In vitro comparison of the effectiveness of various antimicrobial locks with taurolidine in the treatment and prevention of catheter-related bloodstream infections in patients receiving parenteral nutrition.

OBJECTIVES: To evaluate the effectiveness of various taurolidine solutions in the prevention and treatment of catheter-related bloodstream infections (CRBSIs) caused by the entire spectrum of microbes in patients receiving parenteral nutrition in a shorter period of time. METHODS: The in vitro method was used to test for eradication of biofilm. Different locks were used: TauroSept (2%), TauroLock (1.35%), TauroLock half concentration, and 3.5% taurolidine and tested on Staphylococcus (S.) epidermidis, S. aureus, S. hominis, methicillin-resistant S. aureus (MRSA), Pseudomonas (P.) aeruginosa (PSAE), multidrug-resistant P. aeruginosa (MR PSAE), vancomycin-resistant enterococci, Klebsiella pneumoniae producing carbapenemase (KPC), Klebsiella pneumoniae producing extended-spectrum beta-lactamase (KLPN ESBL), Candida (C.) albicans, and C. glabrata. Broviac catheters were incubated for growth of each organism and then incubated in lock solutions. Colony forming units (CFUs) were then counted after 30 min, 60 min, and 120 min of incubation. RESULTS: A statistically significant decrease in CFUs was observed after 30 min of taurolidine exposure for S. hominis, PSAE, KLPN ESBL, KLPN KPC, C. albicans, and C. glabrata; after 60 min of exposure for S. epidermidis, PSAE, MR PSAE, KLPN ESBL, KPC, C. albicans, and C. glabrata; and after 120 min of exposure for S. epidermidis, S. hominis, S. aureus, PSAE, MR PSAE, KLPN ESBL, KPC, C. albicans, C. glabrata. CONCLUSIONS: The application of taurolidine is effective in the treatment of CRBSIs. Taurolidine proved to be more effective against Gram-negative microorganisms during a 30-min exposure. Using 0.675% taurolidine is still effective. To achieve the required antimicrobial effect, the catheter must be sanitized for at least 2 h.

A Rare Case of Osteomyelitis of an Ankle Caused by Mycobacterium chelonae.

Mycobacterium chelonae, a rapidly growing nontuberculous mycobacterium, is usually described as a causative agent of soft tissue infections (postsurgical, posttraumatic, posttransplantation, postinjection, catheter infection, etc.), but only rarely as a cause of osteomyelitis. The authors describe a case report of a 72-year-old man with osteomyelitis of the talus. Initially, the infection was assessed as a soft tissue infection, without any osteolytic changes on the X-ray. After cultivation with subsequent targeted molecular typing of the rpoB gene, M. chelonae was identified from the affected tissue. The bone involvement was subsequently detected on MRI and confirmed histologically with findings of the granulomatous tissue and acid-fast bacilli. The patient was initially treated intravenously with a combination of tigecycline, amikacin, and moxifloxacin for 4 weeks, after which the oral combination of doxycycline and moxifloxacin continued. Identification of the infecting pathogen using molecular typing thus helped to establish the correct diagnosis and represents a rarely described case of osteomyelitis caused by M. chelonae.

Dual Infection of an Open Fracture Caused by Mycobacterium setense and Clostridium celerecrescens.

Infections caused by Mycobacterium setense or Clostridium celerecrescens are extremely rare. In this report, for the first time a dual infection with these two pathogens is described. An 18-year-old female suffered multiple injuries, including an open comminuted fracture of the right humeral diaphysis after falling from a fifth-floor balcony in January 2019. Five months after the accident, a fistula appeared in the scar, reaching the bone tissue. M. setense and C. celerecrescens were cultured from sinus swabs and subsequently from perioperative samples. The patient was initially treated with a combination of intravenous antibiotics (ATBs): imipenem, amikacin, and ciprofloxacin. One month after the fracture fixation with a titanium nail, C. celerecrescens was again detected; therefore, metronidazole was added to the therapy. A triple combination of oral (PO) ATBs (trimethoprim-sulfamethoxazole, moxifloxacin, and metronidazole) followed, 8 weeks after the initial intravenous therapy. C. celerecrescens was cultured again two times, most recently in November 2019, when surgical debridement was supplemented by the topical administration of cancellous bone impregnated with vancomycin. Signs of bone healing were found at follow-ups and ATB treatment was finished in March 2020 after a total of 9 months of therapy. To this day, there have been no signs of reinfection. This case thus illustrates the need for a combination of systemic and individualized local therapy in the treatment of complicated cases of dual infections with rare pathogens.

Mycobacterioses Induced by Mycobacterium abscessus: Case Studies Indicating the Importance of Molecular Analysis for the Identification of Antibiotic Resistance.

Mycobacterioses are less frequently occurring but serious diseases. In recent years, at a global level, the incidence of mycobacterioses induced by the rapidly growing species Mycobacterium abscessus (M. a.), which is considered to be the most resistant to antibiotics and most difficult to treat, has been on the rise. Correct identification to the level of the subspecies (M. a. abscessus, M. a. massiliense, and M. a. bolletii) and determination of its sensitivity to macrolides, which are the basis of combination therapy, are of principal importance for the management of the disease. We describe five cases of mycobacterioses caused by M. a., where the sequencing of select genes was performed to identify the individual subspecies and antibiotic resistance. The analysis of the rpoB gene showed two isolates each of M. a. abscessus and M. a. massiliense and one isolate of M. a. bolletii. The complete (full length) erm(41) gene responsible for the development of inducible resistance to macrolides was demonstrated in both M. a. abscessus and M. a. bolletii isolates. A partially deleted and non-functional erm(41) gene was demonstrated in M. a. massiliense isolates. The subsequent sequencing of the full length erm(41) gene products showed, however, the mutation (T28→C) in both isolates of M. a. abscessus, causing a loss of the function and preserved sensitivity to macrolides. The antibiotic sensitivity testing confirmed that both the isolates of M. a. abscessus and M. a. massiliense were sensitive to clarithromycin even after prolonged 14-day incubation. The inducible resistance to clarithromycin was maintained only in M. a. bolletii. Thus, the sequence analysis of the erm(41) gene can reliably identify the preservation of sensitivity to macrolides and serve as an important tool in the establishment of therapeutic regimens in cases of infections with M. abscessus.

Francisella tularensis caused cervical lymphadenopathy in little children after a tick bite: Two case reports and a short literature review.

Although Francisella (F.) tularensis is a well-described and understood zoonotic pathogen, its importance in Central Europe is relatively minor and, as such, tularaemia may be missed in the differential diagnosis. The annual incidence of tularaemia in the Czech Republic is relatively stable with up to 100 reported cases per year, except in the epidemic years 1998 and 1999 with 225 and 222 reported cases, respectively. It is, however, higher in comparison with the neighbouring countries. The common route of transmission in Central Europe is handling infected animals. Tularaemia is not commonly recognized as a tick-borne disease. Here we report two rare cases of a tick bite-associated ulceroglandular form of tularaemia in 2.5-year-old and 6.5-year-old children presenting with cervical lymphadenopathy. The unusual and interesting features of those cases are the young age and relatively uncommon route of transmission suggesting possible changes in the epidemiology of tularaemia in the Czech Republic. Therefore, the infection with F. tularensis should be considered in the differential diagnosis after a tick bite even in infants.

Effect of Restriction of Fluoroquinolone Antibiotics on Clostridioides difficile Infections in the University Hospital Hradec Králové.

Clostridioides difficile is the most common pathogen responsible for hospital-acquired diarrhea. This complication of antibiotic treatment mainly endangers the health of elder patients. Preventing the development of C. difficile infections (CDI) is still a challenge that needs to be addressed. In our study, the results of 872 C. difficile positive stool samples were used to describe the epidemiological situation affected by a change in the prescription of fluoroquinolone antibiotics. In a total, 93 of strains were typed by polymerase chain reaction (PCR) and capillary gel electrophoresis. Between years 2014 and 2018 the decline in the fluoroquinolones consumption was 69.3 defined daily dose (DDD) per 1000 patient-days (from 103.3 to 34.0), in same period CDI incidence declined by 1.3 cases per 10,000 patient-bed days (from 5.6 to 4.3). Results of epidemiologic and statistical analysis shows that decline in fluoroquinolones consumption has significant influence on CDI incidence and prevalence of hypervirulent strains. In the University Hospital Hradec Králové properly managed antibiotic stewardship policy has reduced CDI incidence by 23.2% and lowered rate of hypervirulent ribotypes 001 and 176.

Evaluation of the risk factors of dental caries in children with very low birth weight and normal birth weight.

BACKGROUND: Health problems of premature infants can affect both general and oral health. The enamel defects, poor dietary and oral hygiene habits may predispose these children to dental caries. This study was conducted to assess the impact of very low birth weight and prematurity on caries risk in early childhood. METHODS: The study cohort consists of 189 of one year old infants. Anamnestic data were obtained from hospital records, feeding practice, bed-time sugar drinks and oral hygiene onset from questionnaires. Saliva samples of children and their mothers were collected for the detection of cariogenic pathogens. RESULTS: VLBW newborns had significantly shorter gestation age (29.6 vs. 38.8)) and lower mean birthweight (1124 g vs 3315 g) compared to NBW ones (p < 0.0001). Statistical significance has been found in the presence of early morbidity (p < 0.0001) and regular medication intake (p = 0.007). VLBW children got more frequently sweetened drinks during the day and night (p = 0.007). Regular oral hygiene practice was more frequent in full term group (p = 0.002). There was statistical difference in the presence of enamel hypoplasia in VLBW children (p = 0.033) but no statistical difference in the presence of hypomineralization (p = 0.0736) in comparison to NBW individuals. Proportional representation and count of S. mutans did not reveal statistical difference neither in both groups of children (p = 0.484) nor in both groups of mothers (p = 0.385). CONCLUSIONS: The study confirmed anamnestic and medical differences between both groups. The proportional representation and count of S. mutans did not reveal statistical difference neither in VLBW and NBW children, nor in their mothers.

The wide-spectrum antimicrobial effect of novel N-alkyl monoquaternary ammonium salts and their mixtures; the QSAR study against bacteria.

Quaternary ammonium salts (QASs) have been widely used for disinfection purposes because of their low price, high efficacy and low human toxicity for decades. However, precise mechanisms of action nor the powerful versatile agent against all antimicrobial species are known. In this study we have prepared 43 novel N-alkyl monoquaternary ammonium salts including 7 N,N-dialkyl monoquaternary ammonium salts differing bearing alkyl chain either of 12, 14 or 16 carbons. Together with 15 already published QASs we have studied the antimicrobial efficacy of all water-soluble compounds together with standard benzalkonium salts against Gram-positive (G+) and Gram-negative (G-) bacteria, anaerobic spore-forming Cl. difficile, yeasts, filamentous fungi and enveloped Varicella zoster virus (VZV). To address the mechanism of action, lipophilicity seems to be a key parameter which determines antimicrobial efficacy, however, exceptions are likely to occur and therefore QSAR analysis on the efficacy against G+ and G- bacteria was applied. We showed that antibacterial activity is higher when the molecule is larger, more lipophilic, less polar, and contains fewer oxygen atoms, fewer methyl groups bound to heteroatoms or fewer hydrogen atoms bound to polarized carbon atoms. In addition, from an application point of view, we have formulated mixtures, on the basis of obtained efficiency of individual compounds, in order to receive wide-spectrum agent. All formulated mixtures completely eradicated tested G+ and G- strains, including the multidrug-resistant P. aeruginosa as well as in case of yeasts. However, effect on A. fumigatus, Cl. difficile and VZV the exposition towards mixture resulted in significant reduction only. Finally, 3 out of 4 formulated mixtures were safer than reference commercial agent based on benzalkonium salts only in the skin irritation test using reconstructed human epidermidis.

The association of a reduced susceptibility to moxifloxacin in causative Clostridium (Clostridioides) difficile strain with the clinical outcome of patients.

OBJECTIVES: To investigate the relationship between Clostridium (Clostridioides) difficile strain characteristics and C. difficile infection (CDI) outcome. METHODS: Between October and December 2017, 16 hospitals collected epidemiological data according to the European Centre for Disease Prevention and Control (ECDC) surveillance protocol for CDI. C. difficile isolates were characterized by ribotyping, toxin genes detection and antibiotic susceptibility testing to metronidazole, vancomycin and moxifloxacin. RESULTS: The overall mean CDI incidence density was 4.5 [95% CI 3.6-5.3] cases per 10,000 patient-days. From the 433 CDI cases, 330 (76.2%) were healthcare-associated, 52 (12.0%) cases were community-associated or of unknown origin and 51 (11.8%) CDI cases recurrent; a complicated course of CDI was reported in 65 cases (15.0%). Eighty-eight (20.3%) of patients died and 59 of them within 30 days after the CDI diagnosis. From the 379 C. difficile isolates, the most prevalent PCR ribotypes were 001 (n = 127, 33.5%) and 176 (n = 44, 11.6%). A total of 186 (49.1%) isolates showed a reduced susceptibility to moxifloxacin (> 4 mg/L) and 96.4% of them had Thr82Ile in the GyrA. Nineteen isolates revealed reduced susceptibility to metronidazole and two isolates to vancomycin (> 2 mg/L). A fatal outcome was associated with a reduced susceptibility to moxifloxacin, the advanced age of the patients and a complicated course of CDI (p<0.05). No association between ribotype, binary toxin and a reduced susceptibility to moxifloxacin and complicated course or recurrent CDI was found. CONCLUSIONS: A reduced susceptibility to moxifloxacin, in causative C. difficile strains was associated with fatal outcome of the patients, therefore it is an important marker in surveillance of CDI.

Worm-like appearance of Listeria monocytogenes brain abscess: presentation of three cases.

Brain abscess caused by Listeria monocytogenes (LM) is a rare, yet serious condition with high mortality if not recognized and treated timely. We present a series of three cases of LM brain abscesses and describe their characteristic radiological appearances which can be considered very typical. LM abscesses frequently present as markedly irregular formations, revealing characteristic worm-like tubular pattern of chaotic curvilinear arrangement. Knowledge of this imaging pattern can be very helpful in early recognition of LM abscesses whose initial differential diagnosis can often be misleading initially.

Isolation of Bordetella trematum from the respiratory tract of a patient with lung cancer: a case report.

We report the case of isolation of Bordetella trematum from the respiratory tract of a patient with lung carcinoma. This gram-negative, opportunistic rod was firstly described in 1996. To date, only several strains of Bordetella trematum have been isolated and reported, mostly from skin and soft tissue infections. The patient was admitted to the ICU of the Pulmonary Department in incipient septic shock with respiratory failure. Intravenous fluid resuscitation and non-invasive ventilation were administered immediately. A broad spectrum antibiotic piperacillin/tazobactam was administered empirically after sampling of material for microbiological examination. The bronchoscopy showed a large cavern of decayed tumour invading into mediastinum. Both sample cultures showed significant quantities of gram-negative non-fermenting bacteria. The isolate was identified using MALDI-TOF MS as Bordetella trematum and the identification was confirmed using 16S ribosomal RNA sequencing. In the last few years, routine bacterial identification using MALDI-TOF MS has enabled correct discrimination of this species. Nevertheless, isolation of Bordetella trematum in clinical samples is still very uncommon, and it is appropriate to confirm the species identification via 16S ribosomal RNA sequencing. To our knowledge, this is the first case of B. trematum isolated from the human respiratory tract since its first description. The clinical significance of Bordetella trematum in the rapid deterioration of the patient's status remains unclear.

A rare case of purulent meningitis caused by Capnocytophaga canimorsus in the Czech Republic - case report and review of the literature.

BACKGROUND: Invasive infections caused by Capnocytophaga canimorsus are rare. Immunocompromised patients, who report being bitten by or having a close contact with an animal, represent a high-risk group for this infection. There are only few dozens of infections by this bacteria manifesting as purulent meningitis reported worldwide. The reported case is a first reported case of purulent meningitis caused by by Capnocytophaga canimorsus in Czech Republic with only a limited risk factor history. CASE PRESENTATION: The patient, a 74 years old man, was referred to the infectious diseases department of a teaching hospital with clear signs of developing purulent meningitis. His anamnestic data did not show any unusual findings. He was treated for compensated diabetes mellitus type II. The blood cultures were negative and the etiological agent did not grow from the cerebrospinal fluid (CSF) on common media. Eventually, it was identified by detecting pan-bacterial DNA and DNA sequencing. Subsequently, the pathogen was confirmed by anaerobic cultivation from CSF. Only after then the patient recalled being bitten by his German shepherd puppy during play. The patient was successfully treated intravenously by ceftriaxone. CONCLUSIONS: Purulent meningitis caused by Capnocytophaga spp. is a rare disease, but it needs to be considered in patients at risk with pre-existing conditions, who report close contact with or being bitten by an animal. It is important to test for this microbe in cases with negative microbiological results for the more common agents.

In vitro comparison of efficacy of catheter locks in the treatment of catheter related blood stream infection.

BACKGROUND & AIMS: Venous access used for parenteral nutrition (PN) application is extremely important for patients with intestinal failure. Potential loss of venous access might be a catastrophy for the patient. Catheter infections are a serious complication of PN application. Systemic administration of antibiotics as well as local antibiotic locks into the catheter to sterilize the catheter are used to treat catheter infections. However, there is no clear recommendation applying use of antibiotic locks, that would specify the type and concentration of antimicrobial medication. Our objective were to compare the efficacy of different types of antimicrobial lock therapy (especially taurolidine) and their concentrations to eradicate infectious agents. METHODS: Bacterial strains of microorganisms (Staphylococcus epidermidis, Staphylococcus aureus, methicillin resistant S. aureus (MRSA), Pseudomonas aeruginosa, multidrug-resistant P. aeruginosa, Candida albicans) were used. Subsequently, the catheter was exposed to the microbes and then was incubated with a specific lock for 2 or 24 h at 37 °C. We used these locks: ethanol 70%, taurolidine, gentamicine in concentrations 0,5, 1 and 10 mg/ml and vancomycine in concentrations 1, 5, and 10 mg/ml. The number of remaining CFU (colony forming units) was compared after incubation. RESULTS: 70% ethanol and taurolidine were most effective for all studied microorganisms. Gentamicine was more effective than vancomycine. CONCLUSIONS: The most effective antimicrobial lock solutions to eradicate selected pathogenic agents were ethanol and taurolidine. Use of antibiotics is often effective after many hours of treatment and there is a risk of inadequate therapy.

Elution kinetics of vancomycin and gentamicin from carriers and their effects on mesenchymal stem cell proliferation: an in vitro study.

BACKGROUND: Musculoskeletal infections remain a major complication in orthopedic surgery. The local delivery of antibiotics provides the high levels required to treat an infection without systemic toxicity. However, the local toxicity of antibiotic carriers to the mesenchymal stem cells, as a result of both the peak concentrations and the type of carrier, may be significant. METHODS: To address this concern, the elution kinetics of vancomycin and gentamicin from several commercially available antibiotic carriers and several carriers impregnated by a surgeon (10 ml of each sterile carrier were manually mixed with a 500 mg vancomycin and an 80 mg gentamicin solution, and the duration of impregnation was 30 min) were assessed. Moreover, the effects of these antibiotic carriers on stem cell proliferation were investigated. The following two types of stem cells were used: bone marrow and dental pulp stem cells. RESULTS: The high eluted initial concentrations from antibiotic impregnated cancellous allogeneic bone grafts (which may be increased with the addition of fibrin glue) did not adversely affect stem cell proliferation. Moreover, an increased dental pulp stem cell proliferation rate in the presence of antibiotics was identified. In contrast to allogeneic bone grafts, a significant amount of antibiotics remained in the cement. Despite the favorable elution kinetics, the calcium carriers, bovine collagen carrier and freeze-dried bone exhibited decreased stem cell proliferation activity even in lower antibiotic concentrations compared with an allogeneic graft. CONCLUSIONS: This study demonstrated the benefits of antibiotic impregnated cancellous allogeneic bone grafts versus other carriers.

The susceptibility to fosfomycin of Gram-negative bacteria isolates from urinary tract infection in the Czech Republic: data from a unicentric study.

BACKGROUND: Against a background of rapid increase of ß-lactamase-producing or multi-resistant pathogenic bacteria and the resulting lack of effective antibiotic treatment, some older antibiotics have been tested for new therapeutic uses. One of these is fosfomycin, to which according to studies these resistant bacteria are very sensitive. Our study was designed because there is no data on the fosfomycin susceptibility rate in the Czech Republic. METHOD: In this study from January 2013 to June 2014 3295 unique isolates of Gram-negative bacteria which had caused urinary tract infections were examined. The antibiotic susceptibility was measured by disk diffusion test. Both EUCAST and CLSI guidelines criteria (for fosfomycin only) were used for the antibiotic susceptibility evaluation. RESULTS: The most frequently tested bacterial isolates were Escherichia coli (51.3%, n = 1703), Klebsiella pneumoniae (19.4%, n = 643) and Proteus spp. (11.8%, n = 392). Among all isolates 29.0% (n = 963) were resistant to fluoroquinolones, 11.3% (n = 374) produced extended spectrum ß-lactamase and 4.2% (n = 141) produced AmpC ß-lactamase. The overall in vitro susceptibility was significantly higher for fosfomycin compared to the other tested per-oral antibiotics (nitrofurantoin, ampicillin, co-trimoxazole, ciprofloxacin and cefuroxime) against all tested Gram-negative rod isolates (excluding Morganella morgani and Acinetobacter spp. isolates). Fosfomycin also remained highly active against those isolates with extended spectrum ß-lactamase (ESBL) production (95.8% in Escherichia coli isolates and 85.3% in Klebsiella pneumoniae isolates), unlike other tested per-oral antibiotics, which showed significant (p < 0.0001) susceptibility decrease. CONCLUSION: We have confirmed in the Czech Republic the very high susceptibility to fosfomycin trometamol of urinary tract infection pathogens, particularly Gram-negative rods including those producing ß-lactamase.

[Outbreak of SARG in a neonatal intensive care unit].

The article provides information on an outbreak of infection with a strain of Staphylococcus aureus resistant to gentamicin (SARG) and a hyperproducer of enterotoxins A and D at a neonatal intensive care unit of the University Hospital in Hradec Králové. A retrospective investigation was carried out that provided an overview of the presence of the SARG strain in the patients, staff and hospital environment. Subsequently, strict infection precautions were implemented and the importance of rational antimicrobial therapy was emphasized.

[Diabetic foot infections]. / Infekce v rámci syndromu diabetické nohy.

Diabetic foot infections (DFIs) are serious problems in persons with diabetes, about 10 to 25 % of patients with dia-betes develop a foot ulcer and 60 % of them are infected. DFIs cause morbidity, limit mobility, worsen patients quality of life. Infections are classified as mild, moderate, or severe. Most DFIs are polymicrobial, with Gram-positive cocci (especially staphylococci), Gram-negative bacilli and obligate anaerobes. Successful therapy of DFI requires proper topical care and often includes surgical interventions but appropriate antibiotic treatment plays a key role. Initial antimicrobial therapy of these infections is usually empirical, the antibiotic regimen should be based on the severity of the infection. Definitive therapy should then be tailored according to the results of culture and susceptibility tests from a reliably obtained specimen.

Pyogenic sacroiliitis: diagnosis, management and clinical outcome.

OBJECTIVE: The purpose of the present study was to evaluate the role of diagnostic tools and management options for patients with pyogenic sacroiliitis, including potential complications. MATERIALS AND METHODS: This retrospective study included 16 patients with pyogenic sacroiliitis who were admitted to a single orthopaedic centre between 2007 and 2012. The following data were collected: demographics, history, radiography, magnetic resonance images (MRI), biological data, type of pathogenic agent, abscess formation, type of management, and clinical outcome. RESULTS: Our study demonstrated that only one-fifth of the patients with lumbogluteal or hip pain had established diagnoses of suspected pyogenic sacroiliitis upon admission. MRIs confirmed this diagnosis in all cases. MRI examinations revealed joint fluid in the sacroiliac joint and significant oedema of the adjacent bone and soft tissues. In 12 of the 16 cases, erosions of the subchondral bone were encountered. Contrast-enhanced MRI revealed that 9 patients had abscesses. All patients received antibiotic therapy. Antibiotic treatment was only successful in 9 cases. The other 7 patients underwent computed tomography (CT)-guided abscess drainage. Drainage was sufficient for 4 patients, but 3 patients required open surgery. One patient required sacroiliac arthrodesis. The clinical outcomes included minimal disability (n = 10), moderate disability (n = 5), and full disability (n = 1) of the spine. CONCLUSIONS: Contrast-enhanced MRI is mandatory for a reliable diagnosis. Abscess formation was observed in approximately half of the MRI-diagnosed sacroiliitis cases and required minimally invasive drainage under CT guidance or frequently open surgery.