Krystyna Gorlach-Lira

Population dynamics and extracellular enzymes activity of mesophilic and thermophilic bacteria isolated from semi-arid soil of Northeastern Brazil

The dynamics of mesophilic and thermophilic bacterial population was studied in the bulk soil and rhizosphere of the grass Aristida adscensionis L. in a caatinga of the semi-arid Brazilian Northeast. Mesophilic heterotrophic bacteria, sporeforming bacteria and actinomycetes were significantly more abundant than thermophiles, and their counts were 106 - 107 CFU g-1 dry soil, showing weak fluctuations over the one-year study period. Thermophiles were below 106 CFU g-1 dry soil and, in general, demonstrated stronger fluctuations over the studied period. Among them, thermophilic actinomycetes showed the greatest decline during the dry season. No significant differences in microbial number were observed between bulk and rhizosphere soil. Mesophilic bacteria were represented mainly by Gram positive, filamentous organisms (actinomycetes) and by Gram positive, endospore-forming, rod-shaped organisms. All isolates of thermophilic bacteria were Gram positive, endospore-forming rods. Most of mesophilic isolates produced amylases and proteases, and 38% of isolates produced all studied extracellular enzymes: amylases, proteases, cellulases and chitinases. Among thermophilic bacteria, all isolates showed proteolytic activity and 77% produced amylases, but no one produced cellulases or chitinases.

Action of nisin and high pH on growth of Staphylococcus aureus and Salmonella sp. in pure culture and in the meat of land crab (Ucides cordatus)

O objetivo deste estudo foi avaliar o potencial de inibicao de nisina e o pH elevado em relacao a multiplicacao de Staphylococcus aureus e Salmonella sp. em culturas puras e inoculadas na carne de caranguejo-uca. Em culturas puras, a multiplicacao de S. aureus foi fortemente inibida por nisina e a de Salmonella sp. por nisina-EDTA (20 mM). A multiplicacao de S. aureus foi inibida ate 8h de incubacao, enquanto que a de Salmonella sp. foi inibida durante todo o experimento (24h). O pH elevado (tampao NaHCO3-NaOH, pH 10 e 11) mostrou-se efetivo na inibicao da multiplicacao de S. aureus e Salmonella sp. Nisina e o pH elevado aplicados na carne contaminada nao apresentaram o mesmo efeito. A nisina nao mostrou eficiencia na inibicao dos patogenos quando inoculados na carne de caranguejo, enquanto que o tampao em pH 10 demonstrou inibicao significante sobre a multiplicacao de Salmonella sp. Estes resultados sugerem que o pH elevado apresenta um potencial como agente antibacteriano, podendo ser util na preservacao quimica da carne de caranguejo.

The abundance of some pathogenic bacteria in mangrove habitats of Paraiba do Norte estuary and crabmeat contamination of mangrove crab Ucides cordatus

Study on mangrove water and sediment of Paraiba do Norte river (Northeastern Brazil) and crabmeat of Ucides cordatus was carried to investigate the relation between the abundance of bacteria in the mangrove habitat and in the crabmeat. Simultaneous collection of samples of water, sediment and crabs (U. cordatus) was done during the rainy and dry seasons at two areas of mangroves that differed in influence of tide and sewage discharge. The numbers of total heterotrophic bacteria in the mangrove water were between 0.04 and 6 log CFU 100mL-1, and in the sediment between 5 and 6 log CFU g-1. The location closer to waste discharge showed higher incidence of total heterotrophic bacteria, coliforms, Staphylococcus aureus and Salmonella spp. The counts of total heterotrophic bacteria in the crabmeat varied from 1.3 to 4.4 log CFU g-1, and did not differ significantly between the locations analysed. No crabmeat sample showed the presence of coliforms, S. aureus and Salmonella spp.

Response surface analysis on the effect of temperature and pH on Growth and proteolytic activity of thermophilic Bacillus sp

Proteolytic activity and cell biomass of thermophilic Bacillus sp. strain were evaluated at various levels of initial pH and temperature by applying response surface methodology. The mineral medium containing yeast extract (0.01%) and starch (1%) was used throughout the experiment. The results of statistical analysis revealed the polynomial model with high coefficient of determination (R2 = 0.8) for the biomass and total proteolytic activity of the strain studied. This model showed a satisfactory adjustment of the statistic model with the experimental data. The p values showed that the temperature and pH had significant effect on biomass and proteolytic activity (P<0.05) of strain tested. The highest proteolytic activity (2.333 U/ml/h) of the Bacillus sp. was predicted at 41o C and pH 4.8. The high biomass values were observed at broad range of temperature and pH.

Production of rhamnolipids and diesel oil degradation by bacteria isolated from soil contaminated by petroleum.

Biosurfactants are microbial secondary metabolites. The most studied are rhamnolipids, which decrease the surface tension and have emulsifying capacity. In this study, the production of biosurfactants, with emphasis on rhamnolipids, and diesel oil degradation by 18 strains of bacteria isolated from waste landfill soil contaminated by petroleum was analyzed. Among the studied bacteria, gram‐positive endospore forming rods (39%), gram positive rods without endospores (17%), and gram‐negative rods (44%) were found. The following methods were used to test for biosurfactant production: oil spreading, emulsification, and hemolytic activity. All strains showed the ability to disperse the diesel oil, while 77% and 44% of the strains showed hemolysis and emulsification of diesel oil, respectively. Rhamnolipids production was observed in four strains that were classified on the basis of the 16S rRNA sequences as Pseudomonas aeruginosa. Only those strains showed the rhlAB gene involved in rhamnolipids synthesis, and antibacterial activity against Escherichia coli, P. aeruginosa, Staphylococcus aureus, Bacillus cereus, Erwinia carotovora, and Ralstonia solanacearum. The highest production of rhamnolipids was 565.7 mg/L observed in mineral medium containing olive oil (pH 8). With regard to the capacity to degrade diesel oil, it was observed that 7 strains were positive in reduction of the dye 2,6‐dichlorophenolindophenol (2,6‐DCPIP) while 16 had the gene alkane mono‐oxygenase (alkB), and the producers of rhamnolipids were positive in both tests. Several bacterial strains have shown high potential to be explored further for bioremediation purposes due to their simultaneous ability to emulsify, disperse, and degrade diesel oil.