scholarly journals POTENCIAL MICROBIOLÓGICO DO SYZYGIUM AROMATICUM (L). SOBRE MICRORGANISMOS DE IMPORTÂNCIA NA MEDICINA VETERINÁRIA

2021 ◽  
Author(s):  
Vanessa Arantes Pinto ◽  
Gabriela Santos Alencar ◽  
Adriano Favero ◽  
Hugo Franciscon ◽  
Fagner Luiz Da Costa Freitas
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Enterococcus Faecalis ◽  
Syzygium Aromaticum ◽  
E Coli ◽  
Streptococcus Uberis

Introdução: O fenômeno de resistência a antimicrobianos está se mostrando um grande problema na saúde, ameaçando tanto a população humana como animal. Devido a isso, a descoberta de novos compostos com capacidade antimicrobiana torna-se imprescindível, e os fitoterápicos têm se mostrado uma alternativa promissora na terapia complementar. O óleo essencial (OE) de Syzygium aromaticum (Cravo da Índia), apresenta potencial capacidade antibacteriana e antifúngica devido às altas concentrações de Eugenol. Objetivos: Avaliar o potencial antimicrobiano do óleo essencial de Syzygium aromaticum (L). contra microrganismos de importância na Medicina veterinária. Materiais e métodos: Foram testadas as bactérias gram positivas: S. aureus LB 25923, S. aureus B24, S. aureus NP 38, Streptococcus uberis, S. agalactiae, Enterococcus faecalis e S. epidermidis; as gram negativas: Pseudomonas aeruginosa ATCC 27853 e E. coli ATCC 25922; e os fungos: C. tropicalis e C. gattii 179. Utilizou-se a técnica de disco difusão (Kirby-bauer) aplicando 10µL de OE, e posteriormente, para os patógenos que apresentaram o halo inibitório, determinou-se, aplicando 15µL de OE, a concentração inibitória mínima (CIM), concentração bactericida mínima (CBM) e concentração fungicida mínima (CFM) através da microdiluição seriada utilizando leitor de ELISA e corante resazurina. Resultados: Todos os microrganismos apresentaram halo inibitório. Com relação aos valores de CIM e CBM para as bactérias, obteve-se, respectivamente: S. aureus LB25923 (18,75% e 18,75%), S. aureus B 24 (4,94% e 4,94%) S. aureus NP 38 (3,64% e 3,64%) Streptococcus uberis (2,60% e 3,125%) S. agalactiae (5,20 %, 5,20%), Enterococcus faecalis (5,20% e 6,25%), S. epidermidis (3,38% e 4,42%), Pseudomonas aeruginosa ATCC 27853 (2,09% e 2,09%), E. coli ATCC 25922 (10,41% e 10,41%). Os resultados de CIM e CFM obtidos para os fungos foram respectivamente: C. tropicalis (1,82% e 1,82%) e C. gattii 179 (5,27% e 5,27%). Conclusão: O óleo essencial de Syzygium aromaticum possui considerável potencial antimicrobiano “in vitro”, e dessa forma, pode ser considerado uma possível alternativa para as terapias sintéticas de tratamento das doenças infecciosas bacterianas e fúngicas encontradas na rotina clínica.

Phänotypische Antibiotikaresistenzen von Bakterienisolaten aus Zier-, Zoo- und falknerisch gehaltenen Greifvögeln

2020 ◽  
Vol 48 (04) ◽  
pp. 260-269
Author(s):  
Leonie Steger ◽  
Monika Rinder ◽  
Rüdiger Korbel
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Klebsiella Pneumoniae ◽  
Enterococcus Faecalis ◽  
Enterobacter Cloacae ◽  
Coli Isolate ◽  
Aureus Isolate ◽  
E Coli ◽  
Klinische Relevanz

Zusammenfassung Gegenstand und Ziel Die Prävalenz von antibiotikaresistenten Bakterien bei Zier-, Zoo- und falknerisch gehaltenen Greifvögeln ist noch weitgehend unbekannt. Daher sollten retrospektiv Antibiogramme schnellwachsender aerober Bakterienarten ausgewertet werden. Material und Methoden Im Auswertungszeitraum von 2007 bis 2016 standen 1036 Antibiogramme zur Verfügung. Die Bakterienisolate stammten vorzugsweise aus Süddeutschland und von 811 Vögeln aus 20 zoologischen Ordnungen (am häufigsten Papageienvögel [61,8 %] und Sperlingsvögel [14,5 %]) sowie aus Proben von klinischen Patienten und Sektionsmaterial. Die phänotypische In-vitro-Empfindlichkeit wurde mittels Plattendiffusionstest ermittelt. Ergebnisse Die meisten Antibiogramme lagen für E. coli (n = 386 Isolate) vor, gefolgt von Staphylococcus (S.). aureus (n = 150), Enterobacter cloacae (n = 122), Klebsiella pneumoniae (n = 86) und Pseudomonas aeruginosa (n = 64). Resistenzen gegen mindestens einen antibiotischen Wirkstoff zeigten 53,1 % der E. coli-Isolate, dabei am häufigsten gegen Doxycyclin (50,3 %) und Ampicillin (46,1 %). Bei 78,0 % der S. aureus-Isolate und bei 95,9 % der Enterococcus faecalis-Isolate wurden Resistenzen gegenüber mindestens einem Wirkstoff nachgewiesen. Multiresistenzen (Resistenz gegenüber ≥ 3 Antibiotikagruppen) traten bei 37,3 % der Isolate von S. aureus auf. Bei Isolaten von Zier- und Greifvögeln wurden höhere Resistenzraten festgestellt als bei Isolaten von Zoovögeln und bei Papageienvögeln höhere Resistenzraten als bei Sperlingsvögeln. Im Untersuchungszeitraum zeigte sich bei E. coli ein tendenzieller Anstieg der Resistenzrate für Fluorchinolone (Minimum von 0 % im Jahr 2013 und Maximum von 27,3 % im Jahr 2015) und bei S. aureus eine tendenzielle Abnahme der Resistenzraten für Tetrazykline (Maximum von 39,4 % im Jahr 2007 und Minimum von 0 % in den Jahren 2014 und 2015). Schlussfolgerung und klinische Relevanz Die Resistenzsituation von Bakterien aus Zier-, Zoo- und falknerisch gehaltenen Greifvögeln ist als problematisch zu bewerten und verdeutlicht die Wichtigkeit der Empfindlichkeitsprüfung für eine gewissenhafte Therapie. Im Fall einer Infektion mit S. aureus bei Zier-, Zoo- oder falknerisch gehaltenen Greifvögeln kann es zu einem Therapienotstand kommen.

ATIVIDADE ANTIMICROBIANA IN VITRO DO EXTRATO AQUOSO, HIDROALCOÓLICO E ALCOÓLICO DE FOLHAS DE ESPÉCIES DA FAMÍLIA LAMIACEAE

2018 ◽  
Vol 4 ◽  
Author(s):  
Karlynne Freire Mendonça ◽  
José Klauber Roger Carneiro ◽  
Maria Auxiliadora Silva Oliveira
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Enterococcus Faecalis ◽  
Streptococcus Pyogenes ◽  
Mentha Arvensis ◽  
Ocimum Gratissimum ◽  
Mueller Hinton ◽  
Plectranthus Amboinicus

Objetivos: avaliar a atividade antimicrobiana em extrato aquoso, hidroalcoólico e alcoólico das folhas de espécies da família Lamiaceae frente a bactérias de interesse. Método: Foram escolhidas quatro espécies: Ocimum gratissimum, Plectranthus amboinicus, Mentha arvensis e Plectranthus barbatus. A partir das folhas foram confeccionados os extratos aquoso, hidroalcoólico e alcoólico nas concentrações 100mg/mL, 50mg/mL e 25mg/mL. Foram selecionadas as bactérias Streptococcus pyogenes, Enterococcus faecalis, Staphylococcus aureus e Pseudomonas aeruginosa para os ensaios de antibiose em Ágar Mueller-Hinton. Resultados: P. barbatus, em seu extrato hidroalcoólico mostrou ativo nas três concentrações para bactéria S. aureus, e ainda foi ativo para P. aeruginosa, demonstrando no extrato alcoólico atividade frente as bactérias. Para M. arvensis e P. amboinicus, seus extratos hidroalcoólico e alcoólico apresentaram atividade para S. aureus. Conclusão: Sugere-se que as espécies em questão apresentem boa atividade antimicrobiana, sendo necessária a realização de mais estudos para melhor entender esse mecanismo.

scholarly journals Influence of nutrient availability on in vitro growth of major bovine mastitis pathogens

2021 ◽  
Vol 88 (1) ◽  
pp. 80-88
Author(s):  
Remo Stürmlin ◽  
Josef J. Gross ◽  
Olga Wellnitz ◽  
Lea A. Wagner ◽  
Camille Monney ◽  
...  
Keyword(s):  
Bacterial Growth ◽  
Nutrient Availability ◽  
Bovine Mastitis ◽  
Milk Composition ◽  
B Vitamins ◽  
E Coli ◽  
Streptococcus Uberis ◽  
Whole Milk ◽  
Mastitis Pathogens

AbstractThe aim of the present study was to investigate the effects of milk composition changes on the in vitro growth of bovine mastitis pathogens. Nutritional requirements of three major bovine mastitis pathogens Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and Streptococcus uberis (S. uberis) were investigated in vitro. We used ultra-high temperature (UHT) treated milk with different contents of fat, protein, and carbohydrates to test the influence of the availability of various milk constituents on pathogen growth characteristics. Additionally, the bacterial growth was investigated under experimentally modified nutrient availability by dilution and subsequent supplementation with individual nutrients (carbohydrates, different nitrogen sources, minerals, and different types of B vitamins) either to milk or to a conventional medium (thioglycolate broth, TB). Varying contents of fat, protein or lactose did not affect bacterial growth with the exception of growth of S. uberis being promoted in protein-enriched milk. The addition of nutrients to diluted whole milk and TB partly revealed different effects, indicating that there are media-specific growth limiting factors after dilution. Supplementation of minerals to diluted milk did not affect growth rates of all studied bacteria. Bacterial growth in diluted whole milk was decreased by the addition of high concentrations of amino acids in S. aureus, and by urea and additional B vitamins in E. coli and S. aureus. The growth rate of S. uberis was increased by the addition of B vitamins to diluted whole milk. The present results demonstrate that growth-limiting nutrients differ among pathogen types. Because reduced bacterial growth was only shown in diluted milk or TB, it is unlikely that alterations in nutrient availability occurring as a consequence of physiological changes of milk composition in the cow's udder would directly affect the susceptibility or course of bovine mastitis.

Cloning the gene for the heat shock response positiwe regulator (sigma 32 homolog) from Pseudomonas aeruginosa

1995 ◽  
Vol 41 (1) ◽  
pp. 75-87 ◽  
Author(s):  
Zerlina M. Naczynski ◽  
Andrew M. Kropinski ◽  
Chris Mueller
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Heat Shock ◽  
Sigma Factor ◽  
Oligonucleotide Probe ◽  
In Vitro Transcription ◽  
Translation System ◽  
Amino Acid Homology ◽  
E Coli ◽  
Transcriptional Start Sites

A 31 base pair synthetic oligonucleotide based on the genes for the Escherichia coli heat shock sigma factor (rpoH) and the Pseudomonas aeruginosa housekeeping sigma factor (rpoD) was employed in conjunction with the Tanaka et al. (K. Tanaka, T. Shiina, and H. Takahashi, 1988. Science (Washington, D.C.), 242: 1040–1042) RpoD box probe to identify the location of the rpoH gene in P. aeruginosa genomic digests. This gene was cloned into plasmid pGEM3Z(f+), sequenced, and found to share 67% nucleotide identity and 77% amino acid homology with the rpoH gene and its product (σ32) of E. coli. The plasmid containing the rpoH gene complemented the function of σ32 in an E. coli rpoH deletion mutant. Furthermore, this plasmid directed the synthesis of a 32-kDa protein in an E. coli S-30 in vitro transcription–translation system. Primer extension studies were used to identify the transcriptional start sites under control and heat-stressed (45 and 50 °C) conditions. Two promoter sites were identified having sequence homology to the E. coli σ70 and σ24 consensus sequences.Key words: heat shock, Pseudomonas aeruginosa, sigma factor, transcription, oligonucleotide probe.

scholarly journals Activity of Ceftazidime-Avibactam against Fluoroquinolone-Resistant Enterobacteriaceae and Pseudomonas aeruginosa

2015 ◽  
Vol 59 (6) ◽  
pp. 3059-3065 ◽  
Author(s):  
C. Pitart ◽  
F. Marco ◽  
T. A. Keating ◽  
W. W. Nichols ◽  
J. Vila
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Resistant Mutant ◽  
Fluoroquinolone Resistance ◽  
Cross Resistance ◽  
Content Type ◽  
E Coli ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
The Impact

ABSTRACTCeftazidime-avibactam and comparator antibiotics were tested by the broth microdilution method against 200Enterobacteriaceaeand 25Pseudomonas aeruginosastrains resistant to fluoroquinolones (including strains with the extended-spectrum β-lactamase [ESBL] phenotype and ceftazidime-resistant strains) collected from our institution. The MICs and mechanisms of resistance to fluoroquinolone were also studied. Ninety-nine percent of fluoroquinolone-resistantEnterobacteriaceaestrains were inhibited at a ceftazidime-avibactam MIC of ≤4 mg/liter (using the susceptible CLSI breakpoint for ceftazidime alone as a reference). Ceftazidime-avibactam was very active against ESBLEscherichia coli(MIC90of 0.25 mg/liter), ESBLKlebsiella pneumoniae(MIC90of 0.5 mg/liter), ceftazidime-resistant AmpC-producing species (MIC90of 1 mg/liter), non-ESBLE. coli(MIC90of ≤0.125 mg/liter), non-ESBLK. pneumoniae(MIC90of 0.25 mg/liter), and ceftazidime-nonresistant AmpC-producing species (MIC90of ≤0.5 mg/liter). Ninety-six percent of fluoroquinolone-resistantP. aeruginosastrains were inhibited at a ceftazidime-avibactam MIC of ≤8 mg/liter (using the susceptible CLSI breakpoint for ceftazidime alone as a reference), with a MIC90of 8 mg/liter. Additionally, fluoroquinolone-resistant mutants from each species tested were obtainedin vitrofrom two strains, one susceptible to ceftazidime and the other a β-lactamase producer with a high MIC against ceftazidime but susceptible to ceftazidime-avibactam. Thereby, the impact of fluoroquinolone resistance on the activity of ceftazidime-avibactam could be assessed. The MIC90values of ceftazidime-avibactam for the fluoroquinolone-resistant mutant strains ofEnterobacteriaceaeandP. aeruginosawere ≤4 mg/liter and ≤8 mg/liter, respectively. We conclude that the presence of fluoroquinolone resistance does not affectEnterobacteriaceaeandP. aeruginosasusceptibility to ceftazidime-avibactam; that is, there is no cross-resistance.

scholarly journals Antibacterial Activity of Leukocyte- and Platelet-Rich Plasma: AnIn VitroStudy

2018 ◽  
Vol 2018 ◽  
pp. 1-8 ◽  
Author(s):  
Agata Cieślik-Bielecka ◽  
Tadeusz Bold ◽  
Grzegorz Ziółkowski ◽  
Marcin Pierchała ◽  
Aleksandra Królikowska ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Enterococcus Faecalis ◽  
Platelet Rich Plasma ◽  
Antibacterial Properties ◽  
Antimicrobial Effect ◽  
Diffusion Method ◽  
Bacterial Strains ◽  
Bovine Thrombin ◽  
Zones Of Inhibition

The aim of the study was to investigate the leukocyte- and platelet-rich plasma (L-PRP) antimicrobial activity. The studied sample comprised 20 healthy males. The L-PRP gel, liquid L-PRP, and thrombin samples were testedin vitrofor their antibacterial properties against selected bacterial strains using the Kirby-Bauer disc diffusion method. Two types of thrombin were used (autologous and bovine). Zones of inhibition produced by L-PRP ranged between 6 and 18 mm in diameter. L-PRP inhibited the growth ofStaphylococcus aureus(MRSA and MSSA strains) and was also active againstEnterococcus faecalisandPseudomonas aeruginosa. There was no activity againstEscherichia coliandKlebsiella pneumoniae. The statistically significant increase of L-PRP antimicrobial effect was noted with the use of major volume of thrombin as an activator. Additionally, in groups where a bovine thrombin mixture was added to L-PRP the zones of inhibition concerning MRSA,Enterococcus faecalis, andPseudomonas aeruginosawere larger than in the groups with autologous thrombin. Based on the conducted studies, it can be determined that L-PRP can evokein vitroantimicrobial effects and might be used to treat selected infections in the clinical field. The major volume of thrombin as an activator increases the strength of the L-PRP antimicrobial effect.

scholarly journals Efeito antimicrobiano do extrato de Cranberry sobre micro-organismos causadores de infecção urinária

2016 ◽  
Vol 11 (31) ◽  
pp. 113-122
Author(s):  
Carla Franco Porto Belmont Souza ◽  
Luiz Eduardo Souza da Silva Irineu ◽  
Renan Silva De Souza ◽  
Renato da Silva Teixeira ◽  
Ivina Sanches Pereira ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Klebsiella Pneumoniae ◽  
Enterococcus Faecalis ◽  
Proteus Mirabilis ◽  
Enterococcus Faecium ◽  
Vaccinium Macrocarpon ◽  
Mueller Hinton

A resistência microbiana tem se mostrado um problema de proporções mundiais, causando estado de morbidade e mortalidade em diversos pacientes. Em vista disso, tem crescido a busca por métodos alternativos naturais de profilaxia. A investigação clínica sugere que o Extrato de Cranberry está entre as melhores propostas de prevenção natural. O Cranberry (Vaccinium macrocarpon) é um fruto que tem crescido comercialmente pelo sabor e propriedades benéficas à saúde. Dentre as formas comercializadas estão: o suco, o chá e as cápsulas contendo o extrato seco. A ação desta planta está relacionada ao tratamento de doenças do trato urinário, por possuir substâncias que inibem a adesão bacteriana ao epitélio do trato urinário, dificultando sua proliferação e reprodução. Dentre todas as infecções relacionadas à assistência a saúde, a Infecção do Trato Urinário é a mais frequentemente associada a procedimentos invasivos. Se não for tratada, pode resultar em complicações como pielonefrite aguda, bacteremia e pionefrose. Portanto, cranberry pode ser uma nova alternativa para o combate das infecções uroepiteliais, por ser um produto natural de preço acessível, e com formas de comercialização diversificada, ao contrário dos antimicrobianos convencionais, que por sua vez são caros e podem acabar causando resistência nos micro-organismos. Este trabalho teve como objetivo avaliar in vitro a atividade antimicrobiana do extrato de Cranberry, adquirido em farmácia de manipulação, sobre 8 micro-organismos isolados de infecções urinárias. As cepas utilizadas, adquiridas da coleção da FIOCRUZ, foram: Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus mirabilis, Serratia marscecens, Staphylococcus aureus, Enterococcus faecalis e Enterococcus faecium. No estudo, foram utilizados o caldo Mueller Hinton (MH), Extrato de Cranberry e as bactérias patogênicas. O ensaio foi realizado em triplicata, com o uso de um controle de crescimento dos micro-organismos e o experimento para avaliação do crescimento bacteriano na presença do extrato. A turbidez foi medida com o auxílio de um espectrofotômetro, no comprimento de onda de 600 nm, antes e após 24 horas de incubação à 37 ºC. O procedimento forneceu a Densidade Ótica, do qual possibilitou a identificação da inibição microbiana. Para análise estatística foi utilizado o Teste t de Student. O Extrato de Cranberry apresentou atividade antimicrobiana sobre as bactérias Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, Serratia marscecens e Enterococcus faecalis (p < 0,05), confirmando seu efeito benéfico em infecções urinárias. No entanto, não teve efeito inibitório significativo sobre Pseudomonas aeruginosa, Proteus mirabilis e Enterococcus faecium (p > 0,05).

scholarly journals Actividad antibacteriana in vitro de aceites esenciales de diferentes especies del género Citrus

2017 ◽  
Vol 46 (2) ◽  
Author(s):  
Miladys Esther Torrenegra Alarcón ◽  
Nerlis Paola Pájaro ◽  
Glicerio León Méndez
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Klebsiella Pneumoniae ◽  
Staphylococcus Epidermidis ◽  
E Coli

Se evaluó la actividad antibacteriana in vitro de aceites esenciales de diferentes especiesdel género Citrus frente a cepas ATCC de Staphylococcus aureus, Staphylococcus epidermidis,Klebsiella pneumoniae, Pseudomonas aeruginosa y Escherichia coli, determinandola concentración mínima inhibitoria (CMI) y la concentración mínima bactericida(CMB). Las bacterias se replicaron en medios de agar y caldos específicos. Se determinóel momento de máxima densidad óptica (DO620) para emplearlo como tiempode incubación; luego se hicieron pruebas de evaluación de sensibilidad con la exposiciónde las cepas a concentraciones a 1000 g/mL del extracto en caldo. Para solubilizarse empleó dimetilsulfóxido (DMSO) al 1%. Posteriormente, se le determinó laconcentración mínima inhibitoria mediante metodologías de microdilución en caldoy la concentración mínima bactericida. Encontrándose una actividad de los aceitesesenciales del género Citrus, con valores de CMI ≥ 600 mg/mL frente a S. aureus,S. epidermidis, K. pneumoniae, P. aeruginosa y E. coli. En función a los resultados obtenidos,se concluye que las diferentes especies del género Citrus son consideradas comopromisorias para el control del componente bacteriano.

scholarly journals In Vitro Activity of Newer and Conventional Antimicrobial Agents, Including Fosfomycin and Colistin, against Selected Gram-Negative Bacilli in Kuwait

Pathogens ◽  
2018 ◽  
Vol 7 (3) ◽  
pp. 75 ◽  
Author(s):  
Wadha Alfouzan ◽  
Rita Dhar ◽  
David Nicolau
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Antimicrobial Agents ◽  
The Other ◽  
In Vitro Activity ◽  
Limited Data ◽  
Gram Negative ◽  
E Coli ◽  
Microbroth Dilution

Limited data are available on susceptibilities of these organisms to some of the recently made accessible antimicrobial agents. The in vitro activities of newer antibiotics, such as, ceftolozane/tazobactam (C/T) and ceftazidime/avibactam (CZA) along with some “older” antibiotics, for example fosfomycin (FOS) and colistin (CL) were determined against selected strains (resistant to ≥ 3 antimicrobial agents) of Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Minimum inhibitory concentrations (MIC) were determined by Clinical and Laboratory Standards Institute microbroth dilution. 133 isolates: 46 E. coli, 39 K. pneumoniae, and 48 P. aeruginosa were tested. Results showed that E. coli isolates with MIC50/90, 0.5/1 μ g / mL for CL; 4/32 μ g / mL for FOS; 0.25/32 μ g / mL for C/T; 0.25/8 μ g / mL for CZA, exhibited susceptibility rates of 95.7%, 97.8%, 76.1%, and 89.1%, respectively. On the other hand, K. pneumoniae strains with MIC50/90, 0.5/1 μ g / mL for CL; 256/512 μ g / mL for FOS; 2/128 μ g / mL for C/T; 0.5/128 μ g / mL for CZA showed susceptibility rates of 92.3%, 7.7%, 51.3%, and 64.1%, respectively. P. aeruginosa isolates with MIC50/90, 1/1 μ g / mL for CL; 128/128 μ g / mL for C/T; 32/64 μ g / mL for CZA presented susceptibility rates of 97.9%, 33.3%, and 39.6%, respectively. Higher MICs were demonstrated against most of the antibiotics. However, CL retained efficacy at low MICs against most of the isolates tested.

scholarly journals Functional Analysis of PvdS, an Iron Starvation Sigma Factor of Pseudomonas aeruginosa

2000 ◽  
Vol 182 (6) ◽  
pp. 1481-1491 ◽  
Author(s):  
Livia Leoni ◽  
Nicola Orsi ◽  
Victor de Lorenzo ◽  
Paolo Visca
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Sigma Factor ◽  
Sigma Factors ◽  
Gene Copy ◽  
Wild Type ◽  
Iron Starvation ◽  
E Coli ◽  
Positive Regulator ◽  
Functional Features

ABSTRACT In Pseudomonas aeruginosa, iron modulates gene expression through a cascade of negative and positive regulatory proteins. The master regulator Fur is involved in iron-dependent repression of several genes. One of these genes, pvdS, was predicted to encode a putative sigma factor responsible for the transcription of a subset of genes of the Fur regulon. PvdS appears to belong to a structurally and functionally distinct subgroup of the extracytoplasmic function family of alternative sigma factors. Members of this subgroup, also including PbrA from Pseudomonas fluorescens, PfrI and PupI from Pseudomonas putida, and FecI from Escherichia coli, are controlled by the Fur repressor, and they activate transcription of genes for the biosynthesis or the uptake of siderophores. Evidence is provided that the PvdS protein of P. aeruginosa is endowed with biochemical properties of eubacterial sigma factors, as it spontaneously forms 1:1 complexes with the core fraction of RNA polymerase (RNAP, α2ββ′ subunits), thereby promoting in vitro binding of the PvdS-RNAP holoenzyme to the promoter region of the pvdA gene. These functional features of PvdS are consistent with the presence of structural domains predicted to be involved in core RNAP binding, promoter recognition, and open complex formation. The activity of pyoverdin biosynthetic (pvd) promoters was significantly lower in E. coli overexpressing the multicopy pvdS gene than in wild-type P. aeruginosa PAO1 carrying the single gene copy, andpvd::lacZ transcriptional fusions were silent in both pfrI (the pvdS homologue) and pfrA (a positive regulator of pseudobactin biosynthetic genes) mutants of P. putida WCS358, while they are expressed at PAO1 levels in wild-type WCS358. Moreover, the PvdS-RNAP holoenzyme purified from E. coli lacked the ability to generate in vitro transcripts from the pvdA promoter. These observations suggest that at least one additional positive regulator could be required for full activity of the PvdS-dependent transcription complex both in vivo and in vitro. This is consistent with the presence of a putative activator binding site (the iron starvation box) at variable distance from the transcription initiation sites of promoters controlled by the iron starvation sigma factors PvdS, PfrI, and PbrA of fluorescent pseudomonads.

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