Tropical soils are a reservoir for fluorescent Pseudomonas spp. biodiversity.

Fluorescent Pseudomonas spp. are widely studied for their beneficial activities to plants. To explore the genetic diversity of Pseudomonas spp. in tropical regions, we collected 76 isolates from a Brazilian soil. Genomes were sequenced and compared to known strains, mostly collected from temperate regions. Phylogenetic analyses classified the isolates in the P. fluorescens (57) and P. putida (19) groups. Among the isolates in the P. fluorescens group, most (37) were classified in the P. koreensis subgroup and two in the P. jessenii subgroup. The remaining 18 isolates fell into two phylogenetic subclades distinct from currently recognized P. fluorescens subgroups, and probably represent new subgroups. Consistent with their phylogenetic distance from described subgroups, the genome sequences of strains in these subclades are asyntenous to the genome sequences of members of their neighbour subgroups. The tropical isolates have several functional genes also present in known fluorescent Pseudomonas spp. strains. However, members of the new subclades share exclusive genes not detected in other subgroups, pointing to the potential for novel functions. Additionally, we identified 12 potential new species among the 76 isolates from the tropical soil. The unexplored diversity found in the tropical soil is possibly related to biogeographical patterns.

Semi-scale production of PHAs from waste frying oil by Pseudomonas fluorescens S48.

The present study aimed at developing a strategy to improve the volumetric production of PHAs by Pseudomonas fluorescens S48 using waste frying oil (WFO) as the sole carbon source. For this purpose, several cultivations were set up to steadily improve nutrients supply to attain high cell density and high biopolymer productivity. The production of PHAs was examined in a 14 L bioreactor as one-stage batch, two-stage batch, and high-cell-density fed-batch cultures. The highest value of polymer content in one-stage bioreactor was obtained after 60 h (33.7%). Whereas, the two-stage batch culture increased the polymer content to 50.1% after 54 h. High-cell-density (0.64 g/L) at continuous feeding rate 0.55 mL/l/h of WFO recorded the highest polymer content after 54 h (55.34%). Semi-scale application (10 L working volume) increased the polymer content in one-stage batch, two-stage batch and high cell density fed-batch cultures by about 12.3%, 5.8% and 11.3%, respectively, as compared with that obtained in 2 L fermentation culture. Six different methods for biopolymer extraction were done to investigate their efficiency for optimum polymer recovery. The maximum efficiency of solvent recovery of PHA was attained by chloroform-hypochlorite dispersion extraction. Gas chromatography (GC) analysis of biopolymer produced by Pseudomonas fluorescens S48 indicated that it solely composed of 3-hydrobutyric acid (98.7%). A bioplastic film was prepared from the obtained PHB. The isolate studied shares the same identical sequence, which is nearly the complete 16S rRNA gene. The identity of this sequence to the closest pseudomonads strains is about 98-99%. It was probably closely related to support another meaningful parsiomony analysis and construction of a phylogenetic tree. The isolate is so close to Egyptian strain named EG 639838.

Semi-scale production of PHAs from waste frying oil by Pseudomonas fluorescens S48

The present study aimed at developing a strategy to improve the volumetric production of PHAs by Pseudomonas fluorescens S48 using waste frying oil (WFO) as the sole carbon source. For this purpose, several cultivations were set up to steadily improve nutrients supply to attain high cell density and high biopolymer productivity. The production of PHAs was examined in a 14 L bioreactor as one-stage batch, two-stage batch, and high-cell-density fed-batch cultures. The highest value of polymer content in one-stage bioreactor was obtained after 60 h (33.7%). Whereas, the two-stage batch culture increased the polymer content to 50.1% after 54 h. High-cell-density (0.64 g/L) at continuous feeding rate 0.55 mL/l/h of WFO recorded the highest polymer content after 54 h (55.34%). Semi-scale application (10 L working volume) increased the polymer content in one-stage batch, two-stage batch and high cell density fed-batch cultures by about 12.3%, 5.8% and 11.3%, respectively, as compared with that obtained in 2 L fermentation culture. Six different methods for biopolymer extraction were done to investigate their efficiency for optimum polymer recovery. The maximum efficiency of solvent recovery of PHA was attained by chloroform-hypochlorite dispersion extraction. Gas chromatography (GC) analysis of biopolymer produced by Pseudomonas fluorescens S48 indicated that it solely composed of 3-hydrobutyric acid (98.7%). A bioplastic film was prepared from the obtained PHB. The isolate studied shares the same identical sequence, which is nearly the complete 16S rRNA gene. The identity of this sequence to the closest pseudomonads strains is about 98-99%. It was probably closely related to support another meaningful parsiomony analysis and construction of a phylogenetic tree. The isolate is so close to Egyptian strain named EG 639838.

Survival of escherichia coli o157: h7 co-cultured with different levels of pseudomonas fluorescens and lactobacillus plantarum on fresh beef

The purpose of this study was to investigate the effect of different levels of Pseudomonas fluorescens (10² and 10(6)log10 cfu/ml)and Lactobacillus plantarum (10² and 10(4)log10 cfu/ml)on the growth of Escherichia coli O157:H7 on beef loins. Beef loins inoculated with E. coli O157:H7 and P. fluorescens were aerobically stored for 7 days at 4 ºC, while those inoculated with E. coli O157:H7 and L. plantarum were vacuum packaged and stored for 8 weeks at 4 ºC. Aerobic Plate Counts (APC), E. coli O157:H7 and either P. fluorescens or L. plantarum counts were determined at different storage intervals. For the aerobically packaged beef loins, E. coli O157:H7 was detected throughout the 7 day storage period regardless of the P. fluorescens level in the inoculum. For the vacuum packaged beef loins, similar inoculum levels of E. coli O157:H7 and L. plantarum allowed E. coli O157:H7 to survive until week 5 of storage, while a higher inoculum level of L. plantarum inhibited E. coli O157:H7 from week 3. Once fresh beef has been contaminated with E. coli O157:H7, the level of P. fluorescens in the background flora does not inhibit its survival and growth. However, under vacuum storage, the application of L. plantarum as a biopreservative inhibits the survival of E. coli O157:H7 on beef. The higher the level of L. plantarum in the system, the earlier the onset of the inhibition. Farmers and abattoirs have to strengthen preventive strategies to eliminate contamination of beef carcasses with E. coli O157:H7.(AU)

Identificación y caracterización de bacilos gramnegativos no fermentados aislados en el medio hospitalario / Identification and characterization of nonfermenting gramnegative bacili isolated from a hospital environment

Se estudio un total de 251 cepas aisladas en el medio hospitalario con diagnóstico presuntivo de bacilos gramnegativos no fermentadores, enviadas por los Laboratorios de Infecciones Nosocomiales de los Centros Provinciales de Higiene y Epidemiología de Ciudad de La Habana, Santiago de Cuba, Holguín, Guantánamo, Camagüey e Isla de la Juventud desde septiembre de 1992 hasta junio de 1993. Se empleó un esquema inicial para corroborar el diagnóstico primario de estas cepas, los medios de producción de pigmentos King A y King B para la identificación de Pseudomonas aeruginosas y pruebas bioquímicas claves para el diagnóstico microbiológico de otros no fermentadores, se utilizó la piocinotipia y la serotipia para la caracterización posterior de la especie aeruginosa. Resultaron confirmadas 238 cepas y de ellas, el 88,23 por ciento correspondió a la especie aeruginosa cuyos piocinotipo y serotipo predominantes fueron al 10a y 011, respectivamente. Otros bacilos encontrados con frecuencia fueron Pseudomonas cepacia (25 por ciento); Pseudomonas fluorescens (17,8 por ciento) y Acinetobacter calcoaceticus var. anitratus 17,8 por ciento

[Utilization by Escherichia coli and Pseudomonas fluorescens of a siderophore from Pseudomonas fluorescens strain PAB]. / Utilización por Escherichia coli y Pseudomonas fluorescens de un sideróforo de Pseudomonas fluorescens cepa PAB.

Pseudomonas fluorescens PAB strain produced pyoverdine in a synthetic medium. Pigment was purified by solvent extraction and ion exchange, and sterilized and used as siderophore for E. coli, P. fluorescens, ATCC 13.525, ATCC 17.400, W and PAB strains. Bacteria were grown in iron-free medium and medium with iron. Free--pyoverdine and pyoverdine bound to Fe+3 were added in different concentrations. P. fluorescens PAB siderophore did not stimulate E. coli growth while it was selective for other P. fluorescens strains. Paper disks impregnated with pyoverdine did not inhibit E. coli growth.

[Metabolism of a psychrophilic bacterium from fresh water]. / Metabolismo de una bacteria criófila de agua dulce

Psychophilic microorganisms, able to grow at 0-1 degrees C, were isolated from water obtained from the Paraná River at Rosario. One of the strains, R-12, was identified as Pseudomonas fluorescens according to the description in Bergey's Manual of Determinative Bacteriology (8th Ed., 1974). The microorganism was able to grow in liquid minimal medium with glucose, acetate, glutamate or casein hydrolysate as sole carbon source, at 20 degrees C. The enzymes of the Entner-Doudoroff pathway were induced in cells grown on glucose. The Krebs cycle was apparently operative in all cases; the lower levels of citrate synthase and isocitrate dehydrogenase were found in glucose-grown cells. Isocitrate lyase was present at a high concentration, and malate synthase considerably increased, in acetate-grown cells, thus suggesting the operation of the glyoxylate cycle. When cells were grown on glucose the anaplerotic function was probably fulfilled by pyruvate carboxylase, although phosphoenolpyruvate carboxylase was also present. The gluconeogenic enzyme phosphoenolpyruvate carboxykinase was repressed by glucose; malic enzyme was repressed by acetate. The regulatory patterns shown by citrate synthase and pyruvate carboxylase were similar to those described for the enzymes from other Pseudomonas. Whole cells of the R-12 strain were able to decarboxylate the aminoacids serine, aspartate and glutamate. Aspartate aminotransferase was present at high levels in aminoacid-grown cells, thus suggesting a catabolic role, whereas glutamate dehydrogenase had increased levels in glucose - or acetate-grown cells, suggesting that it fulfilled a mainly biosynthetic role.