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Food Research International (Ottawa,... Jun 2024The microbiome of surfaces along the beef processing chain represents a critical nexus where microbial ecosystems play a pivotal role in meat quality and safety of end...
The microbiome of surfaces along the beef processing chain represents a critical nexus where microbial ecosystems play a pivotal role in meat quality and safety of end products. This study offers a comprehensive analysis of the microbiome along beef processing using whole metagenomics with a particular focus on antimicrobial resistance and virulence-associated genes distribution. Our findings highlighted that microbial communities change dynamically in the different steps along beef processing chain, influenced by the specific conditions of each micro-environment. Brochothrix thermosphacta, Carnobacterium maltaromaticum, Pseudomonas fragi, Psychrobacter cryohalolentis and Psychrobacter immobilis were identified as the key species that characterize beef processing environments. Carcass samples and slaughterhouse surfaces exhibited a high abundance of antibiotic resistance genes (ARGs), mainly belonging to aminoglycosides, β-lactams, amphenicols, sulfonamides and tetracyclines antibiotic classes, also localized on mobile elements, suggesting the possibility to be transmitted to human pathogens. We also evaluated how the initial microbial contamination of raw beef changes in response to storage conditions, showing different species prevailing according to the type of packaging employed. We identified several genes leading to the production of spoilage-associated compounds, and highlighted the different genomic potential selected by the storage conditions. Our results suggested that surfaces in beef processing environments represent a hotspot for beef contamination and evidenced that mapping the resident microbiome in these environments may help in reducing meat microbial contamination, increasing shelf-life, and finally contributing to food waste restraint.
Topics: Microbiota; Red Meat; Animals; Cattle; Food Microbiology; Food Handling; Bacteria; Metagenomics; Drug Resistance, Bacterial; Abattoirs; Anti-Bacterial Agents; Food Contamination; Drug Resistance, Microbial; Food Packaging
PubMed: 38729711
DOI: 10.1016/j.foodres.2024.114318 -
Food Research International (Ottawa,... Jan 2024Fresh fish is a highly perishable product and is easily spoiled by microbiological activity and chemical oxidation of lipids. However, microbial spoilage is the main...
Fresh fish is a highly perishable product and is easily spoiled by microbiological activity and chemical oxidation of lipids. However, microbial spoilage is the main factor linked with the rapid fish sensorial degradation due to the action of specific spoilage organisms (SSOs) that have the ability to dominate over other microorganisms and produce metabolites responsible for off-flavours. We explored the microbial dynamics in fresh anchovies stored in different packaging (air, modified atmosphere, under vacuum) and temperatures (0, 4 and 10 °C) using shotgun metagenomics, highlighting the selection of different microbial species according to the packaging type. Indeed, Pseudoalteromonas nigrifaciens, Psychrobacter cryohalolentis and Ps. immobilis, Pseudomonas deceptionensis and Vibrio splendidus have been identified as the main SSOs in aerobically stored anchovies, while Shewanella baltica, Photobacterium iliopiscarium, Ps. cryohalolentis and Ps. immobilis prevailed in VP and MAP. In addition, we identified the presence of spoilage-associated genes, leading to the potential production of biogenic amines and different off-flavors (HS, TMA). In particular, the abundance of microbial genes leading to BA biosynthesis increased at higher storage temperature, while those related to HS and TMA production were enriched in aerobically and VP packed anchovies, suggesting that MAP could be an effective strategy in delaying the production of these compounds. Finally, we provided evidence of the presence of a wide range of antibiotic resistance genes conferring resistance to different classes of antibiotic (β-lactams, tetracyclines, polymyxins, trimethoprims and phenicols) and highlighted that storage at higher temperature (4 and 10 °C) boosted the abundance of ARG-carrying taxa, especially in aerobically and MAP packed fish.
Topics: Animals; Food Packaging; Anti-Bacterial Agents; Food Microbiology; Food Preservation; Genomics; Microbiota
PubMed: 38129066
DOI: 10.1016/j.foodres.2023.113788 -
BMC Genomics Dec 2014Microbial communities of traditional cheeses are complex and insufficiently characterized. The origin, safety and functional role in cheese making of these microbial...
BACKGROUND
Microbial communities of traditional cheeses are complex and insufficiently characterized. The origin, safety and functional role in cheese making of these microbial communities are still not well understood. Metagenomic analysis of these communities by high throughput shotgun sequencing is a promising approach to characterize their genomic and functional profiles. Such analyses, however, critically depend on the availability of appropriate reference genome databases against which the sequencing reads can be aligned.
RESULTS
We built a reference genome catalog suitable for short read metagenomic analysis using a low-cost sequencing strategy. We selected 142 bacteria isolated from dairy products belonging to 137 different species and 67 genera, and succeeded to reconstruct the draft genome of 117 of them at a standard or high quality level, including isolates from the genera Kluyvera, Luteococcus and Marinilactibacillus, still missing from public database. To demonstrate the potential of this catalog, we analysed the microbial composition of the surface of two smear cheeses and one blue-veined cheese, and showed that a significant part of the microbiota of these traditional cheeses was composed of microorganisms newly sequenced in our study.
CONCLUSIONS
Our study provides data, which combined with publicly available genome references, represents the most expansive catalog to date of cheese-associated bacteria. Using this extended dairy catalog, we revealed the presence in traditional cheese of dominant microorganisms not deliberately inoculated, mainly Gram-negative genera such as Pseudoalteromonas haloplanktis or Psychrobacter immobilis, that may contribute to the characteristics of cheese produced through traditional methods.
Topics: Bacteria; Cheese; Dairy Products; Databases, Genetic; Fermentation; Genome, Bacterial; Metagenomics; Microbiota; Sequence Analysis
PubMed: 25496341
DOI: 10.1186/1471-2164-15-1101 -
Journal of Clinical Microbiology Mar 2003CDC eugonic oxidizer group 2 (EO-2) is a group of unclassified gram-negative bacterial strains isolated from various human sources. As determined by biochemical tests...
CDC eugonic oxidizer group 2 (EO-2) is a group of unclassified gram-negative bacterial strains isolated from various human sources. As determined by biochemical tests and analyses of fatty acid compositions, these organisms form a homogeneous group that appears to be distinct from but related to other Paracoccus species. Molecular studies were performed on a set of 13 EO-2 strains from various clinical sources and geographic locations in the United States and Canada to determine their relationship to the Paracoccus genus. Control strains were Paracoccus denitrificans ATCC 17741(T), P. versutus ATCC 25364(T), P. aminophilus ATCC 49673(T), P. solventivorans ATCC 700252(T), and Psychrobacter immobilis ATCC 43116(T), which are phenotypically similar to EO-2. Nearly complete (1,500-base) 16S rRNA gene sequencing of eight EO-2 strains showed a high level of sequence similarity (>99.3%) within the group, and a BLAST search of GenBank placed the EO-2 cluster in close proximity to Paracoccus species (95 to 97% similarity). DNA-DNA hybridization studies of 13 of the EO-2 strains showed all to be related at the species level, with >70% relatedness under stringent conditions and a divergence within the group of less than 2%. None of the Paracoccus control strains hybridized at >54% with any of the EO-2 strains. These results indicate that EO-2 represents a new Paracoccus species, the first isolated from human clinical specimens. A new species, Paracoccus yeeii, is proposed for the EO-2 strains. The type strain of P. yeeii is CDCG1212 (ATCC BAA-599 and CCUG 46822), isolated in Pennsylvania from dialysate of a 77-year-old male with peritonitis.
Topics: DNA, Bacterial; Gram-Negative Bacterial Infections; Humans; Molecular Sequence Data; Paracoccus; Phenotype; Phylogeny; RNA, Ribosomal, 16S
PubMed: 12624070
DOI: 10.1128/JCM.41.3.1289-1294.2003 -
Journal of Food Protection Mar 2000Three phenotypic identification systems were employed to identify 106 strains of gram-negative, nonmotile, aerobic bacteria obtained during iced storage of wild (Salmo...
Three phenotypic identification systems were employed to identify 106 strains of gram-negative, nonmotile, aerobic bacteria obtained during iced storage of wild (Salmo trutta and Esox lucius) and farmed (Oncorhynchus mykiss) freshwater fish. Using diagnostic tables and computer-assisted identification, the isolates were Psychrobacter (64 strains), Acinetobacter (24 strains), Moraxella (6 strains), Chryseobacterium (5 strains), Myroides odoratus (2 strains), Flavobacterium (1 strain), Empedobacter (1 strain), and unidentified (3 strains). Overall similarities of all strains were determined for 108 characters by numerical analysis (simple matching coefficient of similarity [S] and clustering by unweighted pair group average linkage [UPGMA]). At the 77% similarity level, 92 strains formed nine major clusters (3 or more strains) and four small clusters (2 strains). Cluster 1 (25 isolates divided into two main subclusters) could be assigned to Psychrobacter phenylpyruvicus, clusters 2 and 3 (26 isolates) were designated as Psychrobacter immobilis, and clusters 4 (3 isolates) and 7 (4 isolates) were identified as Psychrobacter urativorans and Psychrobacter spp., respectively. Clusters 5 (five isolates), 6 (three isolates), and 9 (five isolates) were labeled as Acinetobacter spp., Acinetobacter johnsonii, and Acinetobacter lwoffii, respectively. Cluster 8 (12 isolates), with a high resemblance to Thornley's phenon 4 (a heterogeneous group of bacteria isolated from poultry and related to Acinetobacter), remained unnamed. The restriction pattern was identical for strains grouped into clusters 2 and 3 (P. immobilis) but was different for the remaining Psychrobacter isolates. A large proportion of isolates belonging to the family Moraxellaceae were closely related. Psychrobacters and A. johnsonii were present in freshly caught fish and river water. In the latter stages of storage, P. phenylpyruvicus and acinetobacters tended to decrease, whereas P. immobilis increased.
Topics: Animals; Bacterial Typing Techniques; Cold Temperature; DNA, Ribosomal; Esocidae; Fishes; Food Handling; Fresh Water; Gram-Negative Aerobic Bacteria; Numerical Analysis, Computer-Assisted; Oncorhynchus mykiss; Phenotype; RNA, Ribosomal, 16S; Restriction Mapping; Trout
PubMed: 10716558
DOI: 10.4315/0362-028x-63.3.315 -
European Journal of Biochemistry Feb 1997A heat-labile beta-lactamase has been purified from culture supernatants of Psychrobacter immobilis A5 grown at 4 degrees C and the corresponding chromosomal ampC gene...
A heat-labile beta-lactamase has been purified from culture supernatants of Psychrobacter immobilis A5 grown at 4 degrees C and the corresponding chromosomal ampC gene has been cloned and sequenced. All structural and kinetic properties clearly relate this enzyme to class C beta-lactamases. The kinetic parameters of P. immobilis beta-lactamase for the hydrolysis of some beta-lactam antibiotics are in the same range as the values recorded for the highly specialized cephalosporinases from pathogenic mesophilic bacteria. By contrast, the enzyme displays a low apparent optimum temperature of activity and a reduced thermal stability. Structural factors responsible for the latter property were analysed from the three-dimensional structure built by homology modelling. The deletion of proline residues in loops, the low number of arginine-mediated H-bonds and aromatic-aromatic interactions, the lower global hydrophobicity and the improved solvent interactions through additional surface acidic residues appear to be the main determinants of the enzyme flexibility.
Topics: Adaptation, Physiological; Amino Acid Sequence; Antarctic Regions; Base Sequence; Chromosomes, Bacterial; Cold Temperature; Enzyme Stability; Genes, Bacterial; Gram-Negative Aerobic Bacteria; Hot Temperature; Molecular Sequence Data; beta-Lactamases
PubMed: 9063463
DOI: 10.1111/j.1432-1033.1997.00186.x -
Applied and Environmental Microbiology Dec 1995A class C beta-lactamase has been purified from the culture supernatant of the antarctic psychrophile Psychrobacter immobilis A8. This psychrophilic beta-lactamase...
A class C beta-lactamase has been purified from the culture supernatant of the antarctic psychrophile Psychrobacter immobilis A8. This psychrophilic beta-lactamase displays a low level of thermal stability and a low optimal temperature of activity. In contrast to other cold-adapted enzymes, its level of specific activity is not higher than that of mesophilic class C beta-lactamases.
Topics: Acinetobacter; Amino Acid Sequence; Molecular Sequence Data; Temperature; beta-Lactamases
PubMed: 8534113
DOI: 10.1128/aem.61.12.4474-4476.1995 -
Journal of Clinical Microbiology Sep 1992Location of the double-bond position of monounsaturated fatty acids of various bacteria was accomplished with combined gas chromatography-mass spectrometry analysis of...
Location of the double-bond position of monounsaturated fatty acids of various bacteria was accomplished with combined gas chromatography-mass spectrometry analysis of dimethyl disulfide (DMDS) derivatives. The monoenoic fatty acids from whole cells were converted to methyl esters and then to DMDS adducts and analyzed by capillary gas chromatography-mass spectrometry. The mass spectra of DMDS adducts gave an easily recognizable molecular ion and two major diagnostic ions attributable to fragmentation between the two CH3S groups located at the original site of unsaturation. Twenty-one relatively novel monoenoic fatty acids were identified among the bacteria studied. All Flavobacterium species contained i17:1 omega 8c, Bacillus alvei contained i16:1 omega 11c and i17:1 omega 12c, and Psychrobacter immobilis contained 12:1 omega 9c. Resolution of cis and trans isomers with capillary gas chromatography and subsequent mass spectrometry permitted positive identification of 16:1 omega 7c and 16:1 omega 7t in Arcobacter (Campylobacter) cryaerophila and 16:1 omega 9c and 16:1 omega 9t in Aerococcus viridans.
Topics: Bacteria; Fatty Acids, Monounsaturated; Gas Chromatography-Mass Spectrometry
PubMed: 1401029
DOI: 10.1128/jcm.30.9.2511-2512.1992 -
Applied and Environmental Microbiology Jul 1992A numerical taxonomic study using 75 characters was performed with 132 strains of gram-negative, nonmotile, nonfermentative bacteria selected on the basis of lack of...
A numerical taxonomic study using 75 characters was performed with 132 strains of gram-negative, nonmotile, nonfermentative bacteria selected on the basis of lack of motility and Gram reaction among 1,200 cultures isolated during aerobic storage of lamb carcasses. At the 80% similarity level (SSM), eight clusters were formed. Strains in clusters 1 to 6 could be identified as members of the family Moraxellaceae and, more specifically, as members of the Psychrobacter-[Moraxella] phenylpyruvica subgroup. Of these strains, clusters 1 and 2 (88 strains) were identified as [Moraxella] phenylpyruvica and cluster 3 (15 strains) was identified as Psychrobacter immobilis. Clusters 4, 5, and 6 were not identifiable with any species. Clusters 7 and 8 consisted of 14 strains considered nonmotile variants of Pseudomonas fragi. The highest separation indices corresponded to acid production from certain carbohydrates (melibiose, L-arabinose, and cellobiose). Although strains of Psychrobacter-Moraxella clusters were relatively frequently identified at the completion of slaughter, very few cultures were detected on spoiled carcasses. It appears, therefore, that this group of organisms has only low spoilage potential.
Topics: Animals; Bacteriological Techniques; Cold Temperature; Data Interpretation, Statistical; Food Microbiology; Food Preservation; Gram-Negative Bacteria; Sheep
PubMed: 1637162
DOI: 10.1128/aem.58.7.2245-2249.1992 -
Journal of Clinical Microbiology Sep 1991Psychrobacter immobilis was isolated from the cerebrospinal fluid (CSF) and blood of a 2-day-old infant who appeared well except for a fever and a full anterior...
Psychrobacter immobilis was isolated from the cerebrospinal fluid (CSF) and blood of a 2-day-old infant who appeared well except for a fever and a full anterior fontanelle. The infant was treated with antibiotics intravenously. After 48 h, he became afebrile and CSF and blood cultures were negative; he was then discharged. After 96 h of incubation, CSF and blood cultures yielded a gram-negative organism, P. immobilis. The child was readmitted to the hospital, and the same organism was again isolated from his blood and CSF.
Topics: Humans; Infant, Newborn; Male; Meningitis, Bacterial; Neisseriaceae; Neisseriaceae Infections
PubMed: 1774332
DOI: 10.1128/jcm.29.9.2041-2042.1991