International journal of food microbiology

Publications in this journal

• Article: Effect of pyrimethanil on Cryptococcus laurentii, Rhodosporidium paludigenum, and Rhodotorula glutinis biocontrol of Penicillium expansum infection in pear fruit.

  Chen Yu, Tao Zhou, Kuang Sheng, Lizhen Zeng, Changzhou Ye, Ting Yu, Xiaodong Zheng
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  ABSTRACT: The effect of biocontrol yeasts and pyrimethanil at low concentration on inhibition of blue mold rot caused by Penicillium expansum in pear fruit was investigated. Pyrimethanil at low concentration (40μg/mL) alone had little inhibitory activity against the P. expansum infection in pear fruit wounds although it was effective in inhibiting the survival of P. expansum on Asp-agar medium. Pyrimethanil at this low concentration significantly enhanced the efficacy of Cryptococcus laurentii at 1×10(7)CFU/mL in reducing blue mold rot in vivo compared with C. laurentii at 1×10(7)CFU/mL alone. However, there was no additive inhibitory activity when pyrimethanil was combined for application with biocontrol yeasts Rhodosporidium paludigenum or Rhodotorula glutinis. Combination of pyrimethanil and C. laurentii at low concentration also inhibited blue mold rot when P. expansum was inoculated into fruit wounds 12h before treatment and fruit was stored at low temperature (4°C). Pyrimethanil at 0.04 to 400μg/mL did not influence the survival of C. laurentii in vitro, and it only slightly reduced the population growth of C. laurentii after 48h of incubation in the pear fruit wounds. There was no significant difference in quality parameters including total soluble solids, titratable acidity and ascorbic acid of pear fruit wounds among all treatments after 5days of treatment at 25°C. Integration of C. laurentii and pyrimethanil at low concentration might be an effective and safe strategy to control P. expansum infection in pear fruit, especially in an integrated postharvest disease management strategy.
  International journal of food microbiology 04/2013; 164(2-3):155-160.
• Article: Investigating the efficacy of Bacillus subtilis SM21 on controlling Rhizopus rot in peach fruit.

  Xiaoli Wang, Jing Wang, Peng Jin, Yonghua Zheng
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  ABSTRACT: The efficacy of Bacillus subtilis SM21 on controlling Rhizopus rot caused by Rhizopus stolonifer in postharvest peach fruit and the possible mechanisms were investigated. The results indicated B. subtilis SM21 treatment reduced lesion diameter and disease incidence by 37.2% and 26.7% on the 2nd day of inoculation compared with the control. The in vitro test showed significant inhibitory effect of B. subtilis SM21 on mycelial growth of R. stolonifer with an inhibition rate of 48.9%. B. subtilis SM21 treatment significantly enhanced activities of chitinase and β-1,3-glucanase, and promoted accumulation of H2O2. Total phenolic content and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity were also increased by this treatment. Transcription of seven defense related genes was much stronger in fruit treated with B. subtilis SM21 or those both treated with B. subtilis SM21 and inoculated with R. stolonifer compared with fruit inoculated with R. stolonifer alone. These results suggest that B. subtilis SM21 can effectively inhibit Rhizopus rot caused by R. stolonifer in postharvest peach fruit, possibly by directly inhibiting growth of the pathogen, and indirectly inducing disease resistance in the fruit.
  International journal of food microbiology 04/2013; 164(2-3):141-147.
• Article: Effect of wine-based marinades on the behavior of Salmonella Typhimurium and background flora in beef fillets.

  A Nisiotou, N G Chorianopoulos, A Gounadaki, E Z Panagou, G-J E Nychas
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  ABSTRACT: The aim of this study was to evaluate the wine-based marinades to control the survival of acid-adapted and non-adapted Salmonella Typhimurium and background flora of fresh beef stored aerobically or under modified atmosphere. Beef slices were inoculated with a 3-strain cocktail of acid-adapted or non-adapted Salmonella Typhimurium strains DT 193, 4/74 and DSM 554 and marinated by immersion in wine (W) or wine supplemented with 0.3% thyme essential oil (WEO), for 12h at 4°C. Marinated slices were then stored under air or modified atmosphere conditions at 5°C. S. Typhimurium and background flora were followed for a 19-day period of storage. S. Typhimurium individual strains were monitored by pulsed field gel electrophoresis. Marination with wine significantly (P<0.05) reduced the background flora compared to the control (non-marinated). Furthermore, immersion of fillets in W or WEO marinades for 12h significantly (P<0.05) reduced the levels of S. Typhimurium compared to the non-marinated (control) samples by 1.1 and 1.4logCFU/g or 2.0 and 1.9logCFU/g for acid-adapted and non-adapted cells, respectively. Acid-adapted cells were more susceptible (P<0.05) to the addition of thyme essential oil in the wine marinade. The epidemic multi-drug resistant DT 193, the 4/74 and DSM 554 strains survived marination (for both W and WEO) and were detected at about similar proportions as revealed by PFGE results. Present results indicate that wine-based marinades are efficient, from a safety and shelf life stand point, in reducing pathogen's levels as well as the background beef flora.
  International journal of food microbiology 04/2013; 164(2-3):119-127.
• Article: Effects of nisin-incorporated films on the microbiological and physicochemical quality of minimally processed mangoes.

  Ana Andréa Teixeira Barbosa, Hyrla Grazielle Silva de Araújo, Patrícia Nogueira Matos, Marcelo Augusto Guitierrez Carnelossi, Alessandra Almeida de Castro
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  ABSTRACT: The aim of this study is to examine the effects of nisin-incorporated cellulose films on the physicochemical and microbiological qualities of minimally processed mangoes. The use of antimicrobial films did not affect the physicochemical characteristics of mangoes and showed antimicrobial activity against Staphylococcus aureus, Listeria monocytogenes, Alicyclobacillus acidoterrestris and Bacillus cereus. The mango slices were inoculated with S. aureus and L. monocytogenes (10(7)CFU/g), and the viable cell numbers remained at 10(5) and 10(6)CFU/g, respectively, after 12days. In samples packed with antimicrobial films, the viable number of L. monocytogenes cells was reduced below the detection level after 4days. After 6days, a reduction of six log units was observed for S. aureus. In conclusion, nisin showed antimicrobial activity in mangoes without interfering with the organoleptic characteristics of the fruit. This result suggests that nisin could potentially be used in active packing to improve the safety of minimally processed mangoes.
  International journal of food microbiology 04/2013; 164(2-3):135-140.
• Article: The antimicrobial effects of wood-associated polyphenols on food pathogens and spoilage organisms.

  Carme Plumed-Ferrer, Kati Väkeväinen, Heli Komulainen, Maarit Rautiainen, Annika Smeds, Jan-Erik Raitanen, Patrik Eklund, Stefan Willför, Hanna-Leena Alakomi, Maria Saarela, Atte von Wright
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  ABSTRACT: The antimicrobial effects of the wood-associated polyphenolic compounds pinosylvin, pinosylvin monomethyl ether, astringin, piceatannol, isorhapontin, isorhapontigenin, cycloXMe, dHIMP, ArX, and ArXOH were assessed against both Gram-negative (Salmonella) and Gram-positive bacteria (Listeria monocytogenes, Staphylococcus epidermidis, Staphylococcus aureus) and yeasts (Candida tropicalis, Saccharomyces cerevisiae). Particularly the stilbenes pinosylvin, its monomethyl ether and piceatannol demonstrated a clear antimicrobial activity, which in the case of pinosylvin was present also in food matrices like sauerkraut, gravlax and berry jam, but not in milk. The destabilization of the outer membrane of Gram-negative microorganisms, as well as interactions with the cell membrane, as indicated by the NPN uptake and LIVE/DEAD viability staining experiments, can be one of the specific mechanisms behind the antibacterial action. L. monocytogenes was particularly sensitive to pinosylvin, and this effect was also seen in L. monocytogenes internalized in intestinal Caco2 cells at non-cytotoxic pinosylvin concentrations. In general, the antimicrobial effects of pinosylvin were even more prominent than those of a related stilbene, resveratrol, well known for its various bioactivities. According to our results, pinosylvin could have potential as a natural disinfectant or biocide in some targeted applications.
  International journal of food microbiology 04/2013; 164(1):99-107.
• Article: Comparative genomic analysis of Aspergillus oryzae strains 3.042 and RIB40 for soy sauce fermentation.

  Guozhong Zhao, Yunping Yao, Chunling Wang, Lihua Hou, Xiaohong Cao
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  ABSTRACT: The filamentous fungus Aspergillus oryzae 3.042 (Chinese strain) is a close relative of A. oryzae RIB40 (Japanese strain), which is the important agent used for soy sauce fermentation. The genome of A. oryzae 3.042 was sequenced and compared with A. oryzae RIB40 in an attempt to understand why different soy sauce flavors are produced by these strains. The A. oryzae 3.042 chromosome is 36,547,279bp and contains 11,399 protein-encoding genes. MUMmer analysis revealed that the genomes of A. oryzae 3.042 and RIB40 are mostly collinear. Genome sequence data and comparative analysis of the two strains identified several strain-specific genes that encode putative proteins involved in cell growth, salt tolerance, environmental resistance and flavor formation. A. oryzae 3.042 showed stronger potential for mycelial growth. Some genes unique to A. oryzae RIB40 were related to salt tolerance, especially genes for K(+) transport, while others were associated with ester formation and amino acid metabolism, which likely contribute to flavor formation. In conclusion, comparative genome analysis provided insights into the different genetic traits of the two A. oryzae strains. The unique genes that we found in A. oryzae would make sense to the soy sauce fermentation.
  International journal of food microbiology 04/2013; 164(2-3):148-154.
• Article: Disruption of protein synthesis as antifungal mode of action by chitosan.

  Imelda Galván Márquez, Jones Akuaku, Isabel Cruz, James Cheetham, Ashkan Golshani, Myron L Smith
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  ABSTRACT: The antimicrobial activity of chitosan has been acknowledged for more than 30years and yet its mode-of-action remains ambiguous. We analyzed chemical-genetic interactions of low-molecular weight chitosan using a collection of ≈4600 S. cerevisiae deletion mutants and found that 31% of the 107 mutants most sensitive to chitosan had deletions of genes related primarily to functions involving protein synthesis. Disruption of protein synthesis by chitosan was substantiated by an in vivo β-galactosidase expression assay suggesting that this is a primary mode of antifungal action. Analysis of the yeast gene deletion array and secondary assays also indicate that chitosan has a minor membrane disruption effect - a leading model of chitosan antimicrobial activity.
  International journal of food microbiology 04/2013; 164(1):108-112.
• Article: Fungal profiles in various milk thistle botanicals from US retail.

  V H Tournas, J Rivera Calo, C Sapp
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  ABSTRACT: Milk thistle (MT) dietary supplements are widely consumed due to their possible beneficial effect on liver health. As botanicals, they can be contaminated with a variety of fungi and their secondary metabolites, mycotoxins. This study was conducted in an effort to determine the mycological quality of various MT botanical supplements from the US market. Conventional plating methods were used for the isolation and enumeration of fungi, while conventional microscopy as well as molecular methods were employed for the speciation of the isolated strains. Results showed that a high percentage of the MT samples tested were contaminated with fungi. Total counts ranged between <2.00 and 5.60log10 colony forming units per gram (cfu/g). MT whole seeds carried the highest fungal levels followed by MT cut herb. No live fungi were recovered from MT seed tea bags, liquid extracts, capsules or soft gels. Potentially toxigenic molds from the Aspergillus sections Flavi and Nigri as well as Eurotium, Penicillium, Fusarium and Alternaria species were isolated from MT supplements. The predominant molds were Eurotia (E. repens, E. amstelodami and E. rubrum), A. flavus, A. tubingensis, A. niger and A. candidus. To our knowledge, this is the first study reporting on fungal contamination profiles of MT botanicals.
  International journal of food microbiology 04/2013; 164(1):87-91.
• Article: Indigenous filamentous fungi on the surface of Argentinean dry fermented sausages produced in Colonia Caroya (Córdoba).

  Romina S Canel, Jorge R Wagner, Sebastián A Stenglein, Vanesa Ludemann
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  ABSTRACT: Some producers of dry fermented sausages use fungal starter cultures with the aim to achieve a desirable surface appearance and avoid the growth of mycotoxigenic fungi. These commercial cultures are mainly composed of Penicillium nalgiovense biotype 6. In contrast, in the case of producers who do not use starters, sausages are spontaneously colonized by the house mycobiota, which generally consists of heterogeneous molds corresponding to different genera and species. In this work, the surface mycobiota of dry fermented sausages produced in Colonia Caroya (Córdoba, Argentina) was determined in both summer and winter seasons. All the sausages sampled had been made without the use of surface fungal starters. In the 57 sausages analyzed in the two winter seasons studied (2010 and 2012), we found a total of 95 isolates of filamentous fungi belonging to six genera (Penicillium, Aspergillus, Mucor, Cladosporium, Scopulariopsis and Eurotium) and ten fungal species, whereas in the 36 sausages analyzed in the two summer seasons studied (2011 and 2012), we found 89 isolates belonging to five genera (Penicillium, Aspergillus, Mucor, Cladosporium and Geotrichum) and ten fungal species. Although 16 different species were found in both winter and summer seasons, only 2 of them predominated completely. P. nalgiovense was found in almost 100% of the sausages analyzed, where biotype 4 was the most frequent. This species gives a whitish gray coloration to the sausages. Considering that the factories sampled do not use fungal starter cultures, this predominance is very interesting since mycotoxin production by this fungus has not been reported. Aspergillus ochraceus was isolated with a frequency of 80-90% in the summer seasons, but in none of the winter samples. The presence of this fungus in sausages produced in the summer was attributed to the high environmental temperatures and the uncontrolled temperature in the ripening rooms during the night. In all cases, A. ochraceus was responsible for the undesirable yellowish gold color of the casing. This fungus thus causes significant economic losses to the producers of Colonia Caroya during the months of high temperatures.
  International journal of food microbiology 04/2013; 164(1):81-86.
• Article: In-house validation of a multiplex real-time PCR method for simultaneous detection of Salmonella spp., Escherichia coli O157 and Listeria monocytogenes.

  Alejandro Garrido, María-José Chapela, Belén Román, Paula Fajardo, Juan M Vieites, Ana G Cabado
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  ABSTRACT: A wide variety of qPCR methods currently exist for Salmonella spp., Escherichia coli O157 and Listeria monocytogenes detection. These methods target several genes and use different detection chemistries, either in simplex or in multiplex formats. However, the majority of these methods have not been carefully validated, and the number of validated methods that use multiplex qPCR is even lower. The aim of the present study was to develop and validate a multiplex qPCR method from previously validated simplex qPCR primers and probes. A modified broth medium was selected and primary and secondary enrichment times were further optimized. Efficiency of the newly combined qPCR system was comprised between 91% and 108%, for simplex and multiplex analyses. A total of 152 food and environmental, natural and spiked samples, were analyzed for the evaluation of the method obtaining values above 91% that were reached for all the quality parameters analyzed. A very low limit of detection (5cfu/25g after enrichment) for simultaneous identification of these 3 pathogens was obtained.
  International journal of food microbiology 04/2013; 164(1):92-98.
• Article: Influence of isolation methods on the occurrence of plasmid-carrying Yersinia enterocolitica serotype O:3 in slaughter pig tonsils, faeces and carcass surface swabs.

  I Van Damme, D Berkvens, J Baré, L De Zutter
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  ABSTRACT: Yersinia enterocolitica is an important foodborne pathogen that is primarily transmitted to humans through the consumption of contaminated pork. Different matrices of pigs at slaughter were tested for the presence of human pathogenic types of Y. enterocolitica using direct plating, selective enrichment, and cold enrichment. Y. enterocolitica serotype O:3 was isolated from the tonsils and faeces of 55.3% and 25.6% of pigs, respectively. The pathogen was also recovered from 15.0% of swab samples taken from the carcass surface post-evisceration. Tonsils positive by direct plating revealed an average concentration of 3.99 log10Y. enterocolitica per gram, whereas the majority of positive faecal and carcass samples were contaminated below the detection limit of enumeration. The relative sensitivity of the methods to recover pathogenic Y. enterocolitica varied among the different matrices. Nevertheless, cold enrichment was significantly more efficient than direct plating and selective enrichment for all three sample matrices. From the 2082 recovered Y. enterocolitica isolates, 1742 (83.7%) harboured the virulence plasmid. Isolates obtained from faeces were more likely to contain the virulence plasmid than isolates from tonsils and carcass swabs. To obtain reliable results regarding the presence of plasmid-carrying Y. enterocolitica isolates, sensitive isolation methods should be combined with testing of a sufficient number of isolates.
  International journal of food microbiology 03/2013; 164(1):32-35.
• Article: Isolation and molecular characterization of Salmonella enterica serovar Javiana from food, environmental and clinical samples.

  Ezat H Mezal, Rossina Stefanova, Ashraf A Khan
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  ABSTRACT: A total of 50 Salmonella enterica serovar Javiana isolates, isolated from food, environmental and clinical samples, were analyzed for antibiotic resistance, presence of virulence genes, plasmids and plasmid replicon types. To assess the genetic diversity, pulsed-field gel electrophoresis (PFGE) fingerprinting and plasmid profiles were performed. All of the isolates were sensitive to chloramphenicol, nalidixic acid, and sulfisoxazole, and four isolates showed intermediate resistance to gentamicin or kanamycin. Eleven isolates, including representatives from each of the source types, were resistant to ampicillin. Four isolates from either clinical or environmental sources were resistant to tetracycline, while an additional 20 isolates showed intermediate resistance to this drug. Fourteen isolates, primarily from food sources, showed intermediate resistance to streptomycin. The S. Javiana isolates were screened by PCR for 17 virulence genes (spvB, spiA, pagC, msgA, invA, sipB, prgH, spaN, orgA, tolC, iroN, sitC, IpfC, sifA, sopB, cdtB, and pefA). All isolates were positive for nine to fourteen of these genes, but none were positive for pefA, spvB and lpfC, which are typically present on the Salmonella virulence plasmid. Seven of the virulence genes including cdtB were found in all 50 isolates, suggesting that S. Javiana from food and environmental sources had virulence similar to clinical isolates. Four clinical isolates and two food isolates carried one or more plasmids of approximately 30, 38, and 58kb, with the 58kb plasmids belonging to incompatibility group IncFIIA. Two clinical isolates carried IncI1 type mega plasmid (80kb), and one clinical isolate carried plasmids of 4.5 and 7kb. The PFGE profiles resulted 34 patterns in five clusters at a 90% similarity threshold. Our results indicate that S. Javiana isolates have a diverse clonal population among the clinical, food and environmental samples and this serotype possesses several virulent genes and plasmids that can contribute to the development of salmonellosis in human. This study provides data that support the potential transmission of S. Javiana virulence factors from food and environmental sources to cause infections in humans.
  International journal of food microbiology 03/2013; 164(1):113-118.
• Article: Prevalence and antibiotic resistance profiles of diarrheagenic Escherichia coli strains isolated from food items in northwestern Mexico.

  Adrian Canizalez-Roman, Edgar Gonzalez-Nuñez, Jorge E Vidal, Héctor Flores-Villaseñor, Nidia León-Sicairos
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  ABSTRACT: Diarrheogenic Escherichia coli (DEC) strains are an important cause of intestinal syndromes in the developing world mainly affecting children. DEC strains often infect tourists from developed countries traveling to Mexico, causing so-called "traveler diarrhea". DEC strains are typically transmitted by contaminated food and water; however, the prevalence of these strains in food items that are produced, consumed and sometimes exported in northwestern Mexico has not been evaluated. In this study, we conducted a large microbiological survey of DEC strains in 5162 food items and beverages consumed throughout Sinaloa state during 2008 and 2009. We developed a panel of eight sequential PCR reactions that detected the presence of all DEC categories, including typical or atypical variants. Thermotolerant coliforms (also known as fecal coliforms) and E. coli were detected by conventional bacteriology in 13.4% (692/5162) and 7.92% (409/5162) of food items, respectively. Among 409 E. coli isolates, 13.6% (56/409) belonged to DEC strains. Dairy products (2.8%) were the most contaminated with DEC, while DEC strains were not detected in beverages and ice samples. The pathogenic type that was most commonly isolated was EPEC (78.5%), followed by EAEC (10.7%), STEC (8.9%) and ETEC (1.7%). EHEC, DAEC and EIEC strains were not detected. Approximately 80% of EPEC and EAEC strains were classified as atypical variants; they did not adhere to a culture of HEp-2 cell. Of the isolated DEC strains, 66% showed resistance to at least one commonly prescribed antibiotic. In conclusion, the presence of DEC strains in food items and beverages available in northwestern Mexico is low and may not represent a threat for the general population or those traveling to tourist areas.
  International journal of food microbiology 03/2013; 164(1):36-45.
• Article: Exposure assessment of lovastatin in Pu-erh tea.

  Zhen-Jun Zhao, You-Zhao Pan, Qin-Jin Liu, Xing-Hui Li
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  ABSTRACT: This paper reports the results of an extensive survey on the levels of lovastatin in Pu-erh tea samples. The microbial source of lovastatin was assessed by testing the ability of fungi with higher isolation frequency in the Pu-erh tea samples to produce lovastatin on Czapek yeast extract agar (CYA). Lovastatin was not detected in any of the raw Pu-erh tea samples without storage but was found in almost all the ripe Pu-erh tea samples, with lovastatin contents ranging from 20.61ng/gdw to 226.38ng/gdw. After five years' storage, the lovastatin levels increased obviously in ripe Pu-erh tea samples and 55% of raw Pu-erh tea samples from 2007 were found to contain lovastatin with concentrations ranging between 28.41ng/gdw and 228.61ng/gdw. With increasing storage time, lovastatin concentration in ripe Pu-erh tea, and the occurrence and concentration of lovastatin for raw Pu-erh tea increased significantly. Three genera of fungi: Aspergillus, Penicillium and Trichoderma were often isolated from Pu-erh tea samples. A total of 40 strains from 3 fungal genera were selected to test their ability to produce lovastatin. Only 6 strains, Aspergillus tubingensis, Aspergillus wentii, Aspergillus fumigatus, Penicillium chrysogenum, Trichoderma asperellum and Trichoderma citrinoviride, were able to produce lovastatin reaching concentrations of 9.59±0.42ng/g CYA, 2.33±0.21ng/g CYA, 2.77±0.13ng/g CYA, 3.36±0.69ng/g CYA, 4.8±0.17ng/g CYA, and 1.47±0.36ng/g CYA respectively in Czapek yeast extract agar.
  International journal of food microbiology 03/2013; 164(1):26-31.
• Article: Microbial succession and metabolite changes during fermentation of dongchimi, traditional Korean watery kimchi.

  Sang Hyeon Jeong, Ji Young Jung, Se Hee Lee, Hyun Mi Jin, Che Ok Jeon
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  ABSTRACT: Dongchimi, one of the most common types of watery kimchi in Korea, was prepared using radish and its pH values, microbial cell numbers, bacterial communities, and metabolites were monitored periodically to investigate the fermentation process of watery kimchi. The bacterial abundance increased quickly during the early fermentation period and the pH values concurrently decreased rapidly without any initial pH increase. After 15days of fermentation, the bacterial abundance decreased rapidly with the increase of Saccharomyces abundance and then increased again with a decrease of Saccharomyces abundance after 40days of fermentation, suggesting that bacteria and Saccharomyces have a direct antagonistic relationship. Finally, after 60days of fermentation, a decrease in bacterial abundance and the growth of Candida were concurrently observed. Community analysis using pyrosequencing revealed that diverse genera such as Leuconostoc, Lactobacillus, Pseudomonas, Pantoea, and Weissella were present at initial fermentation (day 0), but Leuconostoc became predominant within only three days of fermentation and remained predominant until the end of fermentation (day 100). Metabolite analysis using (1)H NMR showed that the concentrations of free sugars (fructose and glucose) were very low during the early fermentation period, but their concentrations increased rapidly although lactate, mannitol, and acetate were produced. After 30days of fermentation, quick consumption of free sugars and production of glycerol and ethanol were observed concurrently with the growth of Saccharomyces, levels of which might be considered for use as a potential indicator of dongchimi quality and fermentation time.
  International journal of food microbiology 03/2013; 164(1):46-53.
• Article: Development of novel agar media for isolating guaiacol producing Alicyclobacillus spp.

  S S Chang, S H Park, D H Kang
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  ABSTRACT: The purpose of this study is to develop a selective and differential medium (SK2 agar) for isolating guaiacol producing Alicyclobacillus. Forty-one selected dyes and vanillic acid were incorporated in SK agar for screening selective and differential agents. Two guaiacol producing (1016, 1101) and two non-guaiacol producing (19220, C-GD 1-1) Alicyclobacillus isolates were streaked onto media and color differentiation of the isolates was assessed. Among 41 tested dyes, Chrome Azurol S (CAS) allowed color differentiation of the two types of Alicyclobacillus. Colonies of guaiacol producing Alicyclobacillus isolates appeared as dark purple to royal blue color with yellow background, whereas non-guaiacol producing Alicyclobacillus isolates produced cream colored colonies with yellow background. Vanillic acid not only served as a precursor for guaiacol formation but also inhibited non-guaiacol producing Alicyclobacillus. Non-guaiacol producing isolates did not grow on SK agar containing more than 70ppm vanillic acid, whereas the recovery of guaiacol producing isolates was unaffected. When compared with other Alicyclobacillus isolation media, not only was SK2 agar capable of selectively recovering guaiacol-producing Alicyclobacillus, the degree of growth was also approximately equal if not better than orange serum agar, potato dextrose agar, and K agar. The development of SK2 agar provides the fruit juice industry with an inexpensive, simple to use alternative for the detection of guaiacol producing Alicyclobacillus.
  International journal of food microbiology 03/2013; 164(1):1-6.
• Article: Principles for the risk assessment of genetically modified microorganisms and their food products in the European Union.

  Jaime Aguilera, Ana R Gomes, Irina Olaru
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  ABSTRACT: Genetically modified microorganisms (GMMs) are involved in the production of a variety of food and feed. The release and consumption of these products can raise questions about health and environmental safety. Therefore, the European Union has different legislative instruments in place in order to ensure the safety of such products. A key requirement is to conduct a scientific risk assessment as a prerequisite for the product to be placed on the market. This risk assessment is performed by the European Food Safety Authority (EFSA), through its Scientific Panels. The EFSA Panel on Genetically Modified Organisms has published complete and comprehensive guidance for the risk assessment of GMMs and their products for food and/or feed use, in which the strategy and the criteria to conduct the assessment are explained, as well as the scientific data to be provided in applications for regulated products. This Guidance follows the main risk assessment principles developed by various international organisations (Codex Alimentarius, 2003; OECD, 2010). The assessment considers two aspects: the characterisation of the GMM and the possible effects of its modification with respect to safety, and the safety of the product itself. Due to the existing diversity of GMMs and their products, a categorisation is recommended to optimise the assessment and to determine the extent of the required data. The assessment starts with a comprehensive characterisation of the GMM, covering the recipient/parental organism, the donor(s) of the genetic material, the genetic modification, and the final GMM and its phenotype. Evaluation of the composition, potential toxicity and/or allergenicity, nutritional value and environmental impact of the product constitute further cornerstones of the process. The outcome of the assessment is reflected in a scientific opinion which indicates whether the product raises any safety issues. This opinion is taken into account by the different European regulatory authorities prior to a decision regarding authorisation to commercialise the product.
  International journal of food microbiology 03/2013;
• Article: Prediction of Bacillus weihenstephanensis acid resistance: The use of gene expression patterns to select potential biomarkers.

  N Desriac, F Postollec, L Coroller, D Sohier, T Abee, H M W den Besten
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  ABSTRACT: Exposure to mild stress conditions can activate stress adaptation mechanisms and provide cross-resistance towards otherwise lethal stresses. In this study, an approach was followed to select molecular biomarkers (quantitative gene expressions) to predict induced acid resistance after exposure to various mild stresses, i.e. exposure to sublethal concentrations of salt, acid and hydrogen peroxide during 5min to 60min. Gene expression patterns of unstressed and mildly stressed cells of Bacillus weihenstephanensis were correlated to their acid resistance (3D value) which was estimated after exposure to lethal acid conditions. Among the twenty-nine candidate biomarkers, 12 genes showed expression patterns that were correlated either linearly or non-linearly to acid resistance, while for the 17 other genes the correlation remains to be determined. The selected genes represented two types of biomarkers, (i) four direct biomarker genes (lexA, spxA, narL, bkdR) for which expression patterns upon mild stress treatment were linearly correlated to induced acid resistance; and (ii) nine long-acting biomarker genes (spxA, BcerKBAB4_0325, katA, trxB, codY, lacI, BcerKBAB4_1716, BcerKBAB4_2108, relA) which were transiently up-regulated during mild stress exposure and correlated to increased acid resistance over time. Our results highlight that mild stress induced transcripts can be linearly or non-linearly correlated to induced acid resistance and both approaches can be used to find relevant biomarkers. This quantitative and systematic approach opens avenues to select cellular biomarkers that could be incremented in mathematical models to predict microbial behaviour.
  International journal of food microbiology 03/2013;