V Jihomoravském kraji, který je oblastí s nejnižší prevalencí bacilární tuberkulózy v České repub-lice (6,6 na 100 000 obyvatel v roce 1996), bylo v letech 1993–1996 zjištěno celkem šest lokálnícha rodinných mikroepidemií tuberkulózy. K jejich identifikaci byla použita metoda RFLP fingerprin-tingu založená na průkazu repetitivní sekvence IS6110 v chromosomální DNA vyšetřovaných kmenůMycobacterium tuberculosis, natrávené restrikčním enzymem PvuII. Profily fingerprintů každémikroepidemie byly počítačovým programem zařazeny do hierarchie dendrogramu fingerprintů184 českých kmenů M. tuberculosis, což umožnilo identifikovat případné identické profily kmenůu nemocných ze vzdálených lokalit České republiky.Ve třech rodinných mikroepidermiích složených vždy ze dvou členů nebyly prokázány identicképrofily fingerprintů jiných českých kmenů M. tuberculosis. K čtvrtému clusteru tvořenému šestičleny jedné rodiny se přiřadil identický profil RFLP nemocné žijící v blízké lokalitě. U dalšímikroepidemie zaznamenané u tří bratrů byly nalezeny identické fingerprinty u jiných čtyř nemoc-ných z Jihomoravského kraje a u jednoho ze Středočeského kraje. K poslednímu clusteru dvoubratrů se přiřadilo překvapivě šest identických profilů RFLP nemocných ze Západočeského krajea jednoho z Prahy. Tyto nálezy svědčí o tom, že v oblastech s nízkou prevalencí se tuberkulózaudržuje častěji než v oblastech s vysokou prevalencí formou rodinných a lokálních mikroepidemií.

In the South Moravian region which is the area with the lowest prevalence of bacillary tuberculosisin the Czech Republic (6.6 per 100 000 population in 1996) in 1993–1996 a total of six local and familymicroepidemics of tuberculosis were detected. For their identification the RFLP fingerprintingmethod was used based on evidence of repeated sequence of IS6110 in the chromosomal DNA of theexamined strains of Mycobacterium tuberculosis predigested with restrictive enzyme PvuII. Theprofiles of fingerprints of each microepidemic were included by means of a computer programmeinto the hierarchy of the fingerprint dendrogram of 184 strains of M. tuberculosis which made itpossible to identify possible identical profiles of strains from patients from remote places in theCzech Republic.In three family microepidemics involving always two members no identical fingerprint profiles ofother Czech strains of M. tuberculosis were revealed. To the fourth cluster formed by six membersof one family an identical RFLP profile of a female patient living in a nearby locality was added. Inanother microepidemic recorded in three brothers identical fingerprints were found another fourpatients from the South Moravian region and in one from the Central Bohemian region. The lastcluster of two brothers was surprisingly enlarged by six identical RFLP profiles of patients fromthe West Bohemian region and one from Prague. These findings suggest that in areas with a lowprevalence tuberculosis persists more frequently than in areas with a high prevalence as familialor local microepidemics.

• MeSH
• DNA fingerprinting MeSH
• lidé MeSH
• Mycobacterium tuberculosis genetika   patogenita   MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• tuberkulóza diagnóza   epidemiologie   MeSH
• Check Tag
• lidé MeSH

Genetic diversity among 43 petroleum hydrocarbon-degrading Pseudomonas belonging to four different species and the type strain Pseudomonas aeruginosa MTCC1034 was assessed by using restriction fragment length polymorphism (RFLP) of polymerase chain reaction (PCR)-amplified 16S-23S rDNA intergenic spacer regions (ISRs) polymorphism. PCR amplification from all Pseudomonas species yielded almost identical ISR amplicons of "?" 800 bp and in nested PCR of "?" 550 bp. The RFLP analysis with MboI and AluI revealed considerable intraspecific variations within the Pseudomonas species. The dendrogram constructed on the basis of the PCR-RFLP patterns of 16S-23S rDNA intergenic spacer regions differentiated all the species into seven different clusters.

• MeSH
• DNA bakterií genetika   MeSH
• fylogeneze MeSH
• mezerníky ribozomální DNA genetika   MeSH
• polymerázová řetězová reakce metody   MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• Pseudomonas klasifikace   genetika   izolace a purifikace   metabolismus   MeSH
• RNA ribozomální 16S genetika   MeSH
• RNA ribozomální 23S genetika   MeSH
• ropa mikrobiologie   MeSH
• techniky typizace bakterií metody   MeSH
• uhlovodíky metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH

The Campylobacter species strains (n = 42; isolated from clinical samples and deposited in Czech National Collection of Type Cultures, Prague) originally phenotypically (and biochemically) identified as Campylobacter jejuni were re-classified using molecular biological and mass spectrometric methods. Whole-cell MALDI-TOF MS (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry) separated the isolates into two genetically related strains--C. jejuni (n = 26) and C. coli (n = 16) and, moreover, distinguished the intimate details in the group of tested strains. It also made it possible to create the MALDI-TOF MS dendrogram; similarly, the spectral characteristics were used for the 3D cluster analysis. Polymerase chain reaction (PCR) confirmed the results obtained by mass spectrometry. Both methods (PCR and MALDI-TOF MS) gave the same results which supports their suitability in the rapid and accurate Campylobacter-species determination.

• MeSH
• Campylobacter coli genetika   klasifikace   MeSH
• Campylobacter jejuni genetika   klasifikace   MeSH
• DNA bakterií analýza   izolace a purifikace   MeSH
• druhová specificita MeSH
• feces mikrobiologie   MeSH
• kočky MeSH
• lidé MeSH
• polymerázová řetězová reakce MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice metody   MeSH
• techniky typizace bakterií MeSH
• zvířata MeSH
• Check Tag
• kočky MeSH
• lidé MeSH
• zvířata MeSH

Although Campylobacter jejuni is the pathogen responsible for the most common foodborne illness, tracing of the infection source remains challenging due to its highly variable genome. Therefore, one of the aim of the study was to compare three genotyping methods (MLST, PFGE, and mP-BIT) to determine the most effective genotyping tool. C. jejuni strains were divided into 4 clusters based on strain similarity in the cgMLST dendrogram. Subsequently, the dendrograms of the 3 tested methods were compared to determine the accuracy of each method compared to the reference cgMLST method. Moreover, a cost-benefit analysis has showed that MLST had the highest inverse discrimination index (97%) and required less workflow, time, fewer consumables, and low bacterial sample quantity. PFGE was shown to be obsolete both because of its low discriminatory power and the complexity of the procedure. Similarly, mP‐BIT showed low separation results, which was compensated by its high availability. Therefore, our data showed that MLST is the optimal tool for genotyping C. jejuni. Another aim was to compare the antimicrobial resistance to ciprofloxacin, erythromycin, and tetracycline in C. jejuni strains isolated from human, water, air, food, and animal samples by two gene sequence-based prediction methods and to compare them with the actual susceptibility of C. jejuni strains using the disc diffusion method. Both tools, ResFinder and RGI, synchronously predict the antimicrobial susceptibility of C. jejuni and either can be used.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální léková rezistence genetika   MeSH
• Campylobacter jejuni * genetika   MeSH
• genotyp MeSH
• kampylobakterové infekce * mikrobiologie   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• multilokusová sekvenční typizace MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The capability of Fluorescent Random Amplified Microsatellites (F-RAMS) to profile hallucinogenic mushrooms to species and sub-species level was assessed. Fifteen samples of Amanita rubescens and 22 samples of other hallucinogenic and non-hallucinogenic mushrooms of the genera Amanita and Psilocybe were profiled using two fluorescently-labeled, 5'degenerate primers, 5'-6FAM-SpC3-DD (CCA)5 and 5'-6FAM-SpC3-DHB (CGA)5, which target different microsatellite repeat regions. Among the two primers, 5'-6FAM-SpC3-DHB (CGA)5 provided more reliable data for identification purposes, by grouping samples of the same species and clustering closely related species together in a dendrogram based on amplicon similarities. A high degree of intra-specific variation between the 15 A. rubescens samples was shown with both primers and the amplicons generated for all A. rubescens samples were organized into three classes of amplicons (discriminant, private, and marker) based on their individualizing potential.

• MeSH
• Amanita genetika   MeSH
• DNA fingerprinting metody   MeSH
• DNA primery * MeSH
• fluorescence MeSH
• fluorescenční barviva * MeSH
• mikrosatelitní repetice * MeSH
• polymerázová řetězová reakce MeSH
• Psilocybe genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Cíl práce: Molekulární epidemiologie je oborem, který využívá výsledky typizačních technik k získání informací o detailní charakteristice bakteriálních kmenů umožňujících stanovit identitu, příbuznost nebo odlišnost bakterií stejného rodu, druhu, případně i sérotypu. V současnosti jsou nejvíce používány metody založené na sledování rozdílu v genotypu bakterií. Většina těchto technik je ovšem časově i finančně náročná. Do rutinní praxe mikrobiologických laboratoří se ale rozšiřuje i metoda založená na analýze bakteriálního proteomu - hmotnostní spektrometrie s laserovou desorpcí/ionizací za účasti matrice s použitím průletového analyzátoru (MatrixAssisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry, MALDI-TOF MS). Je využívána zejména k rychlému a přesnému zařazení bakterií do rodu i druhu. Cílem naší práce proto bylo posoudit možnost využití této metody k typizaci kampylobakterů pod úroveň druhu a aplikaci výsledků při epidemiologických šetřeních. Materiál a metody: Do studie bylo zahrnuto 39 kmenů Campylobacter jejuni izolovaných z potravin (16) nebo humánního původu (23). U kmenů bylo provedeno stanovení makrorestrikčních profilů metodou pulzní gelové elektroforézy (PFGE) a současně také analýza proteinových profilů s využitím metody MALDI-TOF MS. Výsledky: Vyhodnocení míry podobnosti pulzních profilů prokázalo shody mezi izoláty pocházejícími ze stejné epidemie nebo z opakovaných odběrů jednoho pacienta. Dále byly detekovány stejné pulzní profily u kmenů bez známé spojitosti, ale se shodným místem původu a rokem izolace. Z porovnání shlukových analýz obou metod vyplývá, že kmeny označené metodou PFGE jako shodné se V dendrogramu MALDI-TOF MS objevují v jedné podskupině s výjimkou kmenů z opakovaného odběru stejného pacienta. Závěr. Naše výsledky ukazují, že potvrzení identity nebo příbuznosti kmenů v souladu se zjištěnými epidemiologickými skutečnostmi se zatím metodou MALDI-TOF nepodařilo jednoznačně prokázat.

Objectives: Molecular epidemiology is a field that uses results of typing techniques to obtain information on detailed characterization of bacterial strains for determining the identity, similarity or difference in bacteria of the same genus, species or serotype. Nowadays, the most commonly used methods are based on monitoring differences in bacterial genotypes. However, most of these techniques are time-consuming and costly. A method increasingly used in routine microbiological testing is matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), which is based on analysis of the bacterial proteome. It is mainly used for rapid and accurate classification of bacteria into genera and species. The aims were to assess the potential use of this method for typing of Campylobacter below the species level and to apply these results in epidemiological investigations. Material and Methods: The study comprised 39 strains of Campylobacter jejuni isolated from food (16) and humans (23). Macrorestriction fragment profiling by pulsed-field gel electrophoresis (PFGE) and simultaneous protein profile analysis using MALDI-TOF MS were performed for all tested strains. Results: Similar pulse profiles were found among isolates originating from the same outbreak or repeatedly collected from a single patient. The same pulse profiles were also detected in strains of unknown relationship but sharing the same place of origin and year of isolation. The comparison of dendrograms from both analyses showed that strains identified as identical by PFGE appeared in the same subgroups in dendrograms obtained by MALDI-TOF MS, the only exception being isolates repeatedly collected from a single patient. Conclusion: The results suggest that confirmation of the identity or similarity of strains in accordance with the established epidemiological facts has not been clearly demonstrated using MALDI-TOF MS.

• Klíčová slova
• MALDI-TOF MS,
• MeSH
• Campylobacter jejuni * izolace a purifikace   MeSH
• lidé MeSH
• pilotní projekty MeSH
• pulzní gelová elektroforéza statistika a číselné údaje   MeSH
• shluková analýza MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice * statistika a číselné údaje   MeSH
• techniky typizace bakterií metody   statistika a číselné údaje   využití   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

Using the means of five cranial indices of three early medieval groups from Austria and four comparative series from neighbouring regions (Southern Germany, CSSR and Hungary) distance measures and similarity relations were computed and the applied methods were compared. In addition, the attempt for a better consideration of the biological conditions was made by combining two known methods at a time (Dendrogram by means of (DD)2 distances; population-composition on the basis of adjacent secured comparative series). The results of this study reveal, that using the above cited simple methods on the only basis of the means of five cranial indices a correspondence with the attributions of the three Austrian series cited in the literature and basing on extensive investigations was obtained. Thus a very strong relation -- especially concerning the males -- resulted between the series from Pitten and the Slavic ones, and between the groups of Zwölfaxing -- more expressed in the females -- and the Avaric series. Of special interest are the results of the mixed population of Zwentendorf were the females resemble quite strikingly the Slavic, the males, however, the Bajuvaric comparative series.

• MeSH
• dějiny středověku MeSH
• kefalometrie MeSH
• lidé MeSH
• paleontologie MeSH
• rasové skupiny * MeSH
• Check Tag
• dějiny středověku MeSH
• lidé MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• historické články MeSH
• Geografické názvy
• Československo MeSH
• Maďarsko MeSH
• Německo západní MeSH
• Rakousko MeSH

Microscopic diagnosis of equine piroplasmoses, caused by Theileria equi and Babesia caballi, is hindered by low parasitaemia during the latent phase of the infections. However, this constraint can be overcome by the application of PCR followed by sequencing. Out of 288 animals examined, the piroplasmid DNA was detected in 78 (27·1%). Multiplex PCR indicated that T. equi (18·8%) was more prevalent than B. caballi (7·3%), while mixed infections were conspicuously absent. Sequences of 69 PCR amplicons obtained by the 'catch-all' PCR were in concordance with those amplified by the multiplex strategy. Computed minimal adequate model analyses for both equine piroplasmid species separately showed a significant effect of host species and age in the case of T. equi, while in the B. caballi infections only the correlation with host sex was significant. Phylogenetic analyses inferred the occurrence of three genotypes of T. equi and B. caballi. Moreover, a novel genotype C of B. caballi was identified. The dendrogram based on obtained sequences of T. equi revealed possible speciation events. The infections with T. equi and B. caballi are enzootic in all ecozones of Jordan and different genotypes circulate wherever dense horse population exists.

• MeSH
• Babesia klasifikace   genetika   izolace a purifikace   MeSH
• babezióza epidemiologie   parazitologie   MeSH
• Equidae parazitologie   MeSH
• fylogeneze MeSH
• genetická variace * MeSH
• genotyp MeSH
• koně MeSH
• molekulární sekvence - údaje MeSH
• multiplexová polymerázová řetězová reakce veterinární   MeSH
• nemoci koní epidemiologie   parazitologie   MeSH
• parazitemie veterinární   MeSH
• prevalence MeSH
• protozoální DNA chemie   genetika   MeSH
• sekvence nukleotidů MeSH
• sekvenční analýza DNA veterinární   MeSH
• skot MeSH
• Theileria klasifikace   genetika   izolace a purifikace   MeSH
• theilerióza epidemiologie   parazitologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Jordánsko MeSH

Of the 19 strains of Rhizopus delemar deposited as Rhizopus oryzae, seven of them, NBRC 4726, NBRC 4734, NBRC 4746, NBRC 4754, NBRC 4773, NBRC 4775, and NBRC 4801, completely hydrolyzed exogenous sucrose and fructooligosaccharides. The sucrose-hydrolyzing enzyme was purified from the culture filtrate of R. delemar NBRC 4754 and classified to β-fructofuranosidase, similar to that of Amylomyces rouxii CBS 438.76. Fragments including β-fructofuranosidase genes (sucA) of seven strains of R. delemar and A. rouxii CBS 438.76 were amplified and sequenced by PCR with degenerated primers synthesized on the basis of the internal amino acid sequences of purified enzymes and successive inverse PCR. Nucleotide sequences of the obtained fragments revealed that open reading frames of 1,569 bp have no intron and encode 522 amino acids. The presumed proteins contained the typical domain of the glycoside hydrolase 32 family, including β-fructofuranosidase, inulinase, levanase, and fructosyltransferases. Amino acid sequences of SucA proteins from the seven strains of R. delemar were identical and showed 90.0 % identity with those of A. rouxii CBS 438.76. A dendrogram constructed from these amino acid sequences showed that SucA proteins are more closely related to yeast β-fructofuranosidases than to other fungal enzymes.

• MeSH
• DNA fungální chemie   genetika   MeSH
• fylogeneze MeSH
• hydrolýza MeSH
• invertasa genetika   izolace a purifikace   metabolismus   MeSH
• molekulární sekvence - údaje MeSH
• Mucorales enzymologie   genetika   MeSH
• oligosacharidy metabolismus   MeSH
• polymerázová řetězová reakce MeSH
• sacharosa metabolismus   MeSH
• sekvenční analýza DNA MeSH
• sekvenční homologie aminokyselin MeSH
• shluková analýza MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

We propose the first comprehensive in-depth study monitoring horses in the Czech Republic. We scanned 9,289 animals from 44 populations for 17 equine STRs. Other equids analysed involved Equus przewalskii and Equus asinus. The total of 228 different alleles were detected, with the mean number of 13.4 per locus. The highest allelic richness (AR) was found in the Welsh Part Bred (6.01), followed by the Camargue (5.93) and Czech Sport Pony (5.91), whereas the Friesian exhibited the lowest AR (3.06). Interpopulation differences explained approximately nine per cent of the total genetic diversity. Reynold's genetic distance ranged from 0.003 between the Czech Warmblood and the Slovak Warmblood to 0.404 between the Friesian and donkeys. Close genetic proximity between the Silesian Noriker and Noriker was revealed. The Moravian Warmblood was better differentiated and more distant from the Czech Warmblood than the Kinsky Horse and retained the original genes of the old Austro-Hungarian tribes. A high gene flow level and a lack of genetic structure were found in the seven studied populations. Despite the historical bottlenecks and previous inbreeding, the Czech-Moravian Belgian Horse, Hucul, Old Kladruber Horse and Silesian Noriker did not suffer a serious loss of genetic diversity due to genetic drift/low effective population size. A NeighborNet dendrogram revealed breeds not classified in their groups according to the nomenclature (the Friesian, Hafling and Merens).

• MeSH
• chov MeSH
• genetická variace MeSH
• genotyp MeSH
• koně genetika   MeSH
• shluková analýza MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH