Publications by authors named "Malgorzata Bulanda"

84 Publications

Physicochemical and microbiological characteristics of urban aerosols in Krakow (Poland) and their potential health impact.

Environ Geochem Health 2021 Apr 29. Epub 2021 Apr 29.

Institute of Geological Sciences, Jagiellonian University, Ul. Gronostajowa 3a, 30-387, Kraków, Poland.

Eight aerosol samples were collected in Krakow using a low-volume sampler in February and March 2019 during variable meteorological conditions and times of the day, to study their single particles' properties (size, morphology and chemical composition analyzed using a scanning electron microscope fitted with an energy-dispersive spectrometer) and microbiological characteristics. The content of particles of different chemical compositions larger than 2.5 μm was low. Considering the number of the particles, submicron particles strongly dominated with a high content of ultrafine particles (nanoparticles). Tar ball-type particles were relatively common in the studied samples, while soot was the dominant component. Soot was present as small agglomerates composed of few particles, but also as bigger agglomerates. Metal-containing particles of various chemical characteristics were abundant, with transition metals commonly occurring in these particles. The physicochemical characteristics of aerosols indicate that despite a relatively low mass concentration, their adverse health impact could be very strong because of the high content of nanoparticles, the abundance of soot and other fuel combustion-related particles, and the high incidence of transition metal-rich particles. Microbiological analysis was based on cultures on both solid and liquid agar. The MALDI-TOF method was used for species identification-for bacteria and fungi. Twelve different species of bacteria were isolated from the collected samples of aerosols. The most frequently isolated species was Gram-positive sporulating Bacillus licheniformis. The isolated mold fungi were of the genus Aspergillus.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s10653-021-00950-xDOI Listing
April 2021

Clonal Dissemination of KPC-2, VIM-1, OXA-48-Producing ST147 in Katowice, Poland.

Pol J Microbiol 2021 Mar 19;70(1):107-116. Epub 2021 Mar 19.

Department of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, Krakow, Poland.

Carbapenem-resistant (CRKP) is an important bacterium of nosocomial infections. In this study, CRKP strains, which were mainly isolated from fecal samples of 14 patients in three wards of the hospital in the Silesia Voivodship, rapidly increased from February to August 2018. Therefore, we conducted microbiological and molecular studies of the CRKP isolates analyzed. Colonized patients had critical underlying diseases and comorbidities; one developed bloodstream infection, and five died (33.3%). Antibiotic susceptibilities were determined by the E-test method. A disc synergy test confirmed carbapenemase production. CTX-Mplex PCR evaluated the presence of resistance genes , , , and the genes , , , , , , and was detected with the PCR method. Clonality was evaluated by Multi Locus Sequence Typing (MLST) and Pulsed Field Gel Electrophoresis (PFGE). Six (40%) strains were of XDR (Extensively Drug-Resistant) phenotype, and nine (60%) of the isolates exhibited MDR (Multidrug-Resistant) phenotype. The range of carbapenem minimal inhibitory concentrations (MICs, μg/mL) was as follows doripenem (16 to >32), ertapenem (> 32), imipenem (4 to > 32), and meropenem (> 32). PCR and sequencing confirmed the , , , and genes in all strains. The isolates formed one large PFGE cluster (clone A). MLST assigned them to the emerging high-risk clone of ST147 (CC147) pandemic lineage harboring the gene. This study showed that the isolates detected in the multi-profile medical centre in Katowice represented a single strain of the microorganism spreading in the hospital environment.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.33073/pjm-2021-010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8008758PMC
March 2021

Analysis of the Gut Mycobiome in Adult Patients with Type 1 and Type 2 Diabetes Using Next-Generation Sequencing (NGS) with Increased Sensitivity-Pilot Study.

Nutrients 2021 Mar 25;13(4). Epub 2021 Mar 25.

Department of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, 18 Czysta Street, 31-121 Krakow, Poland.

The studies on microbiome in the human digestive tract indicate that fungi could also be one of the external factors affecting development of diabetes. The aim of this study was to evaluate the quantitative and qualitative mycobiome composition in the colon of the adults with type 1 (T1D), = 26 and type 2 (T2D) diabetes, = 24 compared to the control group, = 26. The gut mycobiome was characterized in the stool samples using the analysis of the whole internal transcribed spacer (ITS) region of the fungal rDNA gene cluster by next-generation sequencing (NGS) with increased sensitivity. At the L2 (phylum) level, Basidiomycota fungi were predominant in all 3 study groups. Group T1D presented significantly lower number of Ascomycota compared to the T2D group, and at the L6 (genus) level, the T1D group presented significantly lower number of genus compared to control and T2D groups. In the T1D group, a significant positive correlation between total cholesterol and low-density lipoprotein cholesterol (LDL-C) levels and fungi of the genus and in the T2D group, a negative correlation between the total cholesterol level and genus was found. The obtained results seem to be a good foundation to extend the analysis of the relationship between individual genera and species of fungi and the parameters determining the metabolism of carbohydrates and lipids in the human body.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/nu13041066DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8064496PMC
March 2021

Investigation of Activity in Vascular Surgery Units through Epidemiological Management Based on the Analysis of Antimicrobial Resistance, Biofilm Formation and Genotyping.

Int J Environ Res Public Health 2021 02 7;18(4). Epub 2021 Feb 7.

Jagiellonian University Medical College, Faculty of Medicine, Chair of Microbiology, Department of Bacteriology, Microbial Ecology and Parasitology, 31-008 Kraków, Poland.

Background/objectives: The genus demonstrates resistance to antibiotics and has been shown to spread in the hospital environment causing epidemic outbreaks among hospitalized patients. The objectives of the present study was to investigate the antibiotic resistance, biofilm formation, and clonality among strains.

Materials And Methods: The study involved 6 (I Outbreak) and 3 (II Outbreak) strains isolated from patients hospitalized in vascular surgery unit.

Results: All tested strains were extensively drug resistant (XDR) and all the isolates were carbapenem-resistant and among them, all carried the gene, the gene, as well as the gene. All of the investigated strains had the ability to form a biofilm, but all of them produced less biofilm than the reference strain. Multi-locus sequence typing (MLST) showed that all strains belonged to the ST2 clone. Pulsed-field gel electrophoresis (PFGE) divided the tested outbreak strains into two clones (A and B).

Conclusion: This study shows a nosocomial spread of XDR ST2 having the gene, the gene, as well as the gene, low biofilm formers, that was prevalent in the vascular surgery unit. To identify the current situation of vascular surgery departments targeted epidemiological investigation was needed. Effective implementation of infection control prevented the spread of the epidemic outbreaks.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/ijerph18041563DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7915860PMC
February 2021

The Gut Microbiota Profile According to Glycemic Control in Type 1 Diabetes Patients Treated with Personal Insulin Pumps.

Microorganisms 2021 Jan 12;9(1). Epub 2021 Jan 12.

Department of Metabolic Diseases, Jagiellonian University Medical College, 2 Jakubowskiego Street, 30-688 Krakow, Poland.

Recently, several studies explored associations between type 1 diabetes (T1DM) and microbiota. The aim of our study was to assess the colonic microbiota structure according to the metabolic control in T1DM patients treated with insulin pumps. We studied 89 T1DM patients (50.6% women) at the median age of 25 (IQR, 22-29) years. Pielou's evenness ( = 0.02), and Shannon's ( = 0.04) and Simpson's diversity indexes ( = 0.01), were higher in patients with glycosylated hemoglobin (HbA1c) ≥ 53 mmol/mol (7%). There were no differences in beta diversity between groups. A linear discriminant analysis effect size (LEfSe) algorithm showed that one family () was enriched in patients with HbA1c < 53 mmol/mol, whereas one family () and four species (, unclassified species of , , and ) were enriched in patients with HbA1c ≥ 53 mmol/mol. We found that at class level, the following pathways according to Kyoto Encyclopedia of Genes and Genomes were enriched in patients with HbA1c < 53 mmol/mol: bacterial motility proteins, secretion system, bacterial secretion system, ribosome biogenesis, translation proteins, and lipid biosynthesis, whereas in patients with HbA1c ≥ 53 mmol/mol, the galactose metabolism, oxidative phosphorylation, phosphotransferase system, fructose, and mannose metabolism were enriched. Observed differences in alpha diversity, metabolic pathways, and associations between bacteria and HbA1c in colonic flora need further investigation.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/microorganisms9010155DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7826603PMC
January 2021

Metagenomic Analysis of Duodenal Microbiota Reveals a Potential Biomarker of Dysbiosis in the Course of Obesity and Type 2 Diabetes: A Pilot Study.

J Clin Med 2020 Jan 29;9(2). Epub 2020 Jan 29.

Department of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, 31-121 Krakow, Poland.

Numerous scientific studies confirm that, apart from environmental and genetic factors, a significant role is played by gastrointestinal microbiota in the aetiology of type 2 diabetes and obesity. Currently, scientists mainly focus on the distal intestinal microbiota, while the equally important proximal parts of the intestine are overlooked. The aim of the study was a qualitative analysis of the structure of the duodenal mucosa microbiota in groups of patients with obesity and with type 2 diabetes and where obesity qualified for bariatric surgery: sleeve gastrectomy. The microbiological results obtained were compared with some clinical parameters. As a result, it was possible to determine the microbiological core that the treatment and control groups had in common, including phyla: Firmicutes, Proteobacteria, and Actinobacteria. The patients with obesity and with type 2 diabetes and obesity presented a significantly lower number of genus compared to healthy subjects. Furthermore, the numbers of were positively correlated with the high density lipoprotein (HDL) concentration in the groups under study. The obtained results indicate that bacteria of the genus should be considered in the future in the context of a potential biomarker in the progress of type 2 diabetes and obesity.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/jcm9020369DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7074165PMC
January 2020

Evidence for Infections by the Same Strain of Beta 2-toxigenic Type A Acquired in One Hospital Ward.

Pol J Microbiol 2019 Sep 3;68(3):323-329. Epub 2019 Sep 3.

Department of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College , Krakow , Poland.

This study conducts a comparative phenotypic and genetic analysis of strains isolated from two patients hospitalized at the same time in 2017 in the surgical ward of the Provincial Specialist Hospital in Włocławek (Kujawsko-Pomorskie Province) who developed necrotizing soft tissue infections (NSTI). To explain the recurring cases of this infection, a comparative analysis was performed for these strains and the ones originating from infections recorded at the same hospital in three patients with gas gangrene in 2015. The two isolates studied in 2017 (8554/M/17 from patient No. 1 and 8567/M/17 from patient No. 2) had identical biochemical profiles. A comparison of research results using multiplex PCR from 2017 with a genetic analysis of strains from 2015 enabled us to demonstrate that the strains currently studied have the genes encoding the same toxins (α and β2) as the two strains analyzed in 2015: no. 7143 (patient No. 3) and no. 7149 (patient No. 2). A comparative analysis of the strain profiles obtained with pulsed-field gel electrophoresis (PFGE) in 2017 with the results from 2015 has found one identical and genetically unique restriction profile, corresponding to one clone of comprising of two strains: no. 8567/M/17 (patient No. 2 in 2017) and no. 7143 (patient No. 3 in 2015). The epidemiological data and detailed analysis of the course of both events suggest that this clone of possibly survived in adverse conditions of the external environment in the operating block of this hospital for many months.

This study conducts a comparative phenotypic and genetic analysis of strains isolated from two patients hospitalized at the same time in 2017 in the surgical ward of the Provincial Specialist Hospital in Włocławek (Kujawsko-Pomorskie Province) who developed necrotizing soft tissue infections (NSTI). To explain the recurring cases of this infection, a comparative analysis was performed for these strains and the ones originating from infections recorded at the same hospital in three patients with gas gangrene in 2015. The two isolates studied in 2017 (8554/M/17 from patient No. 1 and 8567/M/17 from patient No. 2) had identical biochemical profiles. A comparison of research results using multiplex PCR from 2017 with a genetic analysis of strains from 2015 enabled us to demonstrate that the strains currently studied have the genes encoding the same toxins (α and β2) as the two strains analyzed in 2015: no. 7143 (patient No. 3) and no. 7149 (patient No. 2). A comparative analysis of the strain profiles obtained with pulsed-field gel electrophoresis (PFGE) in 2017 with the results from 2015 has found one identical and genetically unique restriction profile, corresponding to one clone of comprising of two strains: no. 8567/M/17 (patient No. 2 in 2017) and no. 7143 (patient No. 3 in 2015). The epidemiological data and detailed analysis of the course of both events suggest that this clone of possibly survived in adverse conditions of the external environment in the operating block of this hospital for many months.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.33073/pjm-2019-035DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7256698PMC
September 2019

Comparison of PCR, Fluorescent in Situ Hybridization and Blood Cultures for Detection of Bacteremia in Children and Adolescents During Antibiotic Therapy.

Pol J Microbiol 2018;67(4):479-486

Chair of Microbiology, Department of Molecular Medical Microbiology, Faculty of Medicine, Jagiellonian University Medical College , Cracow , Poland.

The gold standard in microbiological diagnostics of bacteremia is a blood culture in automated systems. This method may take several days and has low sensitivity. New screening methods that could quickly reveal the presence of bacteria would be extremely useful. The objective of this study was to estimate the effectiveness of these methods with respect to blood cultures in the context of antibiotic therapy. Blood samples from 92 children with sepsis were analyzed. Blood cultures were carried out in standard automated systems. Subsequently, FISH (Fluorescent In-Situ Hybridization) and nested multiplex-real-time-PCR (PCR) were performed. Blood cultures, FISH and PCR yielded positive results in 18%, 39.1%, and 71.7% of samples, respectively. Significant differences were found between the results obtained through culture before and after induction of antibiotherapy: 25.5% vs. 9.7%. There was no significant difference in FISH and PCR results in relation to antibiotics. The three methods employed demonstrated significant differences in detecting bacteria effectively. Time to obtain test results for FISH and PCR averaged 4-5 hours. FISH and PCR allow to detect bacteria in blood without prior culture. These methods had high sensitivity for the detection of bacteremia regardless of antibiotherapy. They provide more timely results as compared to automated blood culture, and may be useful as rapid screening tests in sepsis.

The gold standard in microbiological diagnostics of bacteremia is a blood culture in automated systems. This method may take several days and has low sensitivity. New screening methods that could quickly reveal the presence of bacteria would be extremely useful. The objective of this study was to estimate the effectiveness of these methods with respect to blood cultures in the context of antibiotic therapy. Blood samples from 92 children with sepsis were analyzed. Blood cultures were carried out in standard automated systems. Subsequently, FISH (Fluorescent In-Situ Hybridization) and nested multiplex-real-time-PCR (PCR) were performed. Blood cultures, FISH and PCR yielded positive results in 18%, 39.1%, and 71.7% of samples, respectively. Significant differences were found between the results obtained through culture before and after induction of antibiotherapy: 25.5% vs. 9.7%. There was no significant difference in FISH and PCR results in relation to antibiotics. The three methods employed demonstrated significant differences in detecting bacteria effectively. Time to obtain test results for FISH and PCR averaged 4–5 hours. FISH and PCR allow to detect bacteria in blood without prior culture. These methods had high sensitivity for the detection of bacteremia regardless of antibiotherapy. They provide more timely results as compared to automated blood culture, and may be useful as rapid screening tests in sepsis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.21307/pjm-2018-056DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7256870PMC
June 2019

Polish infection control nurses - Self-assessment of their duties and professional autonomy in different types of hospitals.

Med Pr 2018 12 23;69(6):605-612. Epub 2018 Nov 23.

Jagiellonian University Medical College, Kraków, Poland (Faculty of Medicine, Chair of Microbiology).

Background: The objective of the study is self-assessment of Polish infection control nurses (ICNs) in terms of the structure of professional tasks and autonomy of decision-making.

Material And Methods: A questionnaire survey was filled out by 208 ICNs (around 21% of all Polish ICNs) in 15 provinces located in Poland. The research encompassed ICNs surveillance healthcare-associated infections (HAIs) in 2014.

Results: The work time that ICNs devote to professional tasks and decision autonomy on the scale of 1-100% was as follows: 34% (67% of decision autonomy) was dedicated to detecting and registering HAIs, 12% (71%) - internal control, 10% (58%) - devising and implementing infection prevention practices, 10% (68%) - staff trainings, 8% (65%) - identification and study of outbreaks, 7% (58%) - promoting hand hygiene, 6% (51%) - consults with infected patients, 4% (57%) - consults on decontamination, 4% (54%) - consults on maintaining cleanliness, 3% (51%) - isolation and application of personal protective measures, 2% - other tasks. Infection prevention and control nurses estimated, on average, that their autonomy of decisions concerning the professional tasks performed amounted to 60%.

Conclusions: Infection control nurses in Poland have difficulty in achieving balance between tasks they perform and the authority they exercise. The ICN professional task structure is dominated by duties associated with monitoring hospital infections, however, the greatest decision autonomy is visible regarding internal control. Decision-making concentrated on internal control may hinder building a positive image of an ICN. We should strive to firmly establish professional tasks and rights of ICNs in legislation concerning performing the duties of a nurse and midwife. Med Pr 2018;69(6):605-612.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.13075/mp.5893.00719DOI Listing
December 2018

Component-resolved diagnosis (CRD) of food anaphylaxis

Folia Med Cracov 2018;58(2):89-102

Department of Clinical and Environmental Allergology, Faculty of Medicine Jagiellonian University Medical College, Śniadeckich 10, Kraków, Poland.

Anaphylaxis is an increasing problem in public health. The food allergens (mainly milk, eggs, and peanuts) are the most frequent cause of anaphylaxis in children and youth. In order to define the cause of anaphylaxis, skin tests, the determination of the concentration of specific IgE in the blood and basophil activation test are conducted. In vitro tests are preferred due to the risk of allergic response during in vivo tests. Component-resolved diagnosis (CRD) is an additional tool in allergology, recommended in the third level of diagnostics when there are diagnostic doubts after the above mentioned tests have been carried out. The paper presents 3 cases of patients with anaphylactic response, and the application of CRD in these patients helped in planning the treatment. Patient 1 is a 4-year-old boy with diagnosed atopic dermatitis and bronchial asthma reported an anaphylactic shock at the age of seven months caused by cow's milk and the exacerbation of bronchial asthma after eating some fruit. Patient 2 is a 35-year-old woman who has had anaphylactic shock three times: in June 2015, 2016, and 2017 and associates these episodes with the consumption of dumplings with a caramel, bun, and the last episode took place during physical exertion few hours after eating waffle. Patient 3 is a 26-year-old man with one-time loss of consciousness after eating mixed nuts and drinking beer. CRD offers the possibility to conduct a detailed diagnostic evaluation of patients with a history of anaphylactic reaction.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.24425/fmc.2018.124661DOI Listing
September 2019

Epitopes of Immunoreactive Proteins of : Enolase, Inosine 5'-Monophosphate Dehydrogenase and Molecular Chaperone GroEL.

Front Cell Infect Microbiol 2018 2;8:349. Epub 2018 Oct 2.

Department of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, Krakow, Poland.

Three (group B streptococci, GBS) immunoreactive proteins: enolase (47.4 kDa), inosine 5'-monophosphate dehydrogenase (IMPDH) (53 kDa) and molecular chaperone GroEL (57 kDa) were subjected to investigation. Enolase protein was described in our previous paper, whereas IMPDH and GroEL were presented for the first time. The aim of our paper was to provide mapping of specific epitopes, highly reactive with umbilical cord blood serum. Bioinformatic analyses allowed to select 32 most likely epitopes for enolase, 36 peptides for IMPDH and 41 immunoreactive peptides for molecular chaperone GroEL, which were synthesized by PEPSCAN. Ten peptides: two in enolase, one in IMPDH and seven in molecular chaperone GroEL have been identified as potentially highly selective epitopes that can be used as markers in rapid immunological diagnostic tests or constitute a component of an innovative vaccine against GBS infections.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fcimb.2018.00349DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6176014PMC
September 2019

The dynamics of vaginal and rectal Lactobacillus spp. flora in subsequent trimesters of pregnancy in healthy Polish women, assessed using the Sanger sequencing method.

BMC Pregnancy Childbirth 2018 Aug 29;18(1):350. Epub 2018 Aug 29.

Department of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, Czysta 18, 31-121, Krakow, Poland.

Background: Lactobacilli play an important role in maintaining vaginal health and protection against bacterial infections in the genital tract. The aim of this study is to show the dynamics of changes of the vaginal and rectal Lactobacillus flora during pregnancy by using the Sanger sequencing method.

Method: The study included 31 healthy pregnant women without clinical signs of genitourinary infections. The material was taken in the three trimesters of pregnancy by vaginal and rectal swabs and grown on the MRS agar quantitatively to estimate the number of Lactobacillus spp. [CFU/ml]. Afterwards, 3 to 8 morphologically different lactobacilli colonies were taken for identification. Bacterial species identification was performed by 16 s rDNA sequence fragment analyses using the Sanger method.

Results: Among the patients tested, the most common species colonizing the vagina in the first trimester were: L. crispatus 29%, L. gasseri 19.4% and L. rhamnosus 16.1%, in the second trimester: L. crispatus 51.6%, L. gasseri 25.8%, L. rhamnosus 19.4% and L. amylovorus 16.1%, and in the third trimester the most common Lactobacillus species were: L. crispatus 25.8%, L. gasseri 25.8% and L. johnsonii 19.4%. In rectal species, the number decreased in the second and third trimesters in comparison to the first trimester (p = 0.003). An analysis of rectal dynamics showed that in the first trimester, the most common species were: L. johnsonii 19.4%, and L. plantarum 9.7%, in the second trimester: L. crispatus 9.7% and L. mucosae 6.5%, and in the third trimester: L. casei 9.7% and L. rhamnosus 9.7%. Individual dynamics of the Lactobacillus species composition showed variability, characterized by continuous, intermittent, or periodic colonization. The patients examined were mostly colonized by three Lactobacillus species in vagina (32.3%), whereas for the rectum, one Lactobacillus species during the whole pregnancy duration was common (32.3%).

Conclusion: This study showed that in the examined group of healthy, pregnant Polish women, the vaginal Lactobacillus flora, both qualitative and quantitative, was stable during the three subsequent trimesters. In contrast, the number of rectal Lactobacillus species dramatically decreased after the first trimester.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s12884-018-1987-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6114255PMC
August 2018

Sternal wound infections following cardiac surgery and their management: a single-centre study from the years 2016-2017.

Kardiochir Torakochirurgia Pol 2018 Jun 25;15(2):79-85. Epub 2018 Jun 25.

Center for Innovative Laboratory Diagnostics, John Paul II Hospital, Krakow, Poland.

Introduction: Despite improvements in surgical management, sternal wound infection (SWI) following cardiac surgery remains a significant complication.

Aim: To evaluate pathogens involved in SWI following median sternotomy.

Material And Methods: We enrolled 164 patients who experienced prolonged sternal wound healing following open-heart surgery. The inclusion criteria were as follows: prolonged sternal wound healing following cardiac surgery via median sternotomy and complete results of microbiological culture obtained from the sternal swab. The exclusion criteria were as follows: partial sternotomy, patients with mechanical sternum dehiscence and incomplete clinical data. Swabs provided information on the type of microorganism present in the wound and the susceptibility of the microorganism to specific antibiotics.

Results: One hundred and fourteen (69.5%) patients aged 68 (interquartile range: 60-76) years developed SWI with positive culture. The in-hospital mortality rate was 21.0% in this group. The most common pathogens included (26.6%), (12.2%), (11.4%), (8.4%), and (6.8%). In most of the cases the isolated was methicillin-resistant ( = 57, 43.5%). was vancomycin-resistant in 11 (8.3%) cases and high-level aminoglycoside resistant (HLAR) in 13 (9.9%) cases. had the extended-spectrum β-lactamase (ESBL) mechanism in 14 (10.6%) cases. Among 114 culture-positive patients, 48 (42.1%) had polymicrobial infection of the sternal wound.

Conclusions: Microorganisms associated with SWI in our study were mainly commensals, with the most common pathogen being
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.5114/kitp.2018.76472DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6066679PMC
June 2018

Epidemiology of healthcare-associated infections in Polish intensive care. A multicenter study based on active surveillance.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2018 Sep 1;162(3):190-197. Epub 2018 Mar 1.

Polish Society of Hospital Infections Team (listed at the end of the article).

Aim: The aim of this study was to evaluate the incidence of health care-associated infections (HAIs) in patients treated in Polish intensive care units (ICUs).

Methods: This retrospective analysis was based on the results of active targeted surveillance, according to the recommendation of the ECDC (European Centre of Disease Control and Prevention, HAI-Net light protocol), conducted in 2013-2015 in seven ICUs for adults located in southern Poland (observational study).

Results: The incidence of HAI was 22.6% and 28.7/1000 person-days (pds). The incidence of pneumonia (PN) was 8.0%, bloodstream infections (BSIs) 7.2% and urinary tract infections (UTIs) 3.7%. The incidence per 1000 pds was as follows: PN 10.2, BSIs 9.2 and UTIs 4.7. PN was the most common source of secondary bloodstream infection (45%); the second was UTIs (22%). Mortality (directly and indirectly) associated with HAI was 10.8% and was related to the presence of PN or primary BSIs. HAIs were usually (69.2%) caused by Gram-negative bacteria; Klebsiella spp. and nonfermenting Gram-negative rods demonstrated very high antibiotic resistance.

Conclusion: Despite the lack of widely implemented active targeted surveillance programmes and top-down incentives, it is possible to carry out effective surveillance of HAIs in ICUs in Poland. The results of this study are comparable with the ECDC data, but the results are alarmingly high in two fields: epidemiology of PN and BSIs and very high antibiotic resistance in Gram-negative rods, which indicate the need for intense control in this area and for further studies to clarify the source of the observed discrepancy.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.5507/bp.2018.006DOI Listing
September 2018

Alarming results of nosocomial bloodstream infections surveillance in Polish intensive care units

Przegl Epidemiol 2018;72(1):33-44

Polish Society of Hospital Infections, Krakow, Poland

Introduction: Nosocomial bloodstream infections (BSI) among patients in intensive care units (ICU) are the most common form of nosocomial infections and they are serious complications of the treatment process. The etiological factors of these infections and the proper antibiotic therapy can influence the effects of patient treatment. The aim of the study was to analyze the epidemiology, microbiology and antibiotic therapy of nosocomial BSI in Polish ICU patients.

Materials And Methods: The retrospective study covered patients hospitalized in seven ICUs for adults with a general profile, located in southern Poland. The studied units participated in a multicentre standardized infection control program in 2013-2015. 2,547 patients hospitalized more than 48 hours in ICUs were qualified to the study. The study was conducted in accordance with the methodology recommended by the European Center for Disease Prevention and Control (ECDC) as part of the European Healthcare-Associated Infections Surveillance Network (HAI-Net): protocols and the criteria for BSI recognition were standardized.

Results: The incidence of BSI was 7.2% per 100 admissions and the incidence density was 9.2 per 1000 persondays of hospitalization. Among 184 BSI cases, 65 cases (35.3%) were primary BSI; 87 cases were secondary BSI (47.3%) (related to an infection occurring elsewhere in the patient’s body), the remaining 32 cases (17.4%) were classified as BSI of unknown origin. Mortality was 20%. The dominant etiological factors in BSI related to central venous catheter were coagulase-negative staphylococci in 20.8% of cases, but in secondary BSI it was Acinetobacter baumannii in 34.5% of cases. Klebsiella pneumoniae showed resistance to third-generation cephalosporins in 96.0% of cases; Acinetobacter baumannii showed resistance to imipenem in 78.8% of cases. Other beta-lactam antibiotics J01D (31.0%) and other antibiotics J01X (26.6%) were most commonly used in the treatment.

Conclusions: The BSI incidence in the studied Polish ICUs was twice as high as in other European countries, and secondary BSI were also more frequent – especially secondary to urinary tract infections. The obtained results indicate the necessity of intervention not only in BSI prevention, but also in the field of prevention of urinary tract infections. In addition, microbial etiology and drug resistance indicate the need for urgent actions for their prevention.
View Article and Find Full Text PDF

Download full-text PDF

Source
July 2019

Analysis of the world epidemiological situation among vancomycin-resistant Enterococcus faecium infections and the current situation in Poland

Przegl Epidemiol 2018;72(1):3-15

Department of Epidemiology of Infections, Chair of Microbiology, Faculty of Medicine Jagiellonian University Medical College, Kraków

Vancomycin-resistant Enterococcus faecium (VREfm) strains have become an important hospital pathogen due to their rapid spread, high mortality rate associated with infections and limited therapeutic options. Vancomycin resistance is predominantly mediated by VanA or VanB phenotypes, which differ as regards maintaining sensitivity to teicoplanin in the VanB phenotype. The majority of VREfm cases in the United States, Europe, Korea, South America and Africa are currently caused by the VanA phenotype. However, the epidemics in Australia and Singapore are chiefly brought about by the VanB phenotype. The rate of VREfm isolate spread varies greatly. The greatest percentage of VREfm is now recorded in the USA, Ireland and Australia. Supervision of VRE is implemented to varying degrees. Therefore, the epidemiological situation in some countries is difficult to assess due to limited data or lack thereof.
View Article and Find Full Text PDF

Download full-text PDF

Source
July 2019

Characteristics of gut microbiota in adult patients with type 1 and type 2 diabetes based on next‑generation sequencing of the 16S rRNA gene fragment.

Pol Arch Intern Med 2018 06 15;128(6):336-343. Epub 2018 Apr 15.

Introduction Scientific data indicate a possible influence of gut microbiota on the development of type 1 and type 2 diabetes mellitus (T1DM and T2DM, respectively). Sequence analysis of 16S ribosomal RNA identified several hundred bacterial species of the intestinal ecosystem, most of which cannot be cultured. Objectives We aimed to evaluate gut microbiota composition in adult patients with T1DM and T2DM and establish a link between microbiological test results and patients' clinical data. Patients and methods We examined DNA isolated from fecal samples in 3 groups: healthy volunteers (n = 23), patients with T1DM (n = 22), and patients with T2DM (n = 23). Next‑generation sequencing was performed on the MiSeq platform. Results At the phylum level, the Firmicutes bacteria prevailed (>77%) in all groups. At the taxonomic levels L2 (phylum) and L6 (genus), significant differences were demonstrated in bacterial profiles, particularly in the T2DM group. A negative correlation was observed between several genera of bacteria and the percentage of glycated hemoglobin A1c in the T2DM group, while a positive correlation was revealed between bacteria belonging to the genus Bifidobacterium and high‑density lipoprotein cholesterol levels in both T1DM and T2DM groups. Conclusions Our results provide grounds for conducting research in the field of gut microbiota in order to develop individualized therapy for patients with diabetes based on modifying the microbiota composition, as a new method for controlling glycemia. Next‑generation sequencing allows a rapid identification of the DNA of all bacteria present in the sample and their taxonomic classification.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.20452/pamw.4246DOI Listing
June 2018

The usefulness of the basophil activation test in monitoring specific immunotherapy with house dust mite allergens.

Postepy Dermatol Alergol 2018 Feb 20;35(1):93-98. Epub 2018 Feb 20.

Department of Experimental Dermatology and Cosmetology, Faculty of Pharmacy, Jagiellonian University, Krakow, Poland.

Introduction: In clinical practice, reliable tools for monitoring specific immunotherapy (SIT) are of utmost importance.

Aim: To assess the usefulness of the basophil activation test (BAT) in monitoring SIT in paediatric patients with allergy to house dust mites (HDM).

Material And Methods: Thirty-one children qualified for SIT with HDM, of whom 21 completed the SIT during the observation period. The BAT was carried out prior to commencing the SIT (time point BAT1) and upon finishing the initial pack of allergy vaccine (cumulative dose of allergen 12487.5 PNU; BAT2), as well as after the second vaccine pack (cumulative dose of allergen 23750.0 PNU; BAT3). Peripheral blood of the patients was stimulated with allergen solutions in five concentrations from 0.00225 ng/ml to 22.5 ng/ml. Basophil activation was measured by CD63 expression in flow cytometry.

Results: For the allergen concentration of 0.225 ng/ml, a statistically significant decrease in median basophil activation was observed, from 51.29% at BAT1 to 8.48% at BAT2 ( = 0.004) and 4.21% at BAT3 ( < 0.001). For the allergen concentration of 0.0225 ng/ml, a statistically significant decrease was seen between BAT1 (1.72%) and BAT3 (0.21%, = 0.01). Median CD-sens index decreased significantly from 1099.02 at BAT1 to 179.31 at BAT2 ( < 0.002) and 168.04 at BAT3 ( < 0.001).

Conclusions: There is a significant decrease in BAT results in the course of specific immunotherapy with HDM allergens in children, with the optimum allergen concentration for monitoring basophil response at 0.225 ng/ml. The CD-sens index seems to be a better monitoring parameter than the plain percentage of CD63-expressing basophils.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.5114/ada.2018.73169DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5872250PMC
February 2018

Practice of hand hygiene and use of protective gloves: Differences in the perception between patients and medical staff.

Am J Infect Control 2018 09 13;46(9):1074-1076. Epub 2018 Mar 13.

Jagiellonian University Medical College, Kraków, Poland.

This study investigated differences in perceptions of hand hygiene and protective glove use among patients and health care workers (HCWs) in Poland. We conducted a survey using an original questionnaire among 462 respondents, including 173 (37.4%) patients and 289 (62.6%) HCWs; HCWs demonstrated poor familiarity with the My 5 Moments for Hand Hygiene. The role of protective gloves in preventing health care-associated infection was overestimated by both patients and HCWs.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.ajic.2018.01.025DOI Listing
September 2018

Typing of Enterococcus spp. strains in 4 hospitals in the Małopolska region in Poland.

Adv Clin Exp Med 2018 Jan;27(1):111-117

Department of Health and Medical Sciences, Andrzej Frycz Modrzewski Krakow University, Poland.

Background: In the Małopolska province, the first case of vancomycin resistant enterococci (VRE) occurrence was an outbreak in 2001 caused by strains of the genus E. faecium carrying the vanA operon.

Objectives: The aim of this study is to determine the antimicrobial resistance and the occurrence of virulence determinants among Enterococcus spp. in patients hospitalized in the Małopolska region in 2015.

Material And Methods: Antimicrobial susceptibility was determined by disc diffusion and the E test. The presence of aminoglycoside and glycopeptide resistance genes and virulence genes (asa1, gelE, cylA, esp, hyl) was investigated using multiplex polymerase chain reaction (PCR). Also, the presence of IS16 was investigated. The activity of gelatinase, cytolysin (hemolysin), and DNase was tested.

Results: All E. faecalis were susceptible to ampicillin, vancomycin, teicoplanin, linezolid and tigecycline. All E. faecium strains were susceptible to quinupristin-dalfopristin. Resistance to ampicillin and vancomycin was detected among all E. faecium isolates from hospitals C and D. 87.32% of E. faecium presented high-level aminoglycoside-resistant (HLAR) phenotype, including 78.33% of strains from hospital C and 100% from hospital D. In hospital C (98.3%) and D (96%), resistance to ciprofloxacin, levofloxacin and norfloxacin was observed. Gene esp was detected in all E. faecium isolates and the majority of E. faecium isolates carried hyl (97%). In E. faecalis, different combinations of virulence genes were detected. All analyzed E. faecium strains showed the presence of IS16 insertion element.

Conclusions: E. faecalis isolates were more susceptible to antimicrobials than E. faecium, which were largely multidrug-resistant. E. faecalis strains have diverse virulence factors. E. faecium showed a high percentage of hyl and esp genes and the presence of IS16.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.17219/acem/68265DOI Listing
January 2018

Antimicrobial effect of copper alloys on species isolated from infections and hospital environment.

Antimicrob Resist Infect Control 2018 22;7:10. Epub 2018 Jan 22.

1Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, ul. Czysta 18, 31-121 Kraków, Poland.

Background: An increased proportion of Gram-negative bacteria have recently been reported among etiologic agents of infection. In Poland, is a big problem for hospitals, especially intensive care units. Touch surfaces made from materials with antimicrobial properties, especially copper alloys, are recommended as a supplementary method of increasing biological safety in the hospital environment.

Aim Of The Study: The objective of this study is to determine the susceptibility to selected copper alloys of three clinical strains, one and an strain isolated from the hospital environment.

Material And Method: The modification of the Japanese Standard, which the ISO 22196:2011 norm was used for testing antimicrobial properties of CuZn37, CuSn6 and CuNi18Zn20 and Cu-ETP and stainless steel as positive and negative control, respectively.

Results: The highest cidal efficiency, expressed as both time and the degree of reduction of the initial suspension density, against all of the tested strains was found for ETP copper. But, the results of our study also confirmed effective activity (bacteriocidal or bacteriostatic) of copper alloys selected for the study, contrary to the stainless steel. The reduction in bacterial suspension density is significantly different depending on the strain and copper alloy composition.

Conslusions: The results of our study confirmed the effective antibacterial activity of copper and its selected alloys against clinical and strains, and strain isolated from the hospital environment.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s13756-018-0300-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5778618PMC
June 2019

Outpatient post-partum antibiotic prescription: method of identification of infection control areas demanding improvements and verification of sensitivity of infection registration.

J Antimicrob Chemother 2018 Jan;73(1):240-245

Chair of Microbiology, Jagiellonian University Medical College, Krakow, Poland.

Objectives: To evaluate data on outpatient antibiotic use in women post-labour as a potential method of monitoring infections in this group of patients.

Methods: Demographic and antibiotic prescription data originated from the registries of the National Health Fund (pol. Narodowy Fundusz Zdrowia). The measure of antibiotic use in this study was the percentage of women who purchased the drugs from prescriptions and DDDs.

Results: Among 67917 females who gave birth in the years 2013-14, 5050 (7.4%) purchased antibiotics prescribed by the obstetrician only. The average number of antibiotics bought per person was equivalent to ∼14 DDDs; in most cases (95.7%) these were β-lactams. Antibiotic use occurred significantly more frequently among younger patients (11.5% patients <18 years of age), those living in rural areas (8.2%) and those who underwent Caesarean section (8.1%). No significant differences were found between the reported day of labour and the post-partum use of antibiotics.

Conclusions: Antibiotic prescribing data can be used to verify/complement the information originating from hospital infection registries to monitor rates of infection in obstetric patients.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1093/jac/dkx369DOI Listing
January 2018

Antibiotic resistance, ability to form biofilm and susceptibility to copper alloys of selected staphylococcal strains isolated from touch surfaces in Polish hospital wards.

Antimicrob Resist Infect Control 2017 14;6:80. Epub 2017 Aug 14.

Faculty of Non-Ferrous Metals, Department of Metal Working and Physical Metallurgy of Non-Ferrous Metals, AGH University of Science and Technology, al. Mickiewicza 30, 30-059 Kraków, Poland.

Background: Despite the employment of sanitary regimes, contact transmission of the aetiological agents of hospital infections is still exceedingly common. The issue of microbe transmission becomes particularly important when facing multidrug-resistant microorganisms such as methicillin-resistant staphylococci. In the case of deficiencies in cleaning and disinfection procedures, hospital equipment made of copper alloys can play an important role, complementing traditional hospital hygiene procedures. The objective of this study was to characterize staphylococcal strains isolated from touch surfaces in Polish hospital wards in terms of their drug resistance, ability to form biofilm and susceptibility to antimicrobial activity of copper alloys.

Methods: The materials for the study were 95 staphylococcal strains isolated from touch surfaces in 13 different hospital wards from Małopolska province (the south of Poland). Phenotypic and genotypic antibiotic resistance were checked for erythromycin, clindamycin, gentamycin, ciprofloxacin, trimethoprim/sulfamethoxazole and mupirocin. Biofilm formation ability for the tested strains was checked with the use of culture on Congo red agar. Susceptibility to copper, tin bronze, brass and new silver was tested using a modification of the Japanese standard.

Results: Over 67% of the analysed staphylococcal strains were methicillin-resistant (MR). Four strains were resistant to all of the tested antibiotics, and 14 were resistant to all except mupirocin. Strains classified as MR had significantly increased resistance to the remaining antibiotic groups. About one-third of the analysed strains revealed biofilm-forming ability. Among the majority of species, biofilm-forming and non-biofilm-forming strains were distributed evenly; in the case of only, negative strains accounted for 92.8%. Susceptibility to copper alloys was different between strains and rather lower than in the case of the SA strain selected for comparison.

Conclusions: Coagulase-negative staphylococci, the most commonly isolated in Polish hospital wards, should not be neglected as an infection risk factor due their high antibiotic resistance. Our experiments confirmed that touch surfaces made of copper alloys may play an important role in eliminating bacteria from the hospital environment.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s13756-017-0240-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5556671PMC
August 2017

Gas Gangrene of Different Origin Associated with Clostridium perfringens Type A in Three Patients Simultaneously Hospitalized in a Single Department of Orthopedics and Traumatology in Poland.

Pol J Microbiol 2017 Jan;65(4):399-406

Department of Infection Epidemiology, Chair of Microbiology, Jagiellonian University Medical College, Krakow, Poland.

The objective of the study was to perform a comparative analysis of phenotypic and genetic similarity, determination of resistance profiles, detection of toxin-encoding genes and molecular typing of Clostridium perfringens isolates originating from patients with gas gangrene. The study encompassed three patients with a clinical and microbiological diagnosis of gas gangrene who were hospitalized in one of the hospitals of the Kujawsko-Pomorskie province in the same period of time between 8 April 2015 and 20 April 2015. The three C. perfringens isolates studied had identical biochemical profiles. Two isolates had identical resistance patterns, while the third presented a different profile. Using the multiplex PCR method, all isolates showed the presence of cpa gene encoding α-toxin; furthermore, the presence of the cpb2 gene encoding β2-toxin was confirmed in two isolates. Genotyping with the use of pulsed field gel electrophoresis (PFGE) indicated that the isolates originating from the three studied patients represent three genetically different restrictive patterns which corresponded to three different clones - clone A, clone B and clone C. As a result of the study, it is possible to conclude that the studied patients simultaneously hospitalized in a single Department of Orthopedics and Traumatology developed three different endogenous infections.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.5604/17331331.1227665DOI Listing
January 2017

Antimicrobial Properties of Selected Copper Alloys on Staphylococcus aureus and Escherichia coli in Different Simulations of Environmental Conditions: With vs. without Organic Contamination.

Int J Environ Res Public Health 2017 07 20;14(7). Epub 2017 Jul 20.

Faculty of Non-Ferrous Metals, Department of Metal Working and Physical Metallurgy of Non-Ferrous Metals, AGH University of Science and Technology, Al. A. Mickiewicza 30, 30-059 Krakow, Poland.

Hospital equipment made from copper alloys can play an important role in complementing traditional methods of disinfection. The aim of this study was to assess the dynamics of the antimicrobial properties of selected copper alloys in different simulations of environmental conditions (with organic contamination vs. without organic contamination), and to test alternatives to the currently used testing methods. A modification of Japanese standard JIS Z 2801 as well as (SA) and (EC) suspended in NaCl vs. tryptic soy broth (TSB) were used in tests performed on seven commonly used copper alloys, copper, and stainless steel. A much faster reduction of the bacterial suspension was observed for the inoculum prepared in NaCl than in TSB. A faster reduction for EC than for SA was observed in the inoculum prepared in NaCl. The opposite results were found for the inoculum based on TSB. A significant correlation between the copper concentration in the copper alloys and the time and degree of bacterial suspension reduction was only observed in the case of EC. This study confirmed the antimicrobial properties of copper alloys, and additionally showed that was more resistant than in the variant of the experiment without organic contamination. However, even for SA, a total reduction of the bacterial inoculum's density took no longer than 2 h. Under conditions simulating organic contamination, all of the tested alloys were shown to have bactericidal or bacteriostatic properties, which was contrary to the results from stainless steel.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/ijerph14070813DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5551251PMC
July 2017

Do bacteria isolated from ICU patients 'ESKAPE' antibiotic treatment? In vitro susceptibility of the Enterobacteriaceae family to tigecycline.

Anaesthesiol Intensive Ther 2017 16;49(3):210-214. Epub 2017 Jul 16.

Department of Mycology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, Cracow, Poland.

Background: Enterobacteriaceae are currently causing the majority of healthcare-associated infections (HAI) and simultaneously expressing increasing levels of antibiotic resistance. The purpose of this study is to assess the in vitro sensitivity of MDR strains from the family Enterobacteriaceae to tigecycline in relation to their origin from patients hospitalized in intensive care units (ICUs) and non-ICUs.

Methods: The study involved 156 clinically significant strains of the Enterobacteriaceae family isolated from patients with complicated intraabdominal infections (cIAIs) and/or complicated skin and skin structure infections (cSSSIs) hospitalized in ICUs and other surgical departments. Tigecycline MICs were determined by Etest.

Results: The highest percentage of tigecycline non-susceptible (intermediate + resistant strains) in vitro strains among the Enterobacteriaceae species were observed for Serratia spp. 77.3%, followed by Citrobacter spp. (76.9%) and Enterobacter spp. (70%); whereas K. pneumoniae and E. coli showed 73-73.8% tigecycline susceptibility rates.

Conclusion: Tigecycline demonstrates a high level of antimicrobial in vitro activity when tested against E. coli and K. pneumoniae, even those with the ESBL-phenotype. Tigecycline retained activity against merely 22-30% of Enterobacter, Citrobacter and Serratia genera.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.5603/AIT.a2017.0034DOI Listing
May 2018

Bacteria contamination of touch surfaces in Polish hospital wards.

Med Pr 2017 Jun 20;68(4):459-467. Epub 2017 Apr 20.

Jagiellonian University Medical College, Kraków, Poland (Chair of Microbiology).

Background: The objective of the study has been to evaluate the pathogenic bacteria contamination of touch surfaces in hospital wards.

Material And Methods: Samples were taken from frequently touched surfaces in the hospital environment in 13 units of various types. Culturing was carried out on solid blood agar and in growth broth (tryptic soy broth - TSB). Species identification was performed using the analytical profile index (API) biochemical testing and confirmed with matrix assisted laser desorption ionization time-flight mass spectrometry (MALDI-TOF-MS) system.

Results: The total of 161 samples were taken for the study. Fifty-two of them, after 24 h of culture on a solid medium, demonstrated bacterial growth and further 60 samples had growth after prior multiplication in TSB. Overall, 69.6% of samples exhibited growth of 19 bacterial species. Pathogenic species - representing indicator organisms of efficiency of hospital cleaning - was demonstrated by 21.4% of samples. Among them Acinetobacter spp., Enterocococci spp. and Staphylococcus aureus were identified. Coagulase-negative staphylococci (CNS) were predominant. The proportion of various groups of bacteria significantly varied in respective hospitals, and in various types of wards. Disturbing observation is a large proportion of resistance of isolated CNS strains as a potential reservoir of resistance genes.

Conclusions: The results show that touch surfaces in hospital units are contaminated by both potentially pathogenic and pathogenic bacterial species. In connection with the reported, also in Poland, frequent omission or incorrect execution of hand hygiene by hospital staff, and probably patients, touch surfaces still constitute important reservoir of pathogenic bacteria. Improving hand hygiene compliance of health-care workers with recommendations is necessary for increasing biological safety of hospital environment. Med Pr 2017;68(3):459-467.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.13075/mp.5893.00575DOI Listing
June 2017

Nested-PCR real time as alternative molecular tool for detection of Borrelia burgdorferi compared to the classical serological diagnosis of the blood.

Med Dosw Mikrobiol 2016;68(1):47-56

Introduction: Lyme disease, caused by Borrelia burgdorferi, is a multisystem disease that often makes difficulties to recognize caused by their genetic heterogenity. Currently, the gold standard for the detection of Lyme disease (LD) is serologic diagnostics based mainly on tests: ELISA and Western blot (WB). These methods, however, are subject to consider- able defect, especially in the initial phase of infection due to the occurrence of so-called serological window period and low specificity. For this reason, they might be replaced by molecular methods, for example polymerase chain reaction (PCR), which should be more sensitivity and specificity. In the present study we attempt to optimize the PCR reaction conditions and enhance existing test sensitivity by applying the equivalent of real time PCR - nested PCR for detection B. burgdorferi DNA in the patient's blood.

Methods: The study involved 94 blood samples of patients with suspected LD. From each sample, 1.5 ml of blood was used for the isolation of bacterial DNA and PCR real time am- plification and its equivalent, in nested version. The remaining part earmarked for serologi- cal testing. Optimization of the reaction conditions made experimentally, using gradient of the temperature and gradient of the magnesium ions concentration for reaction real time in nested-PCR and PCR version.

Results: The results show that the nested-PCR real time, has a much higher sensitivity 45 (47.8%) of positive results for the detection of B. burgdorferi compared to the single- variety, without a preceding pre-amplification 2 (2.1%). Serological methods allowed the detection of infection in 41 (43.6%) samples.

Conclusions: These results support of the nested PCR method as a better molecular tool for the detection of B. burgdorferi infection than classical PCR real time reaction. The nested-PCR real time method may be considered as a complement to ELISA and WB mainly in the early stages of infection, when in the blood circulating B. burgdorferi cells. By contrast, the results of serological and molecular tests should always be carried out tak- ing into account the patient's clinical status.
View Article and Find Full Text PDF

Download full-text PDF

Source
March 2017