Functional synergism of pyoverdine and the S-type pyocins of Pseudomonas aeruginosa / 生物工程学报

Pyocin S2 and S4 in Pseudomonas aeruginosa use the same uptake channels as the pyoverdine does in bacteria, indicating a possible connection between them. In this study, we characterized the single bacterial gene expression distribution of three S-type pyocins (Pys2, PA3866, and PyoS5) and examined the impact of pyocin S2 on bacterial uptake of pyoverdine. The findings demonstrated that the expression of the S-type pyocin genes was highly differentiated in bacterial population under DNAdamage stress. Moreover, exogenous addition of pyocin S2 reduces the bacterial uptake of pyoverdine so that the presence of pyocin S2 prevents the uptake of environmental pyoverdine by non-pyoverdine synthesizing 'cheaters', thereby reducing their resistance to oxidative stress. Furthermore, we discovered that overexpression of the SOS response regulator PrtN in bacteria significantly decreased the expression of genes involved in the synthesis of pyoverdine, significantly decreasing the overall synthesis and exocytosis of pyoverdine. These findings imply a connection between the function of the iron absorption system and the SOS stress response mechanism in bacteria.

Biophysicochemical characterization of Pyocin SA189 produced by Pseudomonas aeruginosa SA189

Abstract Pseudomonas aeruginosa, in spite of being a ubiquitous organism (as it is found in soil, water, and humans), is also an opportunistic pathogen. In order to maintain its diversity in the community, it produces various toxic proteins, known as, bacteriocins. In the present study, pyocin SA189, which is a bacteriocin produced by P. aeruginosa SA189 (isolated from a clinical sample) was characterized. P. aeruginosa SA189, as identified by the conventional and 16S rRNA gene amplification, produced pyocin SA189 of molecular weight of 66 k Da. The pyocin showed antimicrobial activity against several clinically relevant Gram-positive and Gram-negative bacteria and was substantially stable for wide ranges of temperature and pH. Furthermore, the pyocin also retained its biological activity upon treatment with metal ions, organic solvents, and various proteolytic and lipolytic enzymes. The data from the growth kinetics indicated that the maximum bacteriocin production occurred in the late log phase. Overall, our results signify the potential of pyocin SA189 as a bio-control agent.

Comparative study of two bacteriocins produced by representative indigenous soil bacteria

The aim of this research work was to identify and characterized the bacteriocins produced by soil-associated bacteria. Bacillocin from Bacillus brevis Bb and pyocin from Pseudomonas aeruginosa Pa were found bioactive only against gram-positive bacteria tested. Maximum production of both the bacteriocins was observed at 32°C in BHI medium. Production of both the bacteriocins started in the early exponential growth phase while the maximum production was observed during the stationary phase. Bacillocin Bb remained stable during 1-9 pH while pyocin Pa remained stable at pH 1-11. Both of the bacterocins were found resistant to high temperature [100°C for 30 min], detergents [1% solutions of EDTA, Tween 20, Tween 80 and SDS] and organic solvents [1% solutions of Ethanol, Butanol, Methanol, Propanol, Chloroform, and Acetone]. Activity of both was completely lost after proteinase K treatment suggesting their protein nature. Titre of bacillocin Bb was estimated to be 5280 AU/mL while the titre for pyocin Pa was calculated as 640 AU/mL. Both of the bacteriocins showed bacteriolytic mode of action against the indicator Bacillus strain BC31 and were found <10 KDa in their molecular mass

Study of Pseudomonas aeruginosa causing ventilator associated pneumonia.

Ps. aeruginosa is a frequent and prominent cause of nosocomial pneumonia especially in persons on assisted ventilation in the intensive care units. In a year long surveillance of ventilator associated pneumonia (VAP) we isolated 42 strains from broncho alveolar lavage samples collected and processed from 102 patients. By pyocin typing 40 of the 42 strains could be typed into 39 types but this designation changed each time the test was repeated. SDS-PAGE analysis of the whole cell proteins grouped the 42 strains of Ps. aeruginosa into 20 groups. After ribotyping, using an 18 mer DIG labelled oligonucleotide to the conserved region of 16S rRNA gene, the strains were designated into 18 types. The major type contained 8 isolates, but there was no clustering of isolates, indicating that each infecting strain was acquired separately and not from a common source. It would, therefore, appear that cross infection with a single clone was not the predominant mode of Ps. aeruginosa infection causing VAP in our ICU.

Producción y sensibilidad a microcinas en cepas de Pseudomonas aeruginosa / Pseudomonas aeruginosa microcins: production and sensibility

Se aislaron cepas de Pseudomonas aeruginosa de distintos procesos infecciosos con el objeto de estudiar la capacidad de producir microcinas y la sensibilidad a las elaboradas por otras cepas. Se investigaron por el método de estrías cruzadas en condiciones mínimas de crecimiento y en agar nutritivo, siendo más adecuado el primero. Se analizó el efecto de la presencia de metionina en medio mínimo. El tamaño molecular de las bacteriocinas se estimó con membranas capaces de retener moléculas mayores de 8000 daltons. Los resultados obtenidos indicaron una alta incidencia de cepas productoras de microcinas (85,7 por ciento). La sensibilidad a las mismas fue variada desde 0 a 10 cepas susceptible. Al mantener P. aeruginosa en el laboratorio se redujo el espectro de actividad, mientras que las recientemente aisladas inhibieron mayor número de cepas. Se observó también que disminuyó la sensibilidad a microcinas al conservarse las cepas por más de seis meses. Es conveniente realizar tipificación mediante microcinas con bacterias mantenidas por poco tiempo en cultivo

A preliminary study of fingerprinting of Pseudomonas aeruginosa by whole cell protein analysis by SDS-PAGE.

Forty two strains of Pseudomonas aeruginosa isolated from bronchoalveolar lavage fluid from intubated patients admitted to the Intensive care unit in AIIMS between December 1993 to June 1994 were included in the study. After obtaining typical biochemical profile, antimicrobial susceptibility was performed against ceftazidime, amikacin, gentamicin, ampicillin, cefotaxime and ciprofloxacin. Pyocin typing of these 42 strains was performed by scrape and streak method using 22 indicator strains. Forty strains could be typed showing excellent discrimination but on repeated testing the group designation changed indicating that the system had low reproducibility. SDS-PAGE of whole cell protein profile indicated the presence of 45 protein bands of different molecular weights, individual isolates had 37 to 42 protein band ranging in molecular weight from 340 kDa to 14.3 kDa. On the basis of Dice index of similarity the strains could be grouped into 20 types. Since all strains could be typed, the system has adequate typability. Similar results were obtained on repeated testing indicating good reproducibility.

Phenotypic characterization of pseudomonas aeruginosa by pyocin typing of two different methods

No abstract available.

Marcadores de virulencia de Pseudomonas aeruginosa. I - Provas "in vitro" tipos piocinicos e atividade hemolitica. / Virulence markers of Pseudomonas aeruginosa. I. In vitro tests: pyocin type and hemolytic activity

A virulencia de 145 amostras de Pseudomonas aeruginosa, isoladas de quatro servicos do Hospital das Clinicas da UFMG, previamente tipificadas pela producao de piocina, foi investigada quanto a capacidade para produzir "hemolisina". Do Bercario, foram estudadas 57 amostras e 46 destas (80,7%), pertencentes a um mesmo tipo piocinico, apresentaram titulo hemolitico baixa, isto e, igual ou menor que 1:8, enquanto as outras, pertencentes a tipos diferentes, tiveram titulos que variaram de 1:32-1:256. Do Contro de Tratamento Intensivo (CTI), foram estudadas 28 amostras.Duas destas apresentaram titulo hemolitico intermediario (1:16) e os restantes (92,8%), cuja maioria pertence a um grupo nao tipavel pela producao de piocina, mostraram titulos altos. Da Clinica Cirurgica 17 amostras pertencentes a cinco tipos diferentes mostraram titulos variaveis. Da Clinica Urologica, 43 amostras, com predominancia de nao tipaveis, variam quanto a capacidade de produzir "hemolisina"