Distribution of extended-spectrum beta-lactamase TEM and CTX-M resistance genes among Proteus species Isolated in Sudan

Proteus species are found in the human intestinal tract as part of normal flora. Proteus species are also found in multiple environmental habitats, including long-term care facilities and hospitals, and can cause both community and nosocomial infections. For a long time Proteus was known to be susceptible to beta-lactam antibiotics but nowadays they become resistant. The aim of this study was to detect the Extended-spectrum beta-lactamase (ESBL) TEM and CTX-M genes in 90 Proteus species isolated from urine and wound swabs, obtained from different hospitals in Khartoum state, Sudan, from January to August 2018. Antimicrobial sensitivity was carried out using the following set of antibiotics: amoxiclav, ceftazidime, gentamicin, meropenem, cefotaxime, ciprofloxacin, amoxicillin, ceftriaxone and cotrimoxazole. ESBL producing strains were detected by double disc diffusion synergy test and the resistance genes TEM and CTX-M were detected by Polymerase Chain Reaction (PCR). Antibiotic resistance was found: amoxicillin 40 percent, ceftazidime 25.6 percent, ceftriaxone 23.3 percent, gentamicin 22.2 percent, cotrimoxazole 21.1 percent, and cefotaxime 18.9 percent. Most of the isolates were sensitive to meropenem 92.2 percent and ciprofloxacin 86.7 percent. In double-disk diffusion synergy test, 20 isolates (22.2 percent) were found to be positive for ESBL. The PCR demonstrated that TEM gene was present in 18 isolates (90 percent). It was present alone in 11 isolates (55 percent) and in combination with CTX-M gene in seven isolates (35 percent). The percentage of ESBL producing strains of Proteus was 23.5 percent. This percentage is a bit lower than in previous studies in Sudan. In conclusion; it seems that the CTX-M gene is emerging among Proteus species in SudanAU)

Las especies de Proteus se encuentran en el tracto intestinal humano y forman parte de su flora normal. También se localizan en el medio ambiente y otros hábitats, incluyendo hospitales y diversas instituciones de salud, provocando tanto infecciones en la comunidad como nosocomiales. Durante mucho tiempo, las especies de Proteus fueron susceptibles a los antibióticos betalactámicos, pero actualmente se han tornado resistentes. El propósito de este estudio fue detectar genes de resistencia betalactamasas de espectro extendido (BLEE) TEM y CTX-M, en 90 especies de Proteus aisladas en orina y heridas, provenientes de diversos hospitales del estado de Jartum, Sudán, entre enero y agosto de 2018. La sensibilidad antimicrobiana se determinó con el siguiente juego de antibióticos: amoxiclav, ceftazidima, gentamicina, meropenem, cefotaxima, ciprofloxacina, amoxicilina, ceftriaxona y cotrimoxasol. Las cepas productoras de BLEE se detectaron mediante la técnica de sinergia de doble disco, y los genes de resistencia TEM y CTX-M mediante Reacción en Cadena de la Polimerasa (PCR). Se encontró resistencia antibiótica: amoxicilina 40 por ciento, ceftazidima 25,6 por ciento, ceftriaxona 23,3 por ciento, gentamicina 22,2 por ciento, cotrimoxasol 21,1 por ciento y cefotaxima 18,9 por ciento. La mayor parte de los aislamientos fueron sensibles a meropenem (92,2 por ciento) y ciprofloxacina (86,7 por ciento). Con la técnica de sinergia de doble disco se detectó positividad a BLEE en 20 aislamientos (22,2 por ciento). Mediante PCR se demostró que el gen que codifica TEM estaba presente en 18 aislamientos (90 por ciento); de forma aislada en 11 aislamientos (55 por ciento) y combinado con el gen CTX-M en los otros siete (35 por ciento). El porcentaje de cepas de Proteus productoras de BLEE fue de 23,5 por ciento. Este valor es ligeramente inferior que los detectados en estudios previos en Sudán. En conclusión, hay evidencias de que el gen CTX-M está emergiendo entre las especies de Proteus en Sudán(AU)

Urocultivos en infección clínica de vías urinarias / RESULTS OF URINE CULTURE IN PATIENTS WITH CLINICAL URINARY TRACT INFECTION

Introducción: Las infecciones de vías urinarias (IVU) constituyen un problema de salud mundial. El aumento de la resistencia bacteriana a los antimicrobianos limita la administración de antibióticos económicos y de espectro limitado, lo que afecta el costo y el acceso a la atención. El objetivo de este trabajo es determinar la sensibilidad, resistencia y germen causal en urocultivos realizados en pacientes con infección clínica de vías urinarias. Métodos: Estudio transversal. Se analizaron urocultivos de pacientes con infección clínica de vías urinarias, cada urocultivo correspondió a un paciente. Las variables fueron edad, género, microorganismo causal, resistencia y sensibilidad a los antimicrobianos. Se realizó en la Unidad de Medicina Familiar No. 222 del Instituto Mexicano del Seguro Social en Toluca Estado de México. Se evaluaron urocultivos con más de 100000 Unidades formadoras de colonias. Se realizó mediciones descriptivas, frecuencias y porcentajes en el programa SPSS v. 17 para Windows. Resultados: se incluyeron urocultivos de pacientes con infección clínica de vías urinarias. La edad promedio de los pacientes fue de 50.09 ± 19.43 años, con predominio del género femenino (211 pacientes). Los agentes causales más frecuentes fueron: Escherichia Coli (51.91%), Proteus mirabilis (14.70%) y Staphylococcus (11.11 %). Los antibióticos con mayor sensibilidad fueron: imipenem, cefotetan y meropenem (34%). Los antimicrobianos con mayor resistencia fueron: ampicilina (24%), ciprofloxacino (22%) y ampicilina con sulbactam (20%). Conclusiones: los microorganismos más frecuentemente fueron: Escherichia coli y Proteus; y los antimicrobianos a los que mostraron más resistencia bacteriana fueron: ampicilina y quinolonas.

Introduction: Urinary tract infections (UTIs) are a global health problem. Increased bacterial resistance to antimicrobials limits the administration of low-spectrum antibiotics, which affect cost and access to care. The objective of this work is to determine the sensitivity, resistance and causal germ in urine cultures in patients with clinical urinary tract infection Methods: Transversal study. Urine cultures of patients with clinical urinary tract infection were analyzed, each urine culture corresponded to one patient. The variables were age, gender, causal microorganism, resistance and sensitivity to antimicrobials. It was performed at the Family Medicine Unit No. 222 of the Mexican Institute of Social Security in Toluca State of Mexico. Urocultures were evaluated with more than 100,000 colony forming units. Measurements were made frequencies and percentages in the SPSS program version 17 for Windows. Results: there were included 558 urine cultures; the average age was 50.09 ± 19.43 years, female predominance (211 patients). The most common causative microorganisms were Escherichia coli (51.91%), Proteus mirabilis (14.70%) and Staphylococcus (11.11%). Most sensitive antibiotics were: imipenem, meropenem and cefotetan (34%). Most resistance antimicrobial were: ampicillin (24%), ciprofloxacin (22%) and ampicillin with sulbactam (20%). Conclusions: Escherichia coli and Proteus were the most commonly isolated microorganisms; Ampicillin and quinolones showed more bacterial resistence.

Reservorio venoso subcutáneo / Subcutaneous venous reservoir

Objetivo: Analizar el procedimiento, indicaciones, ventajasy las complicaciones asociadas con la instalación de losreservorios venosos subcutáneos. Diseño: Retrospectivo, longitudinal,observacional, no aleatorio. Materiales y métodos: Demarzo 2011 a junio del 2012, fueron instalados un total de 61 reservoriosvenosos subcutáneos por un sólo grupo quirúrgico, en60 pacientes consecutivos y no seleccionados. Resultados: Deltotal de pacientes 57% fueron hombres y 43% mujeres. La indicaciónde instalación del reservorio venoso fue para quimioterapiaen 98% y 2% para tratamiento del dolor. Las enfermedadesoncológicas de colon, recto y hematológicas fueron las causasmás frecuentes. No se registró mortalidad ni morbilidad intraoperatoria.Un caso presentó infección, en otro caso se constatómal funcionamiento del catéter por acodadura proximal del mismo,en ambos se retiraron los reservorios. Conclusión: la colocacióndel reservorio venoso subcutáneo es un procedimientoseguro con mínimas complicaciones y mortalidad nula, ofreceseguridad y bienestar a los pacientes para el manejo terapéuticointravenoso.

Resistencia a cefalosporinas de espectro extendido en enterobacterias sin AmpC inducible: Evaluación de los nuevos puntos de corte / Resistance to extended-spectrum cephalosporins in non-inducible AmpC enterobacteria: Evaluation of the new MIC breakpoints

Los objetivos de este estudio fueron determinar la actividad in vitro de las cefalosporinas de espectro extendido frente a aislamientos clínicos de enterobacterias sin AmpC inducible y evaluar la utilidad de las normativas propuestas por el CLSI 2009 y de los puntos de corte recomendados por el CLSI 2010 y el EUCAST 2010. El análisis incluye la caracterización feno y genotípica de los mecanismos de resistencia. En todos los aislamientos se realizó un antibiograma semicuantitativo y se determinó la CIM por dilución en agar. Asimismo, se realizó la detección fenotípica de p-lactamasas de espectro extendido (BLEE), de AmpC plasmídica (AmpCp) y de carbapenemasas de tipo KPC. En los aislamientos que fueron resistentes a las cefalosporinas de espectro extendido (CEE) se evaluó, mediante PCR múltiple para b/aSHV y b/aCTX-M y PCR con cebadores específicos, el tipo de p-lactamasa pre-valente y la presencia de KPC. Se recuperaron de pacientes 169 aislamientos resistentes a CEE: 95 de K/ebsie//a pneumoniae, 55 de Escherichia co/i y 19 de Proteus mirabi/is. La resistencia a CEE se verificó en el 56,2 %; 32,6 % y 11,2 % de estos conjuntos de aislamientos, respectivamente. Se detectó el fenotipo BLEE en 152 aislamientos (90 %), el fenotipo AmpCp en 12 (7 %) y el KPC en 5 (3 %). Las recomendaciones del CLSI 2009 y los puntos de corte del CLSI 2010 y del EUCAST 2010 para la ceftriaxona permitieron detectar eficientemente las BLEE, mientras que para la ceftacidima, con los puntos de corte del CLSI 2010 solo se detectó el 55 % de las BLEE. Esta discrepancia en los porcentajes de resistencia a ceftriaxona y a ceftacidima se relaciona con la presencia de CTX-M en nuestro medio. Los nuevos puntos de corte detectaron con mayor eficiencia las enzimas de tipo AmpCp.

The aims of this study were to evaluate the in vitro activity of extended-spectrum cephalosporins (ESC) in non-inducible AmpC enterobacteria throµgh phenotypic and genotypic characterization of the mechanisms of resistance (ESBL, plasmid-mediated AmpC and KPC) and to evaluate the interpretation criteria proposed by the existing recommendations and the new breakpoints established by the CLSI and the EUCAST. Susceptibility tests and PCR multiplex for b/aSHV and b/aCTX-M and amplification using specific primers was performed. One hundred sixty nine resistant isolates: K/ebsie//a pneumoniae (95), Escherichia co/i (55), and Proteus mirabi/is (19) were recovered. ESC resistance was 56.2 %, 32.6%, and 11.2 %, respectively. ESBL was detected in 152 (90 %) isolates, plasmid-mediated AmpC in 12 (7 %) and KPC in 5 (3 %). The CLSI 2009 recommendations and the breakpoints sµggested by the CLSI 2010 and the EUCAST for ceftriaxone were efficacious to detect ESBL, whereas the different breakpoints for ceftazidime presented discrepancies. The CLSI 2010 breakpoints only detected 55 % of the ESBL-producing isolates due to the endemic presence of CTX-M ESBLs in our country. Regarding the plasmid-mediated AmpC producers, the recommendations of the CLSI 2010 and the EUCAST 2010 proved to be more efficient than the old ones.

Isolation, identification & characterization of Proteus penneri - a missed rare pathogen.

Background & objectives: Indole negative Proteus species are invariably incorrectly identified as P. mirabilis, missing isolates of Proteus penneri. P. penneri is an invasive pathogen capable of causing major infectious diseases still seldom reported in individual cases. We report here the isolation, differentiation, characterization and typing of P. penneri from patients with different clinical infections. Methods: Urine, pus and body fluids collected from patients in intensive care units, wards and out patients departments of a tertiary health care institute from north India were cultured. A total of 61 indole negative Proteus isolates were subjected to extended biochemical tests to differentiate and identify P. penneri from P. mirabilis including failure to produce ornithine decarboxylase (by 0% strains of P. penneri and 100% strains of P. mirabilis) besides P. penneri being uniformly salicin negative, non-utilizer of citrate but ferments sucrose and maltose. Antibiograms and Dienes phenomenon were performed to characterize and type P. penneri isolates besides screening for β-lactamase production. Results: Eight isolates of P. penneri were identified; four from urine, three from abdominal drain-fluid and one from diabetic foot ulcer. P. penneri was isolated as the sole pathogen in all patients having underlying disease; post-operatively. Swarming was not seen in the first strain on primary isolation and was poor in strain-4. All eight isolates were biochemically homologous but multi-drug resistant (MDR) with resistance to 6-8 drugs (up to 12). β-lactamase production was seen in three of five isolates while Dienes phenomenon found four distinct types and discriminated strains differing in resistance even with a single drug. Interpretation & Conclusions: A few additional biochemical tests identified P. penneri isolates; it infected patients with underlying disease and strains were MDR and heterogenous.

Antimicrobial activities against biofilm formed by Proteus mirabilis isolates from wound and urinary tract infections.

Background: Bacterial species are capable of living as biofilm and/or planktonic forms. There is increasing evidence for the role of bacterial biofilm in various wound and urinary tract infections (UTIs). The aim of the present study was to evaluate the ability of the bacteria, isolated from urinary tract infections (UTIs) and wound infections, to form biofilm and correlate the role of biofilm with their antimicrobial resistance. Materials and Methods: All the isolated bacteria were screened for their ability to form biofilm using the microtitre plate method. Results: Wound isolates of Staphylococcus aureus and Enterobacter sp. had more biofilm forming capacity than the UTI isolates. Proteus mirabilis isolates were among the strongest biofilm forming bacteria and were chosen for antimicrobial study. In sub-MIC concentrations of antimicrobial agents used, ciprofloxacin was found to be the most effective in decreasing biofilm formation. On the other hand, ceftriaxone and ciprofloxacin were effective in partial removal of preformed biofilm biomass. Conclusion: Ciprofloxacin was more effective in killing bacterial cells especially at high antimicrobial concentrations that could be reached in urine levels and can be used in impregenating catheters.

C-reactive protein as an indicator of inflammatory responses to experimentally induced cystitis in dogs

The aim of this study was to demonstrate and assess C-reactive protein (CRP) changes in dogs with induced bacterial cystitis with or without antibiotics. We also evaluated availability of CRP levels to serve as an indicator for monitoring or diagnosing bacterial cystitis. Serial CRP concentrations in dogs with induced bacterial cystitis were higher than those of controls (p < 0.001). CRP concentrations peaked on day 7 and gradually decreased thereafter. In the treatment group, CRP concentrations decreased after medication compared to the untreated group (p = 0.032). CRP levels had a linear correlation with urine white blood cell counts among all groups (r = 0.837, p < 0.001, n = 140). Compared to the negative urine culture group, dogs with positive urine culture results had higher CRP concentrations (median 43.8 mg/L vs. 5.9 mg/L; p < 0.001). Area under the receiver operating characteristic curve was 0.955; when cut-off value was 12.2 mg/L, CRP measurements were found to have a sensitivity of 92.3% and specificity of 86.4%. This result indicates that rapid increases of CRP occurred after inducing bacterial cystitis and CRP may be a useful indicator for monitoring or diagnosing canine bacterial cystitis together with sediment urinalysis and urine bacterial culture.

Caracterización clínica y molecular de bacteriemias causadas por enterobacterias productoras de β -lactamasas de espectro extendido: 2004-2007 / Clinical and molecular characterization of ESBL-producing enterobacteria isolated from bacteremia in a university hospital

Introduction: Extended-spectrum-β-lactamases (ESBL) are plasmid-encoded enzymes that confer resistance to multiple antimicrobials. ESBL-producing enterobacteria that cause bacteremia limit therapeutic options and increase mortality. Objective: To perform a clinical and molecular description of bacteremia caused by ESBL-producing enterobacteria. Method: We retrospectively studied the cases of bacteremia due to ESBL-producing Escherichia coli, Klebsiella pneumoniae and Proteus spp in adults admitted to a university hospital during the years 2004-2007. We reviewed the clinical records and antimicrobial susceptibility patterns. Molecular typing was performed by polymerase chain reaction and study of clonality by pulsed-field electrophoresis. Results: We found a prevalence of 9.8 percent ESBL in enterobacteria causing bacteremia. Decreased susceptibility to quinolones and aminoglycosides was observed, without resistance to carbapenems. The predominant ESBL types were CTX-M (96 percent), TEM (62 percent) and GES (28 percent). 79 percent of the strains presented more than one type of ESBL. Clinical analysis revealed high prevalence of risk factors, previous use of antimicrobials and of invasive devices. There was no significant clonality. Conclusion: The presence of ESBLs in bloodstream infections is a clinical problem that must be considered when choosing empiric therapy.

Introducción: β-lactamasas de espectro extendido (BLEE) son enzimas plasmidiales que confieren resistencia a múltiples antimicrobianos. Las bacteriemias por enterobacterias productoras de BLEE restringen las opciones terapéuticas y aumentan la mortalidad. Objetivo: Realizar una descripción clínica y molecular de las bacteriemias causadas por enterobacterias productoras de BLEE. Método: Se estudiaron retrospectivamente los casos de bacteriemia por Escherichia coli, Klebsiella pneumoniae y Proteus spp. confirmadas para BLEE, en adultos ingresados en un hospital universitario durante los años 2004-2007. Se revisaron los registros clínicos y de susceptibilidad. Se realizó tipificación molecular por reacción de polimerasa en cadena y estudio de clonalidad por electroforesis de campo pulsado. Resultados: Se identificó una prevalencia de BLEE de 9,8 por ciento en enterobacterias causantes de bacteriemias. Se observó susceptibilidad disminuida a quinolonas y aminoglucósidos, sin resistencia a carbapenémicos. Los tipos de BLEE predominantes fueron CTX-M (96 por ciento), TEM (62 por ciento) y GES (28 por ciento). El 79 por ciento de las cepas presentó más de un tipo de BLEE. El análisis clínico reveló alta frecuencia de patologías de riesgo, uso previo de antimicrobianos y uso de dispositivos invasores. No se encontró clonalidad significativa. Conclusión: La presencia de BLEE en bacteriemias constituye un problema clínico que debe ser considerado al elegir la terapia empírica.

Epidemiology of Ciprofloxacin Resistance and Its Relationship to Extended-Spectrum beta-Lactamase Production in Proteus mirabilis Bacteremia

BACKGROUND/AIMS: We evaluated the clinical features of ciprofloxacin-resistant Proteus mirabilis bacteremia and risk factors for ciprofloxacin resistance. METHODS: From October 2000 to July 2009, 37 patients with clinically significant P. mirabilis bacteremia were identified and data from patients with ciprofloxacin-resistant and ciprofloxacin-susceptible P. mirabilis bacteremia were compared. RESULTS: The most common underlying diseases were neurologic disease (37.8%) and solid tumors (29.7%). The most common site of infection was the urinary tract (35.1%). Ten of the 37 patients (27.0%) were infected with ciprofloxacin-resistant isolates, and univariate analysis revealed a significant relationship between ciprofloxacin-resistant P. mirabilis bacteremia and neurologic disease, recent operation, L-tube insertion, percutaneous tube use, and extended-spectrum beta-lactamase (ESBL) production (all p < 0.05). ESBL was detected in six of 10 (60%) ciprofloxacin-resistant isolates, while only three of 27 (11%) ciprofloxacin-susceptible isolates produced ESBL (p = 0.005). In a logistic regression analysis, ESBL production remained a significant factor associated with ciprofloxacin resistance, after adjusting for other variables. CONCLUSIONS: These data indicate a close association between ciprofloxacin resistance and ESBL-production in P. mirabilis bacteremia. This association is particularly troublesome because the therapeutic options for serious infections caused by ESBL-producing P. mirabilis are severely restricted.

Isolation and purification of complex II from proteus mirabilis strain ATCC 29245

A respiratory complex was isolated from plasma membrane of pathogenic Proteus mirabilis strain ATCC 29245. It was identified as complex II consisting of succinate:quinone oxidoreductase (EC 1.3.5.1) containing single heme b. The complex II was purified by ion-exchange chromatography and gel filtration. The molecular weight of purified complex was 116.5 kDa and it was composed of three subunits with molecular weights of 19 kDa, 29 kDa and 68.5 kDa. The complex II contained 9.5 nmoles of cytochrome b per mg protein. Heme staining indicated that the 19 kDa subunit was cytochrome b. Its reduced form showed absorptions peaks at 557.0, 524.8 and 424.4 nm. The á-band was shifted from 557.0 nm to 556.8 nm in pyridine ferrohemochrome spectrum. The succinate: quinone oxidoreductase activity was found to be high in this microorganism.

Formação de biofilme cristalinode Proteus mirabilis em cateterurológico / Formation of crystalline biofilm of Proteus mirabilis in urological catheter

O cateter uretral de longa permanência tem papel de destaque para pacientes com retenção urinária, com obstrução debexiga, com danos neurológicos e outras doenças. A urina contém sais minerais, que em pH alcalino se precipitam,cristalizam-se e bloqueiam o cateter urológico. A cristalização dos componentes iônicos da urina ocorre em presença de urease, enzima produzida por Proteus mirabilis. Esta bactéria tem a capacidade de aderir a superfícies inanimadas e formar biofilme. O objetivo deste estudo foi observar a formação de biofilme cristalino na superfície luminal de cateter urológicode látex siliconizado, por meio de microscópio eletrônico de varredura, após a canalização de urina artificial infectada com Proteus mirabilis. O experimento foi realizado in vitro,em sistema de fluxo dinâmico. A urina artificial, composta de sais de cálcio, magnésio, fosfatos, uréia e albumina deovo, foi infectada com Proteus mirabilis ATCC 25933. O fluxo da urina canalizada foi interrompido após a cristalização dos componentes iônicos. A cristalização foi observada após a alcalinização da urina. A microscopia eletrônica de varredura demonstrou a presença de cristais e morfologiastípicas de bacilos embutidos em massa amorfa. O presente estudo mostrou que a incrustação pode limitar o uso decateter urológico de longa permanência.

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. Thecrystallization 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 artificialurine 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 afteralkalinization 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. Thepresent study showed that catheter encrustation may limit the use of long-term indwelling catheter.

Wound infection; frequency in clean surgical operations

The aim of the study was to evaluate the frequency of infection in clean surgical cases [General and Orthopaedic]. A descriptive study. This descriptive study was conducted at department of surgery and orthopaedic DHQ Teaching Hospital Sargodha from July 2007 to Dec, 2008. In this study 1500 clean surgical cases were included. Wounds were examined on third post operative day and then regularly after removal of stitches. Surgical wounds were examined finally on fifteenth post operative days. Description of wound condition and detailed data of patients were collected on preformed performas. Patients with wound infection developed pain at operation site and fever on third post operative day. Wounds were examined for swelling, redness, discharge; stitch abscess. Routine investigations were done as per protocol ie complete blood examination, complete urine examination, blood sugar, C-reactive proteins etc. Wounds swab was taken for microscopy and culture sensitivity. This study was carried out on fifteen hundred clean surgical cases [General and Orthopaedic]. There were 1064 males and 436 females. Male to Female ratio was 2.4:1. Infection was detected in 110 patients [7.3%] while no infection was found in 1390. Infection was maximum in patients more than 60 yrs of age [10.9%]. Wound infection was minimum in young patients [3.5%].commonest micro organism isolated from the infected wound was staphylococcus areus. Other organism isolated was streptococcus pyogenes, proteus and pseudomonas. No MRSA was detected. In our case study clean cases were found generally free of infection especially young patients. Whereas increased incidence of infection was noted in old patients. Wound infection is associated with significant morbidity in the form of delayed wound heeling, prolonged hospital stay and increased economical pressure on the patient

Soft tissue infections with Arcanobacterium haemolyticum: report of three cases.

We report here three polymicrobial wound infections associated with Arcanobacterium haemolyticum in rural patients aged between 60-65 years. In two patients, one with cellulitis and the other with postoperative wound infection following amputation of the limb, Arcanobacterium haemolyticum was isolated repeatedly along with beta haemolytic streptococci (BHS). The BHS belonged to Lancefield's group G and group C respectively. In another patient, who was a diabetic with chronic osteomyelitis, Arcanobacterium haemolyticum was isolated along with Proteus vulgaris . All the three isolates of Arcanobacterium haemolyticum isolated by us were uniformly resistant to cotrimoxazole and sensitive to penicillin, erythromycin, clindamycin, ciprofloxacin and gentamicin. Erythromycin alone or combined therapy of penicillin with erythromycin or penicillin with ciprofloxacin was effective in treating these infections.

Evaluation of a modified double-disc synergy test for detection of extended spectrum beta-lactamases in AMPC beta-lactamase-producing proteus mirabilis.

The detection of extended-spectrum beta-lactamases (ESBLs) in gram-negative bacteria that produce AmpC beta-lactamases is problematic. In the present study, the performance of modified double-disc synergy test (MDDST) that employs a combination of cefepime and piperacillin-tazobactam for the detection of Proteus mirabilis producing extended spectrum and AmpC beta-lactamases was evaluated and compared with double-disc synergy test (DDST) and NCCLS phenotypic disc confirmatory test (NCCLS-PDCT). A total of 90 clinical isolates of P. mirabilis , which met the CLSI (Clinical and Laboratory Standards Institute) screening criteria that these had broth microdilution (BMD) MIC of > or =2 mg/mL for at least one extended spectrum cephalosporin [ceftazidime (CAZ), cefotaxime (CTX) and cefpodoxime], were selected for the study. MDDST detected ESBLs in 40/90 of the isolates, whereas DDST detected ESBLs in only 25 isolates. NCCLS-PDCT could detect ESBLs in 39 isolates using CAZ and CAZ + clavulanic acid (CLA) combination, whereas CTX and CTX + CLA combination could detect only 37 isolates as ESBL positive. As many as 34/40 ESBL positive isolates were confirmed to be AmpC beta-lactamase positive by the modified three-dimensional test (MTDT). MDDST and NCCLS-PDCT could detect ESBLs in all the 34 AmpC positive isolates, whereas DDST could detect ESBLs in only 19 isolates. The study demonstrated that MDDST is superior to DDST and as sensitive as NCCLS-PDCT. However, MDDST seems to have enhanced potential for the detection of ESBLs in AmpC beta-lactamase-producing P. mirabilis .

Prevalência de microrganismos em infecções do trato urinário de pacientes atendidos no laboratório de análises clínicas da Universidade Paranaense - Umuarama - PR / Prevalence of the microorganisms in urinary tract infections of patients attended in the clinical analysis of the Paranaense University - Umuarama - PR

Infecção urinária é a presença de microorganismo na urina, e sua prevalência varia com o sexo e idade dos pacientes. O presente trabalho objetivou determinar a prevalência de infecção urinaria no laboratório de análises clinicas da UNIPAR no municípiode Umuarama-Pr. Foram analisados 328 prontuários dos pacientes que realizaram exames de cultura de urina no ano de 2005. Durante este período foram identificados 52 casos de infecção urinária e foram encontrados os seguintes agentes etiológicos: Escherichia colii (36,55%), Enterobacter sp. (21,16%), Klebsiella sp (17,30%), Estaphylococcus sp., (7,70%), Citrobacter sp. (5,76%), Pseudomonas aeruginosa (5,76%) Burkholderia cepacia (3,85%) Proteus sp. (1,92%). A maior prevalência de infecção urinária encontrado no presenteestudo foi do gênero feminino com uma prevalência de 14,6%. A Escherichia coli foi o microorganismo prevalente causador desta infecção. Os resultados obtidos neste estudo são de muita valia, pois permitem aplicação de um tratamento mais adequado, evitandodesta forma complicações e recidivas.

Urinary infection is the presence of microorganism in piss, e its prevalence varies with the sex and age of the patients. The present work objectified to determine the prevalence of infection would urinary in the laboratory of clinical analyses of the Unipar in the city of Umuarama-Pr. Had been analyzed 328 handbooks of the patients who had carried through examinations of piss culture in the year of 2005. During this period 52 cases of urinary infection had been identified and had been found the following agents etiologic: Escherichia colli (36.55%), Enterobacter sp. (21,16%), Klebsiella sp. (17.30%), Estaphylococcus sp., (7,70%), Citrobacter sp. (5,76%), Pseudomonas aeruginosa (5.76%) Burkholderia cepacia (3.85%) Proteus sp. (1,92%). The biggest prevalence of found urinaryinfection in the present study was of the feminine sort with a prevalence of 14,6%. The Escherichia coli were the .causing prevalent microorganism of this infection. The results gotten in this study are of much value, therefore they more allow application of an adjustedtreatment, preventing in such a way complications and returns.

Prevalência e perfil de sensibilidade de microrganismos em infecções do trato urinário / Prevalence and antimicrobial susceptibility in urinary tract pathogens

Infecções bacterianas do trato urinário são um problema muito comum tanto em infecções nosocomiais quanto na comunidade. O objetivo deste estudo foi determinar a prevalência de patógenos responsáveis por infecções no trato urinário, relacionando-os com a sua susceptibilidade frente aos antimicrobianos utilizados. Durante o período de janeiro de 1998 a dezembro de 2001 foram analisados resultados de 2378 culturas de urinas positivas de pacientes ambulatoriais, sem restrições de sexo e idade, de Porto Alegre, Rio Grande do Sul, Brasil. Escherichia coli foi o microrganismo isolado mais prevalente: 74,3 porcento, seguido de Proteus mirabilis com 8,0 porcento e Enterobacter aerogenes com 4,88 porcento. Nitrofurantoína (NIT) apresentou altos índices de atividade frente ao Cocos Gram-positivos, enquanto que o Norfloxacin foi o antimicrobiano que apresentou a maior atividade frente aos Bacilos Gram-negativos.

Studies of resistance pattern of bacteria related with urinary tract infections

Six different species of bacteria isolated in previous study from two hundred cases of UTIs were subjected to determine the resistance pattern against seven different groups of antimicrobial agents. The front line antibiotics for treating urinary tract infections due to Escherichia coli, Klebsiella species, Staphyloccus aureus and Proteus species should include Cephradine, Ofloxacin, Cefaclor, Cephalothin and Pipemidic acid. However for treating UTI due to pseudomonas species, the drug of choice must be a member of fluoroquinoline group [norfloxacin, ofloxacin and ciprofloxacin]

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.