The role of the genetic elements bla oxa and IS Aba 1 in the Acinetobacter calcoaceticus-Acinetobacter baumannii complex in carbapenem resistance in the hospital setting

Abstract: INTRODUCTION: Members of the Acinetobacter genus are key pathogens that cause healthcare-associated infections, and they tend to spread and develop new antibiotic resistance mechanisms. Oxacillinases are primarily responsible for resistance to carbapenem antibiotics. Higher rates of carbapenem hydrolysis might be ascribed to insertion sequences, such as the ISAba1 sequence, near bla OXA genes. The present study examined the occurrence of the genetic elements bla OXA and ISAba1 and their relationship with susceptibility to carbapenems in clinical isolates of the Acinetobacter calcoaceticus-Acinetobacter baumannii complex. METHODS: Isolates identified over 6 consecutive years in a general hospital in Joinville, Southern Brazil, were evaluated. The investigation of 5 families of genes encoding oxacillinases and the ISAba1 sequence location relative to bla OXA genes was conducted using polymerase chain reaction. RESULTS: All isolates presented the bla OXA-51-like gene (n = 78), and 91% tested positive for the bla OXA-23-like gene (n = 71). The presence of ISAba1 was exclusively detected in isolates carrying the bla OXA-23-like gene. All isolates in which ISAba1 was found upstream of the bla OXA-23-like gene (n = 69) showed resistance to carbapenems, whereas the only isolate in which ISAba1 was not located near the bla OXA-23-like gene was susceptible to carbapenems. The ISAba1 sequence position of another bla OXA-23-like-positive isolate was inconclusive. The isolates exclusively carrying the bla OXA-51-like gene (n = 7) showed susceptibility to carbapenems. CONCLUSIONS: The presence of the ISAba1 sequence upstream of the bla OXA-23-like gene was strongly associated with carbapenem resistance in isolates of the A. calcoaceticus-A. baumannii complex in the hospital center studied.

Doripenem vs meropenem against Pseudomonas and Acinetobacter.

Recently, doripenem has been approved for the treatment of nosocomial pneumonia (NP), including ventilator-associated pneumonia (VAP). The E-test was performed to determine the MICs of doripenem and meropenem in 203 endotracheal aspirate isolates that consisted of 140 Acinetobacter calcoaceticus-Acinetobacter baumannii complexes and 63 Pseudomonas aeruginosa. Doripenem showed minimum concentration necessary for inhibition of 50% (MIC 50 ) of P. aeruginosa isolates at 0.38 mg/L which is several times (84.2 times) lower than the corresponding MIC 50 value of >32 mg/L for meropenem. The MIC 50 and MIC 90 were similar for both the drugs against A. baumannii. Thus, P. aeruginosa was consistently more susceptible than the A. baumannii.

Screening and characterization of a novel alkaline lipase from Acinetobacter calcoaceticus 1-7 isolated from Bohai Bay in China for detergent formulation

A novel alkaline lipase-producing strain 1-7 identified as Acinetobacter calcoaceticus was isolated from soil samples collected from Bohai Bay, China, using an olive oil alkaline plate, which contained olive oil as the sole carbon source. The lipase from strain 1-7 showed the maximum activity at pH 9.0 under 40ºC. One interesting feature of this enzyme is that it exhibits lipase activity over a broad range of temperatures and good stability. It is also stable at a broad range of pHs from 4.0 to 10.0 for 24 h. Its catalytic activity was highly enhanced in the presence of Ca2+, Mg2+ and K+, but partially inhibited by Cu2+, Al3+, Fe3+ , Ba2+and Zn2+. The fact that it displays marked stability and activity in the presence of TritonX-100, Tween-20, Tween-80, SDS, Hydrogen peroxide, Sodium perborate, Sodium hypochlorite, Sodium citrate, Sodium taurocholate, Glycerine and NaCl suggests that this lipase is suitable as an additive in detergent formulations.

Sensibilidad de Acinetobacter a la colistina evaluada mediante los métodos de predifusión y de concentración inhibitoria mínima: Detección de aislamientos heterorresistentes / Comparative evaluation of the sensitivity of Acinetobacter to colistin, using the prediffusion and minimum inhibitory concentration methods: Detection of heteroresistant isolates

El objetivo de este estudio fue evaluar comparativamente los métodos de predifusión y de concentración inhibitoria mínima para establecer la sensibilidad de aislamientos del complejo Acinetobacter calcoaceticus-baumannii (ABC) a la colistina y detectar a aquellos que presenten heterorresistencia a dicho antibiótico. Se estudiaron 75 aislamientos de ABC recuperados de materiales clínicamente significativos. Se determinó su sensibilidad a la colistina por el método de predifusión y de concentración inhibitoria mínima. Todos los aislamientos resultaron sensibles, con CIM = 2 µg/ml y halos de inhibición en el ensayo de la predifusión = 20 mm. Mediante el método de eficiencia de plaqueo se evaluó la presencia de heterorresistencia a la colistina. Se encontraron 14 aislamientos que originaron colonias heterorresistentes; sus CIM aumentaron en algunos casos en más de 8 veces. Con estas colonias seleccionadas se repitió el ensayo de predifusión. Finalmente se confeccionaron los gráficos de dispersión y se realizaron los análisis de regresión lineal, tanto para el conjunto inicial de todos los aislamientos clínicos como para el subgrupo de los aislamientos resistentes generados durante la evaluación de la heterorresistencia. Se obtuvieron coeficientes de determinación (r²) de 0,2017 y 0,604, respectivamente, lo que indica correlación entre los métodos sólo al evaluar aislamientos preseleccionados por su resistencia a este agente.

The objective of this study is to perform a comparative evaluation of the prediffusion and minimum inhibitory concentration (MIC) methods for the detection of sensitivity to colistin, and to detect Acinetobacter baumanii-calcoaceticus complex (ABC) heteroresistant isolates to colistin. We studied 75 isolates of ABC recovered from clinically significant samples obtained from various centers. Sensitivity to colistin was determined by prediffusion as well as by MIC. All the isolates were sensitive to colistin, with MIC = 2µg/ml. The results were analyzed by dispersion graph and linear regression analysis, revealing that the prediffusion method did not correlate with the MIC values for isolates sensitive to colistin (r² = 0.2017). Detection of heteroresistance to colistin was determined by plaque efficiency of all the isolates with the same initial MICs of 2, 1, and 0.5 µg/ml, which resulted in 14 of them with a greater than 8-fold increase in the MIC in some cases. When the sensitivity of these resistant colonies was determined by prediffusion, the resulting dispersion graph and linear regression analysis yielded an r² = 0.604, which revealed a correlation between the methodologies used.

Caracterização molecular e fenotípica de amostras bacterianas pertencentes ao complexo Acinetobacter calcoaceticus-Acinetobacter baumannii / Molecular and phenotypic characterization of Acinetobacter calcoaceticus-Acinetobacter baumannii isolates

Nos últimos 30 anos, Acinetobacter tornou-se um dos patógenos de maior preocupação clínica pela falta de terapias eficazes em virtude do fenótipo de multirresistência frequentemente apresentado. Dentre as espécies do gênero Acinetobacter, A. baumannii, A. genoespécie 3 e A. genoespécie 13TU são as mais comumente encontradas a partir de amostras biológicas. Estas espécies ao lado de A. calcoaceticus constituem o complexo A. calcoaceticus-A. baumannii (ACB). Este estudo propõe um esquema composto de duas PCRs para a identificação das espécies de interesse médico que fazem parte do complexo ACB. O método é simples, rápido e, além de identificar as espécies, permite pesquisar a presença de genes de resistência. Foram identificadas 515 amostras do complexo ACB, isoladas de pacientes no período de janeiro de 2005 a dezembro de 2010. A identificação das espécies do complexo ACB foi realizada por esquema composto de duas reações de PCR. Foram avaliados os perfis de sensibilidade por disco difusão e a pesquisa da presença dos genes blaOXA-23-like, blaOXA-24-like, blaOXA-51-like, blaOXA-58-like, blaIMP, blaVIM, blaSIM, blaSPM e blaGIM foi realizada por PCR utilizando-se iniciadores específicos. No grupo de amostras estudas, 82,5% são A. baumannii (425), 11,5% A. genoespécie 13TU (59) e 6,0% A. genoespécie 3 (30), sendo A. baumannii mais isolado em pacientes internados em UTIs (p=0,0407) e A. genoespécie 13TU mais isolado em pacientes de outros ambientes hospitalares (p=0,0204). A. baumannii apresentou menor sensibilidade a todos os antimicrobianos quando comparado com A. genoespécie 13TU e A. genoespécie. 3 (p<0,05). Foi possível observar ao longo do período estudado o aumento significativo da resistência aos carbapenêmicos e da sensibilidade a gentamicina por A. baumannii entre os isolados de pacientes de UTIs (p<0.05). Nenhum dos genes codificadores para metalo-lactamases foi detectado nas amostras estudadas Dentre os cepas resistentes aos carbapenêmico...

The genus Acinetobacter has emerged as one of the most troublesome pathogens for health care institutions globally. Its clinical significance, especially over the last 15 years, has been driven by its remarkable ability to up regulate or acquire resistance determinants, making it one of the organisms threatening the current antibiotic era. A. baumannii, A. 3 and A. 13TU are the most commonly species found from biological samples. These species beside A. calcoaceticus are very closely related and difficult to distinguish from each other by phenotypic properties. Therefore, it has been proposed to refer to these species as the A.calcoaceticus-A. baumannii complex(ACB). In the period from 2005 to 2009, the most frequent bacterial isolates among the nosocomial infection at the HU-USP was ACB (18%). Due to the frequency with which species are involved in ACB outbreaks of infection in the HU-USP and the emergency clinic because of expression of the phenotype of resistance to several classes of antibiotics, this study aimed to identify and characterize the species of complex ACB by molecular methods, to study their mechanisms of resistance and to characterize the different clones from patients admitted to different hospital areas. Furthermore, the ability to characterize biofilm formation, adhesion to different cell lines as well as the mechanisms of cell-cell communication were analyzed. From the ACB complex, 515 samples were identified, isolated from patients from January 2005 to December 2010. The identification of clinical species of the ACB was performed by molecular methods that were developed and validated for identification of Acinetobacter sp. include two reactions of PCR. The profiles of sensibility were evaluated by disc diffusion and the detection of the presence of genes blaOXA-23-like, blaOXA-24-like, blaOXA-51-like, blaOXA-58-like, blaIMP, blaVIM, blaSIM, blaGIM, and blaSPM were performed using specific primers. Molecular typing was performed using...

Endophthalmitis caused by Acinetobacter calcoaceticus. A profile.

PURPOSE: To report the clinical and microbiological profile of endophthalmitis caused by Acinetobacter calcoaceticus. METHODS: A retrospective study of case series of Acinetobacter calcoaceticus endophthalmitis. Outcome measures included ability to sterilise the eye, anatomical result (clear media and attached retina) and visual recovery (visual acuity > 6/60). RESULTS: Of the 20 cases studied, 10 were cases of postoperative endophthalmitis, 3 were posttraumatic, 6 were endogenous and one was bleb-related endophthalmitis. Specific features of interest observed were relative chronicity of presentation and absence of any obvious predisposing factor in endogenous endophthalmitis cases. All cases could be sterilised except one, which needed evisceration. Cases with postoperative endophthalmitis had better anatomical outcome (7/10 with attached retina and clear media) and visual outcome (4/10 regained vision > 6/18). Higher smear positivity was seen in vitreous samples (72.2%) compared to aqueous samples (37.5%). Culture positivity was higher from the vitreous cavity compared to aqueous. The organism was sensitive to ciprofloxacin in a high percentage (88.9%) of cases. CONCLUSIONS: Visual recovery in Acinetobacter calcoaceticus endophthalmitis is modest. Ciprofloxacin is the antibiotic of choice.

Characterization of nonfermenters from clinical samples.

OBJECTIVE: Nonfermenters are a group of aerobic non sporing gram-negative bacilli found primarily free in nature and as commensals, whose pathogenic potentials are well established. The current study was conducted to assess the role of these nonfermenters in various infections and to characterize these isolates. METHODS: One hundred nonfermenters isolated from various clinical specimens were grouped according to Weaver-Hollis scheme based on growth on MacConkeys agar, oxidase activity and oxidation/fermentation of glucose. Species level identification was attempted based on a battery of biochemical tests. All isolates were then subjected to antimicrobial sensitivity. RESULTS: Majority of the isolates were encountered from pus and urine (50%). These isolates belonged to six of the seven Weaver-Hollis groups. Fifty six per cent of the isolates belonged to genus Pseudomonas. Multidrug resistance with resistance to more than three antimicrobials was frequently seen. Amikacin and ciprofloxacin were found to be most effective. CONCLUSION: Nonfermenting gram negative organisms are responsible for variety of infective conditions. Amongst them genus Pseudomonas and Acinetobacter calcoaceticus were more frequently encountered. Amikacin or ciprofloxacin (for nonfermenters other than Pseudomonas) appears to be the drug of choice for treatment of such infections.

Identificación y caracterización de bacilos gramnegativos no fermentados aislados en el medio hospitalario / Identification and characterization of nonfermenting gramnegative bacili isolated from a hospital environment

Se estudio un total de 251 cepas aisladas en el medio hospitalario con diagnóstico presuntivo de bacilos gramnegativos no fermentadores, enviadas por los Laboratorios de Infecciones Nosocomiales de los Centros Provinciales de Higiene y Epidemiología de Ciudad de La Habana, Santiago de Cuba, Holguín, Guantánamo, Camagüey e Isla de la Juventud desde septiembre de 1992 hasta junio de 1993. Se empleó un esquema inicial para corroborar el diagnóstico primario de estas cepas, los medios de producción de pigmentos King A y King B para la identificación de Pseudomonas aeruginosas y pruebas bioquímicas claves para el diagnóstico microbiológico de otros no fermentadores, se utilizó la piocinotipia y la serotipia para la caracterización posterior de la especie aeruginosa. Resultaron confirmadas 238 cepas y de ellas, el 88,23 por ciento correspondió a la especie aeruginosa cuyos piocinotipo y serotipo predominantes fueron al 10a y 011, respectivamente. Otros bacilos encontrados con frecuencia fueron Pseudomonas cepacia (25 por ciento); Pseudomonas fluorescens (17,8 por ciento) y Acinetobacter calcoaceticus var. anitratus 17,8 por ciento