Data supporting functional diversity of the marine bacterium Cobetia amphilecti KMM 296

Data is presented in support of functionality of hyper-diverse protein families encoded by the Cobetia amphilecti KMM 296 (formerly Cobetia marina KMM 296) genome (“The genome of the marine bacterium Cobetia marina KMM 296 isolated from the mussel Crenomytilus grayanus (Dunker, 1853)” [1]) providing its nutritional versatility, adaptability and biocontrol that could be the basis of the marine bacterium evolutionary and application potential. Presented data include the information of growth and biofilm-forming properties of the food-associated isolates of Pseudomonas, Bacillus, Listeria, Salmonella and Staphylococcus under the conditions of their co-culturing with C. amphilecti KMM 296 to confirm its high inter-species communication and anti-microbial activity. Also included are the experiments on the crude petroleum consumption by C. amphilecti KMM 296 as the sole source of carbon in the presence of sulfate or nitrate to ensure its bioremediation capacity. The multifunctional C. amphilecti KMM 296 genome is a promising source for the beneficial psychrophilic enzymes and essential secondary metabolites.

Data supports the possibility of inducing of previously described C. amphilecti KMM 296 genes [1] that are known to be involved in different biochemical pathways in response to the change of environmental conditions such as nutrient medium or microbial competitors, leading to change phenotypic characteristics and to significantly higher survival of the marine bacterium.
Data includes information on species-dependent decreasing of bacterial cells and biofilms growth under co-culturing conditions with C. amphilecti KMM 296 whose metabolites may be of interest for food industry and medicine protection approaches particularly against P. aeruginosa.
Data supports hydrocarbon-degrading activity in C. amphilecti KMM 296 in the presence of crude petroleum that may be used in bioremediation of soils and waters.

Data
The mussel-associated isolate of the marine bacterium KMM 296 (formerly C. marina KMM 296) [1][2][3][4] was identified as the species C. amphilecti according to the 16 S rRNA-based analysis ( Fig. 1) with 100% sequence identity with the type strain of recently described species of the genus Cobetia [2]. Although KMM 296 did not exhibit some enzymatic activity in the standard conditions (Table 1), it was able to consume petroleum hydrocarbons as the sole source of carbon in the presence of sulfate or nitrate (4 g/L) after 3-6 days of cultivation at aerobic conditions. The filtrate of C. amphilecti KMM 296 cultivated alone had no any noticeable effect on the bacterial isolates, but their presence in the medium for co-culturing resulted in its inhibition effect, lowering their colony-forming units (CFU mL À 1 ) from 2 to 1000-fold, and degrading completely the B. subtilis and P. aeruginosa biofilms during 96 h of incubation (Figs. 2-4).

Co-culturing assay
The strain KMM 296 and fourteen isolates of Pseudomonas, Bacillus, Listeria, Salmonella and Staphylococcus from ready-to-cook meat foods were used. All bacterial strains were isolated and identified to the species level by standard microbiological methods in the accredited laboratory (ISO/IEC 17025). The food-associated isolates were cultivated on the appropriate strain-specific nutrient mediums. Strain KMM 296 was cultivated for 12 h on a nutrient medium containing (g/L): bacto peptone -2.0 g, hydrolyzed casein -2.0 g, bacto yeast extract -2.0 g, dextrin -1.0 g, KH 2 PO 4 -0.02 g, MgSO 4 Â 7H 2 O -0.005 g, agar -15 g, natural sea water -500 mL, distilled water -500 mL, pH -7.0.

Antibiofilm activity assay
Antibiofilm activity was determined by semi-quantitative adherence assay in sterile U-bottom 96well polystyrene microtiter plates (Greiner Bio-One) [7]. 100 mL suspensions of the overnight bacterial culture of the foodborne isolate grown in tryptic soy broth (TSB, Merc) with the density 1 Â 10 8 CFUmL À 1 were loaded into the wells, and then 100 mL of the filtrate of three-day grown C. amphilecti KMM 296 alone or after co-culturing with the same bacterial isolate was added. In each of the wells, 100 mL 1% peptone was added. The sterile medium for C. amphilecti KMM 296 cultivation and the bacterial cells with 1% peptone were served as controls. The plates were incubated for 2-5 days at the room temperature. The plankton cells were removed and cell number was determined as described above. The biofilm was further assessed by using the crystal violet (CV) assay. The procedure involved washing the plates three times with sterile 0.85% NaCl solution to remove loosely associated cells. Then, 200 mL of 0.5% CV water solution was added in each well and incubated for 15 min at the room  temperature, after that the plates were washed 3-4 times with sterile distilled water to remove unabsorbed stain, and then the plates were air-dried. For the quantities determination of biofilm biomass, 200 mL of 96% ethanol containing 2% acetic acid was added in each well, incubated at the room temperature for 15 min, and the optical density at 600 nm was measured with an automatic plate reader (BioRad). The biofilm adherence or detachment of each isolate was measured in triplicate considering the coefficient of biofilm accretion (OD sample /OD control ).

Statistical analysis
All values presented in this article are representative of at least three independent experiments. Data were analyzed using the Student's t-test of the SigmaPlot 2000 version 6.0 program (SPSS Inc.). Differences from controls were considered significant at P r 0.05. Fig. 4. Effect of C. amphilecti KMM 296 metabolites on the biofilm density of the food-associated bacterial isolates. Bacterial isolates numbers are on the axis X. Coefficient of biofilm density is on the axis Y. Row 1foodborne bacterial biofilms after the treatment by C. amphilecti KMM 296 filtrate from the co-culturing with the same species. Row 2control samples in the same conditions of incubation.