A. Mlynarczyk

Antibiotic resistance in Gram-positive cocci

Gram-positive cocci still predominate as a cause of nosocomial- and community-acquired infections. These organisms frequently reveal a high natural, intrinsic resistance to antimicrobials. Additionally, these bacteria are able to acquire resistance to frequently used drugs rapidly through selective pressure of the environment and via the genetic evolution of bacteria. The wide application of antimicrobials in medical and veterinary practice, usage of antibiotics in agriculture and common usage of antiseptics and disinfectants result in selective pressure. The use of antibiotics directly selects resistant variants to different antibiotics or disinfectants. The same genetic element (e.g. qac or smr) conferring resistance to some disinfectants are often present on the same plasmid conferring resistance to antibiotics. Selection of resistant variants occurs most frequently in the hospital environment. Staphylococcus aureus and enterococci are the most commonly isolated bacteria causing nosocomial infections. Among those giving therapeutic problems are methicillin-resistant staphylococci and vancomycin-resistant enterococci. Resistance to high levels of aminoglycosides or penicillins among hospital enterococcal strains can completely abolish synergism of the drugs. In these cases glycopeptides will be the drugs of choice in the treatment of serious infections. Recently S. aureus strains with decreased susceptibility to vancomycin has appeared. A mechanism for this elevated resistance, although intensively investigated, still remains unknown.

Epidemiological aspects of antibiotic resistance in respiratory pathogens.

Abstract Respiratory infections are the most frequent reason for primary health care consultation. The main causes of respiratory tract infections in children are viruses and the most common types are upper respiratory tract infections: common cold, pharyngitis, otitis media and sinusitis. Pneumonia is much more serious. As well as viruses, bacteria are often involved in respiratory tract infections. Three bacterial species are most commonly isolated: Streptococcus pneumoniae, non-encapsulated Haemophilus influenzae and Moraxella (Branhamella) catarrhalis. The most common bacterial cause of pharyngitis is Streptococcus pyogenes. Bacteria isolated from community-acquired infection usually are sensitive to the majority of suitable drugs, but during the past two decades, significant antibiotic resistance has emerged. Resistance to penicillins has spread among H. influenzae and S. pneumoniae. The mechanism of penicillin resistance in H. influenzae is mainly by production of β-lactamases TEM-1 and ROB-1, whereas in S. pneumoniae resistance is an effect of the changes in penicillin binding proteins. Among respiratory pathogens, resistance to tetracyclines, macrolides, trimethoprim–sulphamethoxazole and fluoroquinolones has also appeared. Several mechanisms depending on changes in target, active efflux and modifying enzymes are involved.

A Threat of the Klebsiella Pneumoniae Carbapenemase–Producing Strains Among Transplant Recipients

BACKGROUND Infections due to Klebsiella pneumoniae carbapenemase (KPC)-producing Enterobacteriaceae are associated with increased therapeutic failure and mortality. Our laboratory recognized several strains producing KPC, most of which originated from transplantation ward patients. MATERIALS AND METHODS All strains of K pneumoniae resistant to at least 1 carbapenem isolated in 2010 were examined for KPC production by disc diffusion and then verified by molecular methods. RESULTS All positive strains originated from 7 patients. Six of them were from transplantation wards. None of the KPC-producing strains was isolated from the patients blood. CONCLUSIONS A quick, accurate diagnosis of KPC-producing strains enabled immediate isolation of carriers or infected persons. Isolation prevented spread of dangerous strains among immunocompromised patients and reduced the possibility of serious infections.

Surgical site infections in the early posttransplant period after simultaneous pancreas-kidney transplantation.

OBJECTIVE Urinary tract infection (UTI) is among the common infection in simultaneous pancreas-kidney transplantation (SPKT). PATIENTS AND METHODS The study included 26 adult patients undergoing SPKT between September 2001 and December 2006. All the patients were followed prospectively for UTI during the first 4 weeks after surgery. Urine samples were investigated for bacteriologic cultures. The micro-organisms were identified in accordance with standard bacteriologic procedures. Susceptibility testing was carried out using Clinical and Laboratory Standards Institute (CLSI) procedures. RESULTS Among 77 urine specimens obtained from all recipients during the first month, there were 30 isolated bacterial strains. The most common were Gram-positive bacteria (53.3%) with predominance of enterococci (75%) associated with high levels of aminoglycoside resistant strains (HLAR; 58.3%) and vancomycin-resistant strains (VRE; 25%). Gram-negative bacteria were detected in 46.7% of positive cultures. CONCLUSIONS In our study, enterococci predominated as 75% of Gram-positive isolates. The increased proportion of multi-drug-resistant bacteria, which can caused severe UTI in patients after SPKT, may be due to the frequent use of prophylaxis of bacterial infections in patients.

Molecular and Phenotypic Characteristics of Methicillin-resistant Staphylococcus aureus Strains Isolated From Hospitalized Patients in Transplantation Wards

OBJECTIVES Hospital-acquired methicillin-resistant Staphylococcus aureus (HA-MRSA) frequently causes therapeutic problems and provides information about the epidemiological condition of the ward. MATERIALS AND METHODS HA-MRSA isolated from patients on transplantation wards in 1991, 1994, 1996, and from 2005 to 2007 were compared using molecular methods such as restriction fragment length polymorphism-pulse field gel electrophoresis, multilocus sequence typing (MLST), multiplex polymerase chain reaction (PCR) for detection type of staphylococcal chromosomal cassette mec, and PCR for detection. RESULTS The analysis covered HA-MRSA strains, each from a different patient. All organisms were typed using molecular methods. MLST results were compared with an international base. The examined strains belonged to five different worldwide known clonal complexes: CC8 (78%), CC5 (12%), CC1 (4%), CC30 (2%), and CC51 (4%). All could be recognized as representatives of a clonal complex CC8 clones: ST239-III (sequence type 239 and SCCmec type III named EMRSA-1, -4, -11, Brasilian, Hungarian) occurred with a frequency of 35.9%, ST254-IV (EMRSA-10, Hannover) occurred in 33.3%, ST247-I (EMRSA-5,-7, Iberian) occurred in 20.5%, ST241-III (Finland-UK) occurred in 5.15%, and ST8-IV (EMRSA-2,-6) occurred in 5.15%. CONCLUSION The predomination of different clones of HA-MRSA in the particular years was observed. In 1991, the EMRSA-10 (Hannover) clone predominated (53.3%). The Brasilian-Hungarian (EMRSA-1, -4, -11) clone predominated in 1994 (50%) as well as from 2005 to 2007 (41.3%), whereas in 1996 the Iberian clone was most frequent (53.9%).

Acinetobacter baumannii Multidrug-Resistant Strain Occurrence in Liver Recipients With Reference to Other High-Risk Groups

INTRODUCTION The increasing clinical significance of Acinetobacter baumannii species is due to its ability to survive in hospital environments, its species-specific multidrug resistance, and its ability to instantly develop various drug-resistance mechanisms through antibiotic pressure. MATERIALS AND METHODS We identified 16 A baumannii strains isolated from patients presenting postoperative infections in 2010. A baumannii isolates were obtained from clinical specimens by standard microbiologic methods. As previously described, we performed polymerase chain reaction (PCR) analysis for carbapenemase-encoding genes (VIM, IMP, SPM, OXA23, OXA24, OXA51, OXA58) in Acinetobacter spp. RESULTS The double-disk synergy test phenotypic method did not detect any A baumannii strains producing metallo-beta-lactamaus cultured from swabs from all the patient groups. No products of PCR amplification with specific starters for VIM, IMP, and SPM (Sao Paulo metallo-β-lactamase) genes were found. All analyzed strains were colistin-sensitive. Among five strains from liver recipients, one was imipenem- and meropenem-resistant. Four among six strains isolated from cancer patients were resistant to imipenem and/or meropenem; 1/5 were imipenem-and meropenem-resistant; 1, meropenem-resistant and imipenem-sensitive; 1, meropenem- and imipenem-resistant; and 1 with intermediate resistance to both meropenem and imipenem among swabs cultured from patients with postoperative complication after bone fracture. Fifteen among 16 analyzed A baumannii strains had an OXA51 gene. Two among five A baumannii strains isolated in liver recipients had only an OXA51 gene; one, OXA51 and OXA24 genes; one, OXA51 and OXA23 genes.

CTX-M and TEM as Predominant Types of Extended Spectrum β-Lactamases Among Serratia marcescens Isolated From Solid Organ Recipients

BACKGROUND Serratia marcescens is an important pathogen in hospital infections since organisms resistant to multiple antimicrobials pose a special threat particularly among transplant patients. The aim of this work was to assess the number of strains producing beta-lactamases with extended spectrum (ESBL) among S. marcescens isolated from our patients. MATERIALS AND METHODS We investigated S. marcescens isolated from 2005 to 2008 for ESBL. The phenotype methods were applied and additionally we chose strains for polymerase chain reactions using primers for the most popular types of ESBL. RESULTS Over the investigated time, 257 patients were infected with S. marcescens with 188 (73%) displaying an ESBL-positive phenotype. A Molecular analysis showed that most of them produced both CTX-M and TEM beta-lactamases. In the last year, the percentage of ESBL-producing strains decreased, but also in the last year, we isolated S. marcescens resistant to carbapenems from three patients. CONCLUSIONS The CTX-M type of ESBL predominated among ESBLs produced by strains of S. marcescens. The appearance of strains resistant to carbapenems is alarming.

Coagulase-negative variants of methicillin-resistant Staphylococcus aureus subsp.aureus strains isolated from hospital specimens

At the Department of Clinical Bacteriology of Medical University of Warsaw, S. aureus strains displaying negative reactions in the coagulase tube-test were isolated with a relatively high frequency from clinical specimens. As many as seventeen of such strains all from different patients were isolated during 1994 among MRSA strains. Despite coagulase negativity all strains were positive in tests for clumping factor (CF), protein A and thermonuclease. Biochemical characteristics, phage patterns and antibiotic resistance characteristics of these strains were tested. Most of the coagulase-negative strains possessed a heterogenous type of methicillin resistance. Apart from methicillin most of them were resistant to many other antimicrobials. All were resistant to gentamicin, tetracyclines and macrolide-lincosamide-streptogramine B. An occurrence of coagulase-negative S. aureus strains may lead to problems in their identification and the necessity of an application of other methods like CF, protein A or biochemical reactions.

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.