Marta Kierzkowska

A comparison of Api 20A vs MALDI-TOF MS for routine identification of clinically significant anaerobic bacterial strains to the species level.

Adequate identification of anaerobic bacteria still presents a challenge for laboratories conducting microbiological diagnostics. The aim of this study was to compare the use of Api 20A and MALDI-TOF MS techniques for identification of obligate anaerobes. The results indicate that MALDI-TOF MS ensures a rapid and accurate identification of the species isolated from patients.

Specific Character of Anaerobic Bacterial Infections in Patients Treated in Transplantation Wards at One of the Clinical Hospitals in Warsaw

Immunocompromised patients and patients undergoing invasive procedures are predisposed to bacterial infections, due to the possibility of micro-organism translocation from their physiological habitat. Infectious complications may occur both in the early and late post-transplantation periods. The purpose of this study was to evaluate the proportion as well as susceptibility profiles of obligatory anaerobes in the etiology of infections in patients hospitalized at transplantation wards of a large clinical hospital in Warsaw. A total of 104 strains of obligatory anaerobes derived from patients hospitalized in two transplantation clinics at a clinical hospital in Warsaw were evaluated. The strains were isolated from 87 clinical samples collected from 84 patients of two transplantation wards between 2007 and 2012. A total of 104 obligatory anaerobic bacterial strains were isolated and identified, with Gram-positive and Gram-negative bacteria constituting 60.6% and 39.4% of the isolates, respectively. Almost exclusively non-spore-forming anaerobes were detected in evaluated samples. The present study showed all isolated Gram-positive bacteria to be susceptible to ß-lactam antibiotics. Metronidazole-resistant bacteria were found among the genera Propionibacterium and Actinomyces. All Gram-negative rods were susceptible to imipenem and metronidazole. Among them, Bacteroides spp. and Parabacteroides distasonis showed resistance to penicillin G (100%). Because of their pathogenicity and altered antibiotic susceptibility profiles, the bacteria of the genera Bacteroides and Parabacteroides are of greatest clinical importance. Approximately 25% of isolates exhibit also resistance to clindamycin. Because of the growing rates of clindamycin resistance, the role of metronidazole in the treatment of Bacteroides spp. is of increasing importance.

Blood Infections in Patients Treated at Transplantation Wards of a Clinical Hospital in Warsaw

Establishment of the etiology in blood infection is always advisable. The purpose of this study was to evaluate the proportion of different bacterial species, including aerobic and anaerobic bacteria in blood cultures of patients hospitalized in transplantation wards of a large clinical hospital between 2010 and 2012. A total of 1994 blood samples from patients who were treated at one of two transplantation wards of a large hospital in Warsaw were analyzed using an automated blood culture system, BacT/ALERT (bioMerieux, France). The 306 bacterial strains were obtained from the examined samples. The highest proportion were bacteria from the family Enterobacteriaceae (112 strains; 36.6%) with Escherichia coli (61 strains), Klebsiella pneumoniae (30 strains), and Enterobacter cloacae (10 strains) most commonly isolated. The non-fermenting bacilli constituted 21.6% (66 strains), with most common Stenotrophomonas maltophilia (31 strains), Pseudomonas aeruginosa (14 strains), Achromobacter spp. (12 strains), and Acinetobacter baumannii (3 strains). Most frequent Gram-positive bacteria were staphylococci (25.2%). Of 77 staphylococcal strains, 56 were coagulase-negative staphylococci and 21 Staphylococcus aureus. Other Gram-positive bacteria included enterococci (14 strains) and Streptococcus pneumoniae (1 strain). Obligatory anaerobic bacteria were represented by 19 strains (6.2% of total isolates). Among all Enterobacteriaceae, 49 isolates (43.7%) produced extended-spectrum ß-lactamases (ESBLs). Resistance to methicillin was detected in 62% of S aureus isolates and in 46% of coagulase-negative staphylococci. Of 14 enterococci cultured from blood samples, 2 strains (14.3%) were resistant to vancomycin. Both were Enterococcus faecium. Resistant strains of Gram-negative and Gram-positive bacteria are significant problems for patients in the transplantation ward.

Synthesis and antimicrobial activity of derivatives of 1H-benzo[de]isoquinoline-1,3(2H)-dione

Abstract Five aminoalkyl 1H-benzo[de]isoquinoline-1,3(2H)-diones were synthesized and evaluated for antimicrobial activity. Microorganisms used in this study included aerobic and facultative anaerobic bacteria Staphylococcus aureus, Escherichia coli and Stenotrophomonas maltophilia, and obligatory anaerobes Bacteroides fragilis, Bacteroides thetaiotaomicron and Propionibacterium acnes. Moreover, Candida albicans yeast was used. The minimum inhibitory concentration (MIC) was determined for the test compounds. Among the tested derivatives, two compounds 1 and 2 have shown the most potent antibacterial as well as antifungal activities.

Participation of Strictly Anerobic Bacteria in Infections among Hospitalized Transplant Patients in a Clinical Hospital in Warsaw

BACKGROUND Many strictly anerobic bacteria are a part of the human commensal microflora. Especially multitudinously they inhabit the skin, mucous membranes, gastrointestinal tract, respiratory, and genital tracts. Infections with these bacteria may occur after escape of the bacteria from their natural habitat. There are often mixed anerobic and aerobic infections. After rupture of the gastrointestinal tract or surgery, the organisms can cause significant pathology including abscesses and bacteremia. OBJECTIVE The aim of this study was to estimate the prevalence of gram-negative strictly anerobic bacteria isolated from different samples collected from patients on transplant wards. MATERIALS AND METHODS Samples from patients with suspected infections. Underwent isolation and identification of microorganisms under anerobic conditions using standard laboratory methods. RESULTS Gram-negative rods were observed in 46% of clinical samples, most frequently Bacteroides genus as well as Fusobacterium necrophorum/F.nucleatum and F.mortiferum (14%). Most species of the genus Bacteroides were accompanied by Escherichia coli, less frequently with other aerobic gram-negative rods or gram-positive cocci. CONCLUSION Gram-negative bacteria were frequently isolated in the samples. Because they are the part of the normal flora, this observation indicated the endogenous nature of infections resulting from bacterial translocation out of their natural habitat.

Difficulties in identifying the bacterial species from the genus Clostridium in a case of injury-related osteitis

Most Clostridium species are part of saprophytic microflora in humans and animals; however, some are well-known human pathogens. We presented the challenges in identifying the Clostridium species isolated from a patient with an infected open dislocation of the proximal interphalangeal joint of the fourth digit of the right hand. The clinical materials were intraoperative samples collected from a patient diagnosed with an injury-related infection, with soft tissue loss and tendon sheath involvement. The available biochemical, molecular, and genetic techniques were used in identifying the isolated bacteria. The isolated bacterium was shown to have low biochemical activity; hence, it was not definitively identified via biochemical tests Api 20A or Rapid 32A. Vitek 2 and mass spectrometry methods were equally inconclusive. Clostridium tetani infection was strongly suspected based on the bacterium’s morphology and the appearance of its colonies on solid media. It was only via the 16S rRNA sequencing method, which is non-routine and unavailable in most clinical laboratories, that this pathogen was excluded. Despite appropriate pre-laboratory procedures, which are critical for obtaining reliable test results, the routine methods of anaerobic bacterium identification are not always useful in diagnostics. Diagnostic difficulties occur in the case of environment-derived bacteria of low or not fully understood biological activity, which are absent from databases of automatic bacterial identification systems.

Orthopedic infections caused by obligatory anaerobic Gram-negative rods: report of two cases

Anaerobic bone and joint infections are uncommon, although the number of anaerobic infections is presumably underestimated because of difficulties with isolation and identification of obligate anaerobes. This study describes two cases of complicated Bacteroides fragilis peri-implant infection of the lumbar spine, infection of the hip and osteomyelitis. Bacteria were identified with the use of a mass spectrometer, VITEK MS system. Drug susceptibility was performed with the use of E-test. The EUCAST breakpoints were used for interpretation with B. fragilis ATCC 25285 as a control. In the two described cases clinical samples were collected for microbiological examination intraoperatively and simultaneously empirical treatment was applied. B. fragilis was isolated in monoculture or in a combination with other bacteria. The treatment was continued according to the susceptibility tests. In a case one clindamycin failure was observed and clindamycin resistance of the isolate was likely due to inadequate time of therapy. Difficulties in collecting an adequate samples and culturing anaerobic bacteria cause that not all infections are properly recognized. In a successful therapy, identification and determination of the susceptibility of the pathogen are essential as well as an appropriate surgical debridement.

The in vitro effect of clindamycin against Bacteroides and Parabacteroides isolates in Poland

OBJECTIVES The aims of this study were (i) to analyse strains of the genera Bacteroides and Parabacteroides isolated from clinical specimens for phenotypic resistance to clindamycin, (ii) to detect erm genes in the isolates and (iii) to determine any correlation between in vitro resistance and the presence of erm genes. METHODS The Bacteroides and Parabacteroides isolates analysed were obtained from patients hospitalised at teaching hospitals in Poland. Antimicrobial susceptibility testing was performed by Etest and the results were interpreted according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. All isolates were analysed by PCR for the presence of the resistance genes ermF, ermB and ermG. RESULTS Resistance to clindamycin was detected in 31.0% (62/200) of all evaluated isolates, with the ermF and ermB genes detected in 31.0% (62/200) and 0.5% (1/200) of isolates, respectively. No isolates with ermG were detected among the evaluated strains. Pearsons test showed an almost perfect correlation between clindamycin minimum inhibitory concentrations (MICs) and the presence of ermF in Bacteroides spp. and Parabacteroides distasonis isolates, although the ermF gene was also present in 10 clindamycin-susceptible isolates of Bacteroides spp. CONCLUSIONS This study demonstrated a substantial proportion of Bacteroides (22.5-100% depending on the species) and 50.0% of Parabacteroides strains exhibiting resistance to clindamycin. The clindamycin MIC for resistant strains in each case was ≥256mg/L. Resistance to clindamycin in Bacteroides and Parabacteroides species is correlated mainly with the presence of the ermF gene.

Synthesis and biological activity of 6-substituted 5-acetyl-4,7-dimethoxybenzofuran derivatives

Abstract In the search for new antimicrobial and anticancer agents, a series of (aryl/heteroaryl-piperazino-alkyl)-substituted derivatives of benzo[b]furans were prepared. All compounds were characterized by 1H NMR, 13C NMR, ESI-MS spectra and elemental analyses. Most of the investigated compounds had no antimicrobial activity (MIC > 512 mg/L) except for 2l, 2m and 2o, which showed activity against Candida albicans. None of the tested compounds showed significant anticancer activity in K562 and HeLa cells.