In vitro antibacterial activity of sulbactam-durlobactam in combination with other antimicrobial agents against Acinetobacter baumannii-calcoaceticus complex.

Combinations of the ß-lactam/ß-lactamase inhibitor sulbactam-durlobactam and seventeen antimicrobial agents were tested against strains of Acinetobacter baumannii in checkerboard assays. Most combinations resulted in indifference with no instances of antagonism. These results suggest sulbactam-durlobactam antibacterial activity against A. baumannii is unlikely to be affected if co-dosed with other antimicrobial agents.

Characterization of Acinetobacter baumannii-calcoaceticus complex isolates and microbiological outcome for patients treated with sulbactam-durlobactam in a phase 3 trial (ATTACK).

Acinetobacter baumannii-calcoaceticus complex (ABC) causes severe, difficult-to-treat infections that are frequently antibiotic resistant. Sulbactam-durlobactam (SUL-DUR) is a targeted ß-lactam/ß-lactamase inhibitor combination antibiotic designed to treat ABC infections, including those caused by multidrug-resistant strains. In a global, pathogen-specific, randomized, controlled phase 3 trial (ATTACK), the efficacy and safety of SUL-DUR were compared to colistin, both dosed with imipenem-cilastatin as background therapy, in patients with serious infections caused by carbapenem-resistant ABC. Results from ATTACK showed that SUL-DUR met the criteria for non-inferiority to colistin for the primary efficacy endpoint of 28-day all-cause mortality with improved clinical and microbiological outcomes compared to colistin. This report describes the characterization of the baseline ABC isolates from patients enrolled in ATTACK, including an analysis of the correlation of microbiological outcomes with SUL-DUR MIC values and the molecular drivers of SUL-DUR resistance.

Acinetobacter baumannii-calcoaceticus Complex-associated Orbital Cellulitis: A Case Report and Literature Review.

Acinetobacter baumannii-calcoaceticus complex is emerging as one of the most common causes of hospital-acquired infection globally. We present a case of orbital cellulitis caused by the A. baumannii-calcoaceticus complex. A 73-year-old Chinese woman with a history of rheumatoid arthritis presented with subacute onset of right upper eyelid swelling for 1 week. Computer tomography revealed a post-septal soft tissue lesion located at the right superior orbit that was enhanced with contrast, compressing on the superior aspect of the globe. Anterior orbitotomy with incisional biopsy of the right superior orbital lesion was performed, and histopathological examination was consistent with nonspecific inflammatory mass. The microbiological culture of the specimen yielded A. calcoaceticus and A. baumannii complex, which was sensitive to ciprofloxacin, ampicillin, and gentamicin. The infection resolved after a 1-week course of intravenous augmentin. Ophthalmologists should be alert to the possibility of patients having A. baumannii and A. calcoaceticus in periorbital cellulitis.

Cadmium and antibiotic-resistant Acinetobacter calcoaceticus strain STP14 reported from sewage treatment plant.

A bacterium designated as strain STP14 was isolated from a sewage treatment plant and identified as Acinetobacter calcoaceticus based on 16S ribosomal RNA gene sequencing. Strain STP14 exhibited resistance to several metals such as mercury, cobalt, copper, nickel, lead, and cadmium. Among these metals, the bacterium showed maximum resistance to cadmium in concentration up to 1200 mg/L. The antimicrobial susceptibility test of A. calcoaceticus strain STP14 showed coresistance to all tested antibiotics except tigecycline and chloramphenicol for which 16 ± 1- and 15 ± 1-mm zone of inhibition was observed, respectively. The protein pattern of the crude cellular extract revealed substantial differences in protein bands of untreated control and cadmium treated A. calcoaceticus strain STP14 suggesting variable protein expression under cadmium stress. Metals and antibiotic resistance are increasing phenomenon and universal concern of public health. This study improves our understanding regarding the bacterial coresistance against metals and antibiotics and the possible emergence of multidrug resistance due to selective pressure and coselection in the metal polluted sewage sludge.

Sepsis from an apheresis platelet contaminated with Acinetobacter calcoaceticus/baumannii complex bacteria and Staphylococcus saprophyticus after pathogen reduction.

BACKGROUND: Strategies to reduce platelet (PLT) bacterial contamination include donor screening, skin disinfection, sample diversion, bacterial culture, pathogen reduction (PR), and day-of-transfusion tests. We report bacterial sepsis following a pathogen-reduced PLT transfusion. CASE REPORT: An adult male with relapsed acute lymphoblastic leukemia was successfully treated for central catheter-associated Staphylococcus aureus bacteremia. A peripherally inserted central catheter (PICC) was placed. Chills, rigors, and flushing developed immediately after PICC-infused pathogen-reduced PLTs, progressing to septic shock requiring intensive care management. METHODS: PICC and peripheral blood (PB), transfused bag saline flushes (TBFs), environmental samples, and the pathogen-reduced untransfused co-component (CC) were cultured. Plasma metagenomic and bacterial isolate whole-genome sequencing; PLT mitochondrial DNA (mtDNA) testing of untransfused CC and TBF; CC testing for amotosalen (S-59)/S-59 photoproducts; isolate PR studies (INTERCEPT); and TBF polymerase chain reaction for recipient Y-chromosome DNA were performed. RESULTS: PB and PICC cultures grew Acinetobacter calcoaceticus/baumannii complex (ACBC). TBF was gram-positive; mass spectrometry identified ACBC and Staphylococcus saprophyticus (SS). CC Gram stain and cultures were negative. Environmental cultures, some done after decontamination, were ACBC/SS negative. Posttransfusion patient plasma and TBF ACBC sequences were genetically identical. No Y-chromosome signal was detected in TBF. S-59 photoproducts and evidence of mtDNA amplification inhibition were found in the CC. Spiking PR studies showed >5.9-log inactivation for both isolates. Donor skin cultures for Acinetobacter were negative. CONCLUSION: CC sterility, PR studies, residual S-59 photoproducts, and mtDNA amplification inhibition suggest successful PR. Unidentified environmental sources and inherent or acquired bag defects may have contributed to postmanufacturing pathogen-reduced PLT contamination.

Enzyme cocktail containing NADH regeneration system for efficient bioremediation of oil sludge contamination.

Oil sludge is one kind of toxic and persistent contamination to ecology system from petroleum industry. In order to recycle contaminated sands and reduce environmental impacts at a lower operating cost, enzyme cocktail 21/CbFDH including NADH regeneration system for oily sludge bioremediation was constructed for the first time. The intracellular enzymes of oil-degrading strain Acinetobacter calcoaceticus 21 were prepared and the formate dehydrogenase gene Cbfdh from Candida boidinii was cloned and functionally expressed in E.coli BL21 induced by lactose. The activity and stability of CbFDH was enhanced through self-induction medium optimization using Box-Behnken design. The CbFDH activity was 12.2 times increased and was only decreased 3.9% upon storage at 30 °C for 5 d. The CbFDH increased the degradation rate of oil in high concentration. For the sludge with 10% oil (w/w), the degradation rate achieved 35.6% after 12 h using enzyme 21/CbFDH with the protein ratio of 1:4. The results will provide novel perspectives for creation and operation of petroleum-degrading enzymes involving formate dehydrogenase with higher efficiency and lower cost comparing to current microbial strains or consortium.

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.

An endophytic bacterium Acinetobacter calcoaceticus Sasm3-enhanced phytoremediation of nitrate-cadmium compound polluted soil by intercropping Sedum alfredii with oilseed rape.

Intensive agricultural system with high input of fertilizer results in high agricultural output. However, excessive fertilization in intensive agricultural system has great potential to cause nitrate and heavy metal accumulation in soil, which is adverse to human health. The main objective of the present study was to observe the effects of intercropping and inoculation of endophytic bacterium Acinetobacter calcoaceticus Sasm3 on phytoremediation of combined contaminated soil in oilseed rape (Brassica napus L.). The results showed that with Sasm3 inoculation, the biomass of rape was increased by 10-20% for shoot, 64% for root, and 23-29% for seeds while the nitrate accumulation in rape was decreased by 14% in root and by 12% in shoot. The cadmium concentration in rape increased significantly with mono-inoculating treatment, whereas it decreased significantly after intercropping treatment. By denaturing gradient gel electrophoresis (DGGE) and real-time quantitative PCR analysis, the diversity of bacterial community and the number of nirS and nirK gene copies increased significantly with inoculation or/and intercropping treatment. In conclusion, the endophytic bacterium Sasm3-inoculated intercropping system not only improved the efficiency of clearing cadmium from soil without obstructing crop production, but also improved the quality of crop.

[Destruction of oil in the presence of Cu2+ and surfactants of Acinetobacter calcoaceticus IMV B-7241, Rhodococcus erythropolis IMV Ac-5017 and Nocardia vaccinii IMV B-7405].

The effect of copper cations (0.01-1.0 mM) and surface-active agents (surfactants) of Acinetobacter calcoaceticus IMV B-7241, Rhodococcus erythropolis IMV Alc-5017 and Nocardia vaccinii IMV B-7405 in the form of culture liquid on the destruction of oil in water (3.0-6.0 g/L) and soil (20 g/kg), including in the presence of Cd2+ and Pb2+ (0.01-0.5 mM), was investigated. It was shown that the degree of oil degradation in water and soil after 20 days in the presence of low concentrations of Cu2+ (0.01-0.05 mM) and culture liquid of strains IMV B-7241, IMV Ac-5017, and IMV B-7405 was 15 - 25% higher than without copper cations. The activating effect of Cu2+ on the decomposition of complex oil and Cd2+ and Pb2+ pollution was established: after treatment with surfactant of A. calcoacelicus IMV B-7241 and R. erythropolis IMV Ac-5017 destruction of oil in water and soil was 85-95%, and after removal of the copper cations decreased to 45-70%. Intensification of oil destruction in the presence of copper cations may be due to their stimulating effect on the activity of alkane hydroxylases as in surfactant-producing strains, and natural (autochthonous) oxidizing microbiota.

Sulbactam treatment for pneumonia involving multidrug-resistant Acinetobacter calcoaceticus-Acinetobacter baumannii complex.

BACKGROUND: Multidrug-resistant (MDR) Acinetobacter calcoaceticus-Acinetobacter baumannii (Acb) complex has become an important cause of nosocomial pneumonia. Sulbactam is a ß-lactamase inhibitor with antimicrobial activity against MDR Acb complex. METHODS: To investigate outcomes of pneumonia involving MDR Acb complex treated with sulbactam or ampicillin/sulbactam for at least 7 days, we conducted a retrospective study of 173 adult patients over a 34 month period. RESULTS: Of 173 patients, 138 (79.8%) received combination therapy, mainly with carbapenems (119/138, 86.2%). The clinical response rate was 67.6% and the 30 day mortality rate was 31.2%. The independent predictors of clinical failure were malignancy, bilateral pneumonia and shorter duration of treatment. In patients with sulbactam-susceptible strains, there was no difference in clinical and microbiological outcome between combination therapy and monotherapy. Compared to the sulbactam-susceptible group, the sulbactam-resistant group had a lower rate of airway eradication, a longer duration of treatment and a higher rate of combination therapy with predominantly carbapenems (p < 0.05). There was no significant difference between the two groups in clinical resolution and 30 day mortality rates. CONCLUSIONS: Sulbactam could be a treatment option for pneumonia involving MDR Acb complex, and combination therapy with carbapenems could be considered for sulbactam-resistant cases.

Risk factors and outcomes for patients with bloodstream infection due to Acinetobacter baumannii-calcoaceticus complex.

Identifying patients at risk for bloodstream infection (BSI) due to Acinetobacter baumannii-Acinetobacter calcoaceticus complex (ABC) and providing early appropriate therapy are critical for improving patient outcomes. A retrospective matched case-control study was conducted to investigate the risk factors for BSI due to ABC in patients admitted to the Detroit Medical Center (DMC) between January 2006 and April 2009. The cases were patients with BSI due to ABC; the controls were patients not infected with ABC. Potential risk factors were collected 30 days prior to the ABC-positive culture date for the cases and 30 days prior to admission for the controls. A total of 245 case patients were matched with 245 control patients. Independent risk factors associated with BSI due to ABC included a Charlson's comorbidity score of ≥ 3 (odds ratio [OR], 2.34; P = 0.001), a direct admission from another health care facility (OR, 4.63; P < 0.0001), a prior hospitalization (OR, 3.11; P < 0.0001), the presence of an indwelling central venous line (OR, 2.75; P = 0.011), the receipt of total parenteral nutrition (OR, 21.2; P < 0.0001), the prior receipt of ß-lactams (OR, 3.58; P < 0.0001), the prior receipt of carbapenems (OR, 3.18; P = 0.006), and the prior receipt of chemotherapy (OR, 15.42; P < 0.0001). The median time from the ABC-positive culture date to the initiation of the appropriate antimicrobial therapy was 2 days (interquartile range [IQR], 1 to 3 days). The in-hospital mortality rate was significantly higher among case patients than among control patients (OR, 3.40; P < 0.0001). BSIs due to ABC are more common among critically ill and debilitated institutionalized patients, who are heavily exposed to health care settings and invasive devices.

Increased lysine content is the main characteristic of the soluble form of the polyamide cyanophycin synthesized by recombinant Escherichia coli.

Cyanophycin, a polyamide of cyanobacterial or noncyanobacterial origin consisting of aspartate, arginine, and lysine, was synthesized in different recombinant strains of Escherichia coli expressing cphA from Synechocystis sp. strain PCC 6308 or PCC 6803, Anabaena sp. strain PCC 7120, or Acinetobacter calcoaceticus ADP1. The molar aspartate/arginine/lysine ratio of the water-soluble form isolated from a recombinant strain expressing CphA6308 was 1:0.5:0.5, with a lysine content higher than any ever described before. The water-insoluble form consisted instead of mainly aspartate and arginine residues and had a lower proportion of lysine, amounting to a maximum of only 5 mol%. It could be confirmed that the synthesis of soluble cyanobacterial granule polypeptide (CGP) is independent of the origin of cphA. Soluble CGP isolated from all recombinant strains contained a least 17 mol% lysine. The total CGP portion of cell dry matter synthesized by CphA6308 from recombinant E. coli was about 30% (wt/wt), including 23% (wt/wt) soluble CGP, by using terrific broth complex medium for cultivation at 30°C for 72 h. Enhanced production of soluble CGP instead of its insoluble form is interesting for further application and makes recombinant E. coli more attractive as a suitable source for the production of polyaspartic acid or dipeptides. In addition, a new low-cost, time-saving, effective, and common isolation procedure for mainly soluble CGP, suitable for large-scale application, was established in this study.

Air-lift reactor system for the treatment of waste-gas-containing monochlorobenzene.

An air-lift bioreactor (ALR) system, applied for the treatment of waste-gas-containing monochlorobenzene (MCB) was seeded with pure culture of Acinetobacter calcoaceticus, isolated from soil as a starter seed. It was found that MCB was biologically converted to chloride as chloride was mineralized in the ALR. After the built up of the biomass in the ALR, the reactor parameters which have major influence on the removal efficiency and elimination capacity were studied using response surface methodology. The data generated by running the reactor for 150 days at varying conditions were fed to the model with a target to obtain the removal efficiency above 95% and the elimination capacity greater than 60%. The data analysis indicated that inlet loading was the major parameter affecting the elimination capacity and removal efficiency of >95%. The reactor when operated at optimized conditions resulted in enhanced performance of the reactor.

Acinetobacter calcoaceticus-baumannii complex strains induce caspase-dependent and caspase-independent death of human epithelial cells.

We investigated interactions of human isolates of Acinetobacter calcoaceticus-baumannii complex strains with epithelial cells. The results showed that bacterial contact with the cells as well as adhesion and invasion were required for induction of cytotoxicity. The infected cells revealed hallmarks of apoptosis characterized by cell shrinking, condensed chromatin, and internucleosomal fragmentation of nuclear DNA. The highest apoptotic index was observed for 4 of 10 A. calcoaceticus and 4 of 7 A. baumannii strains. Moreover, we observed oncotic changes: cellular swelling and blebbing, noncondensed chromatin, and the absence of DNA fragmentation. The highest oncotic index was observed in cells infected with 6 A. calcoaceticus isolates. Cell-contact cytotoxicity and cell death were not inhibited by the pan-caspase inhibitor z-VAD-fmk. Induction of oncosis was correlated with increased invasive ability of the strains. We demonstrated that the mitochondria of infected cells undergo structural and functional alterations which can lead to cell death. Infected apoptotic and oncotic cells exhibited loss of mitochondrial transmembrane potential (ΔΨ(m)). Bacterial infection caused generation of nitric oxide and reactive oxygen species. This study indicated that Acinetobacter spp. induced strain-dependent distinct types of epithelial cell death that may contribute to the pathogenesis of bacterial infection.

Vertical immobilization of viable bacilliform bacteria into polypyrrole films.

We have successfully developed a novel technique for inserting viable bacilliform bacteria into polypyrrole films. All of the five different bacterial cells (Pseudomonas aeruginosa, Acinetobacter calcoaceticus, Serratia marcescens, Bacillus subtilis and Escherichia coli) studied in this work were inserted normal to the film surface, and the viability of P. aeruginosa was unaffected by this immobilization procedure. It was also found that the polypyrrole layer was important to keep the cells alive.

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.

Clinical characteristics of tertiary hospital patients from whom Acinetobacter calcoaceticus-Acinetobacter baumannii complex strains were isolated.

OBJECTIVE: Acinetobacter baumannii is a worldwide nosocomial pathogen that has become increasingly common over the past few decades, and strains of multidrug-resistant A. baumannii have been increasing. The aim of this study was to assess the clinical characteristics of A. calcoaceticus-A. baumannii complex (Acb complex) strains and to determine the risk factors of this infection. METHODS: The medical records of 121 patients at Nagasaki University Hospital from whom Acb complex had been isolated between January 2007 and December 2009 were retrospectively reviewed. Patient backgrounds, sensitivity to antibiotics, risk factors for infection, and prognosis were evaluated. RESULTS: Lower respiratory isolates accounted for 73% (147 strains) of all 201 isolates. Most of the isolates were sensitive to carbapenems. Of the 121 patients (74 males and 47 females; mean age: 62.1 years), 48 (39.7%) had malignancy and 75 (62.0%) were treated with antibiotics prior to isolation. Thirty-seven of the patients in this study (30.6%) were infected by Acb complex and the most frequent clinical manifestation was pneumonia (18 cases; 48.6%). Approximately 60% of infected patients were treated with ß-lactam agent in combination with ß-lactamase inhibitors or carbapenems. The mortality rate of infected patients was significantly higher than that of colonized patients (infected: 24.3%, colonized: 6.0%, p<0.05). Risk factors for Acb complex infection include being over 60 years of age, chronic liver disease, and the use of first-generation cephalosporins prior to isolation. CONCLUSION: Acb complex was relatively sensitive to antibiotics. The appropriate usage of antibiotics should be continued for the prevention of drug resistance in Acb complex.

[Chemiluminescence analysis of oil oxidizing bacteria Actinetobacter calcoaceticus extracts: effects of the extracts on pSoxS-lux biosensor].

A comparative H2O2-luminol- and Fe(II)-induced chemiluminescence analysis of extracts of two strains of marine oil oxidizing bacteria Actinetobacter calcoaceticus cultivated either in the presence or absence of oil was carried out. Effects of these extracts on E. coli MG1655 biosensor (pSoxS-lux) were studied. Activation of H2O2-induced chemiluminescence in the presence of oil was observed. This suggests activation of free radical lipid peroxidation. Aqueous extracts of microorganisms cultivated in the presence of oil were shown to activate reactive oxygen species production (ROS) in Fe(II)-induced chemiluminescence reaction mixture. Acetone-ethanol extracts induced antioxidative systems of both strains. Chemiluminescence analysis in a biological system carried utilizing E. coli MG1655 (pSoxS-lux) revealed that aqueous extracts of the strains cultivated in the absence of oil contained potential antioxidants.

Direct quantification of inorganic polyphosphate in microbial cells using 4'-6-diamidino-2-phenylindole (DAPI).

Inorganic polyphosphate (polyP) is increasingly being recognized as an important phosphorus sink within the environment, playing a central role in phosphorus exchange and phosphogenesis. Yet despite the significant advances made in polyP research there is a lack of rapid and efficient analytical approaches for the quantification of polyP accumulation in microbial cultures and environmental samples. A major drawback is the need to extract polyP from cells prior to analysis. Due to extraction inefficiencies this can lead to an underestimation of both intracellular polyP levels and its environmental pool size: we observed 23-58% loss of polyP using standard solutions and current protocols. Here we report a direct fluorescence based DAPI assay system which removes the requirement for prior polyP extraction before quantification. This increased the efficiency of polyP detection by 28-55% in microbial cultures suggesting quantitative measurement of the intracellular polyP pool. It provides a direct polyP assay which combines quantification capability with technical simplicity. This is an important step forward in our ability to explore the role of polyP in cellular biology and biogeochemical nutrient cycling.

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.