Assessing the safety of Pseudomonas putida introduction in the environment: an overview of ecotoxicological tests.

Risk assessment guidelines for the environmental release of microbial agents are performed in a tiered sequence which includes evaluation of exposure effects on non target organisms. However, it becomes important to verify whether environmental risk assessment from temperate studies is applicable to tropical countries, as Brazil. Pseudomonas putida is a bacteria showing potential to be used for environmental applications as bioremediation and plant disease control. This study investigates the effects of this bacteria exposure on rodents and aquatic organisms (Daphnia similes) that are recommended to be used as non-target organism in environmental risk assessments. Also, the microbial activity in three different soils under P. putida exposure was evaluated. Rats did not show clinical alterations, although the agent was recovered 16 h after the exposure in lung homogenates. The bacteria did not reduce significantly the reproduction and survival of D. similis. The soil enzymatic activities presented fluctuating values after inoculation with bacteria. The measurement of perturbations in soil biochemical characteristics is presented as an alternative way of monitoring the overall effects of the microbial agent to be introduced even in first stage (Tier I) of the risk assessment in tropical ecosystems.

Efeito de Pseudomonas chlororaphis sobre a podridão radicular (Pythium aphanidermatum)e o crescimento vegetal em pimentão hidropônico.

A podridão radicular causada por Pythium spp. é uma das principais doenças em cultivos hidropônicos. O objetivo do trabalho foi avaliar o efeito de Pseudomonas chlororaphis (63-28) sobre a podridão radicular e o crescimento vegetal em pimentão hidropônico. Plantas de pimentão cv. 35-206 RZ foram cultivadas em unidades hidropônicas, e infestadas com P. chlororaphis sete dias antes da inoculação com o patógeno. Os tratamentos foram: testemunha, testemunha inoculada com o patógeno, P. chlororaphis, P. chlororaphis com a adição do patógeno. As variáveis avaliadas foram: severidade da doença, expansão foliar, clorofila foliar e incidência do patógeno. A severidade da doença decresceu em 51% nas plantas inoculadas com a bactéria aos 12 dias após a inoculação, em comparação com a testemunha inoculada. O conteúdo de clorofila decresceu em 12% nas plantas inoculadas com Pythium, em comparação com a testemunha. Plantas tratadas com P. chlororaphis sem a presença do patógeno tiveram maior taxa de expansão foliar. Conclui-se que P. chlororaphis é um promissor agente de controle biológico da podridão radicular e promotor de crescimento de pimentão hidropônico.

Formulação e vida de prateleira de Pseudomonas chlororaphis 63- 28 em fibra de coco, talco e turfa.

2009

Pseudomonas Aeruginosa AmpR Transcriptional Regulatory Network

In Enterobacteriaceae, the transcriptional regulator AmpR, a member of the LysR family, regulates the expression of a chromosomal β-lactamase AmpC. The regulatory repertoire of AmpR is broader in Pseudomonas aeruginosa, an opportunistic pathogen responsible for numerous acute and chronic infections including cystic fibrosis. Previous studies showed that in addition to regulating ampC, P. aeruginosa AmpR regulates the sigma factor AlgT/U and production of some quorum sensing (QS)-regulated virulence factors. In order to better understand the ampR regulon, the transcriptional profiles generated using DNA microarrays and RNA-Seq of the prototypic P. aeruginosa PAO1 strain with its isogenic ampR deletion mutant, PAO∆ampR were analyzed. Transcriptome analysis demonstrates that the AmpR regulon is much more extensive than previously thought influencing the differential expression of over 500 genes. In addition to regulating resistance to β-lactam antibiotics via AmpC, AmpR also regulates non-β-lactam antibiotic resistance by modulating the MexEF-OprN efflux pump. Virulence mechanisms including biofilm formation, QS-regulated acute virulence, and diverse physiological processes such as oxidative stress response, heat-shock response and iron uptake are AmpR-regulated. Real-time PCR and phenotypic assays confirmed the transcriptome data. Further, Caenorhabditis elegans model demonstrates that a functional AmpR is required for full pathogenicity of P. aeruginosa. AmpR, a member of the core genome, also regulates genes in the regions of genome plasticity that are acquired by horizontal gene transfer. The extensive AmpR regulon included other transcriptional regulators and sigma factors, accounting for the extensive AmpR regulon. Gene expression studies demonstrate AmpR-dependent expression of the QS master regulator LasR that controls expression of many virulence factors. Using a chromosomally tagged AmpR, ChIP-Seq studies show direct AmpR binding to the lasR promoter. The data demonstrates that AmpR functions as a global regulator in P. aeruginosa and is a positive regulator of acute virulence while negatively regulating chronic infection phenotypes. In summary, my dissertation sheds light on the complex regulatory circuit in P. aeruginosa to provide a better understanding of the bacterial response to antibiotics and how the organism coordinately regulates a myriad of virulence factors.

Characterization of the poxAB operon encoding a class D carbapenemase in Pseudomonas aeruginosa

Pseudomonas aeruginosa is a dreaded opportunistic pathogen that causes severe and often intractable infections in immunocompromised and critically ill patients. This bacterium is also the primary cause of fatal lung infections in patients with cystic fibrosis and a leading nosocomial pathogen responsible for nearly 10% of all hospital-acquired infections. P. aeruginosa is intrinsically recalcitrant to most classes of antibiotics and has the ability to acquire additional resistance during treatment. In particular, resistance to the widely used β-lactam antibiotics is frequently mediated by the expression of AmpC, a chromosomally encoded β-lactamase that is ubiquitously found in P. aeruginosa strains. This dissertation delved into the role of a recently reported chromosomal β-lactamase in P. aeruginosa called PoxB. To date, no detailed studies have addressed the regulation of poxB expression and its contribution to β-lactam resistance in P. aeruginosa. In an effort to better understand the role of this β-lactamase, poxB was deleted from the chromosome and expressed in trans from an IPTG-inducible promoter. The loss of poxB did not affect susceptibility. However, expression in trans in the absence of ampC rendered strains more resistant to the carbapenem β-lactams. The carbapenem-hydrolyzing phenotype was enhanced, reaching intermediate and resistant clinical breakpoints, in the absence of the carbapenem-specific outer membrane porin OprD. As observed for most class D β-lactamases, PoxB was only weakly inhibited by the currently available β-lactamase inhibitors. Moreover, poxB was shown to form an operon with the upstream located poxA, whose expression in trans decreased pox promoter (Ppox) activity suggesting autoregulation. The transcriptional regulator AmpR negatively controlled Ppox activity, however no direct interaction could be demonstrated. A mariner transposon library identified genes involved in the transport of polyamines as potential regulators of pox expression. Unexpectedly, polyamines themselves were able induce resistance to carbapenems. In summary, P. aeruginosa carries a chromosomal-encoded β-lactamase PoxB that can provide resistance against the clinically relevant carbapenems despite its narrow spectrum of hydrolysis and whose activity in vivo may be regulated by polyamines.^

A non-classical LysR-type transcriptional regulator PA2206 is required for an effective oxidative stress response in Pseudomonas aeruginosa

LysR-type transcriptional regulators (LTTRs) are emerging as key circuit components in regulating microbial stress responses and are implicated in modulating oxidative stress in the human opportunistic pathogen Pseudomonas aeruginosa. The oxidative stress response encapsulates several strategies to overcome the deleterious effects of reactive oxygen species. However, many of the regulatory components and associated molecular mechanisms underpinning this key adaptive response remain to be characterised. Comparative analysis of publically available transcriptomic datasets led to the identification of a novel LTTR, PA2206, whose expression was altered in response to a range of host signals in addition to oxidative stress. PA2206 was found to be required for tolerance to H2O2 in vitro and lethality in vivo in the Zebrafish embryo model of infection. Transcriptomic analysis in the presence of H2O2 showed that PA2206 altered the expression of 58 genes, including a large repertoire of oxidative stress and iron responsive genes, independent of the master regulator of oxidative stress, OxyR. Contrary to the classic mechanism of LysR regulation, PA2206 did not autoregulate its own expression and did not influence expression of adjacent or divergently transcribed genes. The PA2214-15 operon was identified as a direct target of PA2206 with truncated promoter fragments revealing binding to the 5'-ATTGCCTGGGGTTAT-3' LysR box adjacent to the predicted -35 region. PA2206 also interacted with the pvdS promoter suggesting a global dimension to the PA2206 regulon, and suggests PA2206 is an important regulatory component of P. aeruginosa adaptation during oxidative stress.

Resistência de representantes dos gêneros pseudomonas sp e bacillus sp mediante exposição à radiação ultravioleta c.

As mudanças climáticas causadas pela degradação da camada de ozônio e pelo aumento da emissão de gases causadores do efeito estufa, podem ocasionar alterações não somente aos seres humanos, mamíferos, répteis ou aves, mas também aos microrganismos. Dentro deste contexto, o aumento nos níveis de radiação solar incidente na superfície da Terra pode afetar as interações que ocorrem entre bactérias e plantas, alterando tanto a ocorrência de doenças como as interações benéficas entre estes organismos. Muitos são os gêneros de bactérias que interagem com as plantas, colonizando os mais diferentes nichos presentes neste hospedeiro. Dentre os grupos bacterianos destacam-se os gêneros Bacillus sp e Pseudomonas sp, que possuem efeitos benéficos como a promoção de crescimento vegetal e a proteção de plantas contra doenças e pragas. Dessa forma, o atual trabalho tem a princípio, como enfoque, constatar a resistência de isolados dos dois gêneros mencionados, à radiação ultravioleta C (200 a 250 nm). Os resultados obtidos demonstram maior resistência da linhagem de Bacillus sp à radiação ultravioleta C em comparação à linhagem de Pseudomonas sp. fato que possivelmente está relacionado com a capacidade de produzir esporos, característica do gênero Bacillus. Este trabalho fornece dados iniciais para futuras avaliações do comportamento destas bactérias quando em interação com plantas e sob maiores níveis de radiação ultravioleta no ambiente.

Influence of Pseudomonas putida AF7 inoculation on soil enzymes.

Pseudomonas may use as bioremediator and as biopesticide. The use of soil enzymatic assays as biological indicator of possible negative effects in soil functioning was evaluated after P.putida AF7 inoculation. For that, AF7 was originally isolated from the rizosphere of rice and was inoculated on three soils: Rhodic Hapludox (RH), Typic Hapludox (TH); and Arenic Hapludult (AH). Soil characteristics were measured in each plot. Acid phosphatase, ?-glucosidase and protease activities were measured at 7, 14 and 21 days. The enzyme activity waved during the experimental period but there is a significant reduction of ?-glucosidase activity in RH soil on day 14. Corg was positively correlated to the activities of ?-glucosidase and protease. The presented data indicate that soil biochemical properties may be useful as indicator of soil perturbations.

Influence of pseudomonas putida af7 inoculation on soil enzymes activities.

2008

Influence of Pseudomonas putida AF7 inoculation on soil enzymes activities.

2008

Antagonismo de rizobactérias do gênero Pseudomonas à Thielaviopsis sp.

2016

Antifungal activity of Pseudomonas frederiksbergensis CMAA 1323 isolated from the Antarctic hair grass Deschampsia antarctica.

Abstract: Aims: Epiphytic bacteria, isolated from Deschampsia antarctica, were screened for their potential to inhibit the plant pathogen Botrytis cinerea, the causal agent of gray mold disease of strawberry pseudofruits. This phytopathogenic fungus is more active and the disease is more serious in temperate climate where the temperatures are lower.

Desenvolvimento de um meio semi-seletivo para a detecção de Pseudomonas syringae pv. tomato.

Desenvolveu-se um meio semi-seletivo para Pseudomonas syringae pv. tomato (Pst), objetivando seu uso em detecção do patógeno em sementes de tomate. A seletividade do meio foi obtida através de antibiogramas e testes de crescimento do patógeno, representado por 04 isolados procedentes de Guaíra, SP, Botucatu, SP, Coimbra,MG e Planaltina, GO. Também, foram comparadas diversas concentrações do agente antifúngico clorotalonil e do agente halofílico. O meio proposto tem a seguinte constituição, tendo como base o meio B de King: proteose peptona n.3, 10g; KH2PO4, 1,5g; MgSO4, 7H2O, 15g; glicerol, 10 ml; NaCl, 15 g; Clorotalonil, 200 ug/ml; agar, 15g; agua destilada, 1000 ml; acrescido dos antibioticos cefadroxil, 50 ug/ml; cefalexina, 50 ug/ml e clindamicina, 100 ug/ml. Este meio apresenta um índice de repressividade de 1,3%, alto índice de supressividade e sensibilidade de 10(2) UFC/ml de Pst.

Culturable Bacterial Diversity From A Feed Water Of A Reverse Osmosis System, Evaluation Of Biofilm Formation And Biocontrol Using Phages.

Biofilm formation on reverse osmosis (RO) systems represents a drawback in the application of this technology by different industries, including oil refineries. In RO systems the feed water maybe a source of microbial contamination and thus contributes for the formation of biofilm and consequent biofouling. In this study the planktonic culturable bacterial community was characterized from a feed water of a RO system and their capacities were evaluated to form biofilm in vitro. Bacterial motility and biofilm control were also analysed using phages. As results, diverse Protobacteria, Actinobacteria and Bacteroidetes were identified. Alphaproteobacteria was the predominant group and Brevundimonas, Pseudomonas and Mycobacterium the most abundant genera. Among the 30 isolates, 11 showed at least one type of motility and 11 were classified as good biofilm formers. Additionally, the influence of non-specific bacteriophage in the bacterial biofilms formed in vitro was investigated by action of phages enzymes or phage infection. The vB_AspP-UFV1 (Podoviridae) interfered in biofilm formation of most tested bacteria and may represent a good alternative in biofilm control. These findings provide important information about the bacterial community from the feed water of a RO system that may be used for the development of strategies for biofilm prevention and control in such systems.

Evaluation Of The Cytotoxicity And Phototoxicity Of Caryocar Brasiliense Supercritical Carbon Dioxide Extract.

Caryocar brasiliense Camb (Pequi) is a typical Brazilian Cerrado fruit tree. Its fruit is used as a vitamin source for culinary purposes and as a source of oil for the manufacture of cosmetics. C. brasiliense supercritical CO2 extracts exhibit antimicrobial activity against the bacteria Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus and also possess antioxidant activity. This study was designed to evaluate the in vitro cytotoxicity and phototoxicity of the supercritical CO2 extract obtained from the leaves of this species. In vitro cytotoxicity and phototoxicity of C. brasiliense supercritical CO2 extracts were assessed using a tetrazolium-based colorimetric assay (XTT) and Neutral Red methods. We found that the C. brasiliense (Pequi) extract obtained by supercritical CO2 extraction did not present cytotoxic and phototoxic hazards. This finding suggests that the extract may be useful for the development of cosmetic and/or pharmaceutical products.