Characterisation of the spoilage microbiota in raw salmon (Salmo salar) steaks stored under vacuum or modified atmosphere packaging combining conventional methods and PCR-TTGE

In order to characterise the spoilage related to microbiota of raw salmon, a combination of culture-dependent and -independent methods, including PCR–TTGE, was used to analyse 3 raw salmon batches stored for 3 days at chilled temperature in modified atmosphere packaging (MAP) (50% CO₂/50% N₂) or under vacuum. Sensory evaluation, microbiological enumeration and chemical analysis were performed after 3, 7 and 10 days of storage. At the onset of spoilage, 65 bacterial isolates were picked from the plates. Thus, 13 different genera or species were identified by phenotypic and molecular tests: Serratia spp., Photobacterium phosphoreum, Yersinia intermedia, Hafnia alvei, Buttiauxella gaviniae, Pseudomonas sp., Carnobacterium maltaromaticum, Carnobacterium divergens, Lactococcus piscium, Lactobacillus fuchuensis, Vagococcus carniphilus, Leuconostoc gasicomitatum and Brochothrix thermosphacta. The PCR–TTGE profiles and band identification enabled a shift of the dominant populations during the storage to be visualised for all the batches, probably due to the temperature change and the packaging. At the beginning of storage, Pseudomonas sp. dominated the raw salmon microbiota while in the following days (7 and 10), P. phosphoreum and L. piscium were identified as the main bacterial groups. This study enhances the knowledge of MAP and vacuum-packed raw salmon spoilage microbiota.     

Shelf life extension of whole Norway lobster Nephrops norvegicus using modified atmosphere packaging

Once a nuisance by-catch, today the Norway lobster (Nephrops norvegicus) is a valuable UK fisheries commodity. Unfortunately, the species is very susceptible to quality deterioration post harvest as it quickly develops black spots and also spoils rapidly due to bacterial growth. Treatment with chemicals can stop the blackening and carefully monitored cold storage can result in a sensory shelf life of up to 6.5 days. The high susceptibility to spoilage greatly restricts the extent to which N. norvegicus can be distributed to retailers and displayed for sale. The application of modified atmosphere (MA) could be extremely beneficial, allowing the chilled product to stay fresh for a long period of time, thus ensuring higher sales. In the present study, we identified a gas mix for the MA packaging (MAP) of whole N. norvegicus lobster into 200 g retail packs. Our results show that a shelf life extension to 13 days can be achieved when retail packs are stored in MAP at 1 °C. Effectiveness of the MAP was evaluated by using a newly developed QIM for MA-packaged whole N. norvegicus and also by analyzing bacterial plate counts. Changes in the microflora and effects of different storage temperatures on the quality of the MA packs are also presented. The main specific spoilage organism (SSO) of modified atmosphere packaged Norway lobster is Photobacterium phosphoreum.     

冷冻即食海参质构特性指标的相关性研究

目的：全面评价冷冻即食海参的质构特性。方法：采用质构仪(质地剖面、剪切力和应力松弛模式)检测和感官评价方法对4种方式解冻的冷冻即食海参进行质构测定,对仪器检测各指标间、仪器检测指标与感官评价指标间进行相关性分析,并对仪器检测指标进行主成分分析。结果：4种解冻组的仪器检测指标与感官评价指标均具有显著性差异(P<0.05)。除TPA黏聚性与TPA弹性、纵剪切力、粘滞系数η外,9项仪器检测指标间均呈较高的相关性(r≥0.800);同时,仪器检测指标和感官评价指标间存在一定的相关性(r=0.577～0.994)。主成分分析表明感官评价总分最高的冷藏解冻组趋近于主成分1和2的正方向,且能同时被TPA黏聚性、TPA硬度、横剪切力和弹性模量E₁较好地解释。结论：冷冻即食海参质构特性指标间具有很好的相关性,本研究结果为冷冻即食海参的质构评价研究提供参考。     

Diversity of culturable microorganisms and occurrence of Listeria monocytogenes and Salmonella spp. in Tuber aestivum and Tuber melanosporum ascocarps

The aim of this study was to investigate the total mesophilic microorganisms, Pseudomonas genus, Enterobacteriaceae family, mold and yeast counts and the presence of Listeria monocytogenes and Salmonella spp on Tuber aestivum and Tuber melanosporum ascocarps. The results confirmed that the major percentage of the microorganisms, approximately 9.0 log ufc/g, were present in the peridium, the glebas of healthy truffles being practically free of microorganisms. The predominant microbial group was the Pseudomonas averaging 8.3 and 8.4 log cfu/g on T. aestivum and T. melanosporum whole ascocarps, respectively. The Enterobacteriaceae also achieved high populations, especially in T. aestivum truffles, with 6.3 log cfu/g. Molds and yeasts never exceeded 5.0 log cfu/g. The characterization of the isolates revealed that the fluorescens pseudomonads were the most prevalent. Raoultella terrigena and Enterobacter intermedius were the dominant Enterobacteriaceae. The identification of the yeast isolates revealed five species: Debaryomyces hansenii, Issatchenkia scutulata, Rhodotorula aurantiaca, Saccharomyces dairensis and Trichosporon beigelii subspecies A and B. The mold genera detected in both species of truffles were Aspergillus, Cladosporium, Penicillium and Fusarium, Trichoderma being present only in T. aestivum. L. monocytogenes was found in 10% of the samples of T. aestivum analysed but Salmonella spp. was not detected. Knowledge of the microbial population in terms of possible food borne and pathogen microorganisms is very useful for establishing successful disinfection and storage methods to prolong the shelf-life of ascocarps of T. aestivum and T. melanosporum.     

Occurrence of psocids and natural predators on organic rice in Calasparra (Murcia, Spain)

Five Psocoptera species: Lepinotus reticulatus, Liposcelis entomophila, Liposcelis mendax, Liposcelis bostrychophila and Liposcelis paeta were identified on organic paddy stored in south-east Spain. The main natural predator associated with the psocids in the study area was the pseudoscorpion Withius piger, reported for the first time within the Iberian Peninsula.     

Oxidation in HiOx-packaged pork Longissimus muscle predisposes myofibrillar and sarcoplasmic proteins to N-nitrosamine formation in nitrite-curing solution

The effect of meat protein in situ oxidation on the formation of N-nitrosodiethylamine (NDEA) was investigated. Fresh minced pork was untreated (Con) or treated with 700 mg/kg α-tocopherol (Toc) or 300 mg/kg tea polyphenols (PPE), packaged in a HiOx atmosphere (78.8% O₂, 18.8% CO₂, 2.4% N₂), then stored at 2 ± 1 °C for up to 10 days. Crude myofibrillar (MP) or sarcoplasmic (SP) protein (20 mg/mL) extracted from stored meat was reacted with 43 μM sodium nitrite at 80 °C for 1 h. Lipid oxidation was totally inhibited in PPE pork but increased in Con and Toc samples after 10 days. There was significant protein oxidation (losses of sulfhydryls, formation of protein carbonyls) in both MP and SP in all samples during storage. However, the Con group suffered more extensive protein oxidation than Toc and PPE and produced more NDEA (P < 0.05), indicating that protein oxidation promoted nitrosation.     

Isolation and quantitative determination of ergosterol peroxide in various edible mushroom species

Ergosterol peroxide, the steroidal derivative with cytotoxic activity, has been isolated for the first time from the mycelium of edible and medicinal mushroom Hericiumerinaceum (lion’s mane mushroom) together with erinacine A. The new densitometric method was applied for the quantitative determination of ergosterol peroxide in n-hexane extracts of H. erinaceum, Laetiporus sulfureus (chicken mushroom), and Morchella esculenta (common morel) mycelia, as well as in Boletus edulis (king bolete), Suillus bovinus (Jersey cow mushroom), and B. badius (bay bolete) fruiting bodies. The ergosterol peroxide content reached 15.98 ± 0.78, 10.07 ± 0.75, 13.37 ± 0.56, 29.32 ± 1.43, 17.27 ± 0.84, and 12.60 ± 0.59 mg per 100 g, respectively. What is significant was that ergosterol peroxide was identified for the first time, to the best of our knowledge, in edible mushrooms mentioned above.     

Chemical composition and insecticidal properties of some aromatic herbs essential oils from Algeria

The biological effects on Sitophilus granarius were evaluated for three aromatic herbs essential oils: Foeniculum vulgare, Rosmarius officinalis and Lippia citriodora. Stored grain pests were currently controlled by chemical pesticides. This control method leads to pollution of the environment and intoxication of consumers. Essential oils of aromatic plants are more considered as good control alternative tools. Filter papers treated with 5, 50 and 500 μl of test oil were placed in the bottom cover of 1 l plastic bottle. The insects, 50 adults per bottle, were exposed for 1–5 days. Cumulative mortalities were determined 24 and 120 h after treatment. All treatments were replicated five times. The components of the essential oils were identified through GC and GC–MS. The identity of the constituents was confirmed and their relative proportions determined. The results indicate that these natural products may find potential application as useful, environmentally safe insect control and crop protectant agents.     

Purification, immunological properties and molecular cloning of two allergenic parvalbumins from the crimson sea bream, Evynnis japonica

Parvalbumin, a calcium-binding sarcoplasmic protein of 12 kDa, represents a major allergen of fish. Previous immunoblotting experiments suggested the presence of a 14 kDa allergen, together with parvalbumin, in sea breams. In this study, the 14 kDa allergen (PA I) and parvalbumin (PA II) were purified from the white muscle of crimson sea bream, Evynnis japonica, by gel filtration and reverse-phase HPLC. Amino acid sequencing of lysylendopeptidase peptide fragments demonstrated that both PA I and PA II were isoforms of parvalbumin. As analysed by ELISA, PA I showed weaker reactivities with IgG (monoclonal or polyclonal antibody) and serum IgE in fish-allergic patients than did other fish parvalbumins, including PA II. The amino acid sequences of PA I and PA II were elucidated by cDNA cloning. PA I was found to have more specific amino acid residues at several positions compared to the other fish parvalbumins.     

Evaluation of lactic acid bacteria and yeast diversity in traditional white pickled and fresh soft cheeses from the mountain regions of Serbia and lowland regions of Croatia

The goal of this study was the characterisation of indigenous lactic acid bacteria (LAB) and yeasts isolated from nine white pickled (BG) and nine fresh soft (ZG) artisanal cheeses collected in Serbia and Croatia. While LAB were present in all of the cheeses collected, yeasts were found in all BG cheeses but only in three ZG cheese samples. High LAB and yeast species diversity was determined (average H′_L = 0.4 and H′_Y = 0.8, respectively). The predominant LAB species in white pickled (BG) cheeses were Lactococcus lactis, Lactobacillus plantarum, and Leuconostoc mesenteroides, while in fresh soft (ZG) cheeses the most dominant LAB species were L. lactis, Enterococcus faecalis, and Leuconostoc pseudomesenteroides. Among the 20 yeast species found, Debaryomyces hansenii, Candida zeylanoides, and Torulaspora delbrueckii were found to be predominant in BG cheeses, while Yarrowia lipolytica was predominant in ZG cheeses. The characterisation of metabolic and technological potentials revealed that 53.4% of LAB isolates produced antimicrobial compounds, 44.3% of LAB strains showed proteolytic activity, while most of the yeast species possessed either lipolytic or proteolytic activity. In conclusion, the results obtained in this study showed that the composition of LAB and yeast populations in white pickled and fresh soft cheeses is region specific. The knowledge gained in this study could eventually be used to select region specific LAB and yeast strains for the production of white pickled and fresh soft artisanal cheeses with geographically specific origins under controlled conditions.     

Variation in the susceptibility of two Callosobruchus species to essential oils

GC/MS analysis of essential oils extracted from two Cymbopogon species revealed that limonene (23%) and p-mentha-2,8-dien-1-ol in cis (14.3%) and trans (5.6%) forms were the main compounds in Cymbopogon giganteus oil whereas citronellal (31%) and geraniol (24%) were identified in Cymbopogon nardus oil. The toxicity tests performed by fumigation on eggs and adults of Callosobruchus maculatus and Callosobruchus subinnotatus using both essential oils showed a variation in bruchid susceptibility. Essential oil of C. giganteus was more toxic to adults of both bruchid species while essential oil of C. nardus showed the better ovicidal activity. Comparative susceptibility analysis suggested that eggs and adults of C. subinnotatus were two-fold more tolerant to essential oils than C. maculatus in both stages. Oviposition of treated females was strongly reduced in the presence of essential oils. Callosobruchus subinnotatus was more affected than C. maculatus by the essential oil of C. giganteus (oviposition reduction by at least 91% v.s 81% in C. maculatus at 5 μL/L) but the two species were affected similarly by the essential oil of C. nardus.     

Characterization of Longissimus thoracis,Semitendinosus and Masseter muscles and relationships with technological quality in pigs. 1. Microscopic analysis of muscles

Three porcine muscles (Longissimus thoracis, Semitendinosus, Masseter), known to have large differences in biochemical and histological traits, were fully characterized and the link between muscle structure and quality evaluated. The oxidative Masseter had more pigment, higher content of metmyoglobin, haem iron, protein and collagen, and was redder with higher fibre numbers, fibre circularity, pH and water holding capacity than the glycolytic Longissimus. Fibre type distribution showed predominance of type IIB in Longissimus and Semitendinosus white, type I in Semitendinosus red and IIA in Masseter. Type I fibres were larger than type IIB and IIA in Semitendinosus and Masseter, respectively, but not in the Longissimus, indicating that fibre size is muscle dependent. Muscle redness was positively correlated with type I fibre traits, haem iron and metmyoglobin, and negatively associated with type II fibre characteristics, non-haem iron and oxymyoglobin. Expressible juice had positive correlation with fibre size and negative with fibre number and connective tissue.     

Reduction of bacteria on spinach, lettuce, and surfaces in food service areas using neutral electrolyzed oxidizing water

Food safety issues and increases in food borne illnesses have promulgated the development of new sanitation methods to eliminate pathogenic organisms on foods and surfaces in food service areas. Electrolyzed oxidizing water (EO water) shows promise as an environmentally friendly broad spectrum microbial decontamination agent. EO water is generated by the passage of a dilute salt solution ( approximately 1% NaCl) through an electrochemical cell. This electrolytic process converts chloride ions and water molecules into chlorine oxidants (Cl(2), HOCl/ClO(-)). At a near-neutral pH (pH 6.3-6.5), the predominant chemical species is the highly biocidal hypochlorous acid species (HOCl) with the oxidation reduction potential (ORP) of the solution ranging from 800 to 900mV. The biocidal activity of near-neutral EO water was evaluated at 25 degrees C using pure cultures of Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis. Treatment of these organisms, in pure culture, with EO water at concentrations of 20, 50, 100, and 120ppm total residual chlorine (TRC) and 10min of contact time resulted in 100% inactivation of all five organisms (reduction of 6.1-6.7log(10)CFU/mL). Spray treatment of surfaces in food service areas with EO water containing 278-310ppm TRC (pH 6.38) resulted in a 79-100% reduction of microbial growth. Dip (10min) treatment of spinach at 100 and 120ppm TRC resulted in a 4.0-5.0log(10)CFU/mL reduction of bacterial counts for all organisms tested. Dipping (10min) of lettuce at 100 and 120ppm TRC reduced bacterial counts of E. coli by 0.24-0.25log(10)CFU/mL and reduced all other organisms by 2.43-3.81log(10)CFU/mL.     

Integrated management of insect vectors of Aspergillus flavus in stored maize, using synthetic antioxidants and natural phytochemicals

The purpose of this study was to investigate the insecticidal activity of two benzoic acids 2(3)-tert-butyl-4 hydroxyanisole (BHA) and 2,6-di(tert-butyl)-p-cresol (BHT); two phenolic acids 3-phenyl-2-propenoic acid (CA) and trans-4-hydroxy-3-methoxycinnamic acid (FA) and two essential oils of Eugenia caryophyllata (clove tree) and Thymus vulgaris (thyme) against Sitophilus zeamais, Tribolium confusum and Rhyzopertha dominica, vector carriers of aflatoxigenic fungi in stored maize. The susceptibility of insects, the frequency of isolation of Aspergillus section Flavi in insects and maize, and the analysis of aflatoxin B₁ in maize were determined. BHA, BHT, BHA/BHT mixture and the natural phytochemicals AF and AF/AC mixture showed the highest insecticidal activity against S. zeamais, T. confusum and R. dominica after 120 days of incubation. The insecticidal efficacy of the volatile fraction of essential oils of clove and thyme showed less inhibition. There was no contamination of Aspergillus section Flavi in dead and live insects collected from maize treated with BHA. No aflatoxin B₁ accumulation was detected in the control and treatments. The information obtained shows that these substances have the potential to control pest insect vectors of aflatoxigenic fungi in stored maize in microcosms during 120 days.     

Selective enumeration of Lactobacillus delbrueckii ssp. bulgaricus,Streptococcus thermophilus,Lactobacillus acidophilus,bifidobacteria, Lactobacillus casei,Lactobacillus rhamnosus,and propionibacteria

Nineteen bacteriological media were evaluated to assess their suitability to selectively enumerate Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus acidophilus, bifidobacteria, and propionibacteria. Bacteriological media evaluated included Streptococcus thermophilus agar, pH modified MRS agar, MRS-vancomycine agar, MRS-bile agar, MRS-NaCl agar, MRS-lithium chloride agar, MRS-NNLP (nalidixic acid, neomycin sulfate, lithium chloride and paramomycine sulfate) agar, reinforced clostridial agar, sugar-based (such as maltose, galactose, sorbitol, manitol, esculin) media, sodium lactate agar, arabinose agar, raffinose agar, xylose agar, and L. casei agar. Incubations were carried out under aerobic and anaerobic conditions at 27, 30, 37, 43, and 45 degrees C for 24, 72 h, and 7 to 9 d. S. thermophilus agar and aerobic incubation at 37 degrees C for 24 h were suitable for S. thermophilus. L. delbrueckii ssp. bulgaricus could be enumerated using MRS agar (pH 4.58 or pH 5.20) and under anaerobic incubation at 45 degrees C for 72 h. MRS-vancomycine agar and anaerobic incubation at 43 degrees C for 72 h were suitable to enumerate L. rhamnosus. MRS-vancomycine agar and anaerobic incubation at 37 degrees C for 72 h were selective for L. casei. To estimate the counts of L. casei by subtraction method, counts of L. rhamnosus on MRS-vancomycine agar at 43 degrees C for 72 h under anaerobic incubation could be subtracted from total counts of L. casei and L. rhamnosus enumerated on MRS-vancomycine agar at 37 degrees C for 72 h under anaerobic incubation. L. acidophilus could be enumerated using MRS-agar at 43 degrees C for 72 h or Basal agar-maltose agar at 43 degrees C for 72 h or BA-sorbitol agar at 37 degrees C for 72 h, under anaerobic incubation. Bifidobacteria could be enumerated on MRS-NNLP agar under anaerobic incubation at 37 degrees C for 72 h. Propionibacteria could be enumerated on sodium lactate agar under anaerobic incubation at 30 degrees C for 7 to 9 d. A subtraction method was most suitable for counting propionibacteria in the presence of other lactic acid bacteria from a product. For this method, counts of lactic bacteria at d 3 on sodium lactate agar under anaerobic incubation at 30 degrees C were subtracted from counts at d 7 of lactic bacteria and propionibacteria.     

Use of Carnobacterium maltaromaticum cultures and hydroalcoholic extract of Lippia sidoides Cham. against Listeria monocytogenes in fish model systems

Minimally processed refrigerated ready-to-eat fishes may offer health risk of severe infection to susceptible individuals due to contamination by the psychrotolerant bacterium L. monocytogenes. In this work, inhibition of L. monocytogenes by a plant extract and lactic acid bacteria (LAB) was studied in model fish systems kept at 5 °C for 35 days. For that, fillets of tropical fish “surubim” (Pseudoplatystoma sp.) and hydroalcoholic extract of the plant Lippia sidoides Cham. (“alecrim pimenta”) were used. Fish peptone broth (FPB), “surubim” broth and “surubim” homogenate were inoculated with combinations of L. monocytogenes and bacteriocin-producing Carnobacterium maltaromaticum (C2 and A9b⁺) and non bacteriocin-producing C. maltaromaticum (A9b^-), in the presence or absence of extract of “alecrim pimenta” (EAP). In all model systems, monocultures of L. monocytogenes and carnobacteria reached final populations ≥ 10⁸ CFU/ml after 35 days, except for L. monocytogenes in “surubim” homogenate (10⁴ CFU/ml). In FPB, EAP alone and combined with cultures of LAB inhibited L. monocytogenes but carnobacteria without EAP were only weakly antilisterial. In “surubim” broth, EAP alone did not prevent L. monocytogenes growth but cultures of carnobacteria combined or not with EAP inhibited L. monocytogenes, with more pronounced effect being observed for C. maltaromaticum C2, which produced bacteriocin. In “surubim” homogenate, EAP alone and combined with cultures of C. maltaromaticum A9b⁻ and A9b⁺ were strongly inhibitory to L. monocytogenes, while C. maltaromaticum C2 with EAP caused transient inhibition of L. monocytogenes. No significant inhibition of L. monocytogenes was observed for carnobacteria in “surubim” homogenate without EAP. In conclusion, it was observed that the use of EAP and cultures of carnobacteria have potential to inhibit L. monocytogenes in fish systems and the applications should be carefully studied, considering the influence of food matrix.     

Efficacy of powder and essential oil from Chenopodium ambrosioides leaves as post-harvest grain protectants against six-stored product beetles

Powder and essential oil obtained from dry ground leaves of Chenopodium ambrosioides were tested under laboratory conditions (25±1°C, 70-75% r.h.) for their ability to protect grains from damage by six insect pests, Callosobruchus chinensis, C. maculatus, Acanthoscelides obtectus, Sitophilus granarius, S. zeamais and Prostephanus truncatus. The insects were reared and tested on whole maize grain for S. zeamais and P. truncatus, whole wheat for S. granarius, green peas for C. chinensis, mung bean for C. maculatus and white bean for A. obtectus. The powder prepared from dry leaves of C. ambrosioides was mixed with grains at different dosages ranging from 0.05-0.80% (wt/wt) for C. chinensis, C. maculatus and A. obtectus and from 0.8-6.4% (wt/wt) for S. granarius, S. zeamais and P. truncatus. The dosage of 0.4% killed more than 60% of all the bruchids 2 days after treatment, while a dosage of 6.4% induced total mortality of S. granarius and S. zeamais within the same exposure time. All levels of the dry ground leaf concentrations inhibited F1 progeny production and adult emergence of the tested insects. The dosage of 0.2 μl/cm² of the essential oil killed 80-100% of the beetles within 24 h except C. maculatus and S. zeamais, where this dosage induced only 20% and 5% mortality, respectively. These results indicate a scientific rationale for the use of this plant in grain protection by local communities in the western highlands of Cameroon.