Biological Characterization of the Biocontrol Agent Bacillus amyloliquefaciens CPA-8: The Effect of Temperature, pH and Water Activity on Growth, Susceptibility to Antibiotics and Detection of Enterotoxic Genes.

This work focuses on the biological understanding of the biocontrol agent Bacillus amyloliquefaciens CPA-8 in order to accomplish the characterization required in the registration process for the development of a microorganism-based product. The tolerance of CPA-8 to grow under different pH-temperature and water activity (a w)-temperature conditions was widely demonstrated. Regarding the pH results, optimum growth at the evaluated conditions was observed at 37 °C and pH between 7 and 5. On the contrary, the slowest growth was recorded at 20 °C and pH 4.5. Moreover, the type of solute used to reduce a w had a great influence on the minimum a w at which the bacterium was able to grow. The lowest a w values for CPA-8 growth in media modified with glycerol and glucose were 0.950 and 0.960, respectively. Besides, the lowest a w for CPA-8 growth increased when the temperature decreased to 20 °C, at which CPA-8 was not able to grow at less than 0.990 a w, regardless of the type of solute. Antibiotic susceptibility tests were carried out to determine which antibiotic could affect the behavior of the bacteria and revealed that CPA-8 was clearly resistant to hygromycin. Finally, a PCR amplification assay to detect the presence of enterotoxic genes from Bacillus cereus in CPA-8 was also performed. CPA-8 gave negative results for all the genes tested except for nheA gene, which is not enough for the toxicity expression, suggesting that fruit treated with this antagonist will not be a potential vehicle for foodborne illnesses.

Corneal flap thickness and topography changes induced by flap creation during laser in situ keratomileusis.

PURPOSE: To determine the corneal flap thickness profile produced by 3 microkeratomes and the topographic changes induced by flap creation in laser in situ keratomileusis (LASIK). SETTING: Cornea and Refractive Surgery Unit, Instituto de Microcirugia Ocular de Barcelona, Autonoma University, Barcelona, Spain. METHODS: In this prospective consecutive nonrandomized comparative study, patients were divided into 2 groups. In Group 1 (75 eyes), 3 microkeratomes were used: Moria LSX One, Moria M2, and Amadeus (AMO); 25 eyes per microkeratome. Pachymetry was measured with a DGH pachymeter in the center of the cornea and 3.0 mm from the center at 4 cardinal points (superior, inferior, nasal, and temporal; 3 measurements at each point) before and after the cut. The flap thickness in each sector was calculated by subtracting the mean post-flap corneal thickness from the mean pre-flap corneal thickness. In Group 2 (33 eyes), the M2 microkeratome with a 130 microm plate was used to create a superotemporal hinged flap (9 eyes) or a superonasal hinged flap (24 eyes). The topographic change induced by the microkeratome cut was evaluated using 4 sequential data acquisitions by the Keratron Scout topographic unit (Optikon) before and immediately after the cut (before laser ablation). Cardinal and oblique astigmatism and change in the axis were calculated by vectorial analysis of the simulated keratometry. Topographic Zernike analysis was performed in a subgroup. RESULTS: With the LSX One microkeratome, the mean flap thickness was 151.7 microm centrally, 161.9 microm superiorly, 151.4 microm inferiorly, 156.1 microm temporally, and 167.5 microm nasally. There was no statistically significant difference between the areas studied (P<.05). With the M2, the mean flap thickness was 131.7 microm centrally, 155.5 microm superiorly, 146.7 microm inferiorly, 143.7 microm temporally, and 160.5 microm nasally. There was a statistically significant difference between flap thickness centrally and in the other areas (P>.05). With the Amadeus microkeratome, the mean flap thickness was 140.0 microm centrally, 152.5 superiorly, 128.5 microm inferiorly, 145.0 microm temporally, and 147.0 microm nasally. Statistically significant differences (P>.05) were found in the 4 sectors of the flap. With vectorial analysis, there was no statistically significant difference between superonasal and superotemporal hinge placement in the cardinal and oblique components but there was a statistically significant difference in the axis change with both placements (P>.05). CONCLUSIONS: The LSX One microkeratome was the most predictable. A significant difference was noted in all sectors except superiorly with the M2 and in all sectors with the Amadeus. No differences between nasal and superior hinge placement were found with the M2. Topographic Zernike analysis demonstrated a difference in the orientation of the induced coma as a function of hinge position.