Isolation of Cronobacter spp. (formerly Enterobacter sakazakii) from nostrils of healthy stable horse--short communication.

Cronobacter spp. belongs to the family Enterobacteriaceae. It is a motile (peritricha) Gram-negative non-spore forming bacterium. At present, Enterobacter sakazakii is reported as a Cronobacter spp. species with 16 biogroups. It is a ubiquitous organism whose isolation used to be associated with a contaminated powdered infant formula and feed for neonates and infants. Information about the Cronobacter spp. species incidence in the environment, its potential dissemination and its vectors, is very limited. The authors have documented incidence of Cronobacter spp. in the nostril mucous membrane of a healthy stabled horse. The above points out at the absolutely insufficient and unsystematic information about the dissemination of the Cronobacter spp. strain in the environment of animals and the people who are in contact with them.

Prevalence of Campylobacter jejuni and its resistance to antibiotics in poultry in the Czech Republic.

Thermotolerant Campylobacter spp., in particular Campylobacter jejuni, are among the most frequently identified pathogens, found to be causing human gastrointestinal infections in Europe, with the Czech Republic being no exception. The presented work aimed at assessing results of the first nationwide monitoring of prevalence and antibiotic resistance of Campylobacter spp. in broiler flocks in the Czech Republic, including a comparison of antibiotic resistance of C. jejuni isolates collected from poultry and the human community. The monitoring was carried out in poultry slaughterhouses in 2006 and 2007. From broilers, cloacal swabs were collected and examined. The human isolates of C. jejuni were acquired from rectal swabs in community patients with diarrhoeal diseases. Suspected isolates of both animal and human origin were confirmed by the PCR methods. Antibiotic resistance to selected anti-microbial agents was tested by the microdilution method. In the monitored period, the prevalence of thermotolerant Campylobacter spp. in broilers in the Czech Republic reached almost 50%. In 2006, C. jejuni was detected in 46% and Campylobacter coli in 3% of the tested samples. In 2007, C. jejuni was found in 43% and C. coli in 2% of the samples. The results of anti-microbial susceptibility testing of C. jejuni showed higher resistance in animals when compared with humans. The only exception was tetracycline with higher resistance in isolates of human origin. The highest resistance detected was to quinolone antibiotics. Resistance to oxolinic acid was 77% in animal and 60% in human isolates, to ciprofloxacin 72% in isolates from poultry and 55% in those from humans. In ampicillin, 26% of poultry isolates and 16% of human isolates were resistant. Moreover, 9% of animal isolates demonstrated resistance to streptomycin, undetected in human isolates. In erythromycin, resistance was found in 6% of poultry and 1% of human isolates.

Prevalence of genes encoding extracellular virulence factors among meticillin-resistant Staphylococcus aureus isolates from the University Hospital, Olomouc, Czech Republic.

A rather fast and complicated progression of an infection caused by some strains of Staphylococcus aureus could be associated with the expression and co-action of virulence factor complexes in these strains. This study screened the antibiotic susceptibility and prevalence of virulence markers in isolates of meticillin-resistant S. aureus (MRSA) obtained from patients hospitalized at the University Hospital in Olomouc, Czech Republic. A total of 100 isolates was screened for 13 genes encoding extracellular virulence determinants (tst, pvl, eta, etb, sea, seb, sec, sed, see, seg, seh, sei and sej) and for their distribution in sample types. Eighty-nine isolates were positive for at least one of the genes. Genes for etb, pvl, see and seh were not detected in any of the MRSA isolates. No statistically significant differences in the occurrence of the determinants studied among sample types were found.

Genotypic characterisation of vancomycin-resistant Enterococcus faecium isolates from haemato-oncological patients at Olomouc University Hospital, Czech Republic.

This study describes the first molecular characterisation of clinical isolates of vancomycin-resistant enterococci (VRE) in the Czech Republic. Of 2647 patient isolates of Enterococcus spp. from 1997-2002, 121 (4.6%) were identified as VRE. The most common isolates were VanA+ Enterococcus faecium (78%) and VanB+ Enterococcus faecalis (10%). In addition, five VanA+ E. faecium isolates were obtained from environmental and staff sampling. Macrorestriction analysis of SmaI restriction fragment length polymorphism was performed for 54 VanA+ E. faecium clinical isolates and the five VanA+ E. faecium environmental isolates. Thirty-two unique restriction endonuclease patterns were identified, including two predominant clonal types represented by five or more isolates. Two environmental VanA+ E. faecium isolates were closely related to two patient isolates, which had an identical SmaI macrorestriction pattern. The results indicated potential survival of strains in the hospital environment and possible subsequent transmission to hospitalised patients.

The influence of antibiotic use on the occurrence of vancomycin-resistant enterococci.

BACKGROUND: Several studies have documented the influence of antibiotic selective pressure, mainly from the use of glycopeptides, third-generation cephalosporins, quinolones and lincosamides, on the frequency of vancomycin-resistant enterococci (VRE) occurrence in hospitals. The aim of this study was to evaluate the relationship between VRE occurrence and antibiotic use in the Department of Hemato-Oncology of the Teaching Hospital in Olomouc (DHO), Czech Republic, over a 6-year period under standard and unchanged hygienic and epidemiological conditions. METHODS: During the period of 1998-2003, Enterococcus sp. strains and VRE were isolated by standard methods from clinical samples taken from DHO in-patients. The frequency of VRE occurrence was expressed as the number of isolated strains per 100 bed-days/year. DHO antibiotic consumption data were processed according to the anatomical therapeutic chemical (ATC)/defined daily dose (DDD) system valid in 2003 and expressed in defined daily dose per 100 bed-days (DDD/100 bed-days) for each year of the period. RESULTS: Since 1998, the occurrence of VRE decreased significantly (from 0.28 to 0.17 VRE/100 bed-days in 2001). Between 2001 and 2003, a significant (P < 0.05) increase from 0.17 to 0.38 was observed. The antibiotic use decreased from 205.2 in 1998 to 161.0 DDD/100 bed-days in 1999 and after an increase in 2001 (to 181.8 DDD/100 bed-days) it remained relatively stable. A significant decrease was observed in third-generation cephalosporins and quinolones (from 29.5 to 9.7 and from 42.2 to 30.2 DDD/100 bed-days respectively) between 1998 and 1999. In 2002-2003, the use of third-generation cephalosporins and glycopeptides increased substantially (from 10.1 to 13.9 and from 11.3 to 15.2 DDD/100 bed-days respectively). The Pearson correlation value was significantly positive (P < 0.05) for VRE occurrence and the use of glycopeptides and third-generation cephalosporins. CONCLUSIONS: While our study confirms the effect of use of glycopeptides and third-generation cephalosporins on occurrence of VRE, no influence of quinolones and lincosamides over the 6-year period was shown.

Occurrence of variants with temperature-dependent susceptibility (TDS) to antibiotics among Stenotrophomonas maltophilia clinical strains.

Susceptibility to 20 antibiotics was tested in 104 Stenotrophomonas maltophilia strains at 37 and 30 degrees C by means of a dilution micromethod to verify the phenomenon of temperature-dependent susceptibility (TDS). Trimethoprim-sulfamethoxazole, pefloxacin and ofloxacin were the most active preparations at 37 degrees C (93, 90, and 86% of susceptible strains, respectively), whilst trimethoprim-sulfamethoxazole, cefoperazone-sulbactam and pefloxacin performed best at 30 degrees C (94, 94, and 76% of susceptible strains, respectively). Variants 37TDS (minimum inhibitory concentration, MIC, of tested antibiotics at least 4-times lower at 37 than at 30 degrees C) occurred in 60%. Variants 30TDS (at least 4-times lower value of MIC at 30 than at 37 degrees C) were found in 7.7%. Both variants in susceptibility to tested antibiotics appeared in 23.1%, whilst neither of them was observed in 9.6%. The 37TDS phenomenon was recorded most of all with gentamicin (51% of strains), amikacin (47), colistin (44) and tetracycline (34). The 30TDS phenomenon was found particularly with cefoperazone-sulbactam (16.0% of strains) and colistin (10.0%). The above phenomena may be due to changes in membrane permeability, temperature-dependent ribosomal changes, and insufficient adaptation to higher temperatures of some strains of the originally environmental species S. maltophilia.

Influence of an extended incubation period on values of minimum inhibitory concentrations of antibiotics in Stenotrophomonas maltophilia clinical strains.

In 106 Stenotrophomonas maltophilia clinical strains the susceptibility to 19 kinds of antibiotics was tested by the broth dilution micromethod at 24 h and 48 h incubation. Isolated strains demonstrated the lowest frequency of resistance to cotrimoxazole (7.5% of resistant strains at 24 h incubation and 18.9% at 48 h), ofloxacin (13.2% and 30.2%), ciprofloxacin (19.8% and 50.9%) and to cefoperazone/sulbactam (20.8% and 37.7%). The smallest growth of the number of resistant strains after extended incubation was recorded in gentamicin (by 10.4%), ceftazidime (by 11.3%) and cotrimoxazole (by 11.4%). On the contrary, the largest growth of resistance was demonstrated in cefoperazone and ciprofloxacin (by 31.1%). Average values of the growth of minimum inhibitory concentrations (MICs) were lowest in ciprofloxacin and ofloxacin (2.3 times) and highest in piperacillin/tazobactam (4.5 times) and piperacillin (5.0 times). As far as the stability of MIC is concerned, the largest occurrence of strains with the MIC growth doubled as a maximum was found in ceftazidime (78.4%), ofloxacin (76.1%) and ciprofloxacin (75.3%), the smallest in piperacillin/tazobactam (43.2%) and piperacillin (38.9%). The importance of incubation extended to 48 h during the testing of S. maltophilia strains was noted for correctly setting their susceptibility to antibiotics.

[Development of a vaccine for treatment of urinary tract infections]. / Vývoj vakcíny pro lécbu zánetlivých onemocnení mocových cest.

Receiving uroinfections are predominantly treated with antibiotics and chemotherapeutics. An effective alternative treatment can be immunotherapy with inactivated bacteria. The authors have focused on the development of polybacterial vaccine with both specific and nonspecific immunostimulatory activity. The vaccine contains killed bacteria which most frequently occur as an etiological agent in urinary tract infections. Determining for the selection of particular strains for the vaccine preparation were their immunostimulatory properties evaluated after intraperitoneal application of killed cells to mice. The results were evaluated according to weight gain of liver and spleen and verified by histological examination of these organs. The final form of the vaccine URVAKOL contains immunogenically most active strains of Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa and Enterococcus faecalis, representing specific effective part of the preparation, adjuvant activity is provided by Propionibacterium acnes. (Tab. 1, Fig. 2, Ref. 27.)

[Immunomodulation of recurrent urinary tract infections with Urvakol vaccine]. / Imunomodulace recidivujících uroinfekcí vakcínou URVAKOL.

The authors present the results of the URVAKOL vaccine use in clinical practice. The vaccine was administered in the treatment of recurrent cystitis, persistent lower urinary tract infection, chronic pyelonephritis and prostatovesiculitis. The clinical efficacy of the vaccine was assessed by detection of bacteria and leukocytes in the urine, subjective complaints, skin tests and the assessment of selected specific and nonspecific immunity parameters in the urine and saliva. Significantly positive clinical and laboratory responses were observed in patients with uncomplicated cystitis. In the group of persistent urinary tract infections excellent responses were detected in one half of the patients. The remainder half stated disappearance of subjective complaints, but with persistence of pyuria or bacteriuria. Similar results of "improvement" were observed in 10 patients with chronic pyelonephritis. There were no changes of the clinical state observed in patients with chronic prostatitis. (Tab. 5, Fig. 2, Ref. 7.)

[Immunostimulating activity of vaccines in the treatment of recurrent urinary tract infections. I]. / Immunostimulacní aktivita vakcíny pro lécbu recidivujících uroinfekcí. I.

The authors inform about the immunomodulatory properties of the vaccine URVAKOL aimed for the treatment of recidiving urinary infections. The results of immunostimulatory activity of the preparation and its effects on cellular and humoral immunity in mice following intraperitoneal administration of the vaccine are presented. The vaccine markedly increases cytotoxic activity of adhering peritoneal cells and has protective effects in model infection induced by intracellular pathogen Francisella tularensis (strain 15 L). (Tab. 6, Fig. 6, Ref. 9.)

[Immunostimulatory activity of the vaccine used in the treatment of recurrent urinary infections. II]. / Imunostimulacní aktivita vakcíny pro lécbu recidivujících uroinfekcí. II.

The authors describe on the immunostimulatory properties of the vaccine URVAKOL aimed for the treatment of recurrent urinary infections. Detection of immunostimulatory activity of the preparation and its effects on the humoral and cellular immunity were performed after oral administration of the preparation. Important was the evidence of nonspecific immunity of mice against intracellular pathogen Francisella tularensis induced with URVAKOL strain 15L. (Tab. 4, Fig. 1, Ref. 8.)

[Personal experience with classification of yeast microorganisms. I. The combined biotyping method of Mencl and Otcenásek and typing using the "killer" phenomenon]. / Nase zkusenosti s klasifikací kvasinkovitých mikroorganismu. I. Kombinovaná biotypizacní metoda podle Mencla a Otcenáska a typizace s vyuzitím "killer" fenoménu.

Using a group of 150 isolates of Candida albicans, C. glabrata, C. krusei, C. tropicalis, C. parapsilosis a C. kefyr the differentiating capacity of two biotyping systems was tested-the combined method according to Mencl and Otcenásek, and typing using the so-called killer phenomenon. With the combined method comparable results with the original work of the authors were obtained. This applies to the number of biotypes as well as to the ratio of the dominant biotype. As regards the differentiating characteristics of different biotypes the two studies differed fundamentally. As to typing, using the "killer" phenomenon, its practical usefulness was tested, the differentiating capacity of the method was, however, very much influenced by the small number of available killer-positive yeast strains.

[Characteristics of multiresistant gram-negative bacteria and their occurrence at the Medical School Hospital in Olomouc]. / Charakteristika multirezistentních gramnegativních bakterií a jejich frekvence ve Fakultní nemocnici v Olomouci.

Currently, a persistent growth of bacterial resistance to antibiotics takes place. Multiresistant bacterial strains thus represent a serious danger for therapy in practice. The aim of the study was to characterize the multiresistant Gram-negative bacteria and to judge their antibiograms by using the minimum inhibition concentrations (MIC). At the same time the study evaluates the relations to the most frequent diagnoses and epidemiologic data concerning the occurrence of these strains in different clinical cases treated at individual departments of the Faculty Hospital in Olomouc. Out of the total of 4.988 strains of Gram-negative rods, the number of isolated multiresistant strains reached 316 (6.3%). Individual bacterial species were isolated in the following order: Pseudomonas aeruginosa (28.8%), Enterobacter cloacae (23.4%) and Acinetobacter Baumannii complex (Acb complex) (18.7%). The most frequent occurrence of investigated bacteria was registered at the departments in the following order: urology (45.9%), internal medicine (18%), paediatrics (13.6%), and surgery (11.1%). The most frequent diagnoses based on positive cultivation of these bacteria were the infections of the urogenital tract (53.8%), postoperative infections (16.8%), and the infections of the upper respiratory ducts (7.3%). 64.2% of strains were cultivated from urine, 16.1% from pus, and 6.9% from swabs taken from the upper respiratory ducts. (Tab. 8, Ref. 18.).

[Occurrence of Klebsiella pneumonia strains resistant to third-generation cephalosporins at the Pediatric Clinic of the Medical School Hospital in Olomouc]. / Výskyt kmenu Klebsiella pneumoniae rezistentních k cefalosporinum III. generace na Detské klinice Fakultní nemocnice v Olomouci.

The authors describe the high incidence of strains of Klebsiella pneumoniae resistant to cephalosporins of the IIIrd generation at the department of pathological neonates of the Paediatric Clinic of the Faculty Hospital in Olomouc. They discuss possible reasons for this adverse situation and the way how to resolve it. They emphasize the necessity of rational administration of antibiotics and continuous monitoring of resistance in the most frequent causal agents of bacterial infections.

[Peroral vaccine therapy in dermatology]. / Perorální vakcinoterapie v dermatologii.

In a group of 691 patients with different dermatoses the therapeutic effect of two forms of lyophilized vaccines administered orally according to a desensitization (drops) and immunization (tablets) plan. Their effect was as a rule marked already during the third week of treatment. During the final evaluation no signs of the disease were present or great improvement was recorded in 89-100% of the patients suffering from acne, chronic staphylococcal pyoderma or microbial eczema and 62% (drops) or 65% (tablets) of patients with atopic eczema. During treatment the authors did not observe with rare exceptions of focal reactions any undesirable effects. No significant differences were found between the two forms of the drugs as regards therapeutic effects. Contrary to antibiotics, which can cause remission of dermatosis, while early relapses cannot be prevented, oral vaccine treatment leads to long-term (extending over several years) favourable effects on the course of the disease. The immunization pattern used in tablet form vaccines is simpler, packing and storage is simpler, dosage is more accurate and more convenient for the patient.

Resistance to antibiotics in gram-negative rods from clinical material of hospital and community origin.

The authors examined the resistance to 13 antibiotics and chemotherapeutic agents by the diluting micromethod and the routine disk method in the group of 5375 gram-negative strains of 7 genera which prevailed in clinical material of an hospital (FN Olomouc) and in material of community (OHS Olomouc) provenance during the years 1992 and 1993. In the majority of cases, the resistance was more frequent in the strains isolated from the hospital material. The most remarkable differences were found in A cinetobacter sp. (9-76%), Enterobacter sp. (10-60%) and Citrobacter sp. (18-58%). In other examined species, the differences varied in the ranges 3-40% in E.coli, 2-33% in Klebsiella sp., 0-28% in P. vulgaris, 2-28% in P.mirabilis, 0-16% in M.morganii, and 0-13% in P. aeruginosa.

[Occurrence and characteristics of gram-negative rods with marginal sensitivity and resistance to ofloxacin]. / Výskyt a charakteristika gramnegativních tycek s hranicní citlivostí a s rezistencí k ofloxacinu.

In the submitted paper the authors focused their attention on the characteristics of Gram-negative bacteria with a marginal sensitivity and resistance to ofloxacin (MIC > or = 2 mg.l-1) isolated from clinical materials in the Olomouc Faculty Hospital. They give an account of their sensitivity (based on assessment of the minimal inhibitory concentration-MIC) to 20 other antibiotics and chemotherapeutic agents and the percentage ratio of different species according to department, clinical material and diagnosis.

[Systemic administration of antibiotics in therapy of brain abscesses. Present trends and possibilities]. / Celková aplikace antibiotik pri lécbe mozkových abscesu. Soucasné trendy a moznosti.

The authors describe possibilities as regards selection of antibiotics in the general treatment of brain abscesses. They mention the most frequent aetiological agents of the disease in relation to predisposing factors and site. Systemic antibiotherapy along with surgery is considered by the authors decisive in the therapy of these conditions.

Application of Corynebacterium parvum lysate into the brain of rats--morphological study.

The authors investigated the influence of application of the Corynebacterium parvum lysate (CPL) into the brain of rats with aim to define the resulting morphological changes. Variable amount of the CPL (0.1, 0.35, 0.7 mg) was injected under stereotactic control into frontal lobe of the left cerebral hemisphere. Morphological changes were studied after intervals of 3, 7, 14, and 30 days respectively. CPL lead to inflammatory and reparative changes in direct relations to the administered dosage. The most pronounced reaction was observed on days 3 and 7. Morphological changes were still visible on day 14. There were no valid difference between the application of efficient CPL and the control physiological saline solution (PSS) after thirty days. The authors suggested that CPL is likely to be safe also in human malignant brain tumor therapy.

[Personal experience with treatment of recurrent herpes infections using combined nonspecific immunostimulation]. / Nase zkusenosti s lécbou recidivujících herpetických infekcí kombinovanou nespecifickou imunostimulací.

Using combined non-specific immunostimulation by means of a dialyzed leucocyte extract and bacterial vaccine, the authors treated 11 patients with relapsing herpetic infections. They achieved disappearance or marked improvement of clinical manifestations of the disease in the course of 12 months' treatment. The favourable clinical effect was associated with improvement of parameters of cellular immunity. Treatment was well tolerated by the patients. The only adverse factor was pain at the site of administration of the dialyzed leucocyte extract which persisted for 30 to 180 minutes, but due to the very favourable effect of treatment, was evaluated as insignificant by the patients.