The application of genetics methods to differentiation of three Lactobacillus species of human origin

In recent decades, the interest in probiotics as diet supplements or drugs has increased. In order to determine a specific bacterial isolate to be probiotic, it is necessary to describe precisely its probiotic characteristics and taxonomic properties, including the strain level. Most of the well-known genotyping methods were designed for the commonly-found pathogenic bacteria. The objective of this study is to undertake an attempt at standardization of FISH, RAPD and PFGE methods to genotype and identify the bacteria belonging to Lactobacillus fermentum, L. gasseri and L. plantarum species. The FISH probes have been designed and tested for Lactobacillus fermentum, L. gasseri and L. plantarum species and an endeavor has been made at standardization of RAPD and PFGE methods for these bacterial species. Moreover, the MLST method was applied to differentiate Lactobacillus plantarum strains. L. plantarum isolated from humans could not be genetically diversified with the use of RAPD, PFGE or MLST methods; only the strains originating from plants have displayed diversification among themselves and have been different from the strains of human origin

Problemy identyfikacji oraz koszty zakażeń szpitalnych

Identification difficulties and costs of hospital acquired infectionsThe aim of this work is to analyze epidemiological data collected in the active HI programme between 2002 and 2004 in forty Polish hospitals and to assess some of the economic consequences of hospital infections. The average cumulative incidence, incidence density and mortality of nosocomial pneumoniae (PNEU), bloodstream infections (BSI), urinary tract infections (UTI) and surgical site infections were calculated. A separate analysis of some costs of hospital infections was also performed in this study. The extra costs of stay in hospital overextended due to SSI, PNEU, BSI and UTI, was assessed for three hospitals of different kind. It is very difficult to assess the total costs of HIs in throughout Poland without a specific study designed for this particular purpose. While the need for such research is obvious, this would be a costly and time-consuming task, unlikely to be carried out without engaging the state’s authorities and central budget

Age and other risk factors of pneumonia among residents of Polish long-term care facilities

SummaryBackgroundPneumonia is one of the leading causes of morbidity and mortality in the elderly population. Nursing home-acquired pneumonia (NHAP) is probably the largest health problem in long-term care facilities (LTCFs). It is the second most common infection in LTCFs and frequently requires hospitalization. The aim of this study was to investigate the incidence rate of NHAP among LTCF residents, its microbial etiology, and the frequency of multidrug-resistant microorganisms. Risk factors for NHAP were analyzed.MethodsThis was a prospective study conducted on a group of 217 elderly subjects aged ≥65 years, recruited from the inhabitants of LTCFs, with disabled elderly individuals living in the community serving as controls. Continuous surveillance was carried out from December 1, 2009 to November 30, 2010.ResultsThe incidence rate of NHAP in the observed population of Polish residents was 0.6/1000 resident-days. Vulnerability to NHAP was due to the poor general condition of residents, expressed by low Barthel index values (relative risk (RR) 1.6), the activities of daily living (ADL) score (RR 1.7), the Katz scale (RR 1.2), and limited physical activity (RR 1.6). Also significant were malnutrition (RR 2.3), the use of a bladder catheter (RR 1.3), dysphagia (RR 1.7), tracheotomy tube (RR 3.1), and gastric feeding tube (RR 3.5). Enterobacteriaceae were the predominant etiological agents of NHAP (56.3%).ConclusionsThe significance of risk factors for NHAP among residents in LTCFs was confirmed. Unfortunately, we also found that a lack of proper supervision with regard to the microbiology of infections is characteristic of Polish health care and LTCFs. There is an opportunity to improve the medical care of patients with severe disabilities, limit the rise in antimicrobial resistance and the need for hospitalization, and improve the prognosis

Possible role of Escherichia coli in propagation and perpetuation of chronic inflammation in ulcerative colitis

BACKGROUND: This study investigated a possible role of Escherichia coli in propagation and perpetuation of the chronic inflammation in ulcerative colitis (UC). The lesions of UC are located superficially on the rectal and/or colonic mucosa. It is suggested that the commensal bacteria of the digestive tract may play a role in the pathogenesis of UC. Several studies have demonstrated proliferation of E. coli in the gut of UC patients. An increase in the number of E. coli in the inflamed tissue is most probably related to the abundance of iron ions produced by the bacteria. METHODS: Colon mucosal biopsies were collected from 30 patients with acute-phase UC, both from tissues with inflammatory changes (n = 30) and unchanged tissue with no inflammatory changes (n = 30) from the same patient. Biopsies were also taken from 16 patients with irritable bowel syndrome diarrhea who comprised the control group. Quantitative and qualitative analysis of the biopsy specimens was performed using culture methods and real-time polymerase chain reaction (PCR). Genotyping of the E. coli isolates was done using pulsed-field gel electrophoresis. Multiplex PCR was used to compare the E. coli strains for the presence of genes responsible for synthesis of iron acquisition proteins: iroN, iutA, iha, ireA, chuA, and hlyA. RESULTS: We demonstrated that there was a significant increase in the number of E. coli at the sites of inflammation in patients with UC compared to the control group (P = 0.031). Comparative analysis of the restriction patterns of E. coli isolated from inflammatory and unchanged tissues showed that the local inflammatory changes did not promote specific E. coli strains. There was a significant difference in the frequency of the iroN gene in E. coli isolated from patients with UC as compared to the control group. CONCLUSIONS: The increase in the numbers of E. coli in the inflammatory tissues is related to the presence of chuA and iutA genes, which facilitate iron acquisition during chronic intestinal inflammatory processes

Zakażenia szpitalne oraz nadzór po zakończeniu hospitalizacji w wybranych procedurach chirurgii naczyń

Background. Nosocomial infections are an unwanted complication present in almost every hospital. Among patients of highly specialized departments, including vascular surgery wards, the most prevalent form of clinical infection are Surgical Site Infections (SSIs). SSIs significantly extend the patient's stay in the hospital, increase costs and above all can be the direct cause of death after vascular procedures. Material and methods. This paper presents the results of studies on infection epidemiology in vascular surgery in the years 2002-2006 (9,573 surgical procedures) as well as a targeted study which included 413 patients of two highly specialized wards in the year 2005. The registration of infections was based on definitions formulated on the basis of CDC guidelines, and included postdischarge cases. Results. Within the Active Surveillance System, 207 cases of SSI have been identified (incidence 2.2%) and 99 cases of hospital acquired pneumonia (incidence 1.0%); other forms of infection accounted for 21 cases. The overall incidence among patients undergoing surgical procedures (vascular surgery) was 3.4%. The targeted study allowed for a detailed analysis of SSI incidence with consideration of the SSI standardized infection ratio. The cumulated incidence rate of SSIs was 2.6% in site I and 5.6% in site II. Among the etiological factors isolated from materials taken from patients with symptoms of SSI, staphylococci were in the majority, among them Stapylococcus aureus. In the targeted study, almost half of the isolated S. aureus strains (40%) manifested a resistance to methicillin (MRSA) as well as macrolides, lincosamides and streptogramin B (MLSβ). Conclusions. The Active Infection Surveillance System confirmed that in the patients of the studied population the most prevalent form of infection was SSI; however, a high incidence of hospital acquired pneumonia was also noticed. In the targeted study, the SSI incidence in site no. I was within the expected values. The possibility of performing postdischarge registration of infections was confirmed as well as the purposefulness of systematic infection control conducted by experienced personnel. The study showed the need for an improvement in microbiological surveillance in infection control, as well as the necessity of performing further detailed studies regarding the relationship of the operating team experience to the incidence of SSIs within Polish vascular surgery wards. We claim that in certain cases the division, together with the later reconstruction, of the left renal vein may facilitate the course of reconstructive operation of the abdominal aorta and decreases the risk of haemo-rrhage complications. Moreover, this manoeuvre does not influence postoperative renal function.Wstęp. Zakażenia szpitalne są niepożądanym powikłaniem występującym praktycznie w każdym szpitalu. Wśród pacjentów oddziałów wysokospecjalistycznych, w tym w chirurgii naczyń, dominującą formę kliniczną tych zakażeń stanowią zakażenia miejsca operowanego. W istotny sposób przedłużają one pobyt pacjenta w szpitalu, zwiększają koszty, a przede wszystkim mogą być bezpośrednią przyczyną śmierci chorych po zabiegu w zakresie naczyń. Materiał i metody. Zaprezentowano wyniki badań dotyczących epidemiologii zakażeń w chirurgii naczyniowej w latach 2002-2006 (9573 zabiegi operacyjne) oraz badania celowanego, którym objęto łącznie 413 pacjentów dwóch oddziałów wysokospecjalistycznych w 2005 roku. Rejestracja zakażeń opierała się na definicjach opracowanych na podstawie zaleceń Centers for Disease Control and Prevention (CDC) i uwzględniała przypadki wykryte po zakończeniu hospitalizacji. Wyniki. Wśród dominujących zakażeń szpitalnych w ramach Systemu Czynnego Nadzoru rozpoznano 207 przypadków zakażenia miejsca operowanego (zachorowalność 2,2%) i 99 szpitalnych zapaleń płuc (zachorowalność 1,0%), inne formy zakażeń stanowiły 21 przypadków. Zachorowalność w grupie pacjentów operowanych - chirurgia naczyniowa - wyniosła ogółem: 3,4%. Badanie celowane umożliwiło szczegółową analizę zachorowalności zakażeń miejsc operowanych z uwzględnieniem standaryzowanego indeksu ryzyka. Współczynniki zachorowalności skumulowanej zakażeń miejsc operowanych wyznaczono na poziomie 2,6% w ośrodku I oraz 5,6% w ośrodku II. Wśród czynników etiologicznych izolowanych z materiałów pochodzących od pacjentów z objawami zakażenia miejsca operowanego dominowały gronkowce, a wśród nich gatunek Stapylococcus aureus. W badaniu celowanym prawie połowa z izolowanych szczepów S. aureus (40%) to szczepy oporne na metycylinę (MRSA) oraz makrolidy, linkozamidy i streptograminy B (MLSb). Wnioski. System Czynnego Nadzoru potwierdził, że w badanej populacji pacjentów dominującą formą zakażenia jest zakażenie miejsca operowanego, jednak zaobserwowano również wysoką zachorowalność dotyczącą szpitalnych zapaleń płuc. W badaniu celowanym stwierdzona zachorowalność zakażenia miejsca operowanego w przypadku I pierwszego badanego ośrodka odpowiadała oczekiwanym wartościom. Potwierdzono możliwość prowadzenia rejestracji zakażeń manifestujących się po zakończeniu hospitalizacji chorego oraz celowość systematycznej kontroli zakażeń prowadzonej przez doświadczony personel. Wykazano konieczność poprawy wykorzystania nadzoru mikrobiologicznego w kontroli zakażeń oraz dalszych szczegółowych badań w zakresie związku pomiędzy doświadczeniem zespołu operującego a zachorowalnością na oddziałach chirurgii naczyniowej w Polsce

Virulence factors of Enterococcus strains isolated from patients with inflammatory bowel disease

Author contributions: Golińska E performed the majority of the experiments, including the detection of gelatinase activity, measurement of hydrogen peroxide production and the determination of hydrogen peroxide decomposition, and wrote the manuscript; Tomusiak A collected and analysed the data; Gosiewski T performed PCR and multiplex PCR; Więcek G evaluated the adherence to human gut epithelium cells; Machul A and Mikołajczyk D evaluated the biofilm production; Heczko PB and Bulanda M supervised the experiments; and Strus M designed the experiments and supervised the project. Abstract AIM: To determine the features of Enterococcus that contribute to the development and maintenance of the inflammatory process in patients with inflammatory bowel disease (IBD). METHODS: Multiplex polymerase chain reaction (PCR) was applied to assess the presence of genes that encode virulence factors [surface aggregating protein (asa1 ), gelatinase (gelE ), cytolysin (cylA ), extracellular surface protein (esp ) and hyaluronidase (hyl )] in the genomic DNA of 28 strains of Enterococcus isolated from the intestinal tissues of children with IBD (n = 16) and of children without IBD (controls; n = 12). Additionally, strains with confirmed presence of the gelE gene were tested by PCR for the presence of quorum sensing genes (fsrA , fsrB , fsrC ) that control the gelatinase production. Gelatinase activity was tested on agar plates containing 1.6% gelatin. We also analysed the ability of Enterococcus strains to release and decompose hydrogen peroxide (using Analytical Merckoquant peroxide test strips) and tested their ability to adhere to Caco-2 human gut epithelium cells and form biofilms in vitro . RESULTS: A comparison of the genomes of Enterococcus strains isolated from the inflamed mucosa of patients with IBD with those of the control group showed statistically significant differences in the frequency of the asa1 gene and the gelE gene. Furthermore, the cumulative occurrence of different virulence genes in the genome of a single strain of Enterococcus isolated from the IBD patient group is greater than in a strain from the control group, although no significant difference was found. Statistically significant differences in the decomposition of hydrogen peroxide and adherence to the Caco-2 epithelial cell line between the strains from the patient group and control group were demonstrated. The results also showed that profuse biofilm production was more frequent among Enterococcus strains isolated from children with IBD than in control strains. CONCLUSION: Enterococcus strains that adhere strongly to the intestinal epithelium, form biofilms and possess antioxidant defence mechanisms seem to have the greatest influence on the inflammatory process

Late-onset bloodstream infections of Very-Low-Birth-Weight infants: data from the Polish Neonatology Surveillance Network in 2009–2011

BACKGROUND: Late-Onset Bloodstream Infections (LO-BSI) continue to be one of the most important complications associated with hospitalization of infants born with very low birth weight (VLBW). The aims of this study were to assess the epidemiology of LO-BSI together with the risk factors and the distribution of causative pathogens at six Polish neonatal intensive care units that participated in the Polish Neonatology Surveillance Network from January 1, 2009 to December 31, 2011. METHODS: The surveillance covered 1,695 infants whose birth weights were <1501 grams (VLBW) in whom LO-BSI was diagnosed >72 hours after delivery. Case LO-BSI patients were defined according to NeoKISS. RESULTS: Four hundred twenty seven episodes of LO-BSI were diagnosed with a frequency of 25.3% and an incidence density of 6.7/1000 patient-days (pds). Results of our multivariate analysis demonstrated that surgical procedures and lower gestational age were significantly associated with the risk of LO-BSI. Intravascular catheters were used in infants with LO-BSI significantly more frequently and/or for longer duration: Central venous cathters (CVC) (OR 1.29) and Peripheral venous catheters (PVC) (OR 2.8), as well as, the total duration of total parenteral nutrition (13 vs. 29 days; OR 1.81). Occurrence of LO-BSI was significantly associated with increased the length of mechanical ventilation (MV) (OR 2.65) or the continuous positive airway pressure (CPAP) (OR 2.51), as well as, the duration of antibiotic use (OR 2.98). The occurrence of more than one infection was observed frequently (OR 9.2) with VLBW with LO-BSI. Microorganisms isolated in infants with LO-BSI were dominated by Gram-positive cocci, and predominantly by coagulase-negative staphylococci (62.5%). CONCLUSIONS: Independent risk factor for LO-BSI in VLBV infants are: low gestational age and requirement for surgery. The incidence rates of LO-BSI especially CVC-BSI were higher in the Polish NICUs surveillance than those of other national networks, similar to the central- and peripheral utilization ratio