ZapA, a possible virulence factor from Proteus mirabilis exhibits broad protease substrate specificity

The opportunistic bacterium Proteus mirabilis secretes a metalloprotease, ZapA, considered to be one of its virulence factors due to its IgA-degrading activity. However, the substrate specificity of this enzyme has not yet been fully characterized. In the present study we used fluorescent peptides derived from bioactive peptides and the oxidized ß-chain of insulin to determine the enzyme specificity. The bradykinin- and dynorphin-derived peptides were cleaved at the single bonds Phe-Ser and Phe-Leu, with catalytic efficiencies of 291 and 13 mM/s, respectively. Besides confirming already published cleavage sites, a novel cleavage site was determined for the ß-chain of insulin (Val-Asn). Both the natural and the recombinant enzyme displayed the same broad specificity, demonstrated by the presence of hydrophobic, hydrophilic, charged and uncharged amino acid residues at the scissile bonds. Native IgA, however, was resistant to hydrolysis by ZapA.

Development of an operational substrate for ZapA, a metalloprotease secreted by the bacterium Proteus mirabilis

The protease ZapA, secreted by Proteus mirabilis, has been considered to be a virulence factor of this opportunistic bacterium. The control of its expression requires the use of an appropriate methodology, which until now has not been developed. The present study focused on the replacement of azocasein with fluorogenic substrates, and on the definition of enzyme specificity. Eight fluorogenic substrates were tested, and the peptide Abz-Ala-Phe-Arg-Ser-Ala-Ala-Gln-EDDnp was found to be the most convenient for use as an operational substrate for ZapA. A single peptide bond (Arg-Ser) was cleaved with a Km of 4.6 µM, a k cat of 1.73 s-1, and a catalytic efficiency of 376 (mM s)-1. Another good substrate for ZapA was peptide 6 (Abz-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-Gln-EDDnp) which was cleaved at a single bond (Phe-Ser) with a Km of 13.6 µM, a k cat of 3.96 s-1 and a catalytic efficiency of 291 (mM s)-1. The properties of the amino acids flanking the scissile bonds were also evaluated, and no clear requirement for the amino acid residue at P1 was found, although the enzyme seems to have a preference for a hydrophobic residue at P2.

Formação de biofilme cristalino de Proteus mirabilis em cateter urológico

The indwelling urethral catheter has an important role for patients with urinary retention, bladder obstruction, neurological damage and other diseases. Urine contains minerals which precipitate in alkaline pH, crystallize and block the urological catheter. The crystallization of the ionic components of urine occurs in the presence of urease, an enzyme produced by Proteus mirabilis. This bacterium adheres to inanimate surfaces and forms biofilms. The aim of this study was to investigate the formation of crystalline biofilm on the luminal surface of siliconized latex catheters by means of scanning electron microscope, after channeling artificial urine infected with Proteus mirabilis. The experiment was performed in vitro using a dynamic flow system. The artificial urine compounds were salts of calcium, magnesium, phosphates, urea and egg albumin, and it was infected with Proteus mirabilis ATCC 25933. The urine flow was stopped after crystallization of the ionic components. Crystallization was observed after alkalinization of urine. Scanning electron microscopy showed the presence of crystals and morphologies typical of bacilli embedded in an amorphous mass on the internal lumen of the catheter. The present study showed that catheter encrustation may limit the use of long-term indwelling catheter. © SBEB - Sociedade Brasileira de Engenharia Biomédica.

Possíveis ressonâncias nos sistemas de Marte-Phobos e Netuno-Triton-Proteus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação de biofilme de proteus mirabilis em modelo experimental de fluxo dinâmico

Pós-graduação em Biociências e Biotecnologia Aplicadas à Farmácia - FCFAR

Palmistichus elaeisis (Delvare & LaSalle, 1993) (Hymenoptera: Eulophidae), possible agent for the biological control of Urbanus proteus (Linnaeus, 1758) (Lepidoptera: Hesperiidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Orquiepididimite causada por Corynebacterium sp e Proteus mirabilis em cordeiro da raça Lacaune: relato de caso

Orchiepididymitis is the inflammation of both, testis and epididymis and may be caused by microbial agents such as Brucella ovis. Thus, the aim of the present report is to present a case of orchiepididymitis in a Lacaune ram with unilateral testicular enlargement. Ultrasonography of the testis revealed the presence of a circular hypoechogenic structure adjacent to the testicular parenchyma. Corynebacterium sp was isolated from the surgically removed testis. Two months after removal of the affected testis, the animal returned to the Veterinary Hospital due to an enlargement of the remaining testis. A new ultrasonographic exam revealed the presence of abscesses in the testicular parenchyma and periorchitis. After orchiectomy, Proteus mirabilis was isolated from the material.

Índice de resistência múltipla aos antimicrobianos, concentração inibitória e beta-lactamases de espectro estendido em linhagens de Proteus mirabilis e Proteus vulgaris isoladas de diferentes afecções em animais domésticos

Pós-graduação em Medicina Veterinária - FMVZ

Atividade antibacteriana do óleo essencial de Melampodium divaricatum (Rich.) DC., Asteraceae

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Comparison of wildlife and captivity rattlesnakes (Crotalus durissus terrificus) microbiota

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Occurrence of yeasts, pseudomonads and enteric bacteria in the oral cavity of patients undergoing head and neck radiotherapy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evaluation of the Diramic system for urine cultures

O sistema Diramic foi avaliado para o diagnóstico das infecções do trato urinário (ITU). O sistema Diramic foi desenvolvido em Cuba e possibilita resultados de diagnóstico das infecções do trato urinário (ITU) em quatro horas e baseia-se na variação da turvação do crescimento microbiano no meio de cultura após incubação a 37ºC/4 horas. 396 amostras de urinas provenientes de ambulatórios e enfermarias do HC da FMB-UNESP-Botucatu/SP foram analisadas pelo sistema Diramic. O método da alça calibrada (AC) foi adotado como método de referência. A taxa de coincidência entre os dois métodos foi de 96,46% (382 amostras de urina), não havendo diferença significativa entre os resultados obtidos nos dois métodos. Os resultados para sensibilidade e especificidade foram 84,37 e 98,80% respectivamente e 10 resultados no Diramic foram falsos negativos (2,5%) e 4 falso positivos (1,01%). Os microrganismos identificados nas urinas positivas foram Escherichia coli (68,75%), Klebsiella pneumoniae (10,94%), leveduras (6,25%), Pseudomonas aeruginosa (4,69%), Enterobacter cloacae (3,12%) e Proteus mirabilis, Staphyloccocus coagulase negativo, Morganella morganii e Citrobacter freundii também foram identificadas (1,56% para cada espécie). O método Diramic foi eficiente na triagem das urinoculturas, porém verificou-se algumas restrições quanto ao diagnóstico das infecções do trato urinário quando causadas por leveduras e em pacientes submetidos a antibioticoterapia.

Pulmonary, microbiological and hematological changes in Crotalus durissus terrificus (Serpentes, Viperidae) parasitized by nematodes of the genus Rhabdias (Nematoda, Rhabdiasidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Microbiological analyses of poultry litter used for ruminant feeding

The aims of this study were to estimate the changes in total bacterial counts (TBC) in poultry litter samples, consisting of rice hulls, after storage, and to identify pathogenic bacteria. For the countings Plate Count agar (Difco) was used. Enrichment and selective media such as blood agar, MacConkey, Baird Parker, brain and heart agar, and egg yolk solid media, and cooked meat and brain and heart infusion, incubated under aerobic or anaerobic conditions were used to isolate Staphylococcus aureus, Salmonella sp, Clostridium perfringens, C. botulinum, C. chauvoei, Campylobacter sp, Escherichia coli, and Corynebacterium sp. Litter samples were collected from the houses of the Veterinary School experimental aviary. A fully randomized experimental design was used with four treatments and four replications, for a total of 16 samples. A decrease in TBC was detected when treatment T1 (zero days of storage) was compared with treatments T2 (14 days of storage). on the other hand the treatments T3 (28 days of storage) and T4 (42 days of storage) presented significantly superior counting in relation to treatment T1. Some pathogenic bacteria of Enterobacteriaceae such as Escherichia coil, Proteus, Arizona, Providencia, Edwardsiella, as well as Staphylococcus aureus, S. epidermidis, different species of genus Clostridium as C. perfringens, C. sordelli, C. chauvoei, C. tetani and C. novyi as well as some strains of Corynebacterium pyogenes were isolated.

Chemical structure, surface properties and biological activities of the biosurfactant produced by Pseudomonas aeruginosa LBI from soapstock

Pseudomonas aeruginosa LBI isolated from petroleum-contaminated soil produced rhamnolipids (RLLBI) when cultivated on soapstock as the sole carbon source. HPLC-MS analysis of the purified culture supernatant identified 6 RL homologues (%): R-2 C-10 C-10 28.9; R-2 C-10 C-12:1 23.0; R-1 C-10 C-10 23.4; R-2 C-10 C-12 11.3; R-2 C-10 C-12 7.9; R-2 C-10 C-12 C-12 5.5. To assess the potential antimicrobial activity of the new rhamnolipid product, RLLBI, its physicochemical properties were studied. RLLBI had a surface tension of 24 mN m(-1) and an interfacial tension 1.31 mN m(-1); the cmc was 120 mg l(-1). RLLBI produced stable emulsions with hydrocarbons and vegetable oils. This product showed good antimicrobial behaviour against bacteria: MIC for Bacillus subtilis, Staphylococcus aureus and Proteus vulgaris was 8 mg l(-1), for Streptococcus faecalis 4 mg l(-1), and for Pseudomonas aeruginosa 32 mg l(-1). RLLBI was active against phytopathogenic fungal species, MIC values of 32 mg l(-1) being found against Penicillium, Alternaria, Gliocadium virens and Chaetonium globosum. Due to its physicochemical properties and antimicrobial behaviour, RLLBI could be used in bioremediation treatment and in the food, cosmetic and pharmaceutical industries.