The main aim of the study was to evaluate associations between morphological variability of Trichuris females from sheep and roe deer and their rDNA polymorphism in whipworm populations from the Czech Republic. The results introduced the use of new molecular markers based on the internal transcribed spacer (ITS)1-5.8S RNA-ITS2 region polymorphisms, as useful tools for the unambiguous differentiation of congeners Trichuris ovis and Trichuris discolor. These markers revealed both parasites in roe deer and in sheep; however, T. ovis females predominated in sheep while T. discolor females occurred mostly in roe deer. Additional analysis of ITS1-5.8 rRNA-ITS2 discovered the genetic uniformity of the analysed T. discolor but high haplotype variation of T. ovis. Simultaneously, molecularly designated female individuals of both species were categorised into four morphotypes (MT) on the basis of morphology of genital pore area. MT1 and MT4 (vulvar opening on everted vaginal appendage/on visible cuticular bulge) occurred only in T. ovis, MT2 (uneverted vagina-vulvar opening without any elevation) was identified only in T. discolor and MT3 (transient type of vulvar opening on a small swelling) was observed in both species. Statistical analysis of biometric data confirmed that morphology of vulva is not a reliable marker for the species determination. On the basis of the ITS1-5.8S RNA-ITS2 region variability, we carried out a phylogenetic analysis (maximum likelihood method, Hasegawa-Kishino-Yano model) which showed that T. ovis haplotypes from the Czech Republic and Ireland and T. discolor haplotypes from the Czech Republic, Spain, Iran and Japan are sister OTUs.

• MeSH
• DNA, Helminth MeSH
• Phylogeny MeSH
• Molecular Typing MeSH
• Sheep Diseases parasitology   MeSH
• Sheep genetics   parasitology   MeSH
• Polymorphism, Genetic MeSH
• DNA, Ribosomal genetics   MeSH
• RNA, Ribosomal MeSH
• Trichuriasis parasitology   veterinary   MeSH
• Trichuris anatomy & histology   classification   genetics   isolation & purification   MeSH
• Deer parasitology   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Czech Republic MeSH

Francisella tularensis (F.t.) je gram-negativní, fakultativně intracelulární bakterie způsobující tularemii. Díky svým ojedinělým vlastnostem však byla v minulosti pravděpodobně zneužita k vojenským cílům jako biologická zbraň, a je proto Centrem pro kontrolu nemocí (CDC) v USA zařazena mezi nejrizikovější infekční agens v rámci možného vývoje biologických zbraní a bioterorismu. Není proto překvapující, že bylo vyvinuto enormní úsilí k objasnění imunopatogeneze onemocnění s cílem vyvinout účinnou a bezpečnou vakcínu. Přestože již v padesátých létech byla využita u exponovaných jedinců v USA vakcína obsahující oslabený kmen Francisella tularensis LVS (live vaccine strain, F.t. LVS), nebyla doposud schválena pro široké klinické využití. Současný výzkum se opírá o analýzu atenuovaných a virulentních kmenů rodu Francisella se zaměřením na rozdíly v genetické výbavě a proteinové produkci a dále o modelování primární infekce in vitro či o pokusy in vivo na zvířecích modelech. Doposud tak byl zmapován bakteriální genom i proteom, byly definovány genové úseky kódující faktory virulence i obecné zákonitosti v reakci imunitního systému na infekci a některé možné mechanismy úniku F.t. z jeho dosahu. V následující práci jsou shrnuty některé současné názory na imunopatogenezi infekce F.t.

Francisella tularensis (F.t.), a causal agent of tularemia is a gram-negative, facultative intracellular bacteria that has already been abused for military intentions as a biological weapon. As F.t. is highly infectious and easily cultivable it is enrolled at the list of Centre for Disease Control (CDC), USA as a high risk bioterrorism agent, category A. There is a long-term effort to understand to the immunopathogenesis of F.t. infection and to construct an effective and safe vaccine. However, the vaccine used in high-risk population in USA in past derived from atenuated F.t. LVS (Live Vaccine Strain), was never accepted for large scale clinical use. Wild type and atenuated strains of F.t. are now under investigation in the terms of gene transcription and protein production. Many in vitro and in vivo models have been established for this purpose. F.t. bacterial genome and proteome is described in details now and many virulence factors have already been defined. Some basic principles of immune response to F.t. infection and mutual interactions with host have been established. Some recent opinions on F.t. immunopathogenesis are summarized in this review article.

• Keywords
• nitrobuněčný parazitismus, primární infekce,
• MeSH
• Bacteriological Techniques methods   utilization   MeSH
• Bioterrorism prevention & control   MeSH
• Financing, Organized MeSH
• Francisella tularensis isolation & purification   pathogenicity   MeSH
• Genetic Techniques utilization   MeSH
• Immune System physiology   physiopathology   MeSH
• Intracellular Space immunology   drug effects   MeSH
• Disease Attributes MeSH
• Humans MeSH
• Microbiological Phenomena genetics   immunology   drug effects   MeSH
• Molecular Biology methods   MeSH
• Tularemia history   diagnosis   etiology   MeSH
• Vaccination methods   utilization   MeSH
• Virulence genetics   immunology   drug effects   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Review MeSH

First multilocus analysis of the largest Neotropical cichlid genus Crenicichla combining mitochondrial (cytb, ND2, 16S) and nuclear (S7 intron 1) genes and comprising 602 sequences of 169 specimens yields a robust phylogenetic hypothesis. The best marker in the combined analysis is the ND2 gene which contributes throughout the whole range of hierarchical levels in the tree and shows weak effects of saturation at the 3rd codon position. The 16S locus exerts almost no influence on the inferred phylogeny. The nuclear S7 intron 1 resolves mainly deeper nodes. Crenicichla is split into two main clades: (1) Teleocichla, the Crenicichla wallacii group, and the Crenicichla lugubris-Crenicichla saxatilis groups ("the TWLuS clade"); (2) the Crenicichla reticulata group and the Crenicichla lacustris group-Crenicichla macrophthalma ("the RMLa clade"). Our study confirms the monophyly of the C. lacustris species group with very high support. The biogeographic reconstruction of the C. lacustris group using dispersal-vicariance analysis underlines the importance of ancient barriers between the middle and upper Paraná River (the Guaíra Falls) and between the middle and upper Uruguay River (the Moconá Falls). Our phylogeny recovers two endemic species flocks within the C. lacustris group, the Crenicichla missioneira species flock and the herein discovered Crenicichla mandelburgeri species flock from the Uruguay and Paraná/Iguazú Rivers, respectively. We discuss putative sympatric diversification of trophic traits (morphology of jaws and lips, dentition) and propose these species flocks as models for studying sympatric speciation in complex riverine systems. The possible role of hybridization as a mechanism of speciation is mentioned with a recorded example (Crenicichla scottii).

• MeSH
• Bayes Theorem MeSH
• Cichlids anatomy & histology   classification   genetics   MeSH
• Phylogeny MeSH
• Phylogeography MeSH
• Genetic Variation MeSH
• Multilocus Sequence Typing MeSH
• Likelihood Functions MeSH
• RNA, Ribosomal, 16S genetics   MeSH
• Fish Proteins genetics   MeSH
• Sequence Alignment MeSH
• Sympatry MeSH
• Genetic Speciation MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Paraguay MeSH
• Uruguay MeSH

The strain Escherichia coli Nissle 1917 (EcN) is widely used as an efficient probiotic in therapy and prevention of human infectious diseases, especially of the intestinal system. Concurrently, small adult pigs are being used as experimental omnivore models to study human gastrointestinal functions. EcN bacteria were applied to 6 adult healthy female pigs in a 2-week trial. 6 Control animals remained untreated. Altogether, 164 and 149 bacterial strains were isolated from smear samples taken from gastrointestinal mucosa in the experimental and control group, respectively. Each individual E. coli strain was then tested for the presence of 29 bacteriocin-encoding determinants as well as for DNA markers of A, B1, B2 and D phylogenetic groups. A profound reduction of E. coli genetic variance (from 32 variants to 13 ones, P = 0.0006) was found in the experimental group, accompanied by a lower incidence of bacteriocin producers in the experimental group when compared to control (21.3 and 34.9%, respectively; P = 0.007) and by changes in the incidence of individual bacteriocin types. The experimental administration of EcN strain was not sufficient for stable colonization of porcine gut, but induced significant changes in the enterobacterial microbiota.

• MeSH
• Genes, Bacterial MeSH
• Bacteriocins genetics   MeSH
• Escherichia coli classification   isolation & purification   MeSH
• Phylogeny MeSH
• Genetic Variation MeSH
• Molecular Typing MeSH
• Swine MeSH
• Probiotics administration & dosage   MeSH
• Biota MeSH
• Intestinal Mucosa microbiology   MeSH
• Animals MeSH
• Check Tag
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Clinical Trial MeSH
• Research Support, Non-U.S. Gov't MeSH

BACKGROUND: Infections, mostly those associated with colonization of wound by different pathogenic microorganisms, are one of the most serious health complications during a medical treatment. Therefore, this study is focused on the isolation, characterization, and identification of microorganisms prevalent in superficial wounds of patients (n=50) presenting with bacterial infection. METHODS: After successful cultivation, bacteria were processed and analyzed. Initially the identification of the strains was performed through matrix-assisted laser desorption/ionization time-of-flight mass spectrometry based on comparison of protein profiles (2-30kDa) with database. Subsequently, bacterial strains from infected wounds were identified by both matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and sequencing of 16S rRNA gene 108. RESULTS: The most prevalent species was Staphylococcus aureus (70%), and out of those 11% turned out to be methicillin-resistant (mecA positive). Identified strains were compared with patients' diagnoses using the method of artificial neuronal network to assess the association between severity of infection and wound microbiome species composition. Artificial neuronal network was subsequently used to predict patients' prognosis (n=9) with 85% success. CONCLUSIONS: In all of 50 patients tested bacterial infections were identified. Based on the proposed artificial neuronal network we were able to predict the severity of the infection and length of the treatment.

• MeSH
• Time Factors MeSH
• Adult MeSH
• Phylogeny MeSH
• Wound Infection microbiology   MeSH
• Middle Aged MeSH
• Humans MeSH
• Microbiota * MeSH
• Young Adult MeSH
• Neural Networks, Computer MeSH
• RNA, Ribosomal, 16S genetics   MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Severity of Illness Index MeSH
• Bacterial Typing Techniques methods   MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Young Adult MeSH
• Male MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

BACKGROUND: Severe Acinetobacter baumannii infections in immunocompetent patients are uncommon, and the virulence mechanisms of this organism are not fully understood. METHODS: Following an outbreak of fatal A. baumannii infections in a cohort of relatively immunocompetent patients (low comorbidity and illness severity scores), isolates were investigated with comparative genomics and in animal models. RESULTS: Two unrelated A. baumannii clades were associated with the outbreak. The clone associated with the majority of patient deaths, clade B, is evolutionarily distinct from the 3 international clonal complexes, belongs to multilocus sequence type (MLST) 10, and is most closely related to strains isolated from the Czech Republic, California, and Germany in 1994, 1997, and 2003, respectively. In 2 different murine models, clade B isolates were more virulent than comparator strains, including the highly virulent reference strain AB5075. The most virulent clade B derivative, MRSN 16897, was isolated from the patient with the lowest combined comorbidity/illness severity score. Clade B isolates possess a unique combination of putative virulence genes involved in iron metabolism, protein secretion, and glycosylation, which was leveraged to develop a rapid and specific clinical assay to detect this clade that cannot be distinguished by MLST. CONCLUSIONS: Clade B warrants continued surveillance and investigation.

• MeSH
• Acinetobacter baumannii classification   genetics   isolation & purification   pathogenicity   MeSH
• Tertiary Care Centers statistics & numerical data   MeSH
• Adult MeSH
• Disease Outbreaks * MeSH
• Phylogeny MeSH
• Genomics MeSH
• Immunocompetence MeSH
• Acinetobacter Infections epidemiology   genetics   microbiology   mortality   MeSH
• Middle Aged MeSH
• Humans MeSH
• Drug Resistance, Multiple, Bacterial * genetics   MeSH
• Disease Models, Animal MeSH
• Multilocus Sequence Typing MeSH
• Mice MeSH
• Aged, 80 and over MeSH
• Virulence genetics   MeSH
• Animals MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Mice MeSH
• Aged, 80 and over MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Geographicals
• Czech Republic MeSH
• California MeSH
• Germany MeSH
• United States MeSH

Francisella tularensis is the causative agent of tularemia. It is an intracellular pathogen with the ability to survive within phagosomes and induce pyroptotic cell death. In this study, we attempted to prove whether oxidative imbalance plays a significant role in tularemia pathogenesis. In our experimental model, we subcutaneously infected female BALB/c mice (dose 10(5) CFU of F. tularensis LVS). Liver, spleen, and blood were collected from mice at regular intervals from days 1-15 after infection. The bacterial burden was assessed by a cultivation test. The burden was unchanging from the 2(nd) to 6(th) day after infection. The bacterial burden corresponded to the plasmatic level of IFN-γ, IL-6, and liver malondialdehyde. After the phase of acute bacteraemia and the innate immunity reaction, the levels of reduced glutathione and total low molecular weight antioxidants decreased significantly and the activity of caspase-3 increased in the liver. The level of reduced glutathione decreased to 25% of the original level, and the total level of low molecular weight antioxidants was less than 50% of the initial amount. The demonstrated effects of tularemia-induced pathology had a more extensive impact on the liver than on the spleen.

• MeSH
• Antioxidants analysis   MeSH
• Bacterial Load MeSH
• Francisella tularensis pathogenicity   MeSH
• Interferon-gamma blood   MeSH
• Interleukin-6 blood   MeSH
• Liver microbiology   MeSH
• Blood microbiology   MeSH
• Malondialdehyde analysis   MeSH
• Disease Models, Animal MeSH
• Mice, Inbred BALB C MeSH
• Mice MeSH
• Oxidative Stress MeSH
• Spleen microbiology   MeSH
• Tularemia microbiology   pathology   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Analysis of actA gene sequence polymorphism has been shown to be an effective and relatively inexpensive method for subtyping Listeria monocytogenes isolates, allowing the division of the population of this species into two deeply separate lineages. This sequence-based method as well as PCR-mediated fingerprinting were applied here for the differentiation of 49 isolates of food and clinical origin. Correlation between these two typing approaches was high. Both methods divided the isolates into two lineages, designated I (33 isolates) and II (16 isolates). All the 33 lineage I isolates were assigned to the same, or closely related, six clusters by both typing methods. For the lineage II isolates, PCR fingerprinting was found to be more discriminatory. The isolates were characterized by cell invasion assay. All highly invasive isolates were assigned to lineage I, which constituted a heterogeneous group also containing low-invasive isolates. High-invasive isolates were not found in the genetically determined lineage II. A particular actA cluster, designated Ha, contained all the isolates showing the lowest invasiveness. A common trait of the isolates belonging to this cluster was the presence of a threonine-441 of the deduced ActA sequence instead of the alanine-441 present in the remaining isolates. Thirteen human isolates were classified to lineage I and five to lineage II. A PCR-based method can therefore differentiate L. monocytogenes isolates in accordance with the current phylogenetic model of the evolution of this species.

• MeSH
• Bacterial Proteins genetics   MeSH
• Cytological Techniques MeSH
• DNA Fingerprinting methods   MeSH
• Phylogeny MeSH
• Humans MeSH
• Listeria monocytogenes genetics   isolation & purification   classification   MeSH
• Listeriosis microbiology   MeSH
• Membrane Proteins genetics   MeSH
• Cell Line, Tumor MeSH
• Polymerase Chain Reaction MeSH
• Food Microbiology MeSH
• Bacterial Typing Techniques methods   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH

Burkholderia pseudomallei is the environmental bacterium that causes the serious disease melioidosis. Recently, a high prevalence of viable B. pseudomallei was reported from natural groundwater seeps around Castle Hill, a clinical focus of melioidosis in Townsville, Australia. This study sought to expand previous findings to determine the extent of B. pseudomallei in more diverse natural groundwater seeps in northern Queensland to ascertain if the presence of the organism in groundwater on Castle Hill was an isolated occurrence. Analysis of water samples (n = 26) obtained from natural groundwater seeps following an intensive rainfall event in the Townsville region determined the presence of B. pseudomallei DNA in duplicates of 18 samples (69.2 % [95 % CI, 51.5 to 87.0]). From 26 water samples, a single isolate of B. pseudomallei was recovered despite plating of both pre-enriched samples and original water samples onto selective media, indicating that the sensitivity of these molecular techniques far exceeds culture-based methods. Furthermore, the identification of new environments endemic for melioidosis may be more effectively determined by analysing surface groundwater seeps than by the analysis of random soil samples. This study suggests that a higher incidence of melioidosis following monsoonal rains may be partially the result of exposure to groundwater sources carrying B. pseudomallei, and that modifications to public health messages in endemic regions may be warranted. Moreover, these findings have implications for predictive models of melioidosis, effective models requiring consideration of topographical and surface hydrological data.

• MeSH
• Bacteriological Techniques MeSH
• Burkholderia pseudomallei genetics   isolation & purification   MeSH
• DNA, Bacterial genetics   isolation & purification   MeSH
• Weather MeSH
• Groundwater microbiology   MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Queensland MeSH