pressure processing

To quantify the inactivation of Serratia liquefaciens exerted by high pressure processing (HPP), slices of dry-cured ham were inoculated and processed combining different levels of technological parameters: pressure (347–852 MPa), time (2.3–15.8 min) and temperature (7.6–24.4 °C) according to a central composite design. Bacterial inactivation, as logarithmic reduction, indicated that S. liquefaciens was relatively sensitive to HPP. Six log reductions were achieved in a total of 10 trials combining pressures of 600 MPa or above with different holding times and temperatures. The inactivation of S. liquefaciens was analysed through the multiple regression analysis to generate a second order polynomial equation. Pressure and time were the two factors which significantly determined the inactivation of S. liquefaciens on dry-cured ham. Temperature did not significantly affect the lethality of the process. The response surface methodology was used to determine optimum process conditions to maximize the inactivation of S. liquefaciens in the experimental range tested. The maximum inactivation of S. liquefaciens in dry-cured ham was achieved by combining a pressure of 650 MPa with a holding time of 8 min. Combinations above these values (i.e. 750 MPa for 13 min) would not significantly improve the lethality of the process.     

Spoilage characteristics of Brochothrix thermosphacta and campestris in chilled vacuum packaged lamb,and their detection and identification by real time PCR

The spoilage potential of Brochothrix campestris and Brochothrix thermosphacta was investigated in vacuum-packed lamb. Striploins (n = 338) were inoculated and stored for twelve weeks at temperatures − 1.5, 0, 2 and 7 °C. Growth around 5–6 log₁₀ CFU/cm² was recorded after six weeks at 0, 2 and 7 °C, and ~ 3 log₁₀ CFU/cm² after nine weeks at − 1.5 °C. B. campestris was shown to cause spoilage by nine weeks at temperatures above 0 °C by the presence of green drip and unacceptable odours. Molecular based assays for the detection and differentiation of B. thermosphacta and B. campestris were developed and validated. A TaqMan assay was designed to target a unique single-nucleotide polymorphism in the Brochothrix 16 s rRNA gene with a sensitivity of < 7 CFU per reaction. Secondly a specific PCR was designed for B. campestris targeting the structural genes, brcA and brcB. These testing regimes offer a rapid and cost effective method for the detection and screening of Brochothrix species in meat products and processing environments.     

Comparative chemical composition,free radical-scavenging and cytotoxic properties of essential oils of six Stachys species from different regions of the Mediterranean Area

The chemical composition of essential oils of six Stachys species, S. cretica L. ssp. vacillans Rech. fil., S. germanica L., S. hydrophila Boiss., S. nivea Labill., S. palustris L. and S. spinosa L., obtained by hydrodistillation, was studied by GC and GC–MS. All the oils have in common a great percentage of fatty acids and esters (24.2–58.5%) and a high amount of sesquiterpenes (16–35.9%), with the exception of the oil from S. palustris, which consisted mainly of carbonylic compounds (25.4%). The antioxidant activity by DPPH test and the antiproliferative activity on a series of human cancer cell lines (C32, amelanotic melanoma and ACHN, renal cell adenocarcinoma) were investigated for all the oils. S. palustris,S. cretica and S. hydrophila showed the highest antiradical effect, with IC₅₀ values of 0.482, 0.652 and 0.664 mg/ml, respectively. The most antiproliferative essential oil against C32 cell line was the oil of S. germanica with a 77% of inhibition at a concentration of 100 μg/ml. S. germanica, S. palustris and S. spinosa showed the most antiproliferative activity on ACHN cell line, at a concentration of 100 μg/ml,with 81%, 77% and 73% inhibition, respectively.     

Application of Kluyveromyces marxianus, Lactobacillus delbrueckii ssp. bulgaricus and L. helveticus for sourdough bread making

The application of Kluyveromyces marxianus (IFO 288), Lactobacillus delbrueckii ssp. bulgaricus (ATCC 11842) and Lactobacillus helveticus (ATCC 15009) as starter cultures for sourdough bread making was examined. Production of lactic and acetic acids, bread rising, volatile composition, shelf-life and organoleptic quality of the sourdough breads were evaluated. The amount of starter culture added to the flour, the dough fermentation temperature and the amount of sourdough used were examined in order to optimise the bread making process. The use of mixed cultures led to higher total titratable acidities and lactic acid concentrations compared to traditionally made breads. Highest acidity (3.41 g lactic acid/kg of bread) and highest resistance to mould spoilage were observed when bread was made using 50% sourdough containing 1% K. marxianus and 4% L. delbrueckii ssp. bulgaricus. The use of these cultures also improved the aroma of sourdough breads, as shown by sensory evaluations and as revealed by GC–MS analysis.     

Sensory,microbiological and chemical assessment of the freshness of red mullet (Mullus barbatus) and goldband goatfish (Upeneus moluccensis) during storage in ice

The shelf life of red mullet and goldband goatfish during ice storage were studied in terms of sensory, microbiological and chemical changes. The sensory acceptability limit was 8 days for goldband goatfish (Upeneus moluccensis) and 11 days for red mullet (Mullus barbatus) stored in ice. The TVC level was correlated with sensory assessment. The TVC exceeded 7 log cfu g⁻¹ after 8 days for goldband goatfish, and 11 days for red mullet. At the end of storage period, pH, TVB-N, TBA, FFA and PV for red mullet were 7.84, 47.19 mg/100 g, 0.69 mg MA kg⁻¹, 1.17% oleic acid and 1.58 meq O₂/kg and for goldband goatfish they were 7.53, 43.97 mg/100 g, 0.74 mg MA kg⁻¹, 1.62% oleic acid and 1.68 meq O₂/kg, respectively. In red mullet, agmatine, serotonin, histamine and dopamine became the dominant amines, reaching 7.30, 5.97, 2.52 and 2.31 mg/100 g, respectively. Also the dominant amines for goldband goatfish were 4.37, 3.88, 3.38 and 2.00 mg/100 g for histamine, agmatine, dopamine and putrescine, respectively.     

Lipid characteristics in cooked, chill-reheated fillets of Indo-Pacific king mackerel (Scomberomorous guttatus)

Slices of Scomberomorous guttatus cooked by four different methods (microwave cooking, grilling, steaming and shallow fat frying) were kept chilled at 5 °C for 2 days, reheated by microwave and then evaluated for the changes in the lipid characteristics. The total lipid content for raw sample was 6.0 g/100 g which consist of 49.4 g/100 g saturated (SFA), 6.9 g/100 g monounsaturated (MUFA) and 43.8 g/100 g polyunsaturated fatty acids (PUFA). Upon cooking, the total lipids changed to about 7 (microwave), 11 (grilling), 5 (steaming) and 10 g/100 g (shallow fat frying). These changes are significant, with the exception of steam cooking. Cooking did not significantly affect the percentage of NPL, PL, SFA, MUFA and PUFA in the samples. Frying resulted in the change of the SFA/PUFA ratio more than other cooking methods. A significant (P < 0.05) decrease in the content of EPA was only found in the fried samples. A significant (P < 0.05) decrease in the content of DHA in all cooked samples were also obtained. The n-6/n-3 ratio did not change significantly in all cooked and reheated samples, except for shallow fat fried. Reheating did not significantly affect the NPL and PL contents of the fillets, except for microwave cooked samples. Cooking increases the FFA, PV and TBA of samples; reheating enhances the increase.