Cefiderocol - An effective antimicrobial for MDR infections but a challenge for routine antimicrobial susceptibility testing.

PURPOSE: Cefiderocol is a novel cephalosporin-siderophore conjugate antibiotic that holds promise to thwart infections caused by multi-drug-resistant gram-negative bacilli. Its antibacterial activity against normally susceptible species is not affected by most ß-lactamases, including metallo-ß-lactamases. Due to the siderophore-mediated entry into the cell, the activity of cefiderocol is less affected by porin loss or active efflux resistance than many other ß-lactam antibiotics. The aim of this study was to assess in vitro susceptibility to the cefiderocol of carbapenemase-producing gram-negative bacilli from clinical samples of hospitalized patients. MATERIALS AND METHODS: We analyzed 102 clinical strains of carbapenemase-producing Enterobacterales and non-fermentives from hospital centers in Lódz, Poland. Antimicrobial susceptibility to cefiderocol was tested by the minimum inhibitory concentration test strips and disc diffusion methods. RESULTS: The obtained results turned out to be ambiguous, and the area of technical uncertainty made their interpretation very difficult. CONCLUSIONS: The cost of therapy with this antibiotic, and difficulties in interpreting the drug susceptibility are the limitations to the use of cefiderocol. Intensive work should be carried out to finally standardize an easily accessible and reliable method for the determination of susceptibility to cefiderocol.

Aztreonam Combinations with Avibactam, Relebactam, and Vaborbactam as Treatment for New Delhi Metallo-ß-Lactamase-Producing Enterobacterales Infections-In Vitro Susceptibility Testing.

Antimicrobial resistance is a major global health issue. Metallo-ß-lactamases (MBL), in particular, are problematic because they can inactivate all classes of ß-lactams except aztreonam. Unfortunately, the latter may be simultaneously inactivated by serine ß-lactamases. The most dangerous known MBL is New Delhi Metallo-ß-lactamase (NDM). This study aimed to test the in vitro susceptibility to aztreonam in combination with novel ß-lactamase inhibitors (avibactam, relebactam, and vaborbactam) in clinical strains of Enterobacterales NDM which is resistant to aztreonam. We investigated 21 NDM isolates-including Klebsiella pneumoniae, Escherichia coli, and Citrobacter freundii-which are simultaneously resistant to aztreonam, ceftazidime/avibactam, imipenem/relebactam, and meropenem/vaborbactam. MICs for aztreonam combinations with novel inhibitors were determined using the gradient strip superposition method. The most effective combination was aztreonam/avibactam, active in 80.95% strains, while combinations with relebactam and vaborbactam were effective in 61.90% and 47.62%, respectively. In three studied strains, none of the studied inhibitors restored aztreonam susceptibility. Aztreonam/avibactam has the most significant antimicrobial potential for NDM isolates. However, combinations with other inhibitors should not be rejected in advance because we identified strain susceptible only to tested combinations with inhibitors other than avibactam. Standardization committees should, as soon as possible, develop official methodology for antimicrobial susceptibility testing for aztreonam with ß-lactamase inhibitors.

Analysis of Cumulative Antibiogram Reports in Search for Optimal Empirical Urinary Tract Infection Treatment at the Central Teaching Hospital of the Medical University of Lodz, Poland: Results of a 3-Year Surveillance.

Urinary tract infections are among the most common bacterial infections, accounting for about two-fifths of all healthcare-associated infections. Appropriate antimicrobial therapy is crucial, e.g., to avoid prolonged hospitalization and limit antimicrobial resistance spread. This study was performed to analyze the microbiological profiles of urinary tract infections in the Central Teaching Hospital in Lodz, Poland, and develop local empirical therapy guidelines. This study was a 3-year retrospective surveillance of the cumulative antibiograms from urine cultures. The procedures were based on the current EUCAST and CLSI guidelines. In 2020-2022, a total of 4656 urine cultures were performed, of which 1134 were positive. The most common bacterial isolates were Escherichia coli, followed by Klebsiella spp. and Enterococcus spp. High levels of susceptibility (>90%) have been observed for carbapenems, piperacillin/tazobactam, amikacin, and nitrofurantoin. Development of the appropriate empirical antimicrobial is a challenging task with persistently high levels of resistance to commonly used antimicrobials. Eventually, we separated the uncomplicated and complicated urinary tract infections in local guidelines and recommended nitrofurantoin and amikacin, respectively, in empiric therapy. The clinicians should make a decision based on the presented symptoms and then-with the urine culture result-correct or continue the therapy.

Nitrofuran Derivatives Cross-Resistance Evidence-Uropathogenic Escherichia coli Nitrofurantoin and Furazidin In Vitro Susceptibility Testing.

The treatment of urinary tract infections is usually empirical. For example, nitrofuran derivatives, mainly nitrofurantoin (but also furazidin), are used in Eastern Europe. A significant problem is the assessment of the usefulness of furazidin, as there are no standards for susceptibility testing. Additionally, a high percentage of strains resistant to nitrofurantoin should prompt caution when choosing furazidin in therapy. This study aimed to answer the question of whether it is possible to use nitrofurantoin susceptibility for furazidin drug susceptibility analyses and if there is any cross-resistance in the nitrofuran derivatives group. One hundred E. coli clinical isolates, obtained from the Central Teaching Hospital of the Medical University of Lodz, were cultured from positive urine samples. For susceptibility testing, microdilution and disk diffusion methods, following EUCAST guidelines, were used. The results showed that the MICs of furazidin were equal to or lower than those of nitrofurantoin in 89% of the tested strains. The MIC50/90 values for furazidin were two times lower than those for nitrofurantoin. Positive correlations were found between MICs and growth inhibition zones for both antibiotics. Based on the obtained data and previous studies, it was assumed that the transfer of susceptibility testing results from nitrofurantoin to furazidin is acceptable due to cross-resistance in nitrofuran derivatives.

In Vitro Activity of Eravacycline against Carbapenemase-Producing Gram-Negative Bacilli Clinical Isolates in Central Poland.

Eravacycline is a novel antibiotic of the tetracycline class with activity against a broad spectrum of clinically significant bacteria, including multi-drug-resistant organisms. For this reason, it may be an alternative to treating critical infections of this etiology. We aimed to assess the in vitro effectiveness of eravacycline to carbapenemase-producing Gram-negative bacilli clinical isolates identified in hospitals in Lódz, Poland. We analyzed 102 strains producing KPC, MBL, OXA-48, GES, and other carbapenemases. Eravacycline susceptibility was determined following the EUCAST guidelines. The highest susceptibility was found in KPC (73%) and MBL (59%) strains. Our results confirmed in vitro the efficacy of this drug against carbapenem-resistant strains. However, eravacycline has been indicated only for treating complicated intra-abdominal infections, significantly limiting its use. This aspect should be further explored to expand the indications for using eravacycline supported by evidence-based medicine. Eravacycline is one of the drugs that could play a role in reducing the spread of multidrug-resistant microorganisms.

Carbapenem-Resistant Gram-Negative Fermenting and Non-Fermenting Rods Isolated from Hospital Patients in Poland-What Are They Susceptible to?

Gram-negative fermenting and non-fermenting bacteria are important etiological factors of nosocomial and community infections, especially those that produce carbapenemases. Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa are the most frequently-detected carbapenemase-producing microorganisms. The predominant type of resistance is metallo-ß-lactamase (MBL). These bacteria are predominantly isolated from bronchial alveolar lavage, urine, and blood. Carbapenemase-producing Enterobacterales (CPE) strains are always multi-drug-resistant. This significantly limits the treatment options for this type of infection, extends the time of patient hospitalization, and increases the risk of a more severe and complicated disease course. Preventing the transmission of these microorganisms should be a major public health initiative. New antibiotics and treatment regimens offer hope against these infections.

The seroprevalence of Bartonella spp. in the blood of patients with musculoskeletal complaints and blood donors, Poland: a pilot study.

BACKGROUND: Bartonella spp. can cause a variety of diseases, such as lymphadenopathies, cat scratch disease, and trench fever, but can also give rise to many non-specific symptoms. No data exists regarding the prevalence of Bartonella spp. in patients with musculoskeletal complaints, nor among blood donors in Poland. METHODS: The presence of anti-Bartonella IgM and IgG in the serum of blood donors (n = 65) (Lodz, Poland) and in the patients of the Department of Rheumatology Clinic (n = 40) suffering from musculoskeletal symptoms was tested by immunofluorescence. Blood samples were cultured on enriched media. Epidemiological questionnaires were used to identify key potential risk factors, such as sex, age, contact with companion animals, and bites from insects or animals. RESULTS: Altogether, 27 of the 105 tested subjects were seropositive for Bartonella henselae IgG (23%) and three for Bartonella quintana IgG (2.85%); IgMs against B. henselae were found in three individuals (2.85%), and IgMs against B. quintana were found in one (1.54%). No statistically significant difference was found between the prevalence of B. henselae in the blood of donors or patients and the presence of unexplained musculoskeletal complaints (23% vs 30%). Individuals who had kept or been scratched by cats were not more likely to be B. henselae seropositive (p > 0.01). Tick bites were more commonly reported in patients, but insignificantly (p > 0.01). CONCLUSION: This is the first report of a high seroprevalence of anti-Bartonella IgG in patients with musculoskeletal symptoms and in blood donors in Poland. The obtained results indicate that such seroprevalence may have a possible significance in the development of musculoskeletal symptoms, although it should be confirmed on a larger group of patients. Asymptomatic bacteremia might occur and pose a threat to recipients of blood from infected donors. Hence, there is a need for more detailed research, including molecular biology methods, to clarify the potential risk of Bartonella spp. being spread to immunocompromised individuals. KEY POINTS: • This is the first study presenting high seroprevalence of Bartonella spp. in Poland. • IgG and IgM antibodies against B. quintana were found in blood samples of blood donors.

Parainfluenza virus infection enhances NSAIDs-induced inhibition of PGE2 generation and COX-2 expression in human airway epithelial cells.

PURPOSE: Respiratory viral infection and nonsteroidal anti-inflammatory drugs (NSAIDs) may affect arachidonic acid (AA) metabolism in the airway epithelium, however their joint effect has not been studied. We hypothesized, that alternations of AA metabolism in human airway epithelial cells (ECs) - induced by Parainfluenza virus type 3 (PIV3) - may be modified by concomitant treatment with NSAIDs. MATERIALS AND METHODS: Nasal (RPMI 2650) and bronchial (BEAS-2B) epithelial cells were cultured into confluence and then infected with PIV3. Prostaglandin E2 (PGE2) and 15-hydroxyeicosatetraenoic acid (15-HETE) levels in cell supernatants were measured by ELISA and expression of cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), 5-lipoxygenase (5-LO) and 15-lipoxygenase (15-LO) mRNA in cells was evaluated after reverse transcription with real-time polymerase chain reactions. RESULTS: PGE2 generation was decreased by PIV3 infection in the upper airway epithelial cells, and increased in the lower airway epithelial cells. Both naproxen and celecoxib induced significant reduction in PGE2 release in both infected and non-infected upper and lower airway epithelial cells. However, in PIV3-infected epithelial cells celecoxib inhibited PGE2 release and COX-2 expression to significantly higher degree as compared to non-infected cells. 15-HETE generation or COX-1, 5-LO and 15-LO expression were not affected by the virus infection or by NSAIDs. CONCLUSION: Virus infection in airway epithelial cells enhances inhibitory effect of NSAIDs on prostaglandin E2 generation.

Toll-Like Receptor Agonists Modulate Wound Regeneration in Airway Epithelial Cells.

BACKGROUND: Impaired regeneration of airway epithelium may lead to persistence of inflammation and remodelling. Regeneration of injured epithelium is a complex phenomenon and the role of toll-like receptors (TLRs) in the stimulation of respiratory virus products in this process has not been established. OBJECTIVE: This study was undertaken to test the hypothesis that the wound repair process in airway epithelium is modulated by microbial products via toll-like receptors. METHODS: Injured and not-injured bronchial epithelial cells (ECs) (BEAS-2B line) were incubated with the TLR agonists poly(I:C), lipopolisacharide (LPS), allergen Der p1, and supernatants from virus-infected epithelial cells, either alone or in combination with TLR inhibitors. Regeneration and immune response in injured and not-injured cells were studied. RESULTS: Addition of either poly(I:C) or LPS to ECs induced a marked inhibition of wound repair. Supernatants from RV1b-infected cells also decreased regeneration. Preincubation of injured and not-injured ECs with TLR inhibitors decreased LPS and poly(I:C)-induced repair inhibition. TGF-ß and RANTES mRNA expression was higher in injured ECs and IFN-α, IFN-ß, IL-8, and VEGF mRNA expression was lower in damaged epithelium as compared to not-injured. Stimulation with poly(I:C) increased IFN-α and IFN-ß mRNA expression in injured cells, and LPS stimulation decreased interferons mRNA expression both in not-injured and injured ECs. CONCLUSION: Regeneration of the airway epithelium is modulated by microbial products via toll-like receptors.

Innate Immune Response to Viral Infections in Primary Bronchial Epithelial Cells is Modified by the Atopic Status of Asthmatic Patients.

PURPOSE: In order to gain an insight into determinants of reported variability in immune responses to respiratory viruses in human bronchial epithelial cells (HBECs) from asthmatics, the responses of HBEC to viral infections were evaluated in HBECs from phenotypically heterogeneous groups of asthmatics and in healthy controls. METHODS: HBECs were obtained during bronchoscopy from 10 patients with asthma (6 atopic and 4 non-atopic) and from healthy controls (n=9) and grown as undifferentiated cultures. HBECs were infected with parainfluenza virus (PIV)-3 (MOI 0.1) and rhinovirus (RV)-1B (MOI 0.1), or treated with medium alone. The cell supernatants were harvested at 8, 24, and 48 hours. IFN-α, CXCL10 (IP-10), and RANTES (CCL5) were analyzed by using Cytometric Bead Array (CBA), and interferon (IFN)-ß and IFN-λ1 by ELISA. Gene expression of IFNs, chemokines, and IFN-regulatory factors (IRF-3 and IRF-7) was determined by using quantitative PCR. RESULTS: PIV3 and RV1B infections increased IFN-λ1 mRNA expression in HBECs from asthmatics and healthy controls to a similar extent, and virus-induced IFN-λ1 expression correlated positively with IRF-7 expression. Following PIV3 infection, IP-10 protein release and mRNA expression were significantly higher in asthmatics compared to healthy controls (median 36.03-fold). No differences in the release or expression of RANTES, IFN-λ1 protein and mRNA, or IFN-α and IFN-ß mRNA between asthmatics and healthy controls were observed. However, when asthmatics were divided according to their atopic status, HBECs from atopic asthmatics (n=6) generated significantly more IFN-λ1 protein and demonstrated higher IFN-α, IFN-ß, and IRF-7 mRNA expressions in response to PIV3 compared to non-atopic asthmatics (n=4) and healthy controls (n=9). In response to RV1B infection, IFN-ß mRNA expression was lower (12.39-fold at 24 hours and 19.37-fold at 48 hours) in non-atopic asthmatics compared to atopic asthmatics. CONCLUSIONS: The immune response of HBECs to virus infections may not be deficient in asthmatics, but seems to be modified by atopic status.

Impact of low-level laser therapy on the dynamics of pressure ulcer-induced changes considering an infectious agent and cathelicidin LL-37 concentration: a preliminary study.

INTRODUCTION: Low-level laser therapy is used in managing chronic wounds including pressure ulcers. Less is known about its impact on the healing process if an inhibitive agent e.g. bacterial infection takes place. Modulating non-specific immunity processes might eliminate bacteria if laser therapy is applied. AIM: To investigate the impact of low-level laser therapy on pressure ulcer dynamics considering an infectious agent and cathelicidin LL-37 concentration. MATERIAL AND METHODS: The study comprised 6 patients with pressure ulcers ranging from stage II to III in Torrance classification and 12 patients without pressure ulcers. Venous blood sample and decubitus wound swab were taken - in study groups A at baseline and after 2 weeks; in control group B once - at a specific point of time. The swabs served for species identification. Drug susceptibility of isolated pathogens and cathelicidin LL-37 in serum concentration were measured. RESULTS: In study group A, the following bacteria predominantly occurred: S. aureus, E. faecalis, P. mirabilis, P. aeruginosa, while in control group B, excluding one MRSA case, S. hominis, S. epidermidis, D. nishinomiyaensis, A. haemolyticus (physiological flora) were present. HLGR resistance mechanisms were detected when analyzing drug susceptibility panels. Study group A findings demonstrated a statistically significant difference between the levels of cathelicidin LL-37 concentration at baseline and at the end. CONCLUSIONS: There is insufficient information to accurately determine the effect of LLLT on pressure ulcer dynamics considering an infectious agent. These effects may occur if innate immunity processes are modulated so that laser therapy might eliminate bacteria indirectly.

Human parainfluenza virus type 3 (HPIV3) induces production of IFNγ and RANTES in human nasal epithelial cells (HNECs).

BACKGROUND: Human parainfluenza virus type 3 (HPIV3), while infecting lower airway epithelial cells induces pneumonia and bronchiolitis in infants and children, and may lead to asthma exacerbations in children and adults. Respiratory viruses invading the airway epithelium activate innate immune response and induce inflammatory cytokine release contributing to the pathophysiology of upper and lower airway disorders. However, the effects of HPIV3 infection on nasal epithelial cells have not been well defined. The aim of this study was to evaluate the effect of the HPIV3 infection on cultured human nasal epithelial cells (HNECs) and the release of interferon gamma and other cytokines. METHODS: RPMI 2650, a human nasal epithelial cell line was cultured into confluence and was infected with HPIV3 (MOI of 0.1, 0.01 and 0.001). The protein release into supernatants and mRNA expression of selected cytokines were assessed 24, 48 and 72 h after infection. Cytokine concentrations in supernatants were measured by ELISA and expression of cytokine mRNA in RPMI 2650 cells confirmed by real time RT-PCR analysis. RESULTS: HNECs infection with HPIV3 did not induce cytotoxicity for at least 48 hours, but significantly increased IFN-γ protein concentration in the cell supernatants at 24 h and 48 h post infection (by 387% and 485% respectively as compared to mock infected cells). At 24 h a significant increase in expression of mRNA for IFNγ was observed. RANTES protein concentration and mRNA expression were significantly increased at 72 h after infection (mean protein concentration: 3.5 ± 1.4 pg/mL for 0.001 MOI, 10.8 ± 4.6 pg/mL for 0.01 MOI and 61.5 ± 18.4 pg/mL for 0.1 MOI as compared to 2.4 ± 1.3 pg/mL for uninfected cells). No measurable concentrations of TNF-α, IL-10, TSLP, IL-8, GM-CSF or eotaxin, were detected in virus infected cells supernatants. CONCLUSIONS: HPIV3 effectively infects upper airway epithelial cells and the infection is associated with induction of IFN-γ and generation of RANTES.

[Microorganisms isolated from healthy newborns in the first and third twenty-four hours]. / Drobnoustroje izolowane od zdrowych noworodków w pierwszej i trzeciej dobie zycia.

From forty healthy newborns of The Neonatology Ward of H.Jordan Hospital in Lodz, skin and nasal septum-swabs have been sampled immediately after the delivery. Whereas, in the third twenty-four hours of their lives, apart from skin and nasal septum-swabs, it has been sampled also faeces-swabs. The aim of the examination was to establish time of bacterial colonization and kinds of microorganisms responsible for that colonization after children's delivery. Two hundred clinic samples have been entirely taken to microbiological examination. It has been received 119 positive inoculations; 186 Gram-positive bacteria strains as well as 38 strains belonging to Enterobacteriaceae family have been cultured from healthy newborns. Eleven strains of methicillin resistant Staphylococcus aureus have been isolated--each from the skin and the nasal septum, during the first twenty-four hours, eight strains from skin in the third twenty-four hours as well as one from the nasal septum in the third twenty four hours. The results gathered on the basis of the study, allow to conclude that healthy newborns as early as in their first twenty four hours are influenced both by the physiological bacterial flora and pathogenic microorganism that can be an ethiological factor of hospital infection.

[Comparison of bacterial flora in the oral cavity of children and adults]. / Porównanie flory bakteryjnej jamy ustnej u dzieci i u osób doroslych.

The aim of the study was to assess levels of occurrence and number of aerobic bacteria hemolysing and non-hemolysing, anaerobic bacteria, streptococci hemolysing and non-hemolysing, staphylococci, and bacteria responsible for tooth decay (Streptococcus mutans and Lactobacillus spp.) on oral cavity in children and adults. The results obtained indicate the difference of the level of occurrence of, aerobic bacteria hemolysing and non-hemolysing, anaerobic bacteria, streptococci hemolysing and non-hemolysing, staphylococci, and bacteria responsible for tooth decay (Streptococcus mutans and Lactobacillus spp.) was not statistically significant in either group. The counts and average values of the counts for aerobic bacteria non-hemolysing, anaerobic bacteria, streptococci hemolysing and non-hemolysing and Streptococcus mutans turned out to be statistically significantly larger in adults than in children. However for aerobic bacteria hemolysing, staphylococci and Lactobacillus spp. the difference of the counts was not statistically significant in either group.