Bacteria involved in biodegradation of creosote PAH - A case study of long-term contaminated industrial area.

Polycyclic aromatic hydrocarbons (PAH) contained in creosote oil are particularly difficult to remove from the soil environment. Their hydrophobic character and low bioavailability to soil microorganisms affects their rate of biodegradation. This study was performed on samples of soil that were (for over forty years) subjected to contamination with creosote oil, and their metagenome and physicochemical properties were characterized. Moreover, the study was undertaken to evaluate the biodegradation of PAHs by autochthonous consortia as well as by selected bacteria strains isolated from long-term contaminated industrial soil. From among the isolated microorganisms, the most effective in biodegrading the contaminants were the strains Pseudomonas mendocina and Brevundimonas olei. They were able to degrade more than 60% of the total content of PAHs during a 28-day test. The biodegradation of these compounds using AT7 dispersant was enhanced only by Serratia marcescens strain. Moreover, the addition of AT7 improved the effectiveness of fluorene and acenaphthene biodegradation by Serratia marcescens 6-fold. Our results indicated that long-term contact with aromatic compounds induced the bacterial strains to use the PAHs as a source of carbon and energy. We observed that supplementation with surfactants does not increase the efficiency of hydrocarbon biodegradation.

Evaluation of Lactobacillus spp. and yeast based probiotic (Lavipan) supplementation for the reduction of Salmonella Enteritidis after infection of broiler chickens.

The number of human cases of salmonellosis in the EU was 94,625 in 2015. Considering the source of these infections, Salmonella spp. was most frequently detected in broiler chicken meat and Salmonella Enteritidis (SE) was the most commonly reported serovar. The efficacy of probiotics in limiting Salmonella spp. infection in poultry has been demonstrated in numerous papers. The administration of probiotics at the level of primary production reduces the risk of contamination of poultry food products with Salmonella spp. A study was carried out in order to determine the potential for reducing the Salmonella spp. population in broiler chickens with the use of the Lavipan (JHJ, Poland) probiotic that comprised selected stains of lactic acid bacteria and Saccharomyces cervisae. Salmonella spp.-free broiler chickens were divided into two groups and received the same feed with (group L) or without (group C) the probiotic throughout the experiment. All day-old chickens were infected per os with SE. Samples of cecum content were collected 2, 4, and 6 weeks after SE infection and pectoral muscles were collected 6 weeks following SE infection for the evaluation of the SE population number. Serum samples for serological examinations were collected 6 weeks after infection. Six weeks after infection, the number of SE-positive cecal samples was lower in the L group (12.5% positive) in comparison to the C group (87.5%). Similar results were demonstrated for the muscle samples (25% in contrast to 87.5%). At the same time, in both cases, the SE CFU/g was significantly lower in the L group. The results of our study indicate that Lavipan was capable of reducing the population of SE in the gastrointestinal tract, which eventually improved the hygienic parameters of the pectoral muscles. Four weeks after infection, SE was not detected in any of the experimental groups. In both groups, no specific anti-SE antibodies were detected.

Biodegradation of clotrimazole and modification of cell properties after metabolic stress and upon addition of saponins.

Azole fungicides constitute an extensive group of potential emerging pollutants which can be found in natural environment. This study focuses on the biodegradation of clotrimazole and the characterization of cell surface properties of microorganisms capable of degradation of this compound. The influence of long-term contact of bacteria with clotrimazole and the impact of the addition of Saponaria officinalis extract on cell surface modification was also checked. The biodegradation of clotrimazole did not exceed 70%. The presence of plant extract increased biodegradation of fungicide. The cells metabolic activity after one-month exposure to clotrimazole was the highest for each tested strain. Moreover, metabolic stress led to a strong modification of cell surface properties. The results are promising for determining the impact of clotrimazole on environmental microorganisms.

Impact of potent bioremediation enhancing plant extracts on Raoultella ornithinolytica properties.

Long-term contact of microorganisms with different compounds in the environment can cause significant changes in cell metabolism. Surfactants adsorption on cell surface or incorporation in the cell membrane, lead to their modification, which helps microorganisms adopt to the conditions of metabolic stress. The main objective of this study was to investigate the effects of three saponin-reach plant extracts from Hedera helix, Saponaria officinalis and Sapindus mucorossi on growth and adaptation of Raoultella ornithinolytica to high concentrations of these substances. For this purpose we investigated cell surface properties, membrane fatty acids and genetic changes of the microorganisms. The results revealed that prolonged exposure of the microorganisms to high concentrations of these surfactants can induce genetic changes of their genes. Moreover, the adaptation to contact with high concentrations of saponins was also associated with changes in composition of fatty acids responsible for the stabilisation of membrane structure and the increase in membrane permeability. The changes affected also the outer layer of cells. A significant increase (p < 0.05) in the cell surface hydrophobicity of tested strain was also observed. The cells after long-term contact with S. officinalis and S. mucorossi acquire properties that may be favourable in hydrophobic substances bioremediation.

Phenotypic and genotypic antimicrobial susceptibility pattern of Streptococcus spp. isolated from cases of clinical mastitis in dairy cattle in Poland.

Mastitis of dairy cattle is one of the most frequently diagnosed diseases worldwide. The main etiological agents of mastitis are bacteria of the genus Streptococcus spp., in which several antibiotic resistance mechanisms have been identified. However, detailed studies addressing this problem have not been conducted in northeastern Poland. Therefore, the aim of our study was to analyze, on phenotypic and genotypic levels, the antibiotic resistance pattern of Streptococcus spp. isolated from clinical cases of mastitis from dairy cattle in this region of Poland. The research was conducted using 135 strains of Streptococcus (Streptococcus uberis, n = 53; Streptococcus dysgalactiae, n = 41; Streptococcus agalactiae, n = 27; other streptococci, n = 14). The investigation of the antimicrobial susceptibility to 8 active substances applied in therapy in the analyzed region, as well as a selected bacteriocin (nisin), was performed using the minimum inhibitory concentration method. The presence of selected resistance genes (n = 14) was determined via PCR. We also investigated the correlation between the presence of resistance genes and the antimicrobial susceptibility of the examined strains in vitro. The highest observed resistance of Streptococcus spp. was toward gentamicin, kanamycin, and tetracycline, whereas the highest susceptibility occurred toward penicillin, enrofloxacin, and marbofloxacin. Additionally, the tested bacteriocin showed high efficacy. The presence of 13 analyzed resistance genes was observed in the examined strains [gene mef(A) was not detected]. In most strains, at least one resistance gene, mainly responsible for resistance to tetracyclines [tet(M), tet(K), tet(L)], was observed. However, a relationship between the presence of a given resistance gene and antimicrobial susceptibility on the phenotypic level was not always observed.

Hydrocarbon-induced changes in proteins and fatty acids profiles of Raoultella ornithinolytica M03.

Microorganisms can support environmental restoration by biodegradation of hydrocarbons but the mechanism of this process has been not described in detail yet. We present the effect of benzene derivatives on Raoultella ornithinolytica M03 cell composition. Comparison of the cell response after short-term and long-term stress revealed significant differences in surface properties, fatty acid composition and proteins profile. R. ornithinolytica M03 after long-term stress was characterized by lower cell surface hydrophobicity and much higher inner membrane permeability. Also decrease in the content of branched and unsaturated fatty acids was observed. Cells after short- and long-term stress were characterized by analyses of changes related to thirty-nine proteins participating in various metabolic pathways. The presence of benzene derivatives resulted in modifications in the abundance of proteins involved in determination of cell shape and ability to ion transport, lipid biosynthesis, amino-acid biosynthesis, tRNA ligases, chaperone and TCA cycle proteins, gluconeogenesis, transcription and nucleotide synthesis. Uptake and transport associated proteins, cell properties and membrane stability were also found to differ in the cells after short- and long-term stress suggesting the use of different mechanisms for transport and biodegradation of benzene derivatives and modification of cell response depending on the length of exposure to the stressor. BIOLOGICAL SIGNIFICANCE: This is the first comprehensive study whose results may contribute to a better understanding of the changes occurring during short- and long-term contact with benzene derivatives. After long term stress R. ornithinolytica M03 was characterized by lower cell surface hydrophobicity and much higher inner membrane permeability and decrease in the content of branched and unsaturated fatty acids. We identified changes related to multiple proteins engaged in various metabolic pathways such as biogenesis of cell membrane/wall, amino-acid biosynthesis, nucleotide and protein synthesis, gluconeogenesis and tRNA ligases. Changes in proteins participating in uptake and transport associated proteins, cell properties and membrane stability indicate modifications in transport and biodegradation of benzene derivatives, connected with the length of exposure to the stressor. The provided results seem to constitute an important aspect of remediation techniques.

Effect of feed supplementation with kynurenic acid on the morphology of the liver, kidney and gills in rainbow trout (Oncorhynchus mykiss Walbaum, 1792), healthy and experimentally infected with Yersinia ruckeri.

Kynurenic acid (KYNA) is an endogenous substance produced on the kynurenine pathway which is primarily known for its neuroactive properties. Recently, it has been proven that KYNA is a selective ligand for G protein-coupled receptor (GPR 35), presented on immunocompetent cells such as T lymphocytes. This opens up new possibilities of its application as an immunostimulating substance in aquaculture. Thus far, no histopathological investigations in fish have been completed to evaluate influence of KYNA supplementation in feed. This study has been undertaken to determine the effect of feed supplementation with KYNA (2.5, 25, 250 mg kg-1 of feed) for 28 days on the liver, gills and kidney in healthy fish and experimentally infected with Yersinia ruckeri. In a control group were observed a fatty liver, which is natural for this fish species in the autumn and winter season. As the dose of the supplement was increased, the fat liver changed, it decreased or completely disappeared. Additionally, inflammatory changes occurred in all the analysed organs, and their intensification was dose dependent. In the fish experimentally infected, KYNA caused aggravation of the signs in the liver, kidneys and gills, and the effect was dose dependent. The results implicate that KYNA may be a stressor for fish.

The in vitro effect of kynurenic acid on the rainbow trout (Oncorhynchus mykiss) leukocyte and splenocyte activity.

Kynurenic acid (KYNA), an endogenous neuroprotectant formed along the kynurenine pathway of tryptophan degradation, is a selective ligand of the GPR35 receptor, which can be found on the surface of various populations of human immune cells. In infections and inflammations, KYNA produces an anti-inflammatory effect through this receptor, by depressing the synthesis of reactive oxygen species and pro-inflammatory cytokines. However, it is still unrecognized whether receptors for kynurenic acid are also localized on immune cells of poikilothermic animals, or whether KYNA is able to affect these cells. The objective of this study has been to determine the effect of different concentrations of kynurenic acid (12.5 microM to 10 mM) on the viability and mitogenic response of lymphocytes and on the activity of phagocytic cells isolated from blood and the spleen of rainbow trout. The results imply low toxicity of kynurenic acid towards fish immune cells, and the proliferative effect observed at the two lowest concentrations of KYNA (12.5 microM and 25 microM) seems indicative of endogenous kynurenic acid being capable of activating fish lymphocytes. Non-toxic, micromole concentrations of KYNA, however, had no influence on the mitogenic response of lymphocytes nor on the activity of phagocytes in rainbow trout under in vitro conditions. There is some likelihood that such an effect could be observed at lower, nanomole concentrations of KYNA.

The influence of surfactants on cell surface properties of Aeromonas hydrophila during diesel oil biodegradation.

In this study the capacity of the newly isolated environmental strain Aeromonas hydrophila was evaluated. The influence of three surfactants: rhamnolipides, saponins and Triton X-100 on cell surface properties of the A. hydrophila environmental strain and the biodegradation process of diesel oil was studied. The surface activities in water, a mineral salts medium and in the biological system of all considered surfactants were estimated by means of equilibrium surface tension experiments. The obtained results indicated that critical micellar concentration in the biological system is twice higher for saponins and Triton X-100, and three times higher for rhamnolipides. Our results indicated also, that cell surface hydrophobicity (CSH) of bacteria is correlated with carbon sources in broth medium. The mechanism of surfactant action seems to be dependent on the type and concentration of surfactant used in the studies. The best effect of saponins on diesel oil biodegradation was observed using the A. hydrophila strain, diesel oil biodegradation after 21 days was 78%.

Yeast and bacteria cell hydrophobicity and hydrocarbon biodegradation in the presence of natural surfactants: rhamnolipides and saponins.

This study concerns the relation between hydrocarbon biodegradation in the presence of natural surfactants and cell hydrophobicity resulting from the use of these surfactants. The relative capabilities of two bacterial strains (Pseudomonas aeruginosa and Bacillus subtilis) and two yeast strains (Candida maltosa, Yarrowia lipolytica) were investigated. The selected microorganisms were tested separately and in combination in order to achieve the optimal degrading performance. The surface cell hydrophobicity of microorganisms and the degree of hydrocarbon biodegradation were measured. The microbial adhesion to the hydrocarbon (MATH) test was used to denote the surface cell hydrophobicity of the microbial species. The results indicate the correlation between the modification of the surface cell and the degree of hydrocarbon biodegradation; however results for bacteria differ from that obtained for yeast strains. Saponins, as the surfactant, was more effective than rhamnolipides during hydrocarbon biodegradation, though the concentration of this surfactant has no significant influence on the surface cell hydrophobicity.