Epidemiology of neonatal sepsis in two neonatal intensive care units in Krakow, Poland in 2016-2017 years.

BACKGROUND: Sepsis in low-birth-weight neonates remains one of the most significant causes of neonatal morbidity and mortality. Approximately 3 million newborns suffer from sepsis globally every year. The aim of this study was to compare demographic and clinical features, as well as etiology and antibiotic susceptibility, of the main pathogens related to neonatal sepsis in two neonatal intensive units during a two-year period. METHODS: We observed early-onset (EO-BSI) and late-onset bloodstream infections (LO-BSI) cases in two high-reference neonatal intensive care units (NICU) over a 24-month period (2016-2017). Samples of patients' blood were tested for the presence of the microorganisms. All bacterial isolates were tested for susceptibility to antibiotics. RESULTS: The majority of sepsis cases weighed above 1000 g and were born by cesarean section. About 10% of the EO-BSI group died. There were differences in the EO-BSI /LO-BSI ratio in the compared wards due to differences among the admitted children. The most common pathogens isolated from blood were coagulase-negative staphylococci (CoNS) were represented by two dominating species: S. epidermidis and S. haemolyticus, followed by Klebsiella spp. strains and E.coli, which were mostly found in EO-BSI cases. No single S. agalactiae (GBS) strain was isolated. The majority of CoNS strains were resistant to methicillin, half were resistant to aminoglycosides, and one-third were resistant to macrolides and lincosamides. Half of the Gram-negative rods were resistant to beta-lactams. CONCLUSIONS: The epidemiology of sepsis in two observed NICUs is comparable to data obtained from other studies with a predominance of methicillin-resistant CoNS in LO-BSI and beta-lactam resistant E. coli in EO-BSI. It is of importance that the campaign for controlling GBS carriage in pregnant women in Poland resulted in the disappearance of GBS as a cause of sepsis. Unfortunately, there are no such measures to control E.coli related sepsis.

Epidemiology of Clostridium difficile infection: results of a hospital-based study in Krakow, Poland.

Over the past two decades Clostridium difficile infection (CDI) has appeared as a major public health threat. We performed a retrospective study based on the records of patients hospitalized for CDI at the University Hospital in Krakow, Poland, between 2008 and 2014. In the study period, CDI occurred in 1009 individuals. There were 790 (78%) individuals who developed infection only once, whereas 219 (22%) developed infection more than once. The percentage of deaths within 14 days of CDI confirmation was 2·4%, with a mean age of 74·2 ± 15·9 years. Crude mortality was 12·9% in medical wards, 5·6% for surgical wards and 27·7% in the ICU setting. The time span between diagnosis and death was 5·1 days on average. Between 2008 and 2012 a 6·5-fold increase of CDI frequency with a posterior stabilization and even reduction in 2013 and 2014 was observed. According to the data analysed, 2/3 patients in our population developed CDI during their hospitalization even though they were admitted for different reasons. Medical wards pose a significantly higher risk of CDI than the surgical ones. Age is a risk factor for CDI recurrence. In the case of patients who died, death occurred shortly after diagnosis. The first CDI episode poses much higher risk of mortality than the consecutive ones.

Susceptibility testing and resistance phenotype detection in Staphylococcus aureus strains isolated from patients with atopic dermatitis, with apparent and recurrent skin colonization.

BACKGROUND: Staphylococcus aureus colonization is accepted to be an important triggering factor in patients with atopic dermatitis (AD) and antibiotic resistance has been recognized to be a serious problem as a consequence and for the management of AD treatment. OBJECTIVES: To investigate the antibiotic resistance pattern of S. aureus strains isolated from patients with AD with apparent (lesional and nonlesional skin areas) and recurrent skin colonization and strains obtained from healthy nasal carriers. METHODS: Eighty-seven patients (age 23+/-11.5 years) with mild to severe AD (SCORAD 46.9+/-16.6), 21 patients (age 19.8+/-6.7 years) before antistaphylococcal treatment and 177 healthy nasal carriers (age 27.5+/-8.4 years) were microbiologically assessed for carriage of S. aureus. Colonization of lesional and nonlesional skin areas was quantified by counting the number of colony forming units on the skin surface (log(10) CFU cm(-2)). Antimicrobial susceptibility and resistance phenotypes of 179 S. aureus strains were assessed with the agar disc-diffusion method. RESULTS: Staphylococcus aureus was isolated from 87% of lesional and 44% of nonlesional skin samples from patients with AD. The colonization density of S. aureus was markedly higher in lesional than in nonlesional skin (P<0.001), and was positively correlated with AD severity (P<0.001) and total serum IgE (P<0.05). Patients with AD had a significantly higher prevalence of chloramphenicol-resistant S. aureus than nasal carriers (P<0.01). Similar rates of resistance were expressed to tetracycline, erythromycin, mupirocin, clindamycin and penicillin. Nearly 35% of S. aureus strains from the lesional skin demonstrated different antimicrobial sensitivity pattern compared with strains from nonlesional skin of the same patients with AD. The trend of increasing resistance to chloramphenicol, erythromycin and fusidic acid was observed among S. aureus strains recovered from patients after approximately 75 days of antibiotic treatment. Methicillin-resistant S. aureus isolates were cultured from two patients, one during exacerbation and the other after subsequent bacterial recolonization. CONCLUSIONS: Discrepancies in antibiotic sensitivity pattern were observed among S. aureus strains colonizing different sites of AD skin (lesional and nonlesional areas), and also in AD patients with prior antibiotic treatment. Therefore, clinicians should consider repeat microbial susceptibility testing on different body sites of patients with AD when clinically indicated.

Hospital outbreak of vancomycin-resistant enterococci caused by a single clone of Enterococcus raffinosus and several clones of Enterococcus faecium.

A mixed outbreak caused by vancomycin-resistant Enterococcus raffinosus and Enterococcus faecium carrying the vanA gene was analysed. The outbreak occurred in a large hospital in Poland and affected 27 patients, most of whom were colonised, in three wards, including the haematology unit. The E. raffinosus isolates had a high-level multiresistant phenotype and were initially misidentified as Enterococcus avium; their unambiguous identification was provided by multilocus sequence analysis. The molecular investigation demonstrated the clonal character of the E. raffinosus outbreak and the polyclonal structure of the E. faecium isolates. All of the isolates carried the same Tn1546-like element containing an IS1251-like insertion sequence, located on a c. 50-kb conjugative plasmid. One of the E. faecium clones, found previously to be endemic in the hospital, was probably the source of the plasmid. The results of the study suggest that difficulties in identification may have led to an underestimate of the importance of E. raffinosus in vancomycin-resistant enterococci (VRE) control strategies.

[Proportion of fungi among isolated microorganisms from blood of patients treated in the teaching departments of The University Hospital in Cracow in 1993-1998]. / Udzial grzybów wsród drobnoustrojów izolowanych ze krwi chorych leczonych w oddzialach klinicznych Szpitala Uniwersyteckiego w Krakowie w latach 1993-1998.

The study was carried out of 9742 blood cultures obtained in 1993-1998 from patients of selected departments of the University Hospital. The received samples were assessed from the standpoint of the participation of fungi strains of clinical importance as aetiological factors in systemic infections and the resistance status of a selected group of pathogens. Microbiological blood studies were conducted in the system of continuous monitoring of the obtained cultures by means of the automatic ATB system using a commercially available ID 32C set. ATB Fungus tests were used for drug resistance assessment. In all, 95 strains of yeasts (5.1%) were obtained in cultures, and an increasing variety of these pathogenic species was noted in sepsis. A systematic reduction was noted in the proportion of C. albicans and a steep rise of C. parapsilosis were observed among the aetiological factors of generalized nosocomial fungal infections. A tendency was noted for a continuous rise in the frequency of invasive candidemias with a most significant rise in their proportion in patients in general surgery and haematology departments. Among the fungi isolated from septic complications the proportion of strains resistant to antifungal drugs has been rising.

[Evaluation of resistance patterns of gram-negative rods from personal clinical materials]. / Ocena stanu lekoopornosci paleczek Gram-ujemnych wyosobnionych z materialu klinicznego.

1862 clinical specimens from neonates with infection risk factors treated in the Department of Neonatology, University of Cracow were examined. The aim of this study was to investigate the participation of clinically important Gram-negative rods in hospital infections and to check the resistance patterns of these pathogens. The strains were identified in automatic ATB system using ID 32E and ID 32GN strips with biochemical tests. The susceptibility of isolates of antibacterial agents was determined in automatic ATB system using ATB G- and ATB PSE strips. 436 strains of Gram-negative rods were cultured. 289 strains (66.3%) belonging to Entero-bacteriaceae family and 147 strains (33.7%) non-fermenting rods were isolated. Among Gram-negative aerobic fermenting rods (mainly K. pneumoniae and E. cloacae), increasing resistance to aminoglycosides and beta-lactams, due to new broad spectrum and so called inducible beta-lactamases, was observed. The contribution of non-fermenting rods, especially Pseudomonas spp. and Acinetobacter spp. to the aetiology of infections in hospitalized newborns has increased. Carbapenems and fluoroquinolones are highly active in vitro against the examined strains of multiresistant Gram-negative rods.

[Participation of multiresistant gram-negative rods in systemic infections of newborns treated in the Neonatal Clinics of the University Hospital in Krakow in 1993-1997]. / Udzial wieloopornych paleczek gram-ujemnych w zakaeniach uogólnionych u noworodków leczonych w Klinice Neonatologii Szpitala Uniwersyteckiego w Krakowie w latach 1993-1997.

Results of blood cultures in 1187 newborns hospitalized in the Department of Neonatology, University Hospital in Kraków in furing the last five years were analysed. A trial of monitoring the presence of multi-resistant strains of Enterobacteriaceae with new mechanisms of resistance to antibiotics as well as analysed of their antibiotics susceptibility patterns were undertaken. Surprisingly high percentage of Gram-negative rods E. cloacae and K. pneumoniae strains resistant to the 3rd generation of cephalosporins and participating in septic complications in the investigated hospital environment was demonstrated. There were identified 53 strains of Enterobacter sp., 35 strains of Klebsiella sp. and 3 strains of E. coli extremely resistant to ceftazidine, accounting for 72%, 56% and 5% of all isolated of the given kind, respectively. Dynamic increase in a number of multiresistant strains of K. pneumoniae from 11.1% of isolates in 1993 to 83% in 1997 was shown as well as remaining on a very high level frequency of isolation of Enterobacter sp. rods capable of producing chromosomal cephalosporinases was demonstrated. Full efficacy of carbapenems and high usefulness of ciprofloxacine in the treatment of infections were confirmed. The authors emphasize necessity for constant monitoring of susceptibility to antibiotics in resistant strains as well as for liquidating their sources and ways of their transmission.

Current status of fungal cell wall components in the immunodiagnostics of invasive fungal infections in humans: galactomannan, mannan and (1-->3)-beta-D-glucan antigens.

Early diagnosis of fungal infections and the implementation of appropriate treatment represent major issues for clinicians, nowadays. Histopathological demonstration of microorganisms in tissue specimens or growth of fungal agents in culture media is still considered the "gold standard", but obtaining such specimens may be difficult. Several groups have investigated serological assays for cell wall elements unique to fungal organisms in serum or other body fluids to improve diagnostics in patients with haematological malignancies or undergoing haematopoietic stem-cell transplantation. In this review we have concentrated on the currently available assays allowing for detection of highly immunogenic components of fungal cell wall: galactomannan, mannan, and also (1-->3)-beta-D-glucan. Rapid serological tests appear to be useful for screening high-risk haematological patients, since they allow for the early diagnosis of invasive fungal infections, including infections with the most common pathogens such as Aspergillus and Candida. Based on current literature, factors increasing the probability of obtaining false-positive or false-negative results detected by each test were also analysed and tabulated.

Selection of a teicoplanin-resistant Enterococcus faecium mutant during an outbreak caused by vancomycin-resistant enterococci with the vanB phenotype.

Vancomycin-resistant enterococci (VRE) have recently become an increasing problem in hospitals in Poland, being responsible for a growing number of nosocomial outbreaks. In this work, we have analyzed the second outbreak of VRE with the VanB phenotype to be identified in the country. It was caused by clonal dissemination of a single strain of vancomycin-resistant Enterococcus faecalis (VRES) and horizontal transmission of vancomycin resistance genes among several vancomycin-resistant Enterococcus faecium (VREM) strains. Two similar restriction fragment length polymorphism types of the vanB gene cluster characterized VRES and VREM isolates, and they both contained the same vanB2 variant of the vanB gene. Two vancomycin-susceptible E. faecium (VSEM) isolates, recovered from the same wards during the outbreak, proved to be related to certain VREM isolates and could represent endemic strains that had acquired vancomycin resistance. One VSEM and four VREM isolates, all identified in the same patient, belonged to a single clone, although they revealed remarkable diversity in terms of susceptibility, PFGE patterns, plasmid content, and number of vanB gene cluster copies. Most probably they reflected the dynamic evolution of an E. faecium strain in the course of infection of a single patient. One of the VREM isolates turned out to be resistant to teicoplanin, which coincided with the use of this antibiotic in the patient's therapy. Its vanB gene variant differed by a single mutation from that found in other isolates; however, it also lacked a large part of the vanB gene cluster, including the regulatory genes vanR(B) and -S(B), and the vancomycin-inducible promoter P(YB). Expression of the resistance genes vanH(B), -B, and -X(B) was constitutive in the mutant, and this phenomenon was responsible for its unusual phenotype.