The first confirmed detection of Staphylococcus argenteus in the Czech Republic.

Staphylococcus argenteus (S. argenteus) is a novel species of coagulase-positive staphylococci described in 2015. This species is phenotypically highly similar and genetically closely related to Staphylococcus aureus (S. aureus). Until recently, differentiation was only possible by molecular genetic methods, multilocus sequence typing and whole-genome sequencing, which are not generally used in routine laboratories due to time-consumingness and expensiveness. A major improvement in the identification of S. argenteus is the application of MALDI-TOF MS, if the available updated mass spectrum reference database is used. In the short report, we would like to present the first confirmed S. argenteus strain isolated from a patient in the Czech Republic and probably the first published S. argenteus strain in Central and Eastern Europe.

BAL fluid analysis in the identification of infectious agents in patients with hematological malignancies and pulmonary infiltrates.

The present study aims to evaluate the diagnostic yield of bronchoalveolar lavage (BAL) fluid in patients with hematological malignancies and describe the most common pathogens detected in BAL fluid (BALF.) An analysis of 480 BALF samples was performed in patients with hematological malignancies over a period of 7 years. The results of culture methods, PCR, and immunoenzymatic sandwich microplate assays for Aspergillus galactomannan (GM) in BALF were analyzed. Further, the diagnostic thresholds for Aspergillus GM and Pneumocystis jiroveci were also calculated. Microbiological findings were present in 87% of BALF samples. Possible infectious pathogens were detected in 55% of cases; 32% were classified as colonizing. No significant difference in diagnostic yield or pathogen spectrum was found between non-neutropenic and neutropenic patients. There was one significant difference in BALF findings among intensive care units (ICU) versus non-ICU patients for Aspergillus spp. (22% versus 9%, p = 0.03). The most common pathogens were Aspergillus spp. (n = 86, 33% of BAL with causative pathogens) and Streptococcus pneumoniae (n = 46, 18%); polymicrobial etiology was documented in 20% of cases. A quantitative PCR value of > 1860 cp/mL for Pneumocystis jirovecii was set as a diagnostic threshold for pneumocystis pneumonia. The absorbance index of GM in BALF of 0.5 was set as a diagnostic threshold for aspergillosis. The examination of BAL fluid revealed the presence of pathogen in more than 50% of cases and is, therefore, highly useful in this regard when concerning pulmonary infiltrates.

Diagnosis and treatment of Bartonella endocarditis.

The Bartonella genus comprises more than 20 species of Gram-negative rods which are difficult to culture. These are facultative intracellular bacteria. Humans are reservoir hosts for B. quintana and B. bacilliformis or accidental hosts for other species. Bartonella is a cause of zoonosis. Bartonella infection can be completely asymptomatic or can be linked to various conditions. Our experience with Bartonella endocarditis from 2012-2017 is presented. The most effective diagnostic method for Bartonella endocarditis is PCR detection of DNA of the pathogen from excised valve tissue. The European Society of Cardiology (ESC) in the guidelines from 2015 recommends the combination doxycycline gentamycin for the treatment of Bartonella endocarditis.

[Bacteriuria and Symptomatic Urinary Tract Infections during Antimicrobial Prophylaxis in Patients with Short-Term Urinary Catheters - Prospective Randomised Study in Patients after Joint Replacement Surgery]. / Bakteriurie a symptomatický mocový infekt pri antimikrobiální profylaxi krátkodobe zavedených mocových katétru ­ prospektivní randomizovaná studie u pacientu po implantaci kloubní náhrady.

PURPOSE OF THE STUDY A very serious complication following joint replacement surgery is periprosthetic joint infection that can be caused by a urinary tract infection. Insertion of an indwelling urinary catheter constitutes a risk factor that may result in urinary tract infections. The aim of this prospective randomised study was to compare the occurrence of significant bacteriuria and symptomatic urinary tract infections during antibiotic prophylaxis at the time of removal of an indwelling urinary catheter by cotrimoxazole in two doses and with no administration of antibiotics. We also monitored the incidence of potential periprosthetic infection following the endoprosthesis implantation. The findings of preoperative urine tests were compared with the declared negative preoperative examination. MATERIAL AND METHODS The study included patients indicated for a total hip or knee replacement with a negative urine culture as a part of the preoperative testing. Where leukocyteria was detected, urine culture by mid-stream clean catch urine was obtained. The second part included patients, in whom an indwelling urinary catheter had to be inserted postoperatively for urine retention and/or monitoring of fluid balance and who were divided into two groups on a rota basis. No antibiotics were administered to the first group, whereas Cotrimoxazol 960 mg tablets p.o. was administered to the second group, 14 and 2 hours before the removal of the catheter. The urine culture test was performed 4 hours after the removal of the indwelling urinary catheter, in both the groups. The test was repeated after 14 days and a questionnaire was filled in to report urinary tract complications. Considered as significant bacteriuria by urinalysis was the laboratory finding of > 10x4 CFU/ml in case of a single pathogen or > 10x5 in case of multiple pathogens. The results were statistically processed by Fischer's exact test with the level of significance = 0.05. RESULTS In the first part of the study leukocyturia was detected by a test strip in 112 of the total of 478 patients. In 10 women, significant bacteriuria was found. Altogether 50 women and 50 men were randomly assigned to the second part of the study. The indwelling urinary catheter was in place for 4 days on average. In men, no statistically significant difference was detected in significant bacteriuria findings, in women a statistically significant difference of p = 0.00162 was found after the removal and after 14 days the borderline of statistical significance of p = 0.0507 was achieved, but no symptomatic urinary tract infection was present. In the period from 20 to 32 months following the total joint replacement, no periprosthetic infection caused by urinary tract infection was reported. DISCUSSION There is enough evidence to prove the correlation between the symptomatic urinary tract infection and periprosthetic infection. On the other hand, asymptomatic bacteriuria is a common finding in patients before the planned hip of knee joint replacement and its treatment is not recommended. No consensus has been achieved as yet regarding the method of antibiotic prophylaxis for an inserted urinary catheter. Antibiotics are administered throughout the period of catheterisation by an indwelling urinary catheter, during its removal, or are not administered at all. In our study antibiotics were administered during the removal of an indwelling urinary catheter and a statistically significant difference was found in women. It concerned, however, only a higher incidence of asymptomatic bacteriuria not treated by antibiotics, which in the next follow-up period did not lead to periprosthetic infection. CONCLUSIONS Despite the negative pre-operative urine culture, frequent incidence of leukocyturia and symptomatic urinary infections were detected in a fairly high number of cases. Therefore, we recommend asking the patients during the hospital admission process specifically about the urinary infection symptoms. The results of our study show that antibiotic prophylaxis during the removal of indwelling urinary catheters placed for a short-term is unnecessary. Key words: endoprosthesis, urinary catheter, bacteriuria, urinary tract infection.

[Evaluation of the importance of a ready-made, gentamicin-impregnated spacer in relation to bacteriological findings in patients with periprosthetic joint infections]. / Zhodnocení významu ready-made spaceru s gentamicinem ve vztahu k bakteriologickým nálezum u pacientu s infekcí kloubní náhrady.

BACKGROUND: Periprosthetic infection is a serious complication in total hip and knee arthroplasty. The complex therapeutic approach within two-stage reimplantation includes the use of antibiotic-impregnated spacers (temporary joint replacements). The aim of this paper was to evaluate bacteriological findings in selected patients with periprosthetic infection in whom a ready-made gentamicin-impregnated spacer was used to treat the infection. MATERIALS AND METHODS: Between 2008 and 2012, a ready-made, gentamicin-impregnated cement spacer was used in 24 patients to treat periprosthetic hip or knee infection within two-stage reimplantation. All components of the prosthesis and periprosthetic tissue samples were sent for microbiological examination at the first revision surgery, while at the second revision surgery, the spacer and surrounding tissue samples were sent in. In six patients with an inserted knee spacer, the level of gentamicin in the joint fluid was measured. Subsequently, the patients were regularly monitored. RESULTS: Twenty-two (92%) of 24 patients were bacteriologically positive by culture. The most commonly detected causative agents were coagulase-negative staphylococci. Other isolates were Staphylococcus aureus, Corynebacterium, anaerobic bacteria, and Salmonella serotype Enteritidis. Nineteen (76%) of 25 primary pathogens were gentamicin sensitive. Spacers from two patients were culture positive for coagulase-negative staphylococci that tested resistant to gentamicin. During the follow-up of at least two years, none of the patients developed another periprosthetic infection. CONCLUSION: The success rate of two-stage-reimplantation total hip and knee arthroplasty using ready-made, gentamicin-impregnated spacers was 100 % in our cohort of patients; no other periprosthetic infection was reported during the follow-up of at least two years. From the bacteriological results, it appears that the ready-made, gentamicin-impregnated spacer only covers 76 % of the range of the causative agents. The solution would be to use a spacer impregnated with a combination of vancomycin and gentamicin that would be effective against all cultured species.

[Leukocyte esterase testing for examination of exudate associated with skeletal system diseases. Comparison with cytological and microbiological examinations]. / Porovnání vysetrení výpotku pohybového aparátu pomocí leukocytární esterázy s jejich cytologickým a kultivacním vysetrením.

PURPOSE OF THE STUDY: Leukocyte esterase is an enzyme in neutrophils from which it is released into exudate; its detection by colorimetric test strips indicates the presence of neutrophils. This is a rapid method to find whether exudate is of infectious or non-infectious aetiology. The aim of the study was to determine the sensitivity and specificity of leukocyte esterase testing with use of AUTION Sticks (Arkray) for examination of exudates obtained in inflammatory diseases of the skeletal system. MATERIAL AND METHODS: Exudates associated with skeletal system diseases were collected from 45 patients in the period from July 1st to December 31 st , 2012. Aspirates obtained under sterile conditions were examined for leukocyte esterase; cytological and microbiological examinations were also carried out. For the detection of leukocyte esterase, a drop of aspirate was placed on the reagent zone of a test strip and the resulting colour reaction was read after 90 minutes. Changes in colour were compared with a reference strip provided by the manufacturer. The results were assessed on a five-shade scale as follows: 0, no colour change; 1 to 4, gradual change from light pink to deep purple. The results were compared with those of cytological and microbiological examinations. Shade 4 on the strip corresponded to a positive cytological finding of bacterial infection, and shades 3 and 4 correlated with a positive microbial finding. The sensitivity and specificity of leukocyte esterase testing were statistically evaluated for both comparisons. RESULTS: Based on the results of cytological and microbiological examinations, an infectious aetiology of exudate was diagnosed in 21 (44.4%) and non-infectious aetiology in 24 (63.6%) patients. With leukocyte esterase reagent strips when shade 4 was taken as a positive result, the sensitivity and specificity of examination was assessed as 0.6190 and 0.9583, respectively. When taking both shade 3 and shade 4 for a positive result, sensitivity and specificity were 0.8571 and 0.8750, respectively. Shades 0 and 1 corresponded to the number of leukocytes in exudate that was no higher than 2 x 109/ml. DISCUSSION: The detection of leukocyte esterase is a quick and easy examination. It is useful for readily excluding or confirming an infectious aetiology of exudate and can, to some extent, substitute a cytological examination. It can also help to make a quick decision whether one- or two-stage joint reimplantation should be performed and thus eliminate the need of intra-operative histological examination of frozen tissue samples. A drawback of the method was that exudate samples contaminated with blood interfered with an assessment of colour shades. However, this can be avoided by centrifugation of the sample and use of a supernatant free from erythrocytes. CONCLUSIONS: Diagnosing infectious aetiology of joint exudate or exudate from an abscess using leukocyte esterase reagent strips appears, according to our results, to be a promising, semi-quantitative method with high specificity and sensitivity which is rapid, simple and affordable. It can be useful particularly in out-patient institutions for a quick diagnosis of arthritis; intraoperatively, it can serve as an additional method to other exudate examinations.

[Evaluation of bacterial colonization of the conjunctival sac in ranibizumab intravitreal injections treated patients]. / Hodnocení bakteriálni kolonizace spojivkového vaku pacientu lécených intravitreálními injekcemi anibizumabu.

AIM: To establish the conjunctival sac bacterial flora structure in patients with wet form of the age-related macular degeneration (ARMD) indicated for the intravitreal application of Ranibizumab (Lucentis, Novartis Pharma AG). To evaluate the efficacy of combined local preparation with broad-spectrum antibiotic moxifloxacine 0.5% (Vigamox, Alcon) and povidone iodine solution, 5% (Betadine, Egis Pharmaceuticals, LTD.) and to evaluate subjective toleration of moxifloxacin. MATERIALS AND METHODS: In a prospective, non-randomized study were evaluated 20 eyes of 20 patients treated by means of intravitreally-applied ranibizumab. In all patients, the swabs from the conjunctival sac of the treated eye were repeatedly taken in a given time-schedule--before the start of using moxifloxacin, on the day of the intravitreal application of ranibizumab--before the irrigation of the conjunctival sac with povidone iodine solution, 5%, further after the irrigation--immediately before the injection and the control was token three days after the intravitreal injection. At the same time, the moxifloxacine toleration was evaluated by a questionnaire. RESULTS: The samples taken from the conjunctival sac of the treated eye before the application of moxifloxacine had positive bacterial culture in 17 eyes (85%) and negative culture in 3 eyes (15 %). Furthermore, in 2 eyes with positive culture, there was established resistance to moxifloxacine. After 3 days of moxifloxacine application, there was negative culture in 13 eyes (65%), in 7 eyes (35%) was the bacterial cultivation positive. After the irrigation with povidone iodine 5% solution was the cultivation negative in 17 eyes (85%), positive cultivation was in 3 eyes (15%); in all three cases, the cultures were susceptible to moxifloxacine. Three days after the intravitreal injection, the negative cultivation from the conjunctival sac was found in 13 eyes (65%), and in 7 eyes was the cultivation positive; the cultivated bacteria were moxifloxacine susceptible. Subjective symptoms after moxifloxacine application were reported by 10 patients altogether; 5 patients were without symptoms and 5 patients did not return the questionnaire. On average, the symptoms started the second day of moxifloxacine treatment and the average grade of symptoms was 1.6 on the scale from 0 to 5. CONCLUSION: In our group we found a broad spectrum of microorganisms colonizing the conjunctival sac of patients indicated to the ARMD intravitreal treatment. After the prophylaxis with moxifloxacine, the incidence of positive bacterial cultivation decreased and the povidone iodine 5% solution irrigation this effect increased. The most common pathogen species was Staphylococcus coagulasis negative. Although the resistance to moxifloxacine in two different bacteria in two eyes in the beginning of observation was established, after moxifloxacine treatment, the cultivation of these bacteria in both eyes was negative, and in all other cases the cultivated bacteria were susceptible to moxifloxacine.

[Cases of menstrual toxic shock syndrome in the Czech Republic in 1997-2011]. / Prípady menstruální formy syndromu toxického soku v Ceské republice v letech 1997-2011.

AIM: To determine toxigenicity and other basic characteristics of 47 strains of Staphylococcus aureus referred to the National Reference Laboratory for Staphylococci (NRL) as suspected causative agents of menstrual toxic shock syndrome (MTSS). MATERIAL AND METHODS: S. aureus strains were collected from 11 administrative regions of the Czech Republic in 1997-2011. The diagnosis was based on phenotypic (reverse latex agglutination test) and genotypic (polymerase chain reaction) methods. RESULTS: Forty-four S. aureus strains were producers of toxic shock syndrome toxin 1 (TSST-1), either alone or in combination with staphylococcal enterotoxin. Three strains only produced enterotoxin (B, C, and H). CONCLUSION: MTSS is a serious multisystem disease. In this study, MTSS often had a severe course requiring intensive care. All MTSS patients used vaginal tampons that had been identified in the literature as a risk factor for MTSS. The case of MTSS in a 36-year-old woman caused by an enterotoxin H positive strain of S. aureus is probably the first to be reported in the world.