Prevalence study on carbapenemase-producing Escherichia coli and Klebsiella pneumoniae isolates in Czech hospitals--results from Czech Part of European Survey on Carbapenemase--Producing Enterobacteriaceae (EuSCAPE).

OBJECTIVE: One of the most important threats of current medicine is the spread of multiresistant Gram-negative bacteria. We report here data from a six-month prevalence study on carbapenemase-producing K. pneumoniae and E. coli performed in Czech hospitals participating on European Survey on Carbapenemase-Producing Enterobacteriaceae (EuSCAPE). METHODS: Ten hospitals covering all regions of the Czech Republic were selected. During the study period (1st November 2013 to 30th April 2014), first ten carbapenem non-susceptible isolates of K. pneumoniae or E. coli isolated from non-surveillance specimens (i.e., blood, lower respiratory tract secretions, urine, puncture fluids, and wound secretions) of single successive patients were collected. Successive carbapenem-susceptible isolates of the same species were also preserved as controls. Susceptibility to 15 antibiotics was determined using EUCAST recommendations. Carbapenemase activity was detected by MALDI-TOF MS meropenem hydrolysis assay. Positive isolates were subjected for molecular typing (multi-locus sequence typing, identification of carbapenemase gene). RESULTS: During the study period, thirty non-susceptible isolates (K. pneumoniae n=28, E. coli n=2) were identified in 5 hospitals. Only two of them were confirmed to be carbapenemase producers. A NDM-1-producing K. pneumoniae ST11 was recovered from a patient, transferred from Ukraine, being injured during a Maidan revolution. The second isolate, an OXA-48-producing K. pneumoniae, belonging to ST101, was recovered from a patient admitted to a hospital for an ischemic stroke. CONCLUSIONS: This study again confirmed that the Czech Republic still belongs to the countries with low prevalence of carbapenemase-producing Enterobacteriaceae (CPE). Cases of CPE are usually restricted to an import from high-prevalence countries or countries with unknown epidemiological situation.

[Clostridium colitis at a surgical department]. / Problematika klostridiové kolitidy na chirurgickém pracovisti.

INTRODUCTION: The aim of our study was to identify risk factors associated with the development of Clostridium difficile colitis and determining the severity of clostridial colitis in a group of surgical patients. Identification of its predispositions is necessary for preventive interventions and effective treatment. MATERIAL AND METHODS: 51 patients diagnosed with clostridial colitis were included in our study. The inclusion criterion was based on laboratory detection of clostridial toxin in each patient. We analysed age, concomitant malignancy, recent surgical intervention, antibiotic and proton pump inhibitors treatment. The severity of clostridial colitis and its correlation to possible risk factors mentioned above was described and statistically evaluated. Non-parametric Fischer test was used for the statistical evaluation. RESULTS: We confirmed the importance of potential risk factors in the evaluated group of surgical patients with Cl. difficile colitis. The course of the disease was described as serious in 39.2% of patients in the study group. Recurrent attack of colitis was diagnosed in 4 patients, in 2 of them the second recurrence, in 1 patient the third relapse occurred. 2 patients with clostridial colitis evaluated in our group died, but the relation of their death to the clostridial GIT infection was not causal and the primary cause of death in both of them was multiple organ failure caused by serious comorbidities. We did not indicate any surgical intervention and no toxic megacolon developed in our study group. We did not detect any statistical correlation between the described risk factors and the severity of colitis. CONCLUSION: Clostridium difficile colitis needs to be recognised as a serious complication, especially in hospitalised patients. Its increased frequency and severity should not be underestimated. Although we can identify potential risk factors, we cannot exclude some of them completely (e.g. antibiotic and PPI treatment). On the other hand, a rational approach to antibiotic treatment in particular can be helpful in reducing the frequency and severity of clostridium difficile colitis.

[Antibiotic resistance in nontyphoidal salmonellae serovars in the Czech Republic]. / Antibiotická rezistence u netyfových sérovaru Salmonella spp. v Ceské republice.

STUDY AIM: To determine antibiotic resistance and incidence of multidrug resistance among Nontyphoidal salmonellae serovars isolated from humans. MATERIAL AND METHODS: Consecutive Salmonella isolates from patients, recovered in 48 microbiology laboratories in May 2012, were analyzed in the respective reference laboratories at the National Institute of Public Health. Strains were re-identified and differentiated into serovars. Their minimum inhibitory concentrations (MICs) to 11 antibiotics were determined by the microdilution method. RESULTS: Of 25 serovars identified among 637 strains of Salmonella enterica, the most frequent were Enteritidis (87.0 %), Typhimurium (4.9 %), and monophasic Typhimurium 4,[5],12:i:- (2.0 %) and Mbandaka (0.6 %); other serovars were rare. Altogether 558 strains (87.6 %) were susceptible to all antibiotics tested and the remaining 79 strains were resistant to one or more antibiotics. The prevalence rates of resistance to individual antibiotics among 637 study strains were as follows: ampicillin 8.5%, tetracycline 5.7%, sulfamethoxazole 5.2%, cipro-floxacin 3.8%, and chloramphenicol 2.5%. Resistance to gentamicin, trimethoprim, and third and fourth generation cephalosporins was rare ( 0.5%) and none of the study strains showed resistance to meropenem. Three producers of extended spectrum beta-lactamase were multidrug resistant and two of them recovered from twins exhibited a different pattern of resistance. Resistant strains were most often assigned to the following serovars: Enteritidis (49.4%), Typhimurium (26.6%), and monophasic Typhimurium (15.2%). While only 7% (39 of 554 strains) of Enteritidis strains were resistant, the serovars Typhimurium and its monophasic variant 4,[5],12:i:- showed high rates of resistance, i.e. 66.7 and 92.3%, respectively. Furthermore, resistance was revealed in all strains of the serovars Virchow (n = 3), Kentucky (n = 1), and Newport (n = 1), in two of three strains of the serovar Infantis, and in one of two strains of the serovar Stanley. All five blood isolates were assigned to the serovar Enteritidis and one of them showed resistance to ciprofloxacin. Of 79 resistant strains, 26.6% showed resistance to ampicillin only and 24.1% to ciprofloxacin only, with multidrug resistance, i.e. resistance to three or more antibiotics, confirmed in 43.0% of strains. CONCLUSION: Despite a relatively low prevalence of resistance to the antibiotics tested among 637 study strains, the following alarming findings were made: Detection of Salmonella enterica strains resistant to ciprofloxacin as the drug of choice or to higher generation cephalosporins and multidrug resistance revealed in two thirds of the strains of the serovar Typhimurium and in all but one strains of its monophasic variant 4,[5],12:i:-.

Importance of biofilm in Candida parapsilosis and evaluation of its susceptibility to antifungal agents by colorimetric method.

The ability of C. parapsilosis (an important cause of nosocomial infections) to produce biofilm was evaluated in 32 bloodstream isolates and 85 strains isolated from skin. The biofilm formation was found in 19 (59%) blood isolates and only in 33 (39%) isolates from skin. The antifungal susceptibility was assessed for amphotericin B, itraconazole and voriconazole in planktonic and biofilm form of the 19 biofilm-positive bloodstream strains by broth microdilution method according to NCCLS standards. The method was modified by the use of resazurin as a colorimetric indicator of the metabolically active cells which makes the determination of the effect of antifungal agents easier. Biofilm forms of all strains were more resistant than their planktonic form.

[Detection and significance of biofilm formation in yeasts isolated from hemocultures]. / Detekce a význam tvorby biofilmu u kvasinek izolovaných z hemokultur.

BACKGROUND: Yeasts of the genus Candida are important opportunistic pathogens responsible for severe bloodstream infections in immunocompromised patients. An important virulence factor that allows them to colonize plastic implants and to survive in the blood is their ability to form biofilm. The purpose of the study was to assess, how frequently this virulence factor occurred in yeasts of the genus Candida isolated from blood cultures. MATERIALS AND METHODS: Cultures of the various strains in wells of microtitre plates were used to demonstrate the presence of biofilms. In total, we tested 72 isolates of yeasts of the genus Candida from the blood culture: C. albicans (33), C. parapsilosis (19), C. tropicalis (11), C. glabrata (4), C. krusei (3), C. lipolytica (1) and C. lusitaniae (1). RESULTS: The ability to form biofilm was demonstrated in 26 strains. The largest number of biofilm-positive strains was found in C. parapsilosis (13) and C. tropicalis (7). The formation of biofilms was far less frequent in the strains of C. albicans (4). The formation of biofilms was also demonstrated in C. krusei (2) and C. glabrata (1). CONCLUSION: The increasing frequency of fungal bloodstream infections is due to the increasing use of catheters and implants. The biofilm formation in these yeasts is the chief factor of virulence promoting such infections. The demonstration of biofilm formation in an isolated strain is indicative of the probable existence of a biofilm focus and of possible difficulties during antimycotic therapy.

Biofilm detection and the clinical significance of Staphylococcus epidermidis isolates.

The ability of Staphylococcus epidermidis to produce biofilm was compared in 147 clinically significant strains repeatedly isolated from blood cultures of patients with bloodstream infection and in 147 strains isolated from skin. The strains were examined for the presence of ica operone, for the ability to form biofilm by Christensen's test-tube method and for the production of slime by Congo Red agar method. The ica operone was found in 92 (62.6 %) blood isolates and in 44 (29.9) isolates from skin. Christensen's test-tube method was positive in 79 (53.7) and 33 (22.4), Congo Red agar method in 64 (43.5) and 31 (21.1) of blood and skin isolates, respectively. All three methods were more frequently positive in clinically significant isolates from blood than in strains isolated from skin. The detection of ica operone and the Christensen's test-tube method showed better correlation with the clinical significance than the Congo Red agar method.

[Rhodococcus equi--a newly recognized opportunistic pathogen in man]. / Rhodococcus equi--nove poznaný oportunní patogen cloveka.

The review informs about substantial features of Rhodococcus equi with emphasis on the analysis of 115 as yet published and still expanding reports on the isolation of this zoopathogenic nocardioform actinomycete from man. Microbiological laboratories of human medicine have to learn not only how to identify R. equi but also recognize it as an opportunistic pathogen in particular in persons with the deficient immunity. R. equi is a gram-positive, encapsulated diphtheroid coccobacillus, partially acid fast. It grows well on common media, its colonies being after 48 hours characteristically mucoid, coalescing, irregular and mostly lightly pinkish. Biochemically it is little active, nevertheless it causes typical synergic haemolysis of erythrocytes influenced by staphylococcal beta-toxin. R. equi is found in soil and manure, especially in horse manure. It causes above all granulomatous pneumonia in young foals. In humans, it causes mostly pneumonia and lung abscess, more frequently in persons with immunity deficiency incl. AIDS, less often extrapulmonary abscesses, sepsis and wound infections. The disease are commonly chronic and recurrent. The ability of R. equi to persist in macrophages and destroy them is important in the pathogenesis of infection. In resistance to infection the cell-mediated immunity seems to be of major importance. The port of entry are the lungs, less often the alimentary tract or injured skin. About a third of the persons gives a history of contact with animals, manure or soil. The standard treatment is prolonged administration of a combination of rifampin and erythromycin. The isolation of R. equi is easy and if a laboratory suspects the presence of this microorganism, its identification is not difficult.

[Review of 105 cases of isolation of Rhodococcus equi in humans]. / Prehled 105 prípadu izolace Rhodococcus equi u cloveka.

BACKGROUND: Rhodococcus equi (formerly Corynebacterium) has been long considered an exclusively zoopathogenic microbe causing mainly granulomatous pneumonias and lung abscesses in young foals. The aim of this paper was to analyse substantial features of R, equi infections hitherto reported in man. METHODS AND RESULTS: MEDLINE database was searched for relevant reports. When the original source was not obtained the data from reviews were employed. Together, 105 cases of R, equi infection in man were reported. Median age was 35 years with a range from 9 months to 83 years. The male: female ratio was 3.3. Lungs were involved in 72 cases (69%), extrapulmonary abscesses as the only symptom of infection were described in 9 cases, septic state in 7 cases. Clinical outcome was known in 98 cases, being fatal in 41 (42%). Therapy was mentioned in 70 reports, the most often used drugs being erythromycin (30 cases, 12 deaths), rifampicin (19 cases, 7 deaths) and vancomycin (18 cases, 6 deaths). R. equi was isolated from the sputum of 69% patients with the pulmonary involvement. Blood cultures were positive in 35% of cases. Out of total, 49% persons were HIV positive. Median age for HIV positive patients was 32 years with a range from 18 to 71 years, for HIV negative patients 52 years with a range from 9 months to 83 years. There were 97% males in the HIV positive group in contrast to 59% in the HIV negative group (p < 0.01). Lungs were involved in 90% of HIV positive and 48% of HIV negative cases (p < 0.01). Extrapulmonary abscesses as the only sign of infection were seen in 2% of HIV positive persons and in 15% of HIV negative ones (p < 0.05). Outcome was fatal for 60% of the HIV positive hosts and for 28% of the HIV negative individuals (p < 0.01). R. equi was isolated from the sputum of 80% pneumonic HIV positive patients and of 50% of pneumonic patients without HIV infection (p < 0.05). R. equi was detected in the blood of 67% of HIV positive patients and of 33% of HIV negative ones (p < 0.01). CONCLUSIONS: The analysis of published reports shows that whereas R. equi causes mainly pneumonia in persons with HIV infection, in HIV negative individuals extrapulmonary manifestations slightly prevail, most often abscesses, sepsis, eye involvement and wound infections.