Evaluation of the Effect of Different Growth Media and Temperature on the Suitability of Biofilm Formation by Enterobacter cloacae Strains Isolated from Food Samples in South Africa
Abstract
:1. Introduction
2. Results and Discussion
2.1. Microtiter Adherence
B. phenotype | Parameters number (%) | |||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
BHI a | TSB a | BHI b | TSB b | |||||||||||||
24 h | 48 h | 24 h | 48 h | 24 h | 48 h | 24 h | 48 h | |||||||||
25 °C | 37 °C | 25 °C | 37 °C | 25 °C | 37 °C | 25 °C | 37 °C | 25 °C | 37 °C | 25 °C | 37 °C | 25 °C | 37 °C | 25 °C | 37 °C | |
NA | 8(27) | 0 | 0 | 0 | 12(40) | 2(7) | 0 | 0 | 3(10) | 2(7) | 0 | 0 | 8(27) | 2(7) | 0 | 0 |
WA | 18(60) | 10(33) | 10(33) | 5(17) | 14 (47) | 17(57) | 18(60) | 4(13) | 20(67) | 11(37) | 12(40) | 4(13) | 19(63) | 14(47) | 6(20) | 2(7) |
MA | 4(13) | 18(60) | 16(53) | 0 | 4 (13) | 10(33) | 11(37) | 10(33) | 7(23) | 14(47) | 15(50) | 9 (30) | 3 (10) | 11(37) | 16(53) | 11(37) |
SA | 0 | 2(7) | 4(13) | 25(83) | 0 | 1 (3) | 1 (3) | 16(53) | 0 | 3(10) | 3(10) | 17(57) | 0 | 3(10) | 8(27) | 17(57) |
T. adh. (%) | 22 (73) | 30 (100) | 30 (100) | 30 (100) | 18 (60) | 28 (93) | 30 (100) | 30 (100) | 27 (90) | 28 (93) | 30 (100) | 30 (100) | 22 (73) | 28 (93) | 30 (100) | 30 (100) |
Parameters | Biofilm formation | |||||||
---|---|---|---|---|---|---|---|---|
Non-adherent | Weakly adherent | Moderately adherent | Strongly adherent | |||||
OD range | Mean OD ± SD | OD range | Mean OD ± SD | OD range | Mean OD ± SD | OD range | Mean OD ± SD | |
OD, optical density; SD, standard deviation; ND, not determined; a, colonies from Nutrient agar; b, inoculums from Nutrient broth; BHI, brain heart infusion broth; TSB, tryptic soy broth. | ||||||||
25 °C BHI (24 h) a | 0.448–0.486 | 0 0.470 ± 0.02 | 0.572–0.990 | 0.733 ± 0.13 | 1.30–2.14 | 1.406 ± 0.15 | ND | ND |
25 °C BHI (48 h) a | ND | ND | 0.611–1.06 | 0.875 ± 0.13 | 1.12–2.01 | 1.344 ± 0.25 | 2.192–2.769 | 2.543 ± 0.39 |
25 °C TSB (24 h) a | 0.352–0.618 | 0.512 ± 0.08 | 0.626–1.131 | 0.785 ± 0.15 | 1.365–1.458 | 1.405 ± 0.07 | ND | ND |
25 °C TSB (48 h) a | ND | ND | 0.625–1.237 | 0.984 ± 0.18 | 1.625–1.789 | 1.507 ± 0.231 | ND | ND |
37 °C BHI (24 h) a | ND | ND | 0.590–1.074 | 0.885 ± 0.12 | 1.164–2.022 | 1.504 ± 0.34 | 2.224–3.327 | 2.775 ± 0.77 |
37 °C BHI (48 h) a | ND | ND | 0.580–0.784 | 1.872 ± 0.32 | 1.331–2.13 | 1.872± 0.32 | 2.35–4.22 | 3.034 ± 0.62 |
37 °C TSB (24 h) a | 0.421–0.464 | 0.442 ± 0.03 | 0.662–1.081 | 0.861 ± 0.169 | 1.297–1.997 | 1.591 ± 0.199 | ND | ND |
37 °C TSB (48 h) a | ND | ND | 0.988–1.583 | 1.483 ± 0.493 | 1.93–2.485 | 2.149 ± 0.190 | 2.556–3.635 | 2.955 ± 0.503 |
25 °C BHI (24 h) b | 0.359–0.462 | 0.420 ± 0.05 | 0.565–1.083 | 0.832 ± 0.14 | 1.098–1.988 | 1.509 ± 0.340 | ND | ND |
25 °C BHI (48 h) b | ND | ND | 0.729–1.048 | 0.855 ± 0.13 | 1.436–2.144 | 1.728 ± 0.584 | 2.253–3.24 | 2.660 ± 1.13 |
37 °C BHI (24 h) b | 0.44–0.481 | 0.460 ± 0.02 | 0.552–1.111 | 0.803 ± 0.17 | 1.123–2.039 | 1.577 ± 0.36 | 2.405–3.27 | 2.837 ± 0.61 |
37 °C BHI (48 h) b | ND | ND | 1.01–1.159 | 1.074 ± 0.06 | 1.392–2.129 | 1.697 ± 0.24 | 2.247–3.805 | 2.93 ± 0.546 |
25 °C TSB (24 h) b | 0.421–0.559 | 0.508 ± 0.04 | 0.62–1.117 | 0.834 ± 0.16 | 1.319–1.774 | 1.548 ± 0.22 | ND | ND |
25 °C TSB (48 h) b | ND | ND | 0.711–1.066 | 1.034 ± 0.18 | 1.364–2.414 | 1.706 ± 0.36 | 2.568–3.382 | 2.929 ± 0.36 |
37 °C TSB (24 h) b | 0.503–0.535 | 0.519 ± 0.2 | 0.623–1.153 | 0.877 ± 0.2 | 1.561–2.413 | 1.802 ± 0.2 | 2.542–2.66 | 2.601 ± 0.08 |
37 °C TSB (48 h) b | ND | ND | 0.841–1.076 | 0.958 ± 1.66 | 1.25–2.19 | 1.831 ± 0.54 | 2.612–3.938 | 3.261 ± 0.54 |
Isolate (biofilm phenotype) | %Autoagg. | Coaggregation indices (%) | |||||||
---|---|---|---|---|---|---|---|---|---|
Partner strains | |||||||||
Range % | S. aureus NCTC 6571 | S. pyogenes A ATCC 49399 | S. Typhimurium ATCC 13311 | P. aeruginosa ATCC 15442 | P. shigelloides ATCC 51903 | A. hydrophila ATCC 35654 | S. sonnei ATCC 29930 | ||
EC 52-2 (NA) | 27 | 55–66 | 55 | 66 | 63 | 55 | 64 | 61 | 58 |
EC 205 (NA) | 38 | 54–66 | 54 | 63 | 66 | 62 | 62 | 64 | 61 |
EC 12-2 (WA) | 45 | 56–69 | 69 | 69 | 59 | 56 | 68 | 69 | ND |
EC 70-2 (MA) | 89 | 12–70 | 57 | 61 | 29 | 70 | 12 | 64 | 56 |
EC 89-2 (MA) | 30 | 31–62 | 43 | 54 | 62 | 40 | 53 | 31 | 23 |
EC 235 (WA) | 34 | 65–74 | 68 | 74 | 74 | 72 | 72 | 65 | 69 |
2.2. The Capability of E. cloacae Strains to Autoaggregate and Coaggregate with Partner Organisms
3. Experimental
3.1. Bacterial Strains
3.2. Microtiter Plate Assay
3.3. Autoaggregation and Coaggregation Assays
3.4. Characterization of Biofilm Formation Using Scanning Electron Microscope
4. Statistical Analysis
5. Conclusions
Acknowledgments
Conflicts of Interest
References
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Nyenje, M.E.; Green, E.; Ndip, R.N. Evaluation of the Effect of Different Growth Media and Temperature on the Suitability of Biofilm Formation by Enterobacter cloacae Strains Isolated from Food Samples in South Africa. Molecules 2013, 18, 9582-9593. https://doi.org/10.3390/molecules18089582
Nyenje ME, Green E, Ndip RN. Evaluation of the Effect of Different Growth Media and Temperature on the Suitability of Biofilm Formation by Enterobacter cloacae Strains Isolated from Food Samples in South Africa. Molecules. 2013; 18(8):9582-9593. https://doi.org/10.3390/molecules18089582
Chicago/Turabian StyleNyenje, Mirriam E., Ezekiel Green, and Roland N. Ndip. 2013. "Evaluation of the Effect of Different Growth Media and Temperature on the Suitability of Biofilm Formation by Enterobacter cloacae Strains Isolated from Food Samples in South Africa" Molecules 18, no. 8: 9582-9593. https://doi.org/10.3390/molecules18089582