Publications by authors named "Monika Brzychczy-Włoch"

53 Publications

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

Next-Generation Sequencing as a Tool to Detect Vaginal Microbiota Disturbances during Pregnancy.

Microorganisms 2020 Nov 18;8(11). Epub 2020 Nov 18.

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

The physiological microbiota of the vagina is responsible for providing a protective barrier, but Some factors can disturb the balance in its composition. At that time, the amounts of the genus decrease, which may lead to the development of infection and severe complications during pregnancy. The aim of the study was the analysis of the bacterial composition of the vagina in 32 Caucasian women at each trimester of pregnancy using the next-generation sequencing method and primers targeting V3-V4 regions. In the studied group, the dominant species were and . Statistically significant differences in the quantitative composition between trimesters were observed in relation to , , spp. Out of the 32 patients, 20 demonstrated fluctuations within the genus and 9 of them, at different stages of pregnancy, exhibited the presence of potentially pathogenic microbiota, among others: spp., , and The composition of the vaginal microbiota during pregnancy was subject to partial changes over trimesters. Although in one-third of the studied patients, both the qualitative and quantitative composition of microbiota was relatively constant, in the remaining patients, physiological and potentially pathogenic fluctuations were distinguished.
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http://dx.doi.org/10.3390/microorganisms8111813DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7698737PMC
November 2020

Deep learning approach to describe and classify fungi microscopic images.

PLoS One 2020 30;15(6):e0234806. Epub 2020 Jun 30.

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

Preliminary diagnosis of fungal infections can rely on microscopic examination. However, in many cases, it does not allow unambiguous identification of the species due to their visual similarity. Therefore, it is usually necessary to use additional biochemical tests. That involves additional costs and extends the identification process up to 10 days. Such a delay in the implementation of targeted therapy may be grave in consequence as the mortality rate for immunosuppressed patients is high. In this paper, we apply a machine learning approach based on deep neural networks and bag-of-words to classify microscopic images of various fungi species. Our approach makes the last stage of biochemical identification redundant, shortening the identification process by 2-3 days, and reducing the cost of the diagnosis.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0234806PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7326179PMC
September 2020

Electrochemical modification of the Ti-15Mo alloy surface in solutions containing ZnO and Zn(PO) particles.

Mater Sci Eng C Mater Biol Appl 2020 Oct 20;115:111098. Epub 2020 May 20.

Faculty of Chemistry, Silesian University of Technology, B. Krzywoustego 6 Street, 44-100 Gliwice, Poland. Electronic address:

This paper reports on the plasma electrolytic oxidation (PEO) of titanium alloy Ti-15Mo in baths containing zinc to obtain biomaterials with bacteriostatic and antibacterial properties. The Ti-15Mo surface was oxidised in a 0.1 M Ca(HPO) bath containing zinc compound particles: ZnO or Zn(PO). During the PEO process, the applied voltage was 300 V, and the current density was 150 mA∙cm. The surface morphology, roughness and wettability were determined. It has been noted that both roughness and wettability of Ti-15Mo alloy surface increased after PEO. EDX and XPS chemical composition analysis was carried out, and Raman spectroscopy was also performed indicating that Zn has been successfully incorporated into oxide layer. To investigate the antibacterial properties of the PEO oxide coatings, microbial tests were carried out. The bacterial adhesion test was performed using four different bacterial strains: reference Staphylococcus aureus (ATCC 25923), clinical Staphylococcus aureus (MRSA 1030), reference Staphylococcus epidermidis (ATCC 700296) and clinical Staphylococcus epidermidis (15560). Performed zinc-containing oxide coatings did not indicate the bacteria growth inducing effect. Additionally, the cytocompatibility of the formed oxide layers was characterised by MG-63 osteoblast-like live/dead tests. The surface bioactivity and cytocompatibility increased after the PEO process. The zinc was successfully incorporated into the titanium oxide layer. Based on the obtained results of the studies, it can be claimed that zinc-containing PEO layers can be an interesting course of bacteriostatic titanium biomaterials development.
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http://dx.doi.org/10.1016/j.msec.2020.111098DOI Listing
October 2020

Antibacterial and cytocompatible coatings based on poly(adipic anhydride) for a Ti alloy surface.

Bioact Mater 2020 Sep 24;5(3):709-720. Epub 2020 May 24.

Faculty of Chemistry, Silesian University of Technology, B. Krzywoustego Street 6, 44-100, Gliwice, Poland.

This paper describes a formation of hybrid coatings on a Ti-2Ta-3Zr-36Nb surface. This is accomplished by plasma electrolytic oxidation and a dip-coating technique with poly(adipic anhydride) ((CHO)n) that is loaded with drugs: amoxicillin (CHNOS), cefazolin (CHNOS) or vancomycin (CHClNO · xHCl). The characteristic microstructure of the polymer was evaluated using scanning electron microscopy and confocal microscopy. Depending on the surface treatment, the surface roughness varied (between 1.53 μm and 2.06 μm), and the wettability was change with the over of time. X-ray photoelectron spectroscopy analysis showed that the oxide layer did not affect the polymer layer or loaded drugs. However, the drugs lose their stability in a phosphate-buffered saline solution after 6.5 h of exposure, and its decrease was greater than 7% (HPLC analysis). The stability, drug release and concentration of the drug loaded into the material were precisely analyzed by high-performance liquid chromatography. The results correlated with the degradation of the polymer in which the addition of drugs caused the percent of degraded polymer to be between 35.5% and 49.4% after 1 h of material immersion, depending on the mass of the loaded drug and various biological responses that were obtained. However, all of the coatings were cytocompatible with MG-63 osteoblast-like cells. The drug concentrations released from the coatings were sufficient to inhibit adhesion of reference and clinical bacterial strains (). The coatings with amoxicillin showed the best results in the bacterial inhibition zone, whereas coatings with cefazolin inhibited adhesion of the above bacteria on the surface.
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http://dx.doi.org/10.1016/j.bioactmat.2020.04.020DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7248586PMC
September 2020

Physico-chemical and biological evaluation of doxycycline loaded into hybrid oxide-polymer layer on Ti-Mo alloy.

Bioact Mater 2020 Sep 25;5(3):553-563. Epub 2020 Apr 25.

Faculty of Chemistry, Silesian University of Technology, B. Krzywoustego Street 6, 44-100, Gliwice, Poland.

Oxide-polymer coatings were formed on the surface of the vanadium-free Ti-15Mo titanium alloy. The Ti alloy surface was modified by the plasma electrolytic oxidation process, and then, the polymer layer of a poly (D, l-lactide--glycolide) with doxycycline was formed. The polymer evenly covered the porous oxide layer and filled some of the pores. However, the microstructure of the polymer surface was completely different from that of the PEO layer. The surface morphology, roughness and microstructure of the polymer layer were examined by scanning electron microscopy (SEM) and a confocal microscope. The results confirmed the effectiveness of polymer and doxycycline deposition in their stable chemical forms. The drug analysis was performed by high-performance liquid chromatography. The H NMR technique was used to monitor the course of hydrolytic degradation of PLGA. It was shown that the PLGA layer is hydrolysed within a few weeks, and the polyglycolidyl part of the copolymer is hydrolysed to glycolic acid as first and much faster than the polylactide one to lactic acid. This paper presents influence of different microstructures on the biological properties of modified titanium alloys. Cytocompatibility and bacterial adhesion tests were evaluated using osteoblast-like MG-63 cells and using the reference and strains. The results showed that the optimum concentration of doxycycline was found to inhibit the growth of the bacteria and that the layer is still cytocompatible.
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http://dx.doi.org/10.1016/j.bioactmat.2020.04.009DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7191259PMC
September 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.
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http://dx.doi.org/10.33073/pjm-2019-035DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7256698PMC
September 2019

Functionalization of PEO layer formed on Ti-15Mo for biomedical application.

J Biomed Mater Res B Appl Biomater 2020 05 23;108(4):1568-1579. Epub 2019 Oct 23.

Faculty of Chemistry, Department of Inorganic Chemistry, Analytical Chemistry and Electrochemistry, Silesian University of Technology, Gliwice, Poland.

In the present work, deposition of poly(sebacic anhydride) PSBA loaded by amoxicillin, cefazolin, or vancomycin on a previously anodized Ti-15Mo surface is presented. PSBA loaded by the drug was deposited so as not to lose the functionality of the porous oxide layer microstructure. The morphology was evaluated using scanning electron microscopy, surface roughness, and wettability. The drug concentration was evaluated using high-performance liquid chromatography. It was determined that the drugs were loaded into coatings in the range of 35.2-122.87 μg/cm of Ti sample. The drugs released more than 16% after 0.5 hr of the hybrid coating immersion in artificial saliva. After 3 days, the PSBA coatings were degraded by 51.3 mol %, and after 7 days by 77.8 mol %, which makes it possible to load the material by different, biologically active substances. An antimicrobial investigation of Staphylococcus aureus (DSM 24167) and Staphylococcus epidermidis (ATCC 700296) confirmed the activity of the hybrid layers against the pathogens. Hybrid layer with vancomycin best inhibits the adhesion of the bacteria, whereas coatings with amoxicillin and cefazolin showed a much better bactericidal activity. In this article, the difference in the obtained results is discussed, as well as the possibility of the application of this functional material in biomedicine.
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http://dx.doi.org/10.1002/jbm.b.34504DOI Listing
May 2020

"Command" surfaces with thermo-switchable antibacterial activity.

Mater Sci Eng C Mater Biol Appl 2019 Oct 27;103:109806. Epub 2019 May 27.

Smoluchowski Institute of Physics, Jagiellonian University, Łojasiewicza 11, 30-348 Kraków, Poland.

In the presented work "smart" antibacterial surfaces based on silver nanoparticles (AgNPs) embedded in temperature-responsive poly(di(ethylene glycol)methyl ether methacrylate) - (POEGMA188) as well as poly(4-vinylpyridine) - (P4VP) coatings attached to a glass surface were successfully prepared. The composition, thickness, morphology and wettability of the resulting coatings were analyzed using ToF-SIMS, XPS, EDX, ellipsometry, AFM, SEM and CA measurements, respectively. Temperature-switched killing of the bacteria was tested against Escherichia coli ATCC 25922 (representative of Gram-negative bacteria) and Staphylococcus aureus ATCC 25923 (representative of Gram-positive bacteria) at 4 and 37 °C. In general at 4 °C no significant difference was observed between the amounts of bacteria accounted on the grafted brush coatings and within the control sample. In contrast, at 37 °C almost no bacteria were visible for temperature-responsive coating with AgNPs, whereas the growth of bacteria remains not disturbed for "pure" coating, indicating strong temperature-dependent antibacterial properties of AgNPs integrated into brushes.
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http://dx.doi.org/10.1016/j.msec.2019.109806DOI Listing
October 2019

PLGA-amoxicillin-loaded layer formed on anodized Ti alloy as a hybrid material for dental implant applications.

Mater Sci Eng C Mater Biol Appl 2019 Jan 12;94:998-1008. Epub 2018 Oct 12.

Faculty of Chemistry, Silesian University of Technology, B. Krzywoustego Street 6, 44-100 Gliwice, Poland.

In this paper, the preparation of a functional hybrid coating loaded with a drug (amoxicillin) on a promising titanium alloy - Ti-15Mo alloy is presented. The titanium alloy surface was anodized in solution with bioactive compounds to obtain a porous oxide layer favorable for MG-63 osteoblast-like cell adhesion. Then, a poly(lactide-co-glycolide) (PLGA) loaded with amoxicillin layer was formed using a dip-coating technique to cover the oxide layer, without filling in all of the pores. The morphology of the surface was evaluated using scanning electron microscopy supported by 3D Roughness Reconstruction software. The surface treatment of the Ti-15Mo alloy surface caused the surface roughness to increase up to 1.71 μm. The anodization process caused the Ti-15Mo alloy surface to become slightly more hydrophilic; however, the formation of the PLGA layer loaded with drug increased the contact angle to 96.5° ± 2.2°, respectively. After 4 weeks of polymer layer degradation, the registered signals on the H NMR spectrum were identical to the signals registered for lactic acid (LAc), which confirms that the polymer layer was degraded within a short period of time. The concentration of drug released into the artificial saliva was investigated using high-performance liquid chromatography (HPLC) up to 12 h of coatings immersion. During the first hour of coating degradation in artificial saliva, and the concentration of the drug (13 μg/ml) was enough to inhibit bacterial growth of S. aureus and S. epidermidis. These results were confirmed by agar plate diffusion method and evaluation of the minimal inhibitory concentration (MIC). The cytocompatibility of the materials was determined using the osteoblast-like cells MG-63, and the viability and cell morphology (live/dead staining) were also evaluated. The results showed that amoxicillin influences the osteoblast-like MG-63 cells' behavior during cell culture, especially for the first few hours. The influence on the type of surface treatment on MG-63 cell behavior during 7 days of culture is discussed in this paper. To the best of our knowledge, this is the first time that a fast-degrading layer with amoxicillin has been deposited on previously anodized Ti surface. The formation of functional coating may find application as a cytocompatible coating to prevent bacterial adhesion on long-term implant surfaces.
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http://dx.doi.org/10.1016/j.msec.2018.10.049DOI Listing
January 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.
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http://dx.doi.org/10.3389/fcimb.2018.00349DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6176014PMC
September 2019

Epitopes identified in GAPDH from Clostridium difficile recognized as common antigens with potential autoimmunizing properties.

Sci Rep 2018 09 17;8(1):13946. Epub 2018 Sep 17.

Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Department of Immunology of Infectious Diseases, Laboratory of Medical Microbiology, Wroclaw, Poland.

Clostridium difficile (CD) infections are a growing threat due to the strain resistance to antibiotic treatment and the emergence of hypervirulent strains. One solution to this problem is the search for new vaccine antigens, preferably surface-localized that will be recognized by antibodies at an early stage of colonization. The purpose of the study was to assess the usefulness of novel immunoreactive surface proteins (epitopes) as potential vaccine antigens. Such approach might be tough to pursue since pathogens have acquired strategies to subvert adaptive immune response to produce humoral response against non-essential proteins for their survival. In this study CD surface proteins were isolated, immunoreactive proteins identified and mapped to select potential epitopes. The results of the study exclude the use of CD glyceraldehyde 3-phosphate dehydrogenase as a vaccine antigen, especially as a whole protein. Sequences P9 (AAGNIVPNTTGAAKAI) and P10 (KGKLDGAAQRVPVVTG) recognized by patients sera are conserved and widespread among CD strains. They show cross-reactivity with sera of people suffering from other bacterial infections and are recognized by sera of autoimmune disease patients. Our study documents that special care in analyzing the sequence of new epitope should be taken to avoid side effects prior to consider it as a vaccine antigen.
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http://dx.doi.org/10.1038/s41598-018-32193-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6141484PMC
September 2018

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

Epitope Mapping of Elongation Factor Tu Protein Recognized by Human Sera.

Front Microbiol 2018 6;9:125. Epub 2018 Feb 6.

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

The elongation factor Tu has been identified as one of the most immunoreactive proteins that was recognized by human sera of GBS (group B streptococcus) positive patients. In this paper, we present the polypeptide-specific epitopes of the bacterial protein that are recognized by human antibodies: LTAAITTVLARRLP (peptide no. 3) and GQVLAKPGSINPHTKF (peptide no. 21). To determine the shortest amino acid sequence recognized by antibodies, truncation peptide libraries were prepared using the PEPSCAN method. The analysis of immunoreactivity of peptides with sera of GBS positive and negative women revealed that the most immunoreactive sequence was HTKF. Moreover, we observed that this sequence also showed the highest specificity which was based on ratio of reactivity with sera of GBS positive relative to sera of GBS negative patients. Epitope was synthetized on Wang resin with the Fmoc strategy. Our results open the possibility to use HTKF peptide in diagnostic assays to determine infection in humans.
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http://dx.doi.org/10.3389/fmicb.2018.00125DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5808131PMC
February 2018

Deep learning approach to bacterial colony classification.

PLoS One 2017 14;12(9):e0184554. Epub 2017 Sep 14.

Department of Infection Epidemiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, 18 Czysta Street, 31-121 Kraków, Poland.

In microbiology it is diagnostically useful to recognize various genera and species of bacteria. It can be achieved using computer-aided methods, which make the recognition processes more automatic and thus significantly reduce the time necessary for the classification. Moreover, in case of diagnostic uncertainty (the misleading similarity in shape or structure of bacterial cells), such methods can minimize the risk of incorrect recognition. In this article, we apply the state of the art method for texture analysis to classify genera and species of bacteria. This method uses deep Convolutional Neural Networks to obtain image descriptors, which are then encoded and classified with Support Vector Machine or Random Forest. To evaluate this approach and to make it comparable with other approaches, we provide a new dataset of images. DIBaS dataset (Digital Image of Bacterial Species) contains 660 images with 33 different genera and species of bacteria.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0184554PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5599001PMC
October 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.
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http://dx.doi.org/10.5604/17331331.1227665DOI Listing
January 2017

Gentamicin-Loaded Polysaccharide Membranes for Prevention and Treatment of Post-operative Wound Infections in the Skeletal System.

Pharm Res 2017 Oct 21;34(10):2075-2083. Epub 2017 Jun 21.

Department of Biomaterials, AGH University of Science and Technology, Faculty of Materials Science and Ceramics, al. A. Mickiewicza 30, 30-059, Kraków, Poland.

Purpose: To develop polysaccharide-based membranes that allow controlled and localized delivery of gentamicin for the treatment of post-operative bone infections.

Methods: Membranes made of gellan gum (GUM), sodium alginate (ALG), GUM and ALG crosslinked with calcium ions (GUM + Ca and ALG + Ca, respectively) as well as reference collagen (COL) were produced by freeze-drying. Mechanical properties, drug release, antimicrobial activity and cytocompatibility of the membranes were assessed.

Results: The most appropriate handling and mechanical properties (Young's modulus, E = 92 ± 4 MPa and breaking force, F  = 2.6 ± 0.1 N) had GUM + Ca membrane. In contrast, COL membrane showed F  = 0.14 ± 0.02 N, E = 1.0 ± 0.3 MPa and was deemed to be unsuitable for antibiotic delivery. The pharmacokinetic data demonstrated a uniform and sustainable delivery of gentamicin from GUM + Ca (44.4 ± 1.3% within 3 weeks), while for COL, ALG and ALG + Ca membranes the most of the drug was released within 24 h (55.3 ± 1.9%, 52.5 ± 1.5% and 37.5 ± 1.8%, respectively). Antimicrobial activity against S. aureus and S. epidermidis was confirmed for all the membranes. GUM + Ca and COL membranes supported osteoblasts growth, whereas on ALG and ALG + Ca membranes cell growth was reduced.

Conclusions: GUM + Ca membrane holds promise for effective treatment of bone infections thanks to favorable pharmacokinetics, bactericidal activity, cytocompatibility and good mechanical properties.
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http://dx.doi.org/10.1007/s11095-017-2212-5DOI Listing
October 2017

Using of the 16S rDNA sequencing for identification of Lactobacillus species.

Med Dosw Mikrobiol 2016;68(1):5-11

Introduction: Lactobacilli play an important role in maintaining vaginal health and pro- tecting the genital tract from bacterial infections, so very often they are used as probiotics. Despite the scientific consensus on the significance of the genus Lactobacillus, its species identification still poses several difficulties. The aim of this study was to find out if the 16S rDNA sequencing method allows exact genotyping of Lactobacillus species.

Methods: 78 isolates from healthy pregnant women were tested: 57 from the vagina and 21 from the rectum. We also examined seven reference strains: L. acidophilus ATCC 4356, L.fermentum ATCC 20052, L. plantarum ATCC 20174, L. plantarum ATCC 14431, L. delbrueckii ssp. bulgaricus ATCC 20074, L. crispatus ATCC 20225 and L. gasseri ATCC 20243 to confirm the effectiveness sequencing method. A fragment of the 16S rDNA was amplified. After amplification, the amplicons were separated by gel electrophoresis and then sequenced. Furthermore, the received consensus sequences were checked for species specificity in the National Center for Biotechnology Information (NCBI) database with BLAST software. Sequences with a ;> 98% match to a database sequence were considered to be the same species.

Results: We have confirmed the genus of seven tested reference strains of lactobacilli with 100% probability. Of the analyzed isolates, all were identified to the species level. 14 spe- cies were identified in the 78 respondents, 9 of which colonized the vagina and 11 appeared in the rectum. The species colonizing the vagina were: L. gasseri 31.6%, followed by L. crispatus 28.2%, L. rhamnosus 14%, L. amylovorus 14%, L. helveticus 3.5%, L. reuteri 3.5%, L. casei 1.7%, L. salivarius 1.7% and L. delbrueckii 1.7%. The species colonizing the anus were: L. caseil L. paracasei 28.6%, L. plantarum 14.3%, L. crispatus 14.3%, L. gasseri 9.5%, L. reuteri 9.5%, L. salivarius 4.8%, L. rhamnosus 4.8%, L. acidophilus 4.8%, L. ruminis 4.8% and L. sakei 4.8%.

Conclusions: Using the 16S rDNA sequencing method made it possible to genotype 100% of the tested isolates of Lactobacillus.
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March 2017

Ceramic scaffolds enriched with gentamicin loaded poly(lactide-co-glycolide) microparticles for prevention and treatment of bone tissue infections.

Mater Sci Eng C Mater Biol Appl 2016 Dec 26;69:856-64. Epub 2016 Jul 26.

AGH University of Science and Technology, Faculty of Materials Science and Ceramics, Department of Biomaterials, Al. A. Mickiewicza 30, 30-059 Krakow, Poland. Electronic address:

Bone scaffolds are susceptible for bacterial infection when implanted, particularly in compromised bone. Therefore anti-bacterial bone scaffolds are desirable. Here a novel approach to provide bactericidal properties for titanium dioxide scaffolds is proposed. Gentamicin loaded poly(L-lactide-co-glycolide) microparticles were immobilized on the scaffold pore walls by sodium alginate hydrogel. The results show that the microparticles were effectively immobilized on the scaffolds. Desired burst release was observed within the first 8h and gentamicin dose reached 125μg from single scaffold that corresponded to ~25% of total drug introduced in the system. Following the initial burst, the dose was gradually decreasing up to day 10 and afterwards a sustained release of 3μg/day was measured. Cumulatively ~90% of the drug was delivered up to day 50. Above pattern, i.e. burst release with following sustained release, is desired for prevention of perioperative bone infections: burst release stops local infections during post-implantation "decisive period" while further sustained drug release prevents bacterial recolonization. In vitro studies confirmed antimicrobial activity of released gentamicin against Staphylococcus spp. and cytocompatibility of the system with osteoblast-like cells (MG-63). Thus the system is a viable option for the treatment of bone tissue defects.
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http://dx.doi.org/10.1016/j.msec.2016.07.065DOI Listing
December 2016

A study of the effects of therapeutic doses of ionizing radiation in vitro on Lactobacillus isolates originating from the vagina - a pilot study.

BMC Microbiol 2016 May 31;16:99. Epub 2016 May 31.

Department of Bacteriology, Microbial Ecology and Parasitology, Chair of Microbiology, Jagiellonian University Medical College, 18 Czysta Street, 31-121, Krakow, Poland.

Background: Ionizing radiation is used as a therapeutic option in the treatment of certain neoplastic lesions located, among others, in the pelvic region. The therapeutic doses of radiation employed often result in adverse effects manifesting themselves primarily in the form of genital tract infections in patients or diarrhea. The data available in the literature indicate disorders in the microbial ecosystem caused by ionizing radiation, which leads to the problems mentioned above. In the present study, we examined the influence of ionizing radiation on 52 selected strains of bacteria: Lactobacillus crispatus, L. fermentum, L. plantarum, L. reuteri, L. acidophilus L. amylovorus, L. casei, L. helveticus, L. paracasei, L. rhamnosus, L. salivarius and L. gasseri. This collection of Lactobacillus bacteria isolates of various species, obtained from the genital tract and gastrointestinal tract of healthy women, was tested for resistance to therapeutic doses of ionizing radiation.

Results: The species studied, were isolated from the genital tract (n = 30) and from the anus (n = 22) of healthy pregnant women. Three doses of 3 Gy (fractionated dose) and 50 Gy (total dose of the whole radiotherapy cycle) were applied. The greatest differences in survival of the tested strains in comparison to the control group (not subjected to radiation) were observed at the dose of 50 Gy. However, the results were not statistically significant. Survival decrease to zero was not demonstrated for any of the tested strains.

Conclusions: Therapeutic doses of radiation do not affect the Lactobacillus bacteria significantly.
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http://dx.doi.org/10.1186/s12866-016-0716-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4886408PMC
May 2016

Injectable hybrid delivery system composed of gellan gum, nanoparticles and gentamicin for the localized treatment of bone infections.

Expert Opin Drug Deliv 2016 16;13(5):613-20. Epub 2016 Feb 16.

a Department of Biomaterials, Faculty of Materials Science and Ceramics , AGH University of Science and Technology , Krakow , Poland.

Objectives: Bone infections are treated with antibiotics administered intravenously, antibiotic-releasing bone cements or collagen sponges placed directly in the infected area. These approaches render limited effectiveness due to the lack of site specificity and invasiveness of implanting cements and sponges. To address these limitations, we developed a novel polysaccharide hydrogel-based injectable system that enables controlled delivery of gentamicin (GENT). Its advantages are minimal invasiveness, and localized and finely regulated release of the drug.

Methods: GENT was incorporated both directly within the gellan gum hydrogel and into poly(L-lactide-co-glycolide) nanoparticles embedded into the hydrogel.

Results: We confirmed the injectability of the system and measured extrusion force was 15.6 ± 1.0 N, which is suitable for injections. The system set properly after the injection as shown by rheological measurements. Desired burst release of the drug was observed within the first 12 h and the dose reached ~27% of total GENT. Subsequently, GENT was released gradually and sustainably: ~60% of initial dose within 90 days. In vitro studies confirmed antimicrobial activity of the system against Staphylococcus spp. and cytocompatibility with osteoblast-like cells.

Conclusions: Developed injectable system enables minimally invasive, local and sustained delivery of the pharmaceutically relevant doses of GENT to combat bone infections.
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http://dx.doi.org/10.1517/17425247.2016.1146673DOI Listing
September 2016

Gentamicin loaded PLGA nanoparticles as local drug delivery system for the osteomyelitis treatment.

Acta Bioeng Biomech 2015 ;17(3):41-8

AGH University of Science and Technology, Faculty of Materials Science and Ceramics, Department of Biomaterials, Kraków, Poland.

Since there are more and more cases of multiresistance among microorganisms, rational use of antibiotics (especially their systemic vs. local application) is of great importance. Here we propose polymeric nanoparticles as locally applied gentamicin delivery system useful in osteomyelitis therapy. Gentamicin sulphate (GS) was encapsulated in the poly(lactide-co-glycolide) (PLGA 85:15) nanoparticles by double emulsification (water/oil/water, W1/O/W2). The nanoparticles were characterized by dynamic light scattering, laser electrophoresis and atomic force microscopy. UV-vis spectroscopy (O-phthaldialdehyde assay, OPA) and Kirby-Bauer tests were used to evaluate drug release and antimicrobial activity, respectively. Physicochemical characterization showed that size, shape and drug solubilization of the nanoparticles mainly depended on GS content and concentration of surface stabilizer (polyvinyl alcohol, PVA). Laser electrophoresis demonstrated negative value of zeta potential of the nanoparticles attributed to PLGA carboxyl end group presence. Drug release studies showed initial burst release followed by prolonged 35-day sustained gentamicin delivery. Agar-diffusion tests performed with pathogens causing osteomyelitis (Staphylococcus aureus and Staphylococcus epidermidis, both reference strains and clinical isolates) showed antibacterial activity of GS loaded nanoparticles (GS-NPs). It can be concluded that GS-NPs are a promising form of biomaterials useful in osteomyelitis therapy.
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September 2016

Injectable gellan gum-based nanoparticles-loaded system for the local delivery of vancomycin in osteomyelitis treatment.

J Mater Sci Mater Med 2016 Jan 30;27(1). Epub 2015 Nov 30.

Department of Biomaterials, Faculty of Materials Science and Ceramics, AGH University of Science and Technology, al. A. Mickiewicza 30, 30-059, Krakow, Poland.

Infection spreading in the skeletal system leading to osteomyelitis can be prevented by the prolonged administration of antibiotics in high doses. However systemic antibiotherapy, besides its inconvenience and often low efficacy, provokes numerous side effects. Thus, we formulated a new injectable nanoparticle-loaded system for the local delivery of vancomycin (Vanc) applied in a minimally-invasive way. Vanc was encapsulated in poly(L-lactide-co-glycolide) nanoparticles (NPs) by double-emulsification. The size (258 ± 11 nm), polydispersity index (0.240 ± 0.003) and surface potential (-25.9 ± 0.2 mV) of NPs were determined by dynamic light scattering and capillary electrophoresis measurements. They have a spherical morphology and a smooth topography as observed using atomic force microscopy. Vanc loading and encapsulation efficiencies were 8.8 ± 0.1 and 55.2 ± 0.5 %, respectively, based on fluorescence spectroscopy assays. In order to ensure injectability, NPs were suspended in gellan gum and cross-linked with Ca(2+); also a portion of dissolved antibiotic was added to the system. The resulting system was found to be injectable (extrusion force 11.3 ± 1.1 N), reassembled its structure after breaking as shown by rheology tests and ensured required burst release followed by sustained Vanc delivery. The system was cytocompatible with osteoblast-like MG-63 cells (no significant impact on cells' viability was detected). Growth of Staphylococcus spp. reference strains and also those isolated from osteomyelitic joints was inhibited in contact with the injectable system. As a result we obtained a biocompatible system displaying ease of application (low extrusion force), self-healing ability after disruption, adjustable drug release and antimicrobial properties.
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http://dx.doi.org/10.1007/s10856-015-5604-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4666281PMC
January 2016

The use of PFGE method in genotyping of selected bacteria species of the Lactobacillus genus.

Methods Mol Biol 2015 ;1301:225-40

Jagiellonian University Medical College, 18 Czysta St., 31-121, Krakow, Poland,

Bacteria belonging to the genus Lactobacillus are a group of microorganisms in which there are numerous strains with high probiotic potential. In recent years, interest in probiotics as nutritional supplements and as drugs has significantly increased. In order to carry out research on probiotic bacterial strains, their beneficial effects on the human body should be confirmed and molecular typing should be carried out with the aim of marking a specific strain. Pulse-field gel electrophoresis (PFGE) is the reference method employed for bacterial genotyping, but it is commonly used for typing mainly pathogenic bacteria. The following study describes a way of performing PFGE analysis in genotyping of bacteria species belonging to the genus Lactobacillus: L. fermentum, L. gasseri, L. plantarum and L. rhamnosus.
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http://dx.doi.org/10.1007/978-1-4939-2599-5_18DOI Listing
December 2015

Dynamics of colonization with group B streptococci in relation to normal flora in women during subsequent trimesters of pregnancy.

New Microbiol 2014 Jul 1;37(3):307-19. Epub 2014 Jul 1.

Department of Bacteriology, Microbial Ecology and Parasitology, Chair of Microbiology, Jagiellonian University Medical College, Krakow, Poland;

The main objective of the study was to compare the qualitative and quantitative composition of vaginal and rectal flora in GBS-positive (n=15) and GBS-negative (n=27) pregnant women examined in three subsequent trimesters of their pregnancy. Study samples consisted of vaginal and rectal smears and urine samples. GBS numbers were determined by the quantitatively cultured method [cfu/ml] and with the use of qPCR. Five GBS colonies were isolated per each positive sample and genotyped by PFGE and serotyping. The normal flora components: Lactobacillus, Bifidobacterium and Candida were quantitatively cultured. Carriage of GBS in subsequent trimesters in vagina/anus was variable and fluctuated between 17% and 28%. Quantitative GBS analyses showed that the vaginal population was at a constant level with the mean value equal to 3.94×104 cfu/ml, in contrast to the rectal population where the highest values appeared in the third trimester 4.37×105. The use of qPCR gave 7% more positive results for vaginal/rectal swabs. Genetic similarity analysis showed that one GBS clone was present in 73% of carriers during pregnancy, while in 27% of patients, 2 clones were found. H2O2-positive vaginal lactobacilli were detected in all women, while H2O2-negative lactobacilli and Bifidobacterium occurred more frequently in the anus in about 50% of women. Candida was present in the vagina in 30% of women. The analysis of women in three consecutive trimesters of pregnancy on the basis of a study group and control group showed no statistically significant differences in either the species (qualitative) or quantitative composition in vaginal and rectal flora in both of the groups. Therefore, GBS should be considered as a component of the microbiota and an opportunistic microorganism rather than a typical pathogen, because it does not distort the composition of women's normal genital tract flora.
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July 2014

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

BMC Infect Dis 2014 Jun 18;14:339. Epub 2014 Jun 18.

Microbiology, Jagiellonian University Medical College, 18 Czysta Street, Krakow 31-121, Poland.

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.
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http://dx.doi.org/10.1186/1471-2334-14-339DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4074408PMC
June 2014

A novel, nested, multiplex, real-time PCR for detection of bacteria and fungi in blood.

BMC Microbiol 2014 Jun 4;14:144. Epub 2014 Jun 4.

Department of Bacteriology, Microbial Ecology and Parasitology, Chair of Microbiology Jagiellonian University Medical College, Czysta 18 Str, 31-121 Kraków, Poland.

Background: The study describes the application of the PCR method for the simultaneous detection of DNA of Gram-negative bacteria, Gram-positive bacteria, yeast fungi and filamentous fungi in blood and, thus, a whole range of microbial etiological agents that may cause sepsis. Material for the study was sterile blood inoculated with four species of microorganisms (Escherichia coli, Staphylococcus aureus, Candida albicans and Aspergillus fumigatus) and blood collected from patients with clinical symptoms of sepsis. The developed method is based on nested-multiplex real-time PCR .

Results: Analysis of the obtained data shows that sensitivity of nested-multiplex real-time PCR remained at the level of 10(1) CFU/ml for each of the four studied species of microorganisms and the percentage of positive results of the examined blood samples from the patients was 70% and 19% for the microbiological culture method. The designed primers correctly typed the studied species as belonging to the groups of Gram-positive bacteria, Gram-negative bacteria, yeast fungi, or filamentous fungi.

Conclusions: Results obtained by us indicated that the designed PCR methods: (1) allow to detect bacteria in whole blood samples, (2) are much more sensitive than culture method, (3) allow differentiation of the main groups of microorganisms within a few hours.
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http://dx.doi.org/10.1186/1471-2180-14-144DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4049433PMC
June 2014

Multilocus sequence types of invasive and colonizing neonatal group B streptococci in Poland.

Med Princ Pract 2014 7;23(4):323-30. Epub 2014 May 7.

Department of Bacteriology, Microbial Ecology and Parasitology, Jagiellonian University Medical College, Krakow, Poland.

Objectives: The present study aimed to investigate the molecular characterization of Streptococcus agalactiae (group B streptococcus; GBS) strains isolated from newborns with invasive neonatal infections and healthy newborns in Poland.

Materials And Methods: Forty-two GBS isolates were characterized by combining different typing methods, i.e. multilocus sequence typing (MLST), molecular serotyping and protein gene profiling.

Results: Using MLST, a total of 16 sequence types (STs) were identified, and among these, 11 were clustered into the following 5 clonal complexes (CCs): CC23 (20; 49%), CC19 (7; 17%), CC17 (4; 10%), CC10 (4; 10%) and CC1 (1; 2%). A statistically significant relationship between ST-17 and invasive isolates (p = 0.0398) and ST-23 and colonizing strains (p = 0.0034) was detected. Moreover, 2 novel STs were detected (ST-637 and ST-638). Molecular serotyping showed that in the invasive isolates serotype III was predominant (11; 50%), followed by serotypes II (6; 27%), V (3; 14%) and Ia (2; 9%). In healthy newborns, serotype III was also dominant (12; 60%), followed by serotypes Ia (4; 20%), II (2; 10%), V (1; 5%) and Ib (1; 5%). Protein gene profiling indicated that the rib gene was predominant in the invasive strains (11; 59%), followed by bca (5; 22%), alp2 (2; 9%), alp3 (1; 5%) and epsilon (1; 5%), while in colonizing strains the alp2 gene was most common (10; 50%), followed by epsilon (5; 25%), rib (2; 10%), bca (2; 10%) and alp3 (1; 5%). A statistically significant relationship was noted between the rib gene and invasive GBS (p = 0.0329), whereas alp2 was related to the colonizing strains (p = 0.0495).

Conclusions: The investigated GBS isolates originating from infections in newborns and healthy neonates represented serotype III in more than half of the cases and differed from one another in terms of resistance to macrolides, ST type affiliation and the presence of genes encoding surface proteins from the Alp family. Further comparative genetic research on a larger number of strains is necessary for epidemiological investigation and vaccine development.
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http://dx.doi.org/10.1159/000362368DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5586894PMC
May 2015

Analysis of genetic similarities between Clostridium perfringens isolates isolated from patients with gas gangrene and from hospital environment conducted with the use of the PFGE method.

Pol Przegl Chir 2014 Mar;86(3):141-6

The objective of the study was to perform a comparative analysis of genetic similarity, with the use of pulsed field gel electrophoresis (PFGE), of Clostridium perfringens isolates originating from patients with gas gangrene and from the hospital environment. The study encompassed two patients with a clinical and microbiological diagnosis of gas gangrene, who were hospitalized in one of the hospitals of the Małopolska province in the time period between 31st March 2012 and 18th May 2012. Clostridium perfringens isolates genotyping indicated that the isolates originating from the two studied patients did not display genetic similarity and represented two different PFGE types, which corresponded to two different clones (clone A and B). Whereas the strains isolated from the hospital environment were genetically identical with the strain coming from the second patient and represented one PFGE type, which corresponded to one clone (clone A). As a result of the study, it is possible to conclude that both patients developed endogenous infection. Even so, the examination of the hospital environment indicates the possibility of the appearance of exogenous infections. It prompts recommending and following the exact regulations of sanitary regime in the ward and the operating theater if a patient is diagnosed with gas gangrene.
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http://dx.doi.org/10.2478/pjs-2014-0026DOI Listing
March 2014