Antibacterial Therapy by Ag+ Ions Complexed with Titan Yellow/Congo Red and Albumin during Anticancer Therapy of Urinary Bladder Cancer.

According to the World Health Organization report, the increasing antibiotic resistance of microorganisms is one of the biggest global health problems. The percentage of bacterial strains showing multidrug resistance (MDR) to commonly used antibiotics is growing rapidly. Therefore, the search for alternative solutions to antibiotic therapy has become critical to combat this phenomenon. It is especially important as frequent and recurring infections can cause cancer. One example of this phenomenon is urinary tract infections that can contribute to the development of human urinary bladder carcinoma. This tumor is one of the most common malignant neoplasms in humans. It occurs almost three times more often in men than in women, and in terms of the number of cases, it is the fifth malignant neoplasm after prostate, lung, colon, and stomach cancer. The risk of developing the disease increases with age. Despite the improvement of its treatment methods, the current outcome in the advanced stages of this tumor is not satisfactory. Hence, there is an urgent need to introduce innovative solutions that will prove effective even in the advanced stage of the disease. In our study, a nanosystem based on ionic silver (Ag+) bound to a carrier-Titan yellow (TY) was analyzed. The possibility of binding the thus formed TY-Ag system to Congo red (CR) and albumin (BSA) was determined. TY-Ag binding to CR provides for better nanosystem solubility and enables its targeted intracellular transport and binding to immune complexes. The binding of TY-Ag or CR-TY-Ag to albumin also protects the system against the uncontrolled release of silver ions. It will also allow the delivery of silver in a targeted manner directly to the desired site in the case of intravenous administration of such a system. In this study, the MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bactericidal Concentration) values of the TY-Ag or BSA-TY-Ag systems were determined in two reference strains (Escherichia coli and Staphylococcus aureus). The paper presents nanosystems with a size of about 40-50 nm, with an intense antibacterial effect obtained at concentrations of 0.019 mM. We have also discovered that TY-Ag free or complexed with BSA (with a minimal Ag+ dose of 15-20 µM) inhibited cancer cells proliferation. TY-Ag complex diminished migration and effectively inhibited the T24 cell viability and induced apoptosis. On the basis of the obtained results, it has been shown that the presented systems may have anti-inflammatory and antitumor properties at the same time. TY-Ag or BSA-TY-Ag are new potential drugs and may become in future important therapeutic compounds in human urinary bladder carcinoma treatment and/or potent antimicrobial factors as an alternative to antibiotics.

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

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.

Non-IgE Mediated Hypersensitivity to Food Products or Food Intolerance-Problems of Appropriate Diagnostics.

The diagnosis of food intolerance is tricky due to the different etiologies of adverse reactions. There is also a lack of clear rules for interpreting alternative tests used to diagnose these problems. The analyses of IgG4 concentration in serum or cytotoxic tests became the basis of elimination diets. However, it can result in nutritional deficiencies and loss of tolerance to eliminated foods. Our study aimed to assess the necessity of food elimination in four cases with food intolerance symptoms based on alternative diagnostic tests. Four patients without food allergies, who manifested diverse clinical symptoms after food, were presented due to the following factors: clinical history, diagnostic tests, elimination diet, and filaggrin gene (FLG) mutation. It was found that higher IgG4 levels against foods and higher cytotoxic test values are not clinically relevant in each of the studied individuals. They should not be decisive for the elimination of food products. The study of FLG-SNVs revealed the association of some clinical symptoms in patients with hypersensitivity to several food allergens and reported genetic variants in the FLG gene.

Hypersensitivity to polyethylene glycol (PEG).

The need for mass population vaccination against Covid-19 poses a public health problem. Allergic symptoms occurring after the 1st dose of the vaccine may result in resignation from the administration of the 2nd dose. However, the majority of patients with mild and/or non-immediate symptoms may be safely vaccinated. The only absolute contraindication to administration of the vaccine is an anaphylactic reaction to any of its ingredients. Polyethylene glycol (PEG), widely used as an excipient in various vaccines, is considered the primary cause of allergic reactions associated with administration of Comirnaty (Pfizer/BioNTech) and Covid-19 Vaccine (Moderna) vaccines. However, hypersensitivity to PEG reported to date seems very rare, considering its widespread use in multiple everyday products, including medicines and cosmetics. In the paper, current literature data describing mechanisms of hypersensitivity reactions to PEG, their clinical symptoms and diagnostic capabilities are presented. Undoubtedly, the issue of hypersensitivity to PEG warrants further research, while patients with the diagnosis require individual diagnostic and therapeutic approach.

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

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 ß-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.

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

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.

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

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 4­5 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.

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

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.

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

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.

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

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.

Component-resolved diagnosis (CRD) of food anaphylaxis

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.

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

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 (p = 0.004) and 4.21% at BAT3 (p < 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%, p = 0.01). Median CD-sens index decreased significantly from 1099.02 at BAT1 to 179.31 at BAT2 (p < 0.002) and 168.04 at BAT3 (p < 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.

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.

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 8th April 2015 and 20th 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 ß2-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.

Bacteria contamination of touch surfaces in Polish hospital wards.

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.

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

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.

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

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.

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

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.

Nursing care for patients infected or colonized with vancomycin-resistant enterococci (VRE).

Advances in medicine enable many patients to regain their health. But, at the same time, they become susceptible to hospital-acquired infections. The occurrence of vancomycin-resistant enterococci is a considerable problem in the modern health system. In order to limit the risk of VRE infection, proper patient care is vital, which is focused on compliance with relevant procedures (isolation, decontamination, education). Mutual cooperation between charge nurses and the ward sister and epidemiological nurse plays a major role in nursing surveillance of patients with VRE.

Prolonged hospitalization of patients with hospital acquired pneumoniae in the intensive care unit ­ morbidity, mortality and costs of

BACKGROUND: The objective of this study was to determine epidemiological rates of hospital-acquired pneumonia (HAP) in the Polish ICU and direct costs associated with excess length of stay incurred by the hospital. MATERIAL AND METHODS: The studied data concern patients hospitalized in the years 2010­2012 in a 9-bed ICU of a multi-profile hospital in the Malopolskie Province. Centers for Disease Control and Prevention diagnostic criteria were the basis for the development of definitions of infections. RESULTS: HAP developed in 34 patients, of which 94.1% were cases associated with the use of mechanical ventilation. Cumulative incidence of HAP was 5.74%, and incidence density amounted to 3.00‰. Median length of stay of patients with HAP amounted to 36 days and this value was more than twenty percent higher than in the case of other forms of infection (27 days), and almost three times higher than in patients without infections (14 days). Annually, costs associated with the excess hospitalization of patients with this form of infection should be evaluated at over PLN 300,000 (72.290€) per hospital; average per patient is PLN 29,106 (7013€). CONCLUSIONS: HAP is one of the most common hospital-acquired infections in intensive care units. It is connected with the greatest additional costs for hospital comparing to other forms of HAIs. For patients, the occurrence of HAP in ICU patients involves the highest risk of death. Detailed, targeted studies concerning financial burden of HAI in Poland should be performed.

[Work experience and seniority in health care vs. medical students' knowledge of selected hand hygiene procedures]. / Praktyka zawodowa i staz pracy w ochronie zdrowia a wiedza studentów kierunków medycznych o wybranych procedurach higieny rak.

BACKGROUND: Hand hygiene (HH) is the most important element of infection prevention. The aim of the study was to analyze the level of HH knowledge among medical students of Jagiellonian University Medical College in correlation with their clinical experience and the presence and extent of trainings in hospital hygiene prior to internships, as well as with HH practice among medical staff perceived by students. MATERIAL AND METHODS: The study was carried out in a group of 414 students from October to December, 2014. The questionnaire built of 14 questions was used as a study tool. RESULTS: Absolutely correct answers to questions about HH were given by 52.9%, and about HH technique by 6.5% of respondents. The degree of accuracy of answers to questions concerning HH did not correlate with the gender of the respondents or with the fact that work placement had been preceded by training in the field of HH or with its scope. A statistically significant correlation was found between the year, the field, and the type of the study. Students with greater professional practice, significantly less often claimed that medical workers comply with HH. Professional practice of 22.9% of students was not preceded by any training in the field of hospital hygiene and in 28% of cases training did not cover HH. Nearly half of the respondents declared that pre-internship training had not addressed the problem of occupational exposure to biological agents. CONCLUSIONS: The results of the study shows that knowledge gained by students participating in the study was not satisfactory. Moreover, there is a need for improving the educational scheme in the discussed subject at all levels of basic and clinical subjects as well as during internships. Med Pr 2016;67(5):623-633.