Villers-Bocage, France
Villers-Bocage, France

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Coton E.,ADRIA Normandie | Mulder N.,University of Groningen | Coton M.,ADRIA Normandie | Pochet S.,French Institute for Research in Computer Science and Automation | And 2 more authors.
Applied and Environmental Microbiology | Year: 2010

A multiplex PCR method, aimed at the detection of genes associated with biogenic amine production, identified the ode gene encoding ornithine decarboxylase in 1 of 15 strains of Staphylococcus epidermidis. The ability of the positive strain, S. epidermidis 2015B, to produce putrescine in vitro was demonstrated by high-performance liquid chromatography (HPLC). In this strain, the ode gene was detected on plasmid DNA, suggesting that the ability to form putrescine is carried by a mobile element, which explains the fact that the trait is strain dependent within the S. epidermidis species. A 6,292-bp nucleotide sequence harboring the putative ode gene was determined. S. epidermidis ornithine decarboxylase (ODC) showed 60 to 65% sequence identity with known ODCs of Gram-positive as well as Gram-negative bacteria. Downstream of the ode gene, a gene encoding a putative amino acid transporter was found that shared 59% sequence identity with the ornithine/putrescine exchanger (PotE) of Escherichia coli. Cloning and expression of the potE gene of S. epidermis 2015B in Lactococcus lactis demonstrated that the gene product transported ornithine and putrescine into the cells and efficiently exchanged putrescine for ornithine. Analysis of the flanking regions showed high identity levels with different S. epidermidis plasmid sequences, which would confirm the plasmidic location of the ode operon. It follows that the ode and potE gene pair encodes a putrescine-producing pathway in S. epidermis 2015B that was acquired through horizontal gene transfer. Copyright © 2010, American Society for Microbiology. All Rights Reserved.

Coton M.,ADRIA Normandie | Delbes-Paus C.,French National Institute for Agricultural Research | Irlinger F.,French National Institute for Agricultural Research | Desmasures N.,University of Caen Lower Normandy | And 4 more authors.
Food Microbiology | Year: 2012

The goal of this study was to identify at the species level a large collection of Gram-negative dairy bacteria isolated from milks or semi-hard and soft, smear-ripened cheeses (cheese core or surface samples) from different regions of France. The isolates were then assessed for two risk factors, antibiotic resistance and volatile and non-volatile biogenic amine production in vitro. In total, 173 Gram-negative isolates were identified by rrs and/or rpoB gene sequencing. A large biodiversity was observed with nearly half of all Gram-negative isolates belonging to the Enterobacteriaceae family. Overall, 26 different genera represented by 68 species including potential new species were identified among the studied Gram-negative isolates for both surface and milk or cheese core samples. The most frequently isolated genera corresponded to Pseudomonas, Proteus, Psychrobacter, Halomonas and Serratia and represented almost 54% of the dairy collection. After Pseudomonas, Chryseobacterium, Enterobacter and Stenotrophomonas were the most frequently isolated genera found in cheese core and milk samples while Proteus, Psychrobacter, Halomonas and Serratia were the most frequently isolated genera among surface samples. Antibiotic resistance profiles indicated that resistances to the aminosid, imipemen and quinolon were relatively low while more than half of all tested isolates were resistant to antibiotics belonging to the monobactam, cephem, fosfomycin, colistin, phenicol, sulfamid and some from the penam families. Thirty-six% of isolates were negative for in vitro biogenic amine production. Among biogenic amine-producers, cadaverine was the most frequently produced followed by isoamylamine, histamine and putrescine. Only low levels (<75 mg/l) of tyramine were detected in vitro. © 2011 Elsevier Ltd.

Coton M.,ADRIA Normandie | Romano A.,French National Institute for Agricultural Research | Spano G.,University of Foggia | Ziegler K.,French National Institute for Agricultural Research | And 5 more authors.
Food Microbiology | Year: 2010

A collection of 810 lactic acid bacteria (LAB) strains isolated from wine and cider was screened for potential biogenic amine (BA) producers by combining molecular and phenotypic approaches. A newly developed multiplex PCR method allowed for the simultaneous detection of four genes involved in the production of histamine (histidine decarboxylase, hdc), tyramine (tyrosine decarboxylase, tyrdc) and putrescine (via either ornithine decarboxylase, odc, or agmatine deiminase, agdi) while TLC and HPLC analysis allowed for BA-production determination. One hundred and fifty-eight LAB strains were monitored by the molecular/phenotypic double approach and revealed a good correlation between genotypic and phenotypic data. Eighteen per cent of the tested strains were positive for at least one BA target gene with up to three detected simultaneously, in particular amongst Lactobacillus brevis and Lactobacillus hilgardii isolates for the tyrdc and agdi genes. The most frequent gene corresponded to the agdi gene detected in 112 strains (14% of all LAB strains) of 10 different LAB species. The tyrdc gene was detected in 67 strains represented by 7 different LAB species (8% overall), especially those isolated from wine. Lower levels of hdc+ (2% of strains) and especially odc+ (0.5% of strains) strains were observed. Interestingly, species that have never been described to carry BA-producing pathway genes were identified in this study. Furthermore, only one cadaverine-producer was detected and corresponded to Lactobacillus 30a, a collection strain not found in fermented beverages, although cadaverine is commonly detected in wines. © 2010 Elsevier Ltd.

Fraisse A.,Food and Water Virology Unit | Temmam S.,ADRIA NORMANDIE | Deboosere N.,Institute Pasteur Of Lille Ipl | Guillier L.,Modelling of Bacterial Behaviour Unit | And 5 more authors.
International Journal of Food Microbiology | Year: 2011

In recent years, raw fruits and vegetables have frequently been involved in foodborne transmission to humans of enteric viruses, particularly noroviruses and hepatitis A virus (HAV). Although viral contamination can occur during all steps of food processing, primary production is a critical stage on which prevention measures must be focused to minimize the risk of infection to consumers. Postharvest sanitation may be a valid technological solution for decreasing the bacterial load on fresh raw material, but there is a lack of data concerning the effectiveness of this process on enteric viruses. In this study, we compared the survival of two human norovirus surrogates, the feline calicivirus (FCV), and the murine norovirus (MNV-1), and of HAV on lettuce after water washing with bubbles and with or without ultrasound, and washing with bubbles in the presence of active chlorine (15 ppm) or peroxyacetic acid-based disinfectant (100 ppm). Cell culture and quantitative RT-PCR assays were used to detect and quantify the viruses on the surface of the lettuce after the sanitizing treatments. Levels of viral inactivation on the lettuce leaves were not significantly different between washing with bubbles and washing with bubbles plus ultrasound and were not dependant on the quantification method. A simple washing without disinfectant resulted in a decrease of approximately 0.7 log units in the quantity of virus detected for HAV and FCV and of 1.0 log unit for MNV-1.In the experimental set-up including a washing step (with or without ultrasound) followed by washing for 2 min in the presence of disinfectants, 15 ppm of active chlorine was found more effective for inactivating FCV (2.9 log units) than HAV and MNV-1 (1.9 log units and 1.4 log units, respectively) whereas 100 ppm of peroxyacetic-based biocide was found effective for inactivating FCV (3.2 log units) and MNV-1 (2.3 log units), but not HAV (0.7 log units). Quantitative RT-PCR results indicated that the presence of viral RNA did not correlate with the presence of infectious viruses on disinfected lettuce, except for MNV-1 processed with chlorine (15 ppm). In comparison with water washing, a substantial additional decrease of genomic FCV titer (1.1 log units) but no significant reduction of the genomic titers of HAV and MNV-1 were found on lettuce treated with chlorine (15 ppm). No significant effect of the disinfection step of lettuce with peroxyacetic-based biocide (100 ppm peracetic acid) was found by qRT-PCR on all genomic viral titers tested. This study illustrates the necessity of determining the effectiveness of technological processes against enteric viruses, using a relevant reference such as HAV, in order to reduce the risk of hepatitis and gastroenteritis by exposure to vegetables. © 2011.

Bridier J.,University of Bordeaux Segalen | Claisse O.,University of Bordeaux Segalen | Coton M.,ADRIA Normandie | Coton E.,ADRIA Normandie | Lonvaud-Funel A.,University of Bordeaux Segalen
Applied and Environmental Microbiology | Year: 2010

Among the lactic acid bacteria (LAB) present in the oenological microbial ecosystem, Oenococcus oeni, an acidophilic lactic acid bacterium, is essential during winemaking. It outclasses all other bacterial species during malolactic fermentation (MLF). Oenological performances, such as malic acid degradation rate and sensorial impact, vary significantly according to the strain. The genetic diversity of the O. oeni species was evaluated using a multilocus sequence typing (MLST) scheme. Seven housekeeping genes were sequenced for a collection of 258 strains that had been isolated all over the world (particularly Burgundy, Champagne, and Aquitaine, France, Chile, South Africa, and Italy) and in several wine types (red wines, white wines, and champagne) and cider. The allelic diversity was high, with an average of 20.7 alleles per locus, many of them being rare alleles. The collection comprised 127 sequence types, suggesting an important genotypic diversity. The neighbor-joining phylogenetic tree constructed from the concatenated sequence of the seven housekeeping genes showed two major phylogenetic groups, named A and B. One unique strain isolated from cider composed a third group, rooting the phylogenetic tree. However, all other strains isolated from cider were in group B. Eight phylogenetic subgroups were statistically differentiated and could be delineated by the analysis of only 32 mutations instead of the 600 mutations observed in the concatenated sequence of the seven housekeeping genes. Interestingly, in group A, several phylogenetic subgroups were composed mostly of strains coming from a precise geographic origin. Three subgroups were identified, composed of strains from Chile, South Africa, and eastern France. Copyright © 2010, American Society for Microbiology.

Coton M.,ADRIA Normandie | Joffraud J.J.,French Research Institute for Exploitation of the Sea | Mekhtiche L.,ADRIA Normandie | Leroi F.,French Research Institute for Exploitation of the Sea | And 2 more authors.
Food Microbiology | Year: 2013

The microbial biodiversity and dynamics of king scallops meat and coral during cold storage (cold chain rupture: 1/3 storage time at 4 °C followed by 2/3 at 8 °C), was assessed by combining culture-dependant and -independent methods. Products were packaged as follows: aerobic, vacuum packed and 3 different CO2/N2 modified atmospheres and the impact of these conditions on the microbial communities was assessed. Results indicated that under air (current packaging condition), the dominant species corresponded to Brochothrix thermosphacta, Pseudomonas spp. and Shewanella spp. These species have regularly been associated in the literature with food (especially seafood), and product spoilage. Moellerella wisconsensis was the only species detected on VRBG medium, however, its impact on the food product is unclear. Packaging conditions influenced the ecosystem equilibrium and biodiversity. Except for day 8, the lowest counts for all studied flora were observed for modified atmosphere packaging (MAP) containing >80% CO2. Moreover, in these conditions, higher biodiversity by Temporal Temperature Gradient Gel Electrophoresis (TTGE) and the non-detection of specific flora (i.e. Pseudoalteromonas haloplanktis) were observed. At day 8, scallops packaged using these conditions were still acceptable from a sensorial point of view (odour), although the initial load of the king scallop was high (total psychrotrophic flora reached 5.5 log CFU/g). © 2013 Elsevier Ltd.

Buron N.,ADRIA Normandie | Buron N.,University of Caen Lower Normandy | Guichard H.,ADRIA Normandie | Coton E.,ADRIA Normandie | And 2 more authors.
Food Chemistry | Year: 2011

A new HPLC method using a diode array detector was developed and validated to quantify 4-ethylphenol, 4-vinylphenol, 4-ethylguaiacol, 4-vinylguaiacol and 4-ethylcatechol in cider. The procedure was linear up to 150. mg/l for each of the five volatile phenols, precise (RSD. < 2.9%) and sensitive, with limits of detection between 0.03 and 0.10. mg/l; moreover, it did not require any sample preparation. This method was applied to 11 phenolic off-flavour defective ciders. In these ciders, the main volatile phenol corresponded to 4-ethylcatechol. Moreover, the observed concentrations (maximum of 164. mg/l) indicated, for the first time, that this compound is an important phenolic off -flavour marker in cider. Then, volatile phenols concentrations were determined for 47 French commercial ciders and showed mean quantities of 3.2 (4-EC), 0.8 (4-EP), 0.1 (4-EG), 0.2 (4-VP) and 0.3. mg/l (4-VG). The majority of the tested commercial ciders presented low volatile phenol levels. © 2010 Elsevier Ltd.

Ladero V.,Institute Productos Lacteos Of Asturias | Coton M.,Adria Normandie | Fernandez M.,Institute Productos Lacteos Of Asturias | Buron N.,Adria Normandie | And 4 more authors.
Food Microbiology | Year: 2011

Biogenic amines (BA) are low molecular weight nitrogenous bases commonly found in fermented foods and beverages and their consumption can induce undesirable reactions. In this work, the BA content in natural cider from Spain and France was determined. Samples from commercially available cider or obtained during the elaboration process were analyzed. A different profile and BA concentration was observed depending on cider origin. qPCR tools developed for the quantitative detection of BA producers from cheese and wine were tested in the cider samples. A good connection between the BA content and the presence of BA-producing microorganisms was observed. Based on these tools, BA-producing bacteria were isolated from the analyzed cider samples, including new potential histamine- and putrescine-producing . Lactobacillus paracollinoides strains. © 2010 Elsevier Ltd.

Coton M.,ADRIA Normandie | Denis C.,ADRIA Normandie | Cadot P.,ADRIA Normandie | Coton E.,ADRIA Normandie
Food Microbiology | Year: 2011

The microbial quality and safety of surimi seafood products was assessed by studying the prevalence and biodiversity of aerobic spore-forming bacteria at the beginning and end of shelf life in 100 surimi samples. Low levels of total flora and sporulated flora were numerated at the beginning of storage, however, residual spores were detected in the majority of samples during storage. Furthermore, for 34 samples, total flora counts > 104 CFU/g were observed at the end of shelf life which could lead to non-compliance with good practice recommendations or product spoilage. In total, 460 strains were isolated, fingerprinted by M13-PCR and grouped into 98 different clusters. Representative strains were then identified at the species level via 16S rRNA gene sequencing. Overall, dominant species belonged to Bacillus simplex, Bacillus subtilis and Bacillus licheniformis; while B. simplex, B. subtilis as well as Sporosarcina aquimarina were clearly the dominant species found in samples with higher total flora counts. Amylolytic and proteolytic activities were very frequent amongst tested strains (80 and 92.5%, respectively). Heat resistance parameters of 4 strains in a surimi-based medium were determined. B. simplex and B. subtilis strains were the most heat resistant (δ96 °C = 27.6 and 23.3 min and zT = 8.6 and 7.9, respectively) which can explain their dominance in surimi samples exhibiting higher microbial counts. The heat resistance data obtained can now be used to model thermal destruction of strains using predictive microbiology tools (Sym'Previus). © 2010 Elsevier Ltd.

Sanchez A.,University of Oviedo | Coton M.,ADRIA Normandie | Coton E.,ADRIA Normandie | Herrero M.,University of Oviedo | And 2 more authors.
Food Microbiology | Year: 2012

Malolactic fermentation (MLF) is an important step in cider production in order to allowing for improvement of microbiological stability and organoleptic characteristics of cider. Induction of this fermentation by using starter cultures enables a better control over this bioprocess, but although it is a common practice in winemaking, starters specifically focussed for cider MLF are not yet commercially available. Proper starter cultures need to present the ability to degrade l-malic acid conferring pleasing sensory characteristics while avoiding toxicological risks. In this work, lactic acid bacteria (LAB) were first isolated from MLF industrial cider samples, obtained in a cellar in the main cider-producing region of Spain, Asturias. Isolates, identified by molecular tools, belonged to the Lactobacillus brevis and Oenococcus oeni species. After a phylogenetic analysis, representative strains of both identified species were evaluated in order to determine their fermentation capacity, showing O. oeni the best behaviour in this cider fermentation, as previously demonstrated for wine in the literature. Consequently, and with the aim to test the influence at strain level, selection of O. oeni isolates as starters for cider fermentation has been undergone. In order to check the influence of geography over biodiversity, O. oeni strains from six different industrial cellars representing the distinct producing areas in the region (located in a ratio of 30 km) were analyzed by using a specific RAPD method. In this way, isolates were typed in five distinct groups, mainly corresponding to each producing area. All strains isolated from the same cellar showed the same RAPD profile revealing the significance of geographical origin in the indigenous cider LAB. Molecular tools were applied to reject those isolates exhibiting presence of genes related to organoleptic spoilage (exopolysaccharides and acrolein production) or food safety (biogenic amine production), as key selection criteria. Representative strains of each of the five O. oeni RAPD groups were tested as pure cultures to evaluate their technological utility for cider production. Experimental data of malic acid degradation and cell concentration obtained were fitted to previously selected kinetic models aimed to optimization and prediction of bioprocess performance. Four strains revealed as suitable potential starter cultures for conducting MLF in cider production. © 2012.

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