Introduction: The aim of the study was the analysis of occurrence of genetic determinants of multi-drug resistance and the assessment of genetic relationship among Acinetobacter baumannii strains. Methods: Multiplex-PCR method was performed in order to: (1) confirm the phenotypic identification and (2) detect the presence of CHDL oxacillinases in the group of thirty A.baumannii strains. Further PCR studies included the analysis of the occurrence of genetic determinants associated with efflux pump, insertion sequence and biofilm formation. The relationship between bacterial strains was assayed using 6 primers in RAPD-PCR method. Results: Detection of the blaOXA-51-like gene confirmed that the strains belong to the A. baumannii species. In the multiplex-PCR, the presence of the blaOXA-23-like and blaOXA-40-like genes was detected in 3 (10%) and 27 (90%) isolates, respectively. Moreover, some strains showed the coexistence of the blaOXA-51-like and blaOXA-23-like genes (10%, n=3) or blaOXA-51-like and blaOXA-40-like (90%, n=27). In the group of analysed strains the presence of the efflux pump gene (adeA) and the insertion sequence ISAba1 were demonstrated in all tested isolates. Biofilm-related genes (abaI, csuE) were found in 100% and 97% (n=29) tested strains adequately. The RAPD-PCR studies revealed the presence of 10 unrelated genotypes. Conclusions: The obtained results suggest that the phenomenon of multi-drug resistance in the studied A. baumannii strains could be attributed to the occurrence of CHDL oxacillinases, AdeABC efflux pump, insertion sequence ISAba1 and the biofilm formation.
Staphylococcus pettenkoferi was first isolated from blood culture a 25-years-old patient with fever of unknown etiology. Based on the literature analysis, it was found that it is an opportunistic microorganism. It causes blood stream infections and was an causative agent of osteomyelitis. Staphylococcus pettenkoferi is a component of the human microbiome, was found in a hospital environment and was also isolated from animals and their surroundings.
Introduction: Lateral flow assays (LFIA) are the technology behind low-cost, simple, rapid and portable detection devices popular in biomedicine. Lately, they are very common used in serodiagnosis of SARS-CoV-2 infections. The aim of the presented study was to assess the usefulness of selected LFIA in serological diagnosis of COVID-19. Methods: The usefulness of seven lateral flow assays in the serodiagnosis of COVID-19 was evaluated (VAZYME, DIAGNOSIS, PCL, INGEZIM, BIOSENSOR, ACCU-TELL, NOVAtest). The study used 107 serum samples obtained from 74 individuals with current SARS-CoV-2 infection confirmed by RT-PCR. The ELISA-IgG (Euroimmun) was used as the reference assay for sensitivity and specificity testing. Results: The highest percentage of positive results was obtained when searching for IgG antibodies with the NOVAtest (40.6%) and DIAGNOSIS (39.2%) sets and the lowest detection for the PCL set - 25.5%. In the case of searching for IgM antibodies in all sets, significantly lower percentages of positive results compared to the IgG class were recorded. In general, all lateral flow assays showed low sensitivity in relation to the Euroimmun ELISA-IgG. The DIAGNOSIS kit (64.5%) was characterized by the highest sensitivity, and the PCL kit was the lowest (38.7%). On the other hand, the specificity of all kits was very high, almost 100% in almost all cases. Conclusions: Lateral flow assays due to their low sensitivity are not suitable for quick diagnosis of the current SARS-CoV-2 infections and cannot be an alternative to genetic or even antigen tests. They may be used only to retrospectively test the presence of IgG antibodies. However, a negative results of LFIA in suspected disease or after vaccination should be confirmed by more sensitive serological tests.
Escherichia coli is an important pathogen causing nosocomial infections. A significant problem in the treatment of infections caused by these microorganisms is their increasing resistance to β-lactam antibiotics, including third and fourth generation cephalosporins. The production of β-lactamases enzymes such as extended-spectrum β-lactamases (ESBLs) and AmpC β-lactamases is among the main mechanisms for resistance to third generation cephalosporins. The genes encoding AmpC cephalosporinases are chromosomal (cAmpC) or plasmid-mediated (pAmpC). In E. coli, the expression of the ampC genes may be conditioned by the constitutive expression of low level ampC chromosomal genes. These strains remain susceptible to β-lactam antibiotics. However, mutations in the promoter region of the ampC may result in increased level of expression of chromosomal ampC genes. This can leads to resistance to cephalosporins. Resistance to cephalosporins in E. coli can be also associated with plasmid-mediated AmpC β−lactamases (pAmpC). In E. coli the presence of one or more plasmid-mediated AmpC β−lactamases along with the neglible expression of chromosomal encoded AmpC enzyme can leads to resistance to broad-spectrum cephalosporins. This review is focused on a resistance mechanisms associated with the production of AmpC cephalosporinases in clinical E. coli isolates.
Introduction: ELISA-Immunoassays can complement the molecular diagnostic methods, and can be one of the important tools of sero-surveillance and vaccine evaluation. The aim of the presented study was to develop in-house ELISA and evaluate 11 commercial ELISA tests for detection of anti-SARS-CoV-2 antibodies in serum samples collected from COVID patients. Methods: In total, 237 serum samples obtained from 165 people with COVID-19 with RT-PCR confirmed SARS-CoV-2 virus infection were used for the study. The specificity of the developed in-house ELISA kit was tested using 170 serum samples obtained from patients with various bacterial and viral infections. The study used an in-house ELISA and 11 commercial ELISA kits developed by various manufacturers. Results: The presented study showed high sensitivity (81.0%) and specificity (97.2%) of the developed in-house kit in relation to the RT-PCR method. The sensitivity of the inhouse test significantly increased (98.1%) when only convalescents - persons at least 3 weeks after COVID-19 were examined. Commercial ELISA kits most frequently detected IgG antibodies (from 44.9% to 89.4%), especially in samples obtained later in the disease, and the least frequent detection of IgM antibodies (from 4.2% to 42.4%). Conclusions: All the presented ELISA kits may be used in serodiagnosis of COVID-19 however the detection of antibodies in individual tests differed quite significantly and was dependent on the period of the disease, on the class of immunoglobulins and the type of antigen used. The sensitivity of serological tests in the IgG class is clearly higher when examining samples obtained at least 2-3 weeks from the onset of clinical symptoms. Searching for IgA antibodies may be useful mainly in the early phase of the disease while IgM antibodies does not provide significant additional information. In the case of asymptomatic or mild infection, the level of antibodies is low which may be the cause problems with the correct interpretation of epidemiological surveys
The human digestive tract is the living environment for billions of cells of various microorganisms that are part of the human microflora. The use of modern molecular biology techniques, such as NGS (Next Generation Sequencing), made it possible to study the microorganisms inhabiting the intestines and to understand their impact on human health. The gut microbiota plays a significant role in the synthesis and metabolism of many nutrients and metabolites, including short-chain fatty acids (SCFA), amino acids, lipids, bile acids and vitamins. Many factors such as diet, age, climate, and socioeconomic conditions influence the diversity of the microbiota. Rapid changes in the composition of the microbiota (disturbance of homeostasis) can lead to dysbiosis - a condition associated not only with intestinal disorders, but also with numerous extraintestinal diseases. The present work is a review of current reports on: research techniques used to analyze microbiota, the impact of various factors on its diversity and the impact of microbiota on our health.
Introduction: The aim of the study was to analyze the results of mycological cultures obtained in the years 2014-2019. The study included an analysis of the incidence of mycosis with regard to their location, as well as the proportion of individual etiological factors in the infection. Methods: The study included materials from 999 patients who gave a total of 1103 cultures. The material was taken directly from the material and mycological cultures were established. Results: Positive results accounted for 35,8%. Trichophyton rubrum (44.2%) was the most common etiologic agent of dermatophytes. Among the yeast-like fungi, Candida albicans (8.8%) and Candida parapsilosis (7,6%) were the most common. Conclusions: Infectious lesions were mainly caused by dermatophytes, where Trichophyton rubrum and Trichophyton mentagrophytes dominated.
The aim of this review is to report the medicinal plants, that are effective on Trichomonas vaginalis. Trichomonosis, caused by the flagellate protozoan vaginalis, is the most common non-viral sexually transmitted disease worldwide. The mainstay medication for trichomoniasis is metronidazole, but some resistant strains to this treatment have been identified. In addition, metronidazole sometimes causes adverse effects. Plant metabolites containing alkaloids, monoterpenes, sesquiterpenes, phenylpropane derivatives, tannins, flavonoids, glycosides, lipids, saponins and lactones were found to possess antitrichomonas properties. Medicinal plants are still invaluable source of safe, less toxic, available and reliable resources of drugs.
Introduction: Diabetic foot ulcer (DFU) caused by Staphylococcus aureus is one of the most feared complications of diabetes mellitus. The studies reporting the oral cavity as a potential reservoir of S. aureus in diabetic patients are sparse. The aim of the study was to compare the prevalence of methicillin-resistant Staphylococcus aureus strains in the oral and in the diabetic foot specimens from DFU patients. Materials and Methods: A total 80 specimens (40 oral swabs and 40 DFU swabs) were collected from diabetic patients with foot ulcer. The specimens were subcultured and the susceptibility of isolated S. aureus strains to antimicrobial agents was determined. Suspected methicillin-resistant S. aureus (MRSA) strains were further examined for the presence of modified PBP2a protein. Results: Less than one-fifth of patients with DFU had oral S. aureus carriage, however the colonization is significantly associated with S. aureus diabetic foot infection. S. aureus strains were isolated from 52.5% of DFU specimens, 17.5% were resistant to methicillin. S. aureus strains were isolated from 17.5% of oral specimens of diabetic patients; 2.5% were methicillin-resistant. The MRSA strains were isolated sevenfold more frequently from the diabetic foot than from the oral cavity. Conclusions: Although diabetic foot infections caused by an endogenous S. aureus strains colonizing the oral cavity of diabetic patients seems unlikely, it is evidently important to monitor the oral S. aureus carriage in diabetic patients and their resistance to antibiotics.
Introduction: Lyme disease is a multiorgan tick-borne disease which endemically occurs primarily in countries in the Northern Hemisphere. It is caused by the bacteria Borrelia burgdorferi, and the vector of infection are ticks from the Ixodes family. In recent years it has been noted that there is an increase in the number of Lyme disease cases in the world. Due to the varied clinical picture of disease and the possibility of complications, Lyme disease should be diagnosed early and properly treated. The objective of the research was an analysis of the occurrence of Lyme disease among inhabitants of “bielski” district and the city of Bielsko-Biala in years 2008-2017. Methods: The analysis of the occurrence of Lyme disease was based on data collected from the Sanitary Epidemiological Station in Bielsko-Biala. Results: In the analysed period there were 1450 cases of Lyme disease diagnosed, mostly among people living in the city which has a population of more than 10000 inhabitants. The disease most frequently afflicted women 51 to 60 and 61 to 70 years of age. Tick bites occurred most frequently during summer months. The most common symptom of infection was malaise, fever and erythema at the site of the tick bite. Conclusions: In the analysed period covering the years 2008-2017, the number of cases of Lyme disease among people living in Bielsko-Biala and the surrounding area remains on the similar level with a noticeable significant increase in 2009 and a decrease in 2014-2015. Women were suffering from Lyme disease more often than men. Lyme disease affected people of all ages. However, the group at risk consisted of people in their fifties and sixties. Tick bites, which resulted in bacterium Borrelia burgdorferi infection, occurred between July and November with the intensity in the summer months.
Objectives: Diagnostics of anaerobic bacterial infections and determination of drug susceptibility are technically difficult and time-consuming; therefore, the number of studies on Anaerobic Gram-negative bacilli is significantly limited, especially in Europe. The aim of the study was to analyze the antibiotic susceptibility of clinically important anaerobic bacteria Bacteroides spp. and Parabacteroides distasonis. Strains were isolated from infections of patients hospitalized at one Polish hospital as a result of routine microbiological diagnostics. Material and methods: Clinical isolates were identified with MALDI-TOF MS. Antimicrobial susceptibility of 276 strains was carried out by E-test gradient strip to commonly used antibiotics i.e. benzylpenicillin, amoxicillin with clavulanic acid, imipenem, clindamycin and metronidazole. MIC values were determined. The interpretation of antimicrobial susceptibility tests were conducted in accordance with The European Committee on Antimicrobial Susceptibility Testing (EUCAST) recommendations. Results: Susceptibility tests of all isolates yielded the following rates of resistance to the evaluated β-lactam antibiotics: benzylpenicillin (96%), amoxicillin/clavulanic acid (7.6%), imipenem (2.1%). In presented study 38.8% of clindamycin-resistant strains were isolated, among them 18.3% of B. fragilis and 53.85% P. distasonis. All strain were susceptible to metronidazole. Conclusions: Obtained results and analysis of the results of other researchers convinces us that it is necessary to routinely or at least periodically monitor drug susceptibility of clinical isolates of anaerobic bacteria and use targeted therapy based on the result of the antibiogram. Although high percentage of the tested Bacteroides and Parabacteroides strains remained susceptible to metronidazole and β-lactam antibiotics the use of clindamycin in empirical therapy may not be efficacious. Antibiogram results should be consult with the staff responsible for patient treatment and hospital antibiotic policies.
Objectives: The aim of this study was to analyze the resistance, virulence and biofilm production of PA strains from respiratory tract infections. Methods: All strains were tested for antibiotic susceptibility according to current EUCAST guidelines. Several resistance and virulence genes were detected by PCR. Biofilm formation was detected by the crystal violet method. Results: 38 strains (47.5%) were categorized as XDR, and 12 (15%) as MDR. Seven (10%) MBL producers were identified. The VIM gene was detected in two strains. Genes connected with aminoglycoside resistance: aac(6΄)-I and ant(2΄΄)-I were detected in 10 strains each, aac(6′)-Ib in 18 strains (21%), and aph (3’)-IV in 7 strains (8%). The most prevalent virulence genes were exoT (95%), apr (93%), A, phzS (95%), and plcH (95%). All the tested strains were strong biofilm producers. Conclusions: The results demonstrate high frequency and heterogeneity of virulence gene profiles among PA isolates from respiratory tract infections. A high number of MDR and XDR strains poses a serious threat for patients and shows clearly that new antimicrobial agents are needed.
Introduction: N. gonorrhoeae is one of the etiological causative agents of one of the most common sexually transmitted diseases. Gonococci has created many resistance mechanisms, which is associated with bacterial evolution. Natural transformation is the basic method of horizontal gene transfer in bacteria of the genus Neisseria, which can lead to a mutation in the gyrA gene encoding DNA gyrase. The aim of the study was to verify the view on the significance of mutations at positions 91 and 95 of the gyrA protein on the sensitivity of N. gonorrhoeae to antibiotics of the quinolone type. Methods: GyrA gene was introduced into an sensitive isolate of N. gonorrhoeae using genetic transformation. Resistance gene donor, recipient and transform strains were tested for susceptibility and the gyrA gene was sequenced. Results: It has been shown that double mutation in amino acid acid sequence of the GyrA protein at positions 91 and 95 increase the value of MIC from 0,003 mg / L to 0,125 mg / L at CIP sensitive N. gonorrhoeae strain. Conclusions: Mutations in the amino acid sequence at positions 91 and 95 affet the strain’s sensitivity to ciprofloxacin, but it is not the only mechanism which could alter the MIC value of quinolones.
After gastrointestinal infection with Y. enterocolitica and Y. pseudotuberculosis, some patients may experience reactive arthritis and erythema nodosum. In the etiopathogenesis of these autoimmune complications, in addition to infectious agents, genetic factors, gender and age of the patients also play a role. In the study of the etiology of reactive arthritis and erythema nodosum after gastrointestinal infection, it is useful to investigate in the serum of patients specific antibodies for selected intestinal pathogens, including Yersinia antigens.
Introduction: Candida albicans survival tests on PCV carriers are necessary for the proper determination of the effectiveness yeasticidal activity of disinfectants. C. albicans is a pathogenic microorganism that causes fungal diseases and therefore its spread in the medical and non-medical areas should be limited by disinfection. Methods: Survival of C. albicans was estimated with using principles of PN-EN 16615 standard. Suspension of C. albicans with interfering substances (0,3 g/l bovine albumin) was dry on PCV carriers under a laminar chamber without fan, at the ambient temperature for no longer than 60 minutes. C. albicans was recovered from the carriers immediately after drying and after contact time (1; 5, 10, 20, 40; 60 minutes). Results: The drying conditions applied reduced the recovery of C. albicans from 1.21 to 2.07 on a logarithmic decimal scale with respect to the test suspension with interfering substances. The difference between the recovery immediately after drying and the recovery after the tested contact times (up to 60 minutes) was insignificant. Conclusions: Achieving the number of C. albicans after the drying process, as provided for in PN-EN 16615, requires further improvement of the drying conditions or an increase of C. albicans suspension density before drying.
Introduction: Salmonella is a serious cause of gastroenteritis in children. Prevention through proper hygiene to minimize potential exposure is a key factor decreasing the number of morbidity. The aim of this study was the analysis of Salmonellosis incidence rate among children hospitalized in Pediatric Hospital in Bielsko-Biała, Poland, in the years 2014-2015 having regard to Salmonella’s serotype, symptoms at admission to hospital and changes of body weight during hospital stay. Methods: The study was conducted on 79 children with Salmonellosis treated in years 2014-2015 in Pediatric Hospital in Bielsko-Biała, Poland. It was based on retrospective analysis of data from children’s medical history. Results: In all age groups the most common serotype was Salmonella Entertidis. Analysis of children’s clinical symptoms at admission to hospital showed that all children, both male and female, had abdominal pains before defecation. The major symptoms included also diarrhea with blood and fever. Among half of observed group emesis were observed. The analysis of body weight at the hospital admission and discharge demonstrated that in majority of children the body weight increased during their stay in the hospital – the weight gain occurred in 68,8% of male and in 48,4% of female children. Conclusions: The cases of Salmonella appeared comparatively often in children at the age of 0-2 years old and 3-5 years old. The most commonly isolated serotype of Salmonella was Salmonella Entertidis. In all age groups the most common symptoms were abdominal pain before defecation and diarrhea with blood. Due to high diarrhea prevalence it is advisable to introduce the education directed both to children and parents.
Introduction: Silver and copper nanoparticles (AgNPs, CuNPs) applied as hydronanocolloids are known to produce strong antibacterial and antifungal activities. They are extensively used in a number of applications including pharmacy, medicine and cosmetology (especially for surface-applied treatment of skin lesions) as well as agriculture, industry (paint, construction, etc.) and home or office (mainly disinfection applications). Moreover, there is a promising perspective of an intra-systemic NP use, especially to optimize targeted drug delivery. For the above reasons NPs cause risk of penetrating human body and exerting toxic effects and/or stress reactions. This issue has inspired the authors to launch studies on the influence of colloidal AgNPs and CuNPs on physiological potential of neutrophils, blood-cells acting as the first line of immunological defense. Methods: Physiological activity of neutrophils was evaluated by measuring their ability to generate oxygen radicals (radical oxygen species, ROS) due to the respiratory (oxidative) burst mechanism. Human, peripheral blood-isolated neutrophils were stimulated with a standard activating agent (polystyrene latex particles) to develop high physiological potential revealed by enhanced ability to produce oxygen radicals. The cells were treated with silver and copper hydronanocolloids (each applied at concentrations ranging from 0.4 to 50 mg/kg) alternatively: in the absence and presence of the mentioned activator. The level of generated ROS upon oxidative burst was monitored chemiluminometrically. Results: The tested Ag and Cu-nanocolloids were not toxic against neutrophils although they hampered mitochondrial dehydrogenase activities when applied at higher levels. At lower concentrations they tended to stimulate ROS generation; however the treatment did not launch the oxidative burst. In the case of the latex-stimulated neutrophils, both types of nanoparticles in all experimental variants did not influence the levels of produced ROS. Conclusions: The obtained results indicate that the exposure of human neutrophils to colloidal AgNPs and CuNPs does not lead to an enhanced ROS generation, which may enable direct intra-blood application of the tested nanostructures, provided further necessary toxicological studies are carried out.
Introduction: Tularemia is a highly infectious zoonotic disease caused by Gram-negative bacterium Francisella tularensis. The microbiological diagnosis of tularemia is based mainly on serological investigations. The present study was undertaken to determine the avidity of IgG class antibodies to Francisella tularensis in the course of tularemia in humans and to evaluate its value for estimation of the phase of diseases. Methods: Fifty two serum samples obtained from 40 patients with tularemia were tested by in-house ELISA in duplicate in the same plate, without and after the 0.5 h incubation with 8M urea. The age of the subjects was between 6 and 77 years. From one patient, a 9-years-old girl with oculoglandular form of tularemia, five serum samples were taken, respectively after 0.5, 1.5, 3, 6 and 12 months from the beginning of the first clinical symptoms. Results: The results of the study showed higher values of the avidity index (AI) of IgG antibodies for F. tularensis, often exceeding the value of 0.9, in children and adolescents than in adults. The examination of serum samples obtained 2-3 times in the course of tularemia from few patients did not show significant differences in the level of avidity index depending on the period of the disease. However, in five serum samples obtained from a 9-years-old girl in the different phases of tularemia the avidity index showed increasing values (0.51, 0.80, 0.92, 0.90 and 0.94, respectively). Conclusions: The avidity index of IgG may be helpful in excluding recent infection, but its usefulness in detecting an active phase of invasion requires further research.
Introduction. Photodynamic therapy is supportive method of non-surgical treatment of periodontitis. The aim of the study was the assessment of photodynamic therapy impact on periodontal disease clinical parameters with a particular emphasis on microbiological aspects during periodontal treatment. Material and methods. The research was conducted on seven healthy patients aged 26- 55 years. All the patients were treated with a non-surgical (SRP) periodontal treatment. Then, the assisted photodynamic therapy was applied in selected pockets. Samples of the material obtained from periodontal pockets before and after the treatment were cultured in a microbiological laboratory. Results. After the introduced therapy the clinical improvement of periodontal disease was observed in the majority (6/7) of patients taking part in the study. There obtained the elimination of inflammation to 85.7% of the patients. Periodontal pocket depths have deteriorated from 0.5 to 3.0 mm in comparison to the pre-treatment values. The overall number of bacteria compared to the pre-treatment value was reduced. In addition, there was noticed a decrease in the number of periopathogens. Control samples showed a vivid dominance of the physiological flora. Conclusions. The non-surgical treatment of periodontitis in combination with the supportive action of photodynamic therapy has promising therapeutic effects by reducing the inflammation and decreasing the depth of periodontal pockets. The change in the ratio of periopathogens to physiological flora may indicate the restoration and maintenance of the state balance in periodontal pockets for patients with periodontitis.
Introduction: Candida parapsilosis and Candida glabrata are another yeasts that form complexes of crypospecies. Although these species have been described more than a decade ago, knowledge about them is still limited. The reason for this is the large phenotypic similarity that unables them from being differentiated by classical diagnostic methods. The aim of the study was to identify species of clinical strains within C. glabrata and C. parapsilosis complexes. Material and methods: Standard PCR-RFLP of the secondary alcohol dehydrogenase gene (SADH) with BanI restriction enzyme served to determine species affiliation within the C. parapsilosis complex. The internal transcribed spacer was used to confirm the identification of C. glabrata sensu stricto. The D1/D2 domain of the 26S rDNA gene was sequenced in order to identify C. nivariensis and C. bracarensis strains. Results: As a result of the molecular analysis, 24 Candida nivariensis isolates and 4 C. metapsilosis strains and 9 C. orthopsilosis strains were detected. Conclusions: Prevalence of new cryptic species was relatively low.
