In vitro activity of plazomicin and other aminoglycosides against Klebsiella pneumoniae multidrug-resistant strains.

Plazomicin is a new aminoglycoside with broad-spectrum activity against multidrug-resistant strains. The aim of this study was to assess the susceptibility of the K. pneumoniae strains to plazomicin and other aminoglycosides. The activity of plazomicin in combination with ceftazidim-avibactam or meropenem with selected strains was evaluated. The study involved 60 ESßL-positive K. pneumoniae isolates and 50 carbapenemase-positive. The susceptibility to aminoglycosides was tested using the gradient strip. The in vitro activities of plazomicin and ceftazidim-avibactam or meropenem were evaluated using the MTSTM cross synergy method. Plazomicin exhibited high activity against K. pneumoniae with MICs ranging from 0.19 to 4 µg ml-1 for ESßL-positive strains and from 0.25 to 256 µg ml-1 for carbapenemase-positive strains. No antagonism was identified with any combinations. Plazomicin demonstrated excellent in vitro activity against analyzed strains, suggesting that this antibiotic may be an effective therapeutic option in the treatment of infections caused by MDR K. pneumoniae strains.

A New 4-Thiazolidinone Derivative (Les-6490) as a Gut Microbiota Modulator: Antimicrobial and Prebiotic Perspectives.

A novel 4-thiazolidinone derivative Les-6490 (pyrazol-4-thiazolidinone hybrid) was designed, synthesized, and characterized by spectral data. The compound was screened for its antimicrobial activity against some pathogenic bacteria and fungi and showed activity against Staphylococcus and Saccharomyces cerevisiae (the Minimum Inhibitory Concentration (MIC) 820 µM). The compound was studied in the rat adjuvant arthritis model (Freund's Adjuvant) in vivo. Parietal and fecal microbial composition using 16S rRNA metagenome sequences was checked. We employed a range of analytical techniques, including Taxonomic Profiling (Taxa Analysis), Diversity Metrics (Alpha and Beta Diversity Analysis), Multivariate Statistical Methods (Principal Coordinates Analysis, Principal Component Analysis, Non-Metric Multidimensional Scaling), Clustering Analysis (Unweighted Pair-group Method with Arithmetic Mean), and Comparative Statistical Approaches (Community Differences Analysis, Between Group Variation Analysis, Metastat Analysis). The compound significantly impacted an increasing level of anti-inflammatory microorganisms (Blautia, Faecalibacterium prausnitzii, Succivibrionaceae, and Coriobacteriales) relative recovery of fecal microbiota composition. Anti-Treponemal activity in vivo was also noted. The tested compound Les-6490 has potential prebiotic activity with an indirect anti-inflammatory effect.

In Vitro Activity of "Old" and "New" Antimicrobials against the Klebsiella pneumoniae Complex.

The Klebsiella pneumoniae complex is a commonly isolated bacteria in human infections. These opportunistic pathogens pose a serious threat to public health due to their potential transmission to the human population. Resistance to carbapenems is a significant antimicrobial resistance mechanism, leading to limited therapeutic options. Therefore, the aim of this study was to evaluate the in vitro activity of fosfomycin, colistin, ceftazidime-avibactam, and meropenem-vaborbactam against multidrug-resistant K. pneumoniae complex strains. This study involved 160 strains of Gram-negative rods, comprising 138 K. pneumoniae and 22 K. variicola. The minimal inhibitory concentration of fosfomycin was estimated using the agar dilution method, and for colistin, the microdilution method was employed. Susceptibility to ceftazidime-avibactam and meropenem-vaborbactam was determined using the gradient strip method. All analyzed K. pneumoniae complex isolates produced extended-spectrum ß-lactamases, and 60.0% exhibited carbapenemases. The majority of the analyzed strains were susceptible to fosfomycin and colistin (62.5%). Among pandrug-resistant K. pneumoniae complex isolates, the highest susceptibility was observed with colistin (43.9%). Fosfomycin demonstrated good activity against ESßLs- and VIM-positive isolates from this complex. Colistin also exhibited satisfactory in vitro activity against VIM- and KPC-positive isolates from the K. pneumoniae complex. Ceftazidime-avibactam displayed good activity against K. pneumoniae complex strains producing ESßLs, KPC, and OXA enzymes. Additionally, meropenem-vaborbactam showed satisfactory in vitro activity against ESßLs- and KPC-positive isolates from this complex.

Satisfactory In Vitro Activity of Ceftolozane-Tazobactam against Carbapenem-Resistant Pseudomonas aeruginosa But Not against Klebsiella pneumoniae Isolates.

Background: Gram-negative rods are one of the most commonly isolated bacteria within human infections. These microorganisms are typically opportunistic pathogens that pose a serious threat to public health due to the possibility of transmission in the human population. Resistance to carbapenems is one of the most important antimicrobial resistance mechanisms amongst them. The aim of this study was to evaluate ceftolozane-tazobactam in vitro activity against carbapenem-resistant Pseudomonas aeruginosa and Klebsiella pneumoniae clinical strains. Information on the antimicrobial activity of this antimicrobial against Gram-negative rods was also supplemented with a brief review of the relevant literature. Methods: The research involved 316 strains of Gram-negative rods: P. aeruginosa-206 and K. pneumoniae-110. Results: Of the tested strains, 86.0% P. aeruginosa and 30.0% K. pneumoniae remained susceptible to ceftolozane-tazobactam. Conclusions: Therefore, ceftolozane-tazobactam might be a good option in the treatment of infections caused by carbapenem-resistant P. aeruginosa strains, including those in ICU patients. Meanwhile, due to dissemination of ESBLs among K. pneumoniae strains, infections with this etiology should not be treated with the ceftolozane-tazobactam combination.

Evaluation of the RESIST-4 O.K.N.V. K-SeT for detection of carbapenemases in Gram-negative bacilli.

Enterobacterales as opportunistic pathogens are commonly associated with nosocomial infections. With increasing frequency, Gram-negative bacilli, especially Klebsiella pneumoniae strains, are mul- tidrug-resistant or pandrug-resistant. Carbapenems were used as the drugs of choice for the treat- ment of infections caused by multidrug-resistant Gram-negative bacilli. The aim of this study was to assess the usefulness of the RESIST-4 O.K.N.V. K-SeT for the rapid detection and identification of the most important carbapenemases (OXA-48, KPC, NDM, VIM) in Enterobacterales bacilli. The study involved the isolates of 97 Enterobacterales strains. The ability to produce carbapenemases was determined by the immunochromatographic RESIST-4 O.K.N.V. K-SeT test. This test detected carbapenemases OXA-48, KPC, NDM, and VIM. For the RESIST-4 O.K.N.V. K-SeT test, a positive result was obtained for 93 strains (95.9%). Four strains negative in the RESIST-4 O.K.N.V. K-SeT were positive in the Eazyplex®SuperBugCRE and PCR. These strains produce VIM enzymes. RE- SIST-4 O.K.N.V. K-SeT test is rapid, simple to perform and can be used for fast detection of the most important carbapenemases (OXA-48, KPC, NDM, VIM) among Gram-negative bacilli.

The Evaluation of Eazyplex® SuperBug CRE Assay Usefulness for the Detection of ESBLs and Carbapenemases Genes Directly from Urine Samples and Positive Blood Cultures.

Increasing antimicrobial resistance of Gram-negative rods is an important diagnostic, clinical and epidemiological problem of modern medicine. Therefore, it is important to detect multi-drug resistant strains as early on as possible. This study aimed to evaluate Eazyplex® SuperBug CRE assay usefulness for beta-lactamase gene detection among Gram-negative rods, directly from urine samples and positive blood cultures. The Eazyplex® SuperBug CRE assay is based on a loop-mediated isothermal amplification of genetic material and allows for the detection of a selection of genes encoding carbapenemases, KPC, NDM, VIM, OXA-48, OXA-181 and extended-spectrum beta-lactamases from the CTX-M-1 and CTX-M-9 groups. A total of 120 clinical specimens were included in the study. The test gave valid results for 58 (96.7%) urine samples and 57 (95.0%) positive blood cultures. ESBL and/or carbapenemase enzymes genes were detected in 56 (93.3%) urine and 55 (91.7%) blood samples, respectively. The Eazyplex® SuperBug CRE assay can be used for a rapid detection of the genes encoding the most important resistance mechanisms to beta-lactams in Gram-negative rods also without the necessity of bacterial culture.

The Differences in the Level of Anti-SARS-CoV-2 Antibodies after mRNA Vaccine between Convalescent and Non-Previously Infected People Disappear after the Second Dose-Study in Healthcare Workers Group in Poland.

(1) Background: In many infections, antibodies play a crucial role in controlling infection. In COVID-19, the dynamics of the immune system response to SARS-CoV-2 is not fully understood. (2) Methods: The study was conducted on 120 healthcare workers from Dr. Antoni Jurasz University Hospital No. 1 in Bydgoszcz, between June and December 2020. In all participants, IgA and IgG antibody serum concentrations were measured using the semi-quantitative Anti-SARS-CoV-2 ELISA test (Euroimmun). After vaccination, in January and February 2021, antibody levels were examined using the quantitative IgG Anti-SARS-CoV-2 Quantivac ELISA test (Euroimmun). (3) Results: During the whole study period, the SARS-CoV-2 infection was confirmed in 29 (24.2%) participants. In all infected participants, IgA and IgG antibodies were detectable after infection by semi-quantitative serological tests. Levels of antibodies were higher one month after the first dose in the convalescents than in the non-previously infected participants. In this second group, the level of antibodies increased significantly after the second dose of vaccines compared to the first dose. (4) Conclusions: The level of antibodies after the first dose of vaccine in the convalescents' group is higher than in the SARS-CoV-2 non-infected group, but the differences disappear after the second vaccination.

Klebsiella pneumoniae infection in a liver transplant patient: A case report and review of the literature.

The share of Klebsiella pneumoniae in infections has been recently increasing. Multidrug-resistant strains that produce more than one antibiotic resistance mechanism are also increasingly isolated. Contamination of the organs preservation fluid occurs quite often, but the isolated microorganisms are mainly saprophytic bacteria that are part of the skin microbiota (coagulase-negative Staphylococcus, Corynebacterium spp). The following case describes a K. pneumoniae blood infection in a patient after liver transplantation. Susceptibility of the strains to chosen antimicrobials was determined using the automated method. For strain isolated from blood, it was confirmed by loop-mediated isothermal amplification of genetic material.

Vibrio metschnikovii: Current state of knowledge and discussion of recently identified clinical case.

Vibrio metschnikovii is a widespread opportunistic pathogen that rarely causes disease in human. It caused graft infection in our case. It is important to differentiate it from another water-transmitted pathogens.

Raoultella terrigena: Current state of knowledge, after two recently identified clinical cases in Eastern Europe.

Raoultella terrigena is a rarely found opportunistic pathogen that can cause healthcare-associated infections with high mortality. It is important to differentiate it from Klebsiella species.

Evaluation of the usefulness of selected methods for the detection of carbapenemases in Klebsiella strains.

Introduction. Klebsiella rods, belonging to the family Enterobacteriaceae, are generally opportunistic pathogens commonly associated with nosocomial infections, especially in intensive care units. Interestingly, strains of this genus also show multi-drug resistance. In recent years, multiple studies have indicated that the prevalence of carbapenem resistance has increased rapidly among Klebsiella representatives.Aim. The aim of this study was to assess the usefulness of selected phenotypic and genotypic methods for the detection of the most important carbapenemases in Klebsiella strains.Methodology. The study involved 51 Klebsiella strains. The ability to produce carbapenemases was determined by phenotypic methods (double disc synergy test, test with four discs and three inhibitors, CarbaNP test, culture on chromogenic medium, panels of automatic method - Phoenix, CIM test and modified Hodge test). The potential for carbapenemase synthesis was also evaluated using real-time PCR, detecting bla VIM/IMP, bla KPC, bla NDM and bla OXA-48 genes.Results. Using the phenotypic methods, positive results were obtained for all of the analysed strains. Using PCR, carbapenemase synthesis potential was confirmed on the molecular level; the bla VIM gene was detected in 23 strains, the bla NDM gene in 26 strains and the bla OXA-48 gene in two strains.Conclusion. There was complete agreement between the carbapenemases detected by the genetic method and the results obtained with phenotypic methods.

Raoultella spp. - reliable identification, susceptibility to antimicrobials and antibiotic resistance mechanisms.

Introduction. Raoultella spp. representatives are Gram-negative rod-shaped bacteria of the Enterobacteriaceae family. These bacteria are commonly found in the natural environment.Aim. The aim of the study was to indicate the reliable method for Raoultella spp. strains identification, evaluate the susceptibility of Raoultella spp. strains to selected antimicrobials and to detect their resistance mechanisms to beta-lactams.Methodology. Susceptibility of the strains to chosen antimicrobials was determined using the automatic method. The presence of particular antimicrobial resistant mechanism and genes encoding ESBLs and MBLs was determined respectively with double-disc synergy test and commercially available kit - eazyplex SuperBug CRE test (Amplex Diagnostics) and standard PCR. For the selected strains, DNA sequencing was performed.Results. Amongst 105 of the examined Raoultella spp. strains, majority were sensitive to: imipenem (99.0 %), meropenem (98.1 %), gentamicin (93.3 %) and ciprofloxacin (92.4 %). Of the tested Raoultella strains, thirteen (12.4 %) produced ESBLs and one strain simultaneously ESBLs and MBLs. The DNA sequencing results were as follows: for all the reference strains the correct species identification was achieved, for the analysed strains two were identified as R. planticola and one as R. ornithinolytica.Conclusion. Although Raoultella spp. strains remain sensitive to antibiotics, there is a constant need to monitor the sensitivity of these bacteria to selected antimicrobials. Isolation of a multi-drug resistant R. ornithinolytica strain indicates that even the less frequently isolated species of Enterobacteriaceae family should be precisely identified because they might be of clinical importance and the particular strain can also produce enzymes that pose the greatest threat today.

Evaluation of eazyplex® SuperBug CRE Test for Beta-Lactamase Genes Detection in Klebsiella spp. and P. aeruginosa Strains.

The multi-drug resistance of Gram-negative rods is one of the most important issues of present medicine. In recent years, more and more strains resistant to the majority or to all possible therapeutic options have been isolated-especially Klebsiella spp. and Pseudomonas spp. representatives. It is very important to detect strains with these phenotypes as quickly and reliably as possible. The aim of the study was to evaluate the usefulness of eazyplex® SuperBug CRE test (Amplex Diagnostics) for the detection of the most important beta-lactam resistance genes. eazyplex® SuperBug CRE test is based on the loop-mediated isothermal amplification (LAMP) method, and detects genes for the following beta-lactamases: KPC, NDM-1, VIM, OXA-48, CTX-M1, CTX-M9 and OXA-181. The study involved 87 strains. For all of the positive strains in the LAMP method, additional PCR were performed to increase the spectrum of ESBLs detected by the genes encoding for enzymes belonging to TEM and SHV families. The results obtained by the tested method and standard PCR were consistent for all Klebsiella spp. strains. The discrepancy between the evaluated test and PCR results was observed for one P. aeruginosa strain. The eazyplex® SuperBug CRE test can be used for quick detection of the most important beta-lactam resistance mechanisms amongst Gram-negative rods.

Emergence of colistin-resistant Klebsiella pneumoniae in Poland.

In recent years, colistin has been the drug of choice for treatment of nosocomial infections, especially in bloodstream infections, lower respiratory tract infections, or urinary tract infections. In this study, 65 multidrug-resistant Klebsiella pneumoniae isolated from different clinical samples were included. Minimum inhibitory concentration (MIC) of colistin was detected by broth microdilution method in three different ways. For selected K. pneumoniae strains, eazyplex SuperBug mcr-1 test was performed. This test detects mcr-1 gene, which encodes a colistin-resistance determinant. Most of the analyzed K. pneumoniae strains were resistant to colistin in all applied methods. The exception was two strains, where MIC of colistin was 2 mg/L in SensiTest Colistin and MIC-Strip Colistin tests. In MIC COL test, MIC for these strains was 4 mg/L. All analyzed strains produced extended-spectrum beta-lactamases and 11 (16.9%) metallo-beta-lactamases. Eleven (16.9%) K. pneumoniae strains were resistant to all antibiotics, whereas 17 (26.1%) were susceptible to only one drug. Colistin MIC values varied from 2 to >64 mg/L in MIC-Strip Colistin test; from 2 to >16 mg/L in SensiTest Colistin and from 4 to >16 mg/L in MIC COL test. None of the analyzed K. pneumoniae strains carried mcr-1 gene. Data of this work suggest that resistance to colistin emerged among multidrug-resistant K. pneumoniae strains. The tests allowed for reliable estimation of susceptibility to colistin and could be used in microbiological diagnostics.

Comparison of the effectiveness of dipping agents on bacteria causing mastitis in cattle.

INTRODUCTION: Mastitis may result in physical, chemical and microbiological changes in milk and pathological lesions in the glandular tissue. Milk derived from cows with mastitis may become a cause of infections in humansw and animals. OBJECTIVES: The aim of this study was to assess the effectiveness of selected dipping agents in the inactivation of several bacteria that may cause mastitis in cattle. MATERIAL AND METHODS: Three strains of each of the following species: Escherichia coli, Klebsiella pneumoniae, Serratia marcescens, Staphylococcus aureus and Listeria monocytogenes, isolated from milk, were used in the study. Identification of isolates was carried out using the automatic system VITEK2 Compact. Evaluation of the genetic similarity between the tested strains was made using the RAPD technique. Drug susceptibility of strains was evaluated with the disc diffusion method. Assessment of the effectiveness of iodine, stabilized iodine, povidone iodine and chlorhexidine was performed using fragments of skin from cow teats. RESULTS: All the tested strains were genetically different. Most of them were susceptible to the studied antibiotics. Only two strains of L. monocytogenes were resistant to all the studied antibiotics. The percentage rate of reduction in the number of bacteria after using of dipping agents was very high (>90%). The most susceptible to the dipping preparations used were L. monocytogenes (99.6 - 99.9%). Stabilized iodine was the most effective dipping agent for all tested bacteria, causing a reduction rate in the number of bacteria from 99.80% (E. coli) - 99.99% (S. aureus, L. monocytogenes). CONCLUSIONS: The results obtained may contribute to a reduction in udder infections in cows, especially mastitis, and improve the quality of the milk.

Fulminant mucormycosis after a traffic accident: a case report.

Mucormycosis is a rare fungal infection in immunocompetent patients, whereas in immunocompromised, it may be systemic and disseminated infection associated with high mortality. Mucormycosis is one of the most rapidly progressing and fulminant forms of fungal infections; Mucor circinelloides is rarely isolated species, also from immunocompromised patients. The reported case of mucormycosis after a traffic accident indicates that it may be the result of a contamination of wound by M. circinelloides coming from the environment. The fungal strain was identified by phenotypic methods and confirmed by molecular methods. Etest method was used for susceptibility testing of the fungal strain. No mycotoxins were detected in the analyzed sample. The infection was successfully treated with amphotericin B, but amputation of the lower limb was necessary.

Identification of Raoultella spp.: Comparison of three methods.

Background: Raoultella is a Gram-negative bacteria, which commonly occur in the natural environment such as water, soil and on plants. In recent years, Raoultella spp. gained more interest. There is also an increasing number of publications describing mainly clinical cases involving these bacteria. Identification of Raoultella spp. is difficult due to a phylogenetic relationship with Klebsiella spp. Purpose: Available biochemical tests do not always allow for their identification to species. Thus, the aim of this study was to evaluate selected methods of identification of Raoultella spp. and their differentiation from genus Klebsiella. Materials and Methods: In this evaluation three methods were used such as manual test ID32E (bioMérieux), automatic test VITEK2 Compact (bioMérieux) and matrix-assisted laser desorption ionisation-time of flight mass spectrometry (MALDI-TOF MS) method (Bruker). Results: Good identification of the species was obtained for 81.4% of the strains in the ID32E test, 93.3% in VITEK2 Compact test, and 97.4% in MALDI-TOF MS method, respectively. Conclusion: It was established that MALDI-TOF MS method is reliable in identifying genus Raoultella.

Raoultella spp.-clinical significance, infections and susceptibility to antibiotics.

The genus Raoultella belongs to the family of Enterobacteriaceae. Raoultella spp. are Gram-negative, aerobic, non-motile rods. This genus can be distinguished from the genus Klebsiella, in that genus use histamine as the only source of carbon in the medium. Also, Raoultella grow at 4 °C and do not produce gas from lactose at 44.5 °C. Raoultella sp. is known to inhabit natural environments (water, soil, plants). The reservoir of Raoultella is the gastrointestinal tract and upper respiratory tract. Raoultella spp. are opportunistic bacteria, which usually cause infections of the biliary tract, pneumonia and bacteraemia in oncologic and with lower immunity patients. Raoultella planticola and Raoultella ornithinolytica are the most frequently encountered human pathogens among the genus Raoultella. In this review, the current knowledge on Raoultella infections is summarized.

Catheter-related blood stream infection caused by Raoultella ornithinolytica.

Raoultella spp. representatives are Gram-negative capsulated, nonmotile rods. These bacteria are found in the natural environment: plants, water, soil and insects. R. ornithinolytica is one of the three species of Raoultella. R. ornithinolytica is the only species within the genus which has the ability to produce ornithine decarboxylase. Human infections related to R. ornithinolytica are exceedingly rare. The present case report describes catheter-related blood stream infection caused by R. ornithinolytica and successfully treated with antibiotic therapy.

The prevalence of infections and colonisation with Klebsiella pneumoniae strains isolated in ICU patients.

BACKGROUND: Klebsiella spp. are among the bacteria most commonly isolated from patients with infections in ICUs. The source of these infections may be the microflora of the patient or the hospital environment. Increasingly, Klebsiella strains are also being isolated from epidemic outbreaks. This situation is largely the result of widespread, irrational antibiotic use, the virulence of the bacterial strains and their ability to survive in the hospital environment. The purpose of this dissertation was to estimate the prevalence of Klebsiella pneumoniae strains isolated from patients hospitalised in a single ICU. METHODS: Seventy-eight isolates of K. pneumoniae were studied. The identification and the susceptibility to selected antibiotics were tested by an automated system, VITEK2 Compact. For the analysed strains, the production of different beta-lactamases was noted. RESULTS: Production of ESBL was detected in 64.1% of the K. pneumoniae strains isolated from infections and 74.4% from rectal swabs. Most of the strains were susceptible to imipenem (97.7%) and meropenem (96.1%). Sixty-nine (57.0%) of the analysed strains were identified as multidrug resistant. CONCLUSION: Most of the analysed Klebsiella pneumoniae strains produced ESBL-beta-lactamases. The frequency of colonisation and infection with multidrug resistant strains of K. pneumoniae in patients hospitalised in the ICU is very high.