Transmission and Persistence of Salmonella enterica in Nutrient Solution of Hydroponic Greenhouse Grown Tomatoes

Hydroponic greenhouse tomato production’s popularity has grown in the United States to meet the demands for year-round availability of fresh tomatoes. Although Salmonella has been the cause of several foodborne illness outbreaks linked to tomatoes, the potential for contamination in hydroponic production is not well understood. The objectives of this study were to determine whether biweekly inoculation of Salmonella in a hydroponic tomato nutrient solution would lead to Salmonella survival and contamination of the tomato fruit and plants, hydroponic nutrient film technique (NFT) troughs, and water receptacles. An avirulent strain of Salmonella Typhimurium was used to contaminate the nutrient solution at a concentration of 105 colony-forming units (CFU)/mL. Inoculation was conducted on day 0 and biweekly until the termination of project at 12 weeks; samples were filtered and plated on selective media. Leaves and biofilm coupons were collected on day 0 and every 2 weeks postinoculation. Leaf samples were analyzed using culture methods. The biofilm coupons were analyzed using tape fluorescence in situ hybridization (FISH) method. Fruit samples were collected 6 weeks postinoculation until termination of project and analyzed using culture methods. Typical Salmonella morphology of colonies on plates streaked from overnight cultures from plant samples were confirmed by automated ribotyping. A 2-log10 reduction of cells was observed in water samples 2 days post initial inoculation. Reduction continued over the 2-week period with few cells surviving until the next inoculation. S. Typhimurium was observed on the surface of the root systems. However, a splash incident resulted in low-level contamination of selected leaves and fruit samples. The results of the study indicate that although contaminated hydroponic nutrient solution led to surface contamination of roots, such an event may not pose a high risk of contamination of hydroponically grown fruit.

Classification of strain CCM 4446T as Rhodococcus degradans sp. nov.

Strain CCM 4446T, with notable biodegradation capabilities, was investigated in this study in order to elucidate its taxonomic position. Chemotaxonomic analyses of quinones, polar lipids, mycolic acids, polyamines and the diamino acid of the cell-wall peptidoglycan corresponded with characteristics of the genus Rhodococcus. Phylogenetic analysis, based on the 16S rRNA gene sequence, assigned strain CCM 4446T to the genus Rhodococcus and placed it in the Rhodococcus erythropolis 16S rRNA gene clade. Further analysis of catA and gyrB gene sequences, automated ribotyping with EcoRI restriction endonuclease, whole-cell protein profiling, DNA–DNA hybridization and extensive biotyping enabled differentiation of strain CCM 4446T from all phylogenetically closely related species, i.e., Rhodococcus baikonurensis, Rhodococcus qingshengii, Rhodococcus erythropolis and Rhodococcus globerulus. The results obtained show that the strain investigated represents a novel species within the genus Rhodococcus, for which the name Rhodococcus degradans sp. nov., is proposed. The type strain is CCM 4446T ( = LMG 28633T).

Comparison between Salmonella enterica Serotype Enteritidis Genotyping Methods and Phage Type

A quantitative comparison between discriminatory indexes and concordance among multilocus variable-number tandem-repeat analysis (MLVA), pulsed-field gel electrophoresis (PFGE), automated ribotyping, and phage typing has been performed, testing 238Salmonella entericaserotype Enteritidis isolates not epidemiologically correlated. The results show that MLVA is the best choice, but each typing method provides a piece of information for establishing clonal relationships between the isolates.

Staphylococcus petrasii subsp. pragensis subsp. nov., occurring in human clinical material

Seven coagulase-negative, oxidase-negative and novobiocin-susceptible staphylococci assigned tentatively as Staphylococcus petrasii were investigated in this study in order to elucidate their taxonomic position. All strains were initially shown to form a genetically homogeneous group separated from remaining species of the genus Staphylococcus by using a repetitive sequence-based PCR fingerprinting with the (GTG)5 primer. Phylogenetic analysis based on 16S rRNA gene, hsp60, rpoB, dnaJ, gap and tuf sequences showed that the group is closely related to Staphylococcus petrasii but separated from the three hitherto known subspecies, S. petrasii subsp. petrasii, S. petrasii subsp. croceilyticus and S. petrasii subsp. jettensis. Further investigation using automated ribotyping, MALDI-TOF mass spectrometry, fatty acid methyl ester analysis, DNA–DNA hybridization and extensive biotyping confirmed that the analysed group represents a novel subspecies within S. petrasii, for which the name Staphylococcus petrasii subsp. pragensis subsp. nov. is proposed. The type strain is NRL/St 12/356T ( = CCM 8529T = LMG 28327T).

The Use of MALDI-TOF Mass Spectrometry, Ribotyping and Phenotypic Tests to Identify Lactic Acid Bacteria from Fermented Cereal Foods in Abidjan (Côte d’Ivoire)

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) protein analysis, automated ribotyping, and phenotypic tests (e.g., cell morphology, gas production from glucose, growth and acid production on homofermemtative-heterofermentative differential (HHD) agar medium, sugar fermentation patterns) were used to identify 23 lactic acid bacteria (LAB) isolated from fermented cereal foods available in Abidjan, Côte d’Ivoire.Pediococcusacidilactici(56.5%),Lactobacillusfermentum(30.4%),L.salivarius(4.3%),P.pentosaceus(4.3%) andL. plantarumsubsp.plantarum(4.3%) were the species and subspecies identified. Protein based identification was confirmed by automated ribotyping for selected isolates and was similar to that provided by the phenotypic characterization. MALDI-TOF MS protein analysis provided a high level of discrimination among the isolates and could be used for the rapid screening of LAB starter cultures.

Enterococcus alcedinis sp. nov., isolated from common kingfisher (Alcedo atthis)

Two Gram-positive, catalase-negative bacterial strains were isolated from the cloaca of common kingfishers (Alcedo atthis). Repetitive sequence-based PCR fingerprinting using the (GTG)5 primer grouped these isolates into a single cluster separated from all known enterococcal species. The two strains revealed identical 16S rRNA gene sequences placing them within the genus Enterococcus with Enterococcus aquimarinus LMG 16607T as the closest relative (97.14 % similarity). Further taxonomic investigation using sequencing of the genes for the superoxide dismutase (sodA), phenylalanyl-tRNA synthase alpha subunit (pheS) and the RNA polymerase alpha subunit (rpoA) as well as application of whole-cell protein fingerprinting, automated ribotyping and extensive phenotyping confirmed that both strains belong to the same species. Based on data from this polyphasic study, these strains represent a novel species of the genus Enterococcus , for which the name Enterococcus alcedinis sp. nov. is proposed. The type strain is L34T ( = CCM 8433T = LMG 27164T).

Isolation of human pathogen Escherichia albertii from faeces of seals (Leptonychotes weddelli) in James Ross Island, Antarctica

A set of nine gram-negative fermenting rods biochemically identified as Escherichia coli was isolated from faeces of seals. These bacteria were characterized by phenotypic classification, 16S rDNA sequence analyses, automated ribotyping, study of whole-cell protein profiles by SDS-PAGE and finally by bacteriocin production. The results of our polyphasic taxonomic study supported the recognition of P4652, P4653 and P4740 isolates as true members of Escherichia albertii species – probably a major enteric human pathogen. To our best knowledge, this is the first evidence showing that E. albertii produces bacteriocin, colicin D. Obtained data unambiguously showed incon-venience of commercial identification systems to distinguish both Escherichia species due to missing data of E. albertii in the commercial databases. The results of Escherichia isolates taxonomy suggest seals as a novel source of human and animal pathogen,E. albertii in the Antarctic region.

Enterococcus ureilyticus sp. nov. and Enterococcus rotai sp. nov., two urease-producing enterococci from the environment

A set of 25 urease-producing, yellow-pigmented enterococci was isolated from environmental sources. Phenotypic classification divided the isolates into two phena. Both phena were characterized using 16S rRNA gene sequence analysis, DNA base composition, rep-PCR fingerprinting and automated ribotyping. The obtained data distinguished the isolates from all members of the genus Enterococcus with validly published names and placed them in the Enterococcus faecalis species group. DNA–DNA hybridization experiments, pheS and rpoA sequencing and whole-cell protein electrophoresis provided conclusive evidence for the classification of each phenon as a novel species of the genus Enterococcus , for which the names Enterococcus ureilyticus sp. nov. (type strain CCM 4629T = LMG 26676T = CCUG 48799T), inhabiting water and plants, and Enterococcus rotai sp. nov. (type strain CCM 4630T = LMG 26678T = CCUG 61593T), inhabiting water, insects (mosquitoes) and plants, are proposed.

International Life Science Institute North America Cronobacter (Formerly Enterobacter sakazakii) Isolate Set

Foodborne pathogen isolate collections are important for the development of detection methods, for validation of intervention strategies, and to develop an understanding of pathogenesis and virulence. We have assembled a publicly available Cronobacter (formerly Enterobacter sakazakii) isolate set that consists of (i) 25 Cronobacter sakazakii isolates, (ii) two Cronobacter malonaticus isolates, (iii) one Cronobacter muytjensii isolate, which displays some atypical phenotypic characteristics, biochemical profiles, and colony color on selected differential media, and (iv) two nonclinical Enterobacter asburiae isolates, which show some phenotypic characteristics similar to those of Cronobacter spp. The set consists of human (n = 10), food (n = 11), and environmental (n = 9) isolates. Analysis of partial 16S rDNA sequence and seven-gene multilocus sequence typing data allowed for reliable identification of these isolates to species and identification of 14 isolates as sequence type 4, which had previously been shown to be the most common C. sakazakii sequence type associated with neonatal meningitis. Phenotypic characterization was carried out with API 20E and API 32E test strips and streaking on two selective chromogenic agars; isolates were also assessed for sorbitol fermentation and growth at 45°C. Although these strategies typically produced the same classification as sequence-based strategies, based on a panel of four biochemical tests, one C. sakazakii isolate yielded inconclusive data and one was classified as C. malonaticus. EcoRI automated ribotyping and pulsed-field gel electrophoresis (PFGE) with XbaI separated the set into 23 unique ribotypes and 30 unique PFGE types, respectively, indicating subtype diversity within the set. Subtype and source data for the collection are publicly available in the PathogenTracker database (www.pathogentracker.net), which allows for continuous updating of information on the set, including links to publications that include information on isolates from this collection.