Tenderization and fragmentation of myofibrillar proteins in bovine longissimus dorsi muscle using proteolytic extract from Sarcodon aspratus

The objective of this study was to examine the Sarcodon aspratus extract including protease how to affect tenderness of the bovine longissimus dorsi muscle. In addition, we investigated myofibrillar protein fragmentation, particularly in myosin, and its influence on meat tenderness. Beef loin chunks were marinated with 0.5 g/100 g, 1 g/100 g, and 2 g/100 g powdered S. aspratus extract, 0.2 g/100 mL papain, and distilled water (control), respectively. Although tenderness of meat is increased by adding S. aspratus extract, differences in meat quality traits, such as muscle pH and meat color, were small and not considered to have practical importance between the control and enzyme-treated samples. Furthermore, the S. aspratus extract influenced the myofibril fragmentation index (MFI) as well as protein solubility. The changes in MFI and protein solubility were due to the myofibrillar protein degradation. Through Western blotting, we found that the S. aspratus extract, as well as papain, caused fragmentation of the myosin heavy chain, but the mushroom extract induced more fragmentations of myofibrillar proteins, and caused more tender meat.     

Advantages of a proteolytic extract by Aspergillus oryzae from fish flour over a commercial proteolytic preparation

Proteolytic extract (CPE) was produced by Aspergillus oryzae 2095 under solid state fermentation using a mix of fish flour with polyurethane foam (70:30, w/w) of particle size of 0.5 mm. The proteolytic activity from CPE was compared to a commercial protease (Flavourzyme 500 MG^®). The maximal activity for both extracts was found at pH 8 and 50 °C. The half-lives of CPE and Flavourzyme 500 MG^® were 52 and 25 min at 50 °C, respectively. Furthermore, the hydrolytic activity for both extracts was tested on muscle of giant sea bass (Epinephelus morio), CPE produced a higher degree of hydrolysis (DH) than Flavourzyme 500 MG^® (22.2 and 9.1%, respectively) under optimal experimental conditions for each extract.     

Multiplex fiber optic biosensor for detection of Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella enterica from ready-to-eat meat samples

Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella enterica are the most common foodborne bacterial pathogens and are responsible for many outbreaks. Therefore, multiplex detection of these three using a single assay platform is highly desirable. The objective was to develop and optimize a fiber optic sensor for simultaneous detection of these three from food. The streptavidin coated optical waveguides were immobilized with biotinylated polyclonal antibodies and exposed to the bacterial suspensions or enriched food samples for 2 h. Pathogens were detected after reacting with Alexa-Fluor 647-labeled monoclonal antibodies. Ready-to-eat beef, chicken and turkey meats were inoculated with each pathogen (∼100 cfu/25 g), enriched in SEL (Salmonella, E. coli, Listeria), a multipathogen selective enrichment broth for 18 h and tested with the biosensor. The biosensor was able to detect each pathogen, individually or in a mixture with very little cross-reactivity. The limit of detection for the sensor was ∼10³ cfu/ml for all three pathogens. Furthermore, the biosensor successfully detected each pathogen, grown in a mixture from enriched meat samples under 24 h. The pathogen presence was further verified by PCR and immunofluorescence assay. The multiplex fiber optic sensor shows promise for detection of the three pathogens if present in the same sample eliminating the use of multiple single pathogen detection platforms.     

Anti-melanogenesis properties of quercetin- and its derivative-rich extract from Allium cepa

In an effort to find a new whitening agent, we have found that the methanol extract of the dried skin of Allium cepa showed inhibition of melanin formation. Bioassay-guided fractionation led to the isolation of quercetin (1) and quercetin 4’-O-β-glucoside (3) from A. cepa as the inhibitors of melanin formation in B16 melanoma cells with IC₅₀ values of 26.5 and 131 μM, respectively. In addition, we evaluated the effect of some quercetin derivatives, such as isoquercitrin (2), quercetin 3,4’-O-diglucoside (4), rutin (5) and hyperin (6) on B16 melanoma cells. These quercetin derivatives did not show any inhibition of melanin formation. Furthermore, the ORAC values of compounds 1–6 were 7.64, 8.65, 4.82, 4.32, 8.17 and 9.34 μmol trolox equivalents/μmol, respectively. Dried skin of red onion showed inhibitory activity against melanin formation in B16 melanoma cells, as well as antioxidant properties.     

Satureja horvatii essential oil: In vitro antimicrobial and antiradical properties and in situ control of Listeria monocytogenes in pork meat

The dominant compounds in Satureja horvatii oil were p-cymene (33.14%), thymol (26.11%) and thymol methyl ether (15.08%). The minimum inhibitory concentration (MIC) varied from 0.03 to 0.57 mg/mL for bacteria, and from 0.56 to 2.23 mg/mL for yeast strains, while minimum bactericidal/yeast-cidal concentration (MBC/MYC) varied from 0.07 to 1.15 mg/mL and 1.11 to 5.57 mg/mL for bacteria and yeasts, respectively. The antiradical potential of the essential oil was evaluated using hydroxyl radical (•OH) generated in Fenton reaction. The meat preserving potential of essential oil from Satureja horvatii was investigated against L. monocytogenes. Essential oil successfully inhibited development of L. monocytogenes in pork meat. Sensorial evaluation on flavor and color of meat was performed. The color and flavor of meat treated with essential oil improved after 4 days of storage. S. horvatii essential oil can act as a potent inhibitor of food spoiling microorganisms, in meat products and also can be a useful source of natural antioxidants.     

Iron-chelating ability and antioxidant properties of phycocyanin isolated from a protean extract of Spirulinaplatensis

The invitro scavenger activities of different reactive oxygen species (superoxide radical, hydroxyl radical, hydrogen peroxide, hypochlorous acid and peroxyl radical), the effects on lipid peroxidation and the iron-chelating ability of a Spirulinaplatensis protean extract and the biliprotein, phycocyanin, isolated from this microalga were studied. S. platensis protean extract inhibited the generation of hydroxyl radical (IC₅₀ = 537 μg/ml for the system with EDTA and 1500 μg/ml without EDTA), the production of peroxyl radical (IC₅₀ = 230 μg/ml), and the lipid peroxidation process (IC₅₀ = 2320 μg/ml for the enzymatic system and 2180 μg/ml for the non-enzymatic system). Besides, phycocyanin inhibited hydroxyl and peroxyl radicals and the lipid peroxidation process. The iron ions decreased the maximum fluorescence emission spectra of S. platensis protean extract and phycocyanin and it was an indicator of the metal-chelating activity. The antioxidant properties of S. platensis and phycocyanin may arise from both radical-scavenging and metal chelation. Our results suggest that S. platensis could be used as a dietary supplement to prevent some diseases where free radicals are involved.     

Physicochemical and functional properties of makal (Xanthosoma yucatanensis) starch

Physicochemical and functional properties of makal (Xanthosoma yucatanensis) starch were determined. Granules were oval in shape and 12.4 μm average diameter. Starch purity was high (96.7%) with low protein (0.1%), fat (0.2%), fibre (0.4%) and ash (0.1%) contents. Amylose content was 22.4%. The gelatinization temperature was 78.5 °C and transition enthalpy was 15 J/g. At 90 °C, solubility was 32.9%, swelling power was 28.6 g water/g starch and water absorption capacity was 19.2 g water/g starch. Pasting characteristics were: temperature 75 °C, maximum viscosity 280 BU, breakdown −8 BU, setback 180 BU and consistency 172 BU. Clarity, expressed as transmittance, was 35.8%. Gel deformation was 20.8% with a 0.03 kgf maximum load. Makal starch’s high gelification temperature and firmness make it appropriate for use in high temperature food systems, but its low stability in refrigeration and freezing cycles make it inadequate for use in foods subject to those conditions.     

Physicochemical characterization of chitosan extracted from Metapenaeus stebbingi shells

In this study, chitosan was extracted from Metapenaeus stebbingi shells. In order to determine physicochemical characteristics of the extracted chitosan, the yield, moisture and ash contents, degree of deacetylation, molecular weight, water and fat binding capacities, apparent viscosity and colour properties were measured using a variety of techniques including Fourier transform infrared spectroscopy, scanning electron microscopy and X-ray diffraction. In addition, the physicochemical characteristics of the chitosan extracted from M. stebbingi shells were compared to commercial chitosan. The degree of deacetylation was calculated by the titration method and elemental analysis. The molecular weight was determined by viscosimetric methods. The results of the study indicate that shrimp shells are a rich source of chitosan as 17.48% of the shell’s dry weight is consisted of this material. Extracted chitosan exhibited a lower molecular weight, higher degree of deacetylation, higher viscosity and higher water and fat binding capacities compared to the commercial chitosan.     

Heat shock and structural proteins associated with meat tenderness in Nellore beef cattle,a Bos indicus breed

Nellore beef cattle, a Bos indicus (Zebu) breed, is well adapted to tropical conditions and has allowed Brazil to become one of the largest producers of red meat. Nevertheless, B. indicus breeds are reported to have less tender meat than Bos taurus. This study was designed to identify genes associated with meat tenderness and thus provides important information for breeding programs. A group of 138 animals was evaluated for longissimus thoracis muscle shear force (SF). Animals with the highest and lowest SF values (six animals each) were then selected for protein abundance studies. Samples were subjected to two-dimensional gel electrophoresis (2-DE) followed by peptide sequencing through mass spectrometry (MS) to identify differentially expressed proteins associated with SF values. Seventeen differentially expressed spots were observed (p < 0.05) between the two groups. The 13 proteins identified included structural proteins (alpha actin-1, MLC1, MLC3, MLC2F and tropomyosin), related to cell organization (HSPB1 and HSP70), metabolism (beta-LG, ACBD6 and Complex III subunit I) and some uncharacterized proteins. Results confirm the existence of differentially expressed proteins associated with SF, which can lead to a better understanding of mechanisms involved in meat tenderness.     

Physicochemical characteristics and production of whole soymilk from Monascus fermented soybeans

Monascus fermented soybeans (MFS) were prepared by solid state fermentation. MFS showed higher solubility than the unfermented control within the acidic pH region but showed the opposite trend at pH 7 and 9. Although the emulsifying activity index of MFS was lower than that of the control, MFS had significantly higher emulsion stability at pH 7 and 9. Significant protein hydrolysis took place during fermentation, and the proportion of peak area less than 1.35 kDa was greatly increased in MFS. Whole soymilk prepared from MFS contained monacolin K (475 μg/g), and was enriched in isoflavone aglycones. Neither homogenisation nor pasteurisation caused significant changes in the isoflavone and monacolin K contents of the Monascus fermented soymilk. Total phenol content and ABTS radical scavenging activity were significantly higher in the Monascus fermented soymilk than in the control soymilk.     

Antimicrobial and antioxidant properties of the essential oils and methanol extract from Mentha longifolia L. ssp. longifolia

This study was designed to evaluate antimicrobial and antioxidant activities of the essential oil and methanol extract from Mentha longifolia ssp. longifolia. The essential oil showed strong antimicrobial activity against all 30 microorganisms tested whereas the methanol extract almost remained inactive. In contrast, the extract showed much better activity than the essential oil in antioxidant activity assays employed, e.g. in the inhibition of free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene/linoleic acid systems. In the former, the extract was able to reduce the stable free radical DPPH with an IC₅₀ of 57.4 μg/ml while that of the oils was 10 700 μg/ml. When compared to BHT, a synthetic antioxidant, both showed weaker antioxidative potential. Similarly, in β-carotene/linoleic acid assay, these samples were not effectively able to inhibit the linoleic acid oxidation; exhibiting only 24% and 36% inhibitions at 2 mg/ml, respectively; both were far below than that of BHT. Total phenolic constituent of the extract was 4.5 g/100 g as gallic acid equivalent. GC–MS analysis of the oil resulted in the identification of 45 constituents, cis-piperitone epoxide, pulegone and piperitenone oxide being the main components.     

Lactic acid resistance of Shiga toxin-producing Escherichia coli and multidrug-resistant and susceptible Salmonella Typhimurium and Salmonella Newport in meat homogenate

This study compared lactic acid resistance of individual strains of wild-type and rifampicin-resistant non-O157 Shiga toxin-producing Escherichia coli (STEC) and of susceptible and multidrug-resistant (MDR) and/or MDR with acquired ampC gene (MDR-AmpC) Salmonella against E. coli O157:H7. After inoculation of sterile 10% beef homogenate, lactic acid was added to a target concentration of 5%. Before acid addition (control), after acid addition (within 2 s, i.e. time-0), and 2, 4, 6 and 8 min after addition of acid, aliquots were removed, neutralized, and analyzed for survivors. Of wild-type and of rifampicin-resistant non-O157 STEC strains, irrespective of serogroup, 85.7% (30 out of 35 strains) and 82.9% (29 out of 35 strains), respectively, reached the detection limit within 0–6 min. Of Salmonella strains, 87.9% (29 out of 33 isolates) reached the detection limit within 0–4 min, irrespective of antibiotic resistance phenotype. Analysis of non-log-linear microbial survivor curves indicated that non-O157 STEC serogroups and MDR and susceptible Salmonella strains required less time for 4D-reduction compared to E. coli O157:H7. Overall, for nearly all strains and time intervals, individual strains of wild-type and rifampicin-resistant non-O157 STEC and Salmonella were less (P < 0.05) acid tolerant than E. coli O157:H7.     

Physicochemical properties of a fibrous fraction from chia (Salvia hispanica L.)

An evaluation was done of some physicochemical properties of a fiber-rich fraction (FRF) obtained by dry processing of defatted chia (Salvia hispanica) flour. The fiber-rich fraction (FRF) had 29.56 g/100 g crude fiber content and 56.46 g/100 g total dietary fiber (TDF) content, of which 53.45 g/100 g was insoluble dietary fiber (IDF) and 3.01 g/100 g was soluble dietary fiber (SDF). The FRF water-holding capacity was 15.41 g/g, its water absorption capacity 11.73 g/g, and its organic molecule absorption capacity 1.09 g/g. The FRF also had low oil-holding (2.02 g/g) and water adsorption (0.3 g/g) capacities. Emulsifying activity in this fraction was 53.26% and emulsion stability was 94.84%. Its evaluated antioxidant activity was 488.8 μmol/L Trolox equivalents/g FRF, which is higher than for many cereals and similar to drinks such as wine, tea, coffee and orange juice. The chia FRF values for the evaluated properties, particularly for water-holding, oil-holding and organic molecule absorption capacity, suggest it could be a useful ingredient in dietetic products such as baked and fried foods, among others.     

Multiple cases of methicillin-resistant CC130 Staphylococcus aureus harboring mecC in milk and swab samples from a Bavarian dairy herd

The discovery of a new mecA homolog, mecC, necessitates a modification of diagnostic procedures for the identification of methicillin-resistant Staphylococcus aureus (MRSA), as most assays used for the genotypic and phenotypic mecA detection cannot currently recognize mecC. Although the prevalence, distribution, and importance of mecC are not yet completely understood, an exchange of mecC-MRSA between humans and animals seems possible. All previously reported observations of mecC-positive strains have been sporadic. To the best of our knowledge, this is the first report about multiple cases of mecC-positive Staph. aureus in 1 dairy herd. Clonal complex 130 Staph. aureus harboring mecC were found in milk samples from 16 of 56 lactating cows kept in a herd in Bavaria, Germany. Almost all quarter milk samples positive for mecC-MRSA had the lowest possible California Mastitis Test score; composite somatic cell counts obtained from monthly milk recordings showed a mean of 51,600 cells/mL in mecC-MRSA affected cows. Additionally, mecC-positive clonal complex 130 Staph. aureus were detected in swab samples from the mammary skin and a teat lesion of 1 cow from this herd. This report suggests that mecC-carrying strains are able to spread among livestock, and that they have the ability to cause multiple cases in single herds. Therefore, future studies targeting MRSA in dairy cows need to consider mecC.     

Physicochemical characterisation of β-chitosan from Sepioteuthis lessoniana gladius

β-Chitin and its chitosan from the gladius of Sepioteuthis lessoniana have been isolated, purified, characterised and compared with the commercial chitosan. Ash, moisture, mineral, metal and elemental content were analyzed using standard techniques. The optical activity of chitin was found to be levorotatory. The degree of deacetylation was calculated by potentiometric titration and ¹H NMR. Viscosity average molecular weight of β-chitosan was calculated by viscometry and size average molecular weight by GPC. The structure of β-chitosan was elucidated with FT-IR and NMR. Thermal nature, crystalline structure and morphology of β-chitosan were characterised through DSC, XRD and SEM, respectively. The water and fat binding capacity of β-chitosan presently studied was significantly higher than that of the commercial chitosan. The result of the present study adds that S. lessoniana gladius is also an additional source of β-chitin and chitosan of higher yield, lower molecular weight and higher degree of deacetylation.     

Short communication: Presence of neutral metallopeptidase (npr) gene and proteolytic activity of Bacillus cereus isolated from dairy products

The control of proteolytic microorganisms is one of the main challenges of the dairy industry, due to their spoilage activity that jeopardizes the quality of their products. Seventy-four Bacillus cereus strains isolated from powdered, UHT, and pasteurized milks were tested for the presence of the neutral metallopeptidase (npr) gene and proteolytic activity at 7, 10, 25, 30, and 37°C. All strains had the npr gene, and proteolytic activity increased with the incubation temperature. The obtained results highlight the relevance of B. cereus as a spoiling agent in the dairy industry in terms of its genetic predisposition for proteolytic capacity, especially at room temperature.     

Antioxidant properties of Agaricus blazei, Agrocybe cylindracea, and Boletus edulis

Three mushrooms are currently available in Taiwan, including Agaricus blazei, Agrocybe cylindracea, and Boletus edulis. Their ethanolic and hot water extracts were prepared and antioxidant properties studied. Ethanolic extracts from three mushrooms were more effective than hot water extracts in antioxidant activity using the conjugated diene method and scavenging ability on 1,1-diphenyl-2-picrylhydrazyl radicals whereas hot water extracts were more effective in reducing power, scavenging ability on hydroxyl radials and chelating ability on ferrous ions as evidenced by their lower EC₅₀ values. Overall, for both extracts, B. edulis was more effective among antioxidant properties assayed. Naturally occurring antioxidant components including total tocopherols (3.18-6.18 mg/g) and total phenols (5.67-5.81 mg/g) were found in the extracts and their contents were associated (r=0.636-0.907) with EC₅₀ value of antioxidant properties. Based on the results obtained, both extracts from these three mushrooms were effective in antioxidant properties.     

Salmonella surveillance and control at post-harvest in the Belgian pork meat chain

Salmonella remains the primary cause of reported bacterial food borne disease outbreaks in Belgium. Pork and pork products are recognized as one of the major sources of human salmonellosis. In contrast with the primary production and slaughterhouse phases of the pork meat production chain, only a few studies have focussed on the post-harvest stages. The goal of this study was to evaluate Salmonella and Escherichia coli contamination at the Belgian post-harvest stages. E. coli counts were estimated in order to evaluate the levels of faecal contamination. The results of bacteriological analysis from seven cutting plants, four meat-mincing plants and the four largest Belgian retailers were collected from official and self-monitoring controls. The prevalence of Salmonella in the cutting plants and meat-mincing plants ranged from 0% to 50%. The most frequently isolated serotype was Salmonella typhimurium. The prevalence in minced meat at retail level ranged from 0.3% to 4.3%. The levels of Salmonella contamination estimated from semi-quantitative analysis of data relating to carcasses, cuts of meat and minced meat were equal to −3.40 ± 2.04 log CFU/cm², −2.64 ± 1.76 log CFU/g and −2.35 ± 1.09 log CFU/g, respectively. The E. coli results in meat cuts and minced meat ranged from 0.21 ± 0.50 to 1.23 ± 0.89 log CFU/g and from 1.33 ± 0.58 to 2.78 ± 0.43 log CFU/g, respectively. The results showed that faecal contamination still needs to be reduced, especially in specific individual plants.