Search

Enterococci are part of the normal intestinal flora of humans and animals. Under certain circumstances, they are capable of extraintestinal conversion to opportunistic pathogens. They cause endogenous as well as exogenous community and nosocomial infections. The gastrointestinal tract of mammals provides them with favorable conditions for acquisition and spread of resistance genes, for example to vancomycin (van), from other symbiotic bacteria. Thus, vancomycin-resistant enterococci (VRE) become potential reservoirs and vectors of the van genes. Their occurrence in the population of the Czech Republic was first reported by Kolář et al. in 1997. Some variants of the vanA gene cluster carried on Tn1546 which encode resistance to vancomycin are identical in humans and in animals. It means that animals, especially cattle, poultry and pigs, could be an important reservoir of VRE for humans. Kolář and Bardoň detected VRE in animals in the Czech Republic for the first time in 2000. In Europe, the glycopeptide antibiotic avoparcin, used as a growth stimulator, is responsible for selection of VRE strains in animals. Strains of Enterococcus faecium from animals may offer genes of antimicrobial resistance to other enterococci or they can be directly dangerous to human. This is demonstrated by finding isolates of E. faecalis from human patients and from pigs having very similar profiles of resistance and virulence genes. The goal of the paper was to point out the similarity between isolates of human and animal strains of enterococci resistant to vancomycin, and the possibility of their bilateral transfer between humans and animals.

The past two decades have witnessed increasing infections due to multidrug-resistant bacteria. Therefore, transmission of these pathogens could limit the antibiotic therapy options. Many reports suggest that initiation of appropriate antimicrobial therapy can be lifesaving. Physicians rely on combination of clinical, epidemiological and demographic data to guide empirical therapy because results of culture and antimicrobial susceptibility testing may require 48 hours or longer. Therefore, an ongoing effort for the development of earlier and more sensitive detection of resistant bacteria is inevitable. This review presents a summary of the most advanced methods (e.g. PCR-based techniques, flow cytometry, mass spectrometry, microarrays and others) that are able to rapidly detect antibiotic resistance in bacterial pathogens which have the potential to become valuable alternatives to the existing methods (standard phenotypic resistance testing) in the very near future.

Toxoplasmosis is a parasitic disease associated with high mortality in immunocompromised patients. It may lead to life-threatening conditions, usually neuroinfections, pneumonia or disseminated disease. It may be potentially dangerous, especially for patients with prolonged lymphopenia or those treated with immunosuppressive drugs. In our centre, we have observed 3 cases of toxoplasmosis in patients after allogeneic haematopoietic stem cell transplantation (HSCT) (2.6% of 116 allo-HSCT patients since 2000) and one case after autologous HSCT (0.3% of 395 auto-HSCT patients since 1997). Toxoplasmosis is manifested by neurological symptoms including hemiparesis and paraparesis, cerebral salt-wasting syndrome (hyponatraemia and hypoosmolality), psychoorganic syndrome and signs of respiratory infection. The diagnosis was made by combining clinical signs and results of PCR and CT examinations. The patients were treated with high-dose pyrimethamine, clindamycin, co-trimoxazole and folic acid. Three of the four patients have survived with no signs of the disease. One patient died prior to treatment. The increasing use of highly immunosuppressive chemotherapy and conditioning regimens (including rituximab, fludarabine and anti-thymocyte globulin) is associated with a significant risk of toxoplasmosis. Variable manifestations, non-specific results of MRI or CT examinations and possibility of PCR negativity are the main obstacles to successful diagnosis.

Background: Currently, important bacterial beta-lactamases of increasing clinical significance include AmpC enzymes. The aim was to assess their occurrence in Klebsiella pneumoniae strains isolated from patients with haematological malignancies in a prospective study., Material and Methods: Over a 2-month period, strains of the species were isolated from clinical material obtained from patients hospitalized at the Department of Haemato-Oncology of the University Hospital Olomouc. The strains were identified using standard microbiological techniques and the Vitek 2 automated system. Production of AmpC beta-lactamases was roughly determined by phenotypic tests and subsequently confirmed by PCR detection of genes encoding these enzymes., Results: During the above-mentioned period, a total of 35 K. pneumoniae isolates were collected. In 7 of them, production of AmpC beta-lactamases was preliminarily detected by phenotypic test. The multiplex PCR method confirmed phenotyping and determined DHA types in all the isolates., Conclusions: All AmpC-positive isolates were false-susceptible to at least one of the tested third-generation cephalosporins. In one patient, clinically apparent infection caused by this strain was documented. The reported results suggest the possibility of occurrence of AmpC-beta-lactamases in K. pneumoniae strains with clinical significance.

Objectives: The study aimed at the assessment of the prevalence of ESBL-positive isolates of Klebsiella pneumoniae in intensive care patients and their molecular biology analysis., Material and Methods: Over a 5-month period, Klebsiella pneumoniae strains were isolated from patients hospitalized at the Department of Anaesthesiology and Resuscitation of the University Hospital in Olomouc. For each isolate, an antibiogram was performed by the standard microdilution method and the production of ESBL was determined by the modified double-disk synergy test. PCR was used to demonstrate the presence of the blaTEM and blaSHV genes. The isolates producing SHV- and TEM-types of beta-lactamases were typed using the restriction fragment length polymorphism (RFLP) method to identify the most common mutations responsible for the development of an ESBL phenotype. Similar or identical isolates were determined by pulsed-field gel electrophoresis (PFGE) of DNA fragments cleaved by the XbaI restriction endonuclease., Results: A total of 67 isolates of Klebsiella pneumoniae were obtained. In 13 of them, the production of ESBL was detected and the presence of the blaSHV gene was confirmed by PCR. Restriction cleavage by NheI revealed mutations at position 238 in all SHV-positive PCR products. The restriction analysis did not confirm the presence of the gene encoding TEM-type extended-spectrum beta-lactamase. Molecular biology typing by PFGE detected the presence of 11 different strains., Conclusions: In the observed group of intensive care patients, the prevalence of ESBL-positive strains of Klebsiella pneumoniae reached 19.4 %. The analysis of SHV and TEM products of PCR by the RFLP method showed the prevalence of SHV-type ESBL. Overall, 84.6 % of the strains had unique restriction profiles. The results suggest both high levels of hygienic and epidemiological measures at the monitored department and rational antibiotic policy.

Viral hepatitis D (HDV) is a serious acute inflammatory liver damage, which frequently is transmitting to the chronic phase, cirrhosis and hepatocellular carcinoma. This disease can be found in the Czech Republic only rarely. The authors present the case of viral hepatitis D imported to the Czech Republic from Moldavia in 28-year old man. According to the examinations of the markers of HBV and HDV, the most probably it is a HDV superinfection in patient after acute HBV with HBsAg carriers.

Background: Achalasia (ACHL) is not a very frequent disease and its etiology is still unclear. In addition to the oldest therapeutical approach represented by myotomy, two conservative methods are commonly used--balloon dilatation and application of botulotoxin. So far, both methods have been used only separately, and their effects have been compared. Literature provides no evidence of the renewal of oesophageal propulsive peristalsis in result of conservative treatment., Objectives: The aim is to use both approaches subsequently within a short period of time, in order to potentiate their effects and at the same time to reduce the risk of possible complications., Methods: The group was formed by 9 patients. Achalasia was diagnosed by flow manometry. Pseudoachalasia was excluded endoscopically and endosonographically. The treatment included application of 250 J of botulotoxin (Dysport) into the region of the lower oesophageal sphincter, and balloon dilatation which was applied 7 days later. Following this treatment, patients were observed for 24 hours. The clinical and manometric examinations were performed in 3-month intervals., Results: All patients felt significantly better after treatment. The clinical state in two patients required the performance of Heller's myotomy. After 1 year, evident clinical and manometric improvement was observed in 7 patients. The longest improvement so far lasted for 36 months. The treatment has renewed the propulsive oesophageal peristalsis in two women., Conclusion: The treatment of primary achalasia by means of combining the application of botulotoxin (Dysport) and balloon dilatation is effective, and it is possible to assume that the clinical remission will last longer than that in result of separate use of just one of the methods. The synchronous treatment of functional blocks in the cervical and thoracic regions of the spine and continuous rehabilitation can participate in the favourable clinical effect. The renewal of primary peristalsis of the oesophagus was achieved in two out of 9 patients. No such change has been either manometrically verified or described in literature. (Fig. 5, Tab. 1, Ref. 39.)

Both degradative and limited proteolysis are involved in physiological processes. Once activated, proteinases are controlled by endogenous inhibitors (antiproteinases). A variety of genetic disorders and exogenous microbial proteinases disturb the balance between proteinases and cognate inhibitors. alpha 1-Antitrypsin deficiency is a model disorder resulting in an insufficient control of leukocyte elastase with a subsequent alveolar tissue damage. It appears that the manifold molar excess of inhibitor required to compensate the deficiency, is the consequence of both the local relative inaccessibility of the serpin, and the mode of proteinase-serpin interaction, in accord with the "branched pathway" mechanism. Slow-binding kinetics and the leak of native proteinase from the complex is illustrated by the peptidolytic action of porcine elastase in the presence of human alpha 1-antitrypsin and by gelatin zymography, respectively. (Tab. 2, Fig. 5, Ref. 44.)

Four out of a group of 5 patients with a colloid cyst located within the III cerebral ventricle were surgically treated by means of a classical microsurgical technique and one of them was treated stereotactically. The preference was given to the frontodorsal transcortical approach. The current CT diagnosis is reliable and by use of MR it is to a certain extent possible to distinguish even the origin of the cyst. Prior to examination we had found out that the cyst was a neuroenteral inclusion and the final surgical result was good in all patients who had been subdued to operation during the initial phase of foramen Monroi enclosure. (Fig. 2, Ref. 10.)

Concentrations of some hazardous pollutants were studied in dust deposit samples (n = 30) taken from the indoor equipment of two feed mixture plants (FMP) in Hodonín district in 1995 and 1996; the pollutants involved mercury, cadmium, lead, hexachlorobenzene (HCB), alpha and beta hexachlorocyclohexane (HCH), lindane (gamma HCH), DDT sum, DDT isomers, polychlorinated biphenyls (Delor 103, Delor 106, PCB indicator congeners). The statistically significantly higher (P < 0.01) average concentration of mercury (0.028 mg/kg) was recorded in dust depositions (n = 15) in FMP K. in comparison with FMP R. Increased concentrations of cadmium (more than 0.5 mg/kg) were determined in dust depositions in FMP K. in 1 sample (0.582 mg/kg), of lead (more than 5.0 mg/kg) in 1 sample (7.540 mg/kg), of HCB (more than 0.01 mg/kg) in two samples (0.034 and 0.036 mg/kg), of beta HCH (more than 0.005 mg/kg) in 4 samples (0.021; 0.022; 0.023 and 0.046 mg/kg), higher DDT sum (more than 0.05 mg/kg) were determined in FMP K. in 3 samples (0.072; 0.085 and 0.190 mg/kg) and of PCB sum (more than 0.05 mg/kg) in 4 samples (0.054; 0.059; 0.074 and 1.085 mg/kg). In comparison with FMP K., dust depositions (n = 15) in FMP R. showed the statistically significantly higher (P < 0.05) average concentration of lead (5.241 mg/kg). Increased concentrations of lead were determined in dust depositions in FMP R. in 7 samples (5.040; 5.230; 6.760; 7.590; 7.620; 8.450 and 15.650 mg/kg), of beta HCH in 7 samples (0.006; 0.008; 0.022; 0.024; 0.026; 0.029 and 0.049 mg/kg), higher DDT sum were recorded in 3 samples (0.053; 0.074 and 0.126 mg/kg) and higher PCB sum in 8 samples (0.057; 0.066; 0.101; 0.134; 0.164; 0.426; 0.821 and 1.202 mg/kg). The highest average concentration of hazardous pollutants in dust depositions (n = 30) in both feed mixture plants was determined in lead (4.122 mg/kg), in a descending order followed by PCB sum (0.151 mg/kg), cadmium (0.127 mg/kg), DDT sum (0.030 mg/kg), mercury (0.019 mg/kg), lindane (0.013 mg/kg), beta HCH (0.010 mg/kg), HCB (0.004 mg/kg and alpha HCH (0.001 mg/kg). Out of PCB indicator congeners, dust depositions contained the highest quantities of congener 138 (0.022 mg/kg), congener 180 (0.018 mg/kg) and congener 153 (0.015 mg/kg). Isomer p,p-DDT was dominant in dust depositions among DDT isomers (0.019 mg/kg). Dust depositions sampled from the indoor equipment of feed mixture plants are a promising indicator of produced feed mixtures contamination with hazardous pollutants. To seek sources of feed mixtures contamination is a permanent task of veterinary service.

During the direct handling with mouflon the ivermectin (IVM) is usually parenterally administered. The used dose is equal to dose recommended to domestic sheep (0.20 mg/kg of body weight). The qualitative and quantitative parasithological differences between domestic and wild sheep (mouflon) exist (Vyslouzil, 1985; Lochman et al., 1979; Boch and Supperer, 1992). The verification of real anthelmintic efficacy in IVM treatment of mouflon was not realized up to now. The aim of this study was to describe anthelmintic efficacy of IVM administered in different doses and to select the optimum dose. The study was realized during winter season in game herd in 23 mouflons of 10-11 months old. Before treatment larvae of Muellerius capillaries (strong infection) and eggs of Nematodirus spp. and Oesophagostomum spp. (mild infection) in common faecal samples were found. The mouflons were randomly divided in four groups control group (3 head) and 3 experimental groups (6-8 animals in group). Before experimental treatment each animal in experimental groups was weighted and marked. The doses of IVM 0.20, 0.60, and 1.00 mg/kg of body weight were used. The drug was administered subcutaneously. All mouflons were placed into enclosured area of 4 hectare. 28-60 days after treatment mouflon were hunted and detail helminthological examinations (lung macroscopical evaluation, larvoscopy of lung verminous spots, larvoscopy and ovoscopy of individual faeces, necropsy of gastrointestinal organs and quantification of nematode adults) were carried out, LPG and EPG values were determined. The study was realized as a simple blind experiment--all helminthological examinations were done without knowledge of the used therapy (control x experimental animals) or IVM dose (experimental animals). Macroscopical examinations of lungs showed symptoms of strong M. capillaries infection (numerous verminous spots) in all 23 mouflons. Larvoscopical examinations (lung tissue, individual faeces) after treatment became negative in 19 treated animals, in the rest of animals the mild larvoscopical finding was determined (Tab. I). Pretreatment common LPG values (ca 2100-2230) in control animals were confirmed, findings in verminous spots of these animals were highly positive (massive presence of live larvae, eggs, and rests of nematode adults). The qualitative findings (quantitatively were findings under the detection limit for the used method) were minimum in 4 mouflons of all treated groups. Pretreatment ovoscopical findings in control animals were confirmed, too. Necropsy examinations of gastrointestinal organs were highly positive in all control animals (mean finding of 95 adults of determined nematodes), in treated animals the findings were positive in 3 animals of different groups (up to 2 adults only). In respect to reached results, pathogenicity of found nematodes, age of experimental mouflons, and economy of treatment, a dose of 0.60 mg/kg of body weight IVM dose as a optimum for parenteral treatment of mouflon nematodoses is recommended.

Since 1968 when bovine tuberculosis was eliminated in the Czech Republic, the epidemiological situation of bovine tuberculosis has been stabilized. At present the incidence of the disease in men and animals caused by conditionally pathogenic mycobacteria is worldwide increasing. In human population, especially people with impaired immunity are affected. In farm animals infections caused by conditionally pathogenic mycobacteria may often result in complications in intravital and postmortal diagnosis of bovine and avian tuberculosis. Those infections are then often incorrectly diagnosed which could have a great negative impact on health, economy and breeding. Therefore the objective of the study was to summarize data from literature and our own experience concerning the occurrence of atypical mycobacteria in environment. The study is divided into 5 summarizing chapters, supplemented with 13 Tables.