Wastewaters belong among the most important sources of environmental pollution, including antibiotic-resistant bacteria. The aim of the study was to evaluate treated wastewaters as a possible transmission pathway for bacterial colonisation of gulls occupying the receiving river. A collection of antibiotic-resistant Escherichia coli originating both from treated municipal wastewaters discharged to the river Svratka (Czech Republic) and nestlings of Black-headed Gull (Chroicocephalus ridibundus) living 35 km downstream of the outlet was obtained using selective cultivation. Isolates were further characterised by various phenotyping and genotyping methods. From a total of 670 E. coli isolates (450 from effluents, 220 from gulls), 86 isolates (41 from effluents, 45 from gulls) showed identical antibiotic resistance phenotype and genotype and were further analysed for clonal relatedness using pulsed-field gel electrophoresis (PFGE). Despite the overall high diversity of the isolates, 21 isolates from both sources showed similar PFGE profiles. Isolates belonging to epidemiologically important sequence types (ST131, 15 isolates; ST23, three isolates) were subjected to whole-genome sequencing. Subsequent phylogenetic analysis did not reveal any close clonal relationship between the isolates from the effluents and gulls' nestlings with the closest strains showing 90 SNPs difference. Although our study did not provide direct evidence of transmission of antibiotic-resistant E. coli to wild gulls via treated wastewaters, we observed gull chicks as carriers of diverse multi-resistant E. coli, including high-risk clones, posing risk of further bacterial contamination of the surrounding environment.
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Conjugative plasmids play a major role in the dissemination of antibiotic resistance genes. Knowledge of the plasmid characteristics and behaviour can allow development of control strategies. Here we focus on the IncX group of plasmids carrying genes conferring quinolone resistance (PMQR), reporting their transfer and persistence within host bacteria of various genotypes under distinct conditions and levels of induced stress in form of temperature change and various concentrations of ciprofloxacin supplementation. METHODS: Complete nucleotide sequences were determined for eight qnr-carrying IncX-type plasmids, of IncX1 (3), IncX2 (3) and a hybrid IncX1-2 (2) types, recovered from Escherichia coli of various origins. This data was compared with further complete sequences of IncX1 and IncX2 plasmids carrying qnr genes (n = 41) retrieved from GenBank and phylogenetic tree was constructed. Representatives of IncX1 (pHP2) and IncX2 (p194) and their qnrS knockout mutants, were studied for influence of induced stress and genetic background on conjugative transfer and maintenance. RESULTS: A high level of IncX core-genome similarity was found in plasmids of animal, environmental and clinical origin. Significant differences were found between the individual IncX plasmids, with IncX1 subgroup plasmids showing higher conjugative transfer rates than IncX2 plasmids. Knockout of qnr modified transfer frequency of both plasmids. Two stresses applied simultaneously were needed to affect transfer rate of wildtype plasmids, whereas a single stress was sufficient to affect the IncX ΔqnrS plasmids. The conjugative transfer was shown to be biased towards the host phylogenetic proximity. A long-term cultivation experiment pointed out the persistence of IncX plasmids in the antibiotic-free environment. CONCLUSIONS: The study indicated the stimulating effect of ciprofloxacin supplementation on the plasmid transfer that can be nullified by the carriage of a single PMQR gene. The findings present the significant properties and behaviour of IncX plasmids carrying antibiotic resistance genes that are likely to play a role in their dissemination and stability in bacterial populations.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence genetika MeSH
- ciprofloxacin farmakologie MeSH
- Escherichia coli * genetika MeSH
- fylogeneze MeSH
- genomika MeSH
- konjugace genetická MeSH
- plazmidy genetika MeSH
- proteiny z Escherichia coli * genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The Philopterus Complex includes several lineages of lice that occur on birds. The complex includes the genera Philopterus (Nitzsch, 1818; Psocodea: Philopteridae), Philopteroides (Mey, 2004; Psocodea: Philopteridae), and many other lineages that have sometimes been regarded as separate genera. Only a few studies have investigated the phylogeny of this complex, all of which are based on morphological data. Here we evaluate the utility of nuclear and mitochondrial loci for recovering the phylogeny within this group. We obtained phylogenetic trees from 39 samples of the Philopterus Complex (Psocodea: Philopteridae), using sequences of two nuclear (hyp and TMEDE6) and one mitochondrial (COI) marker. We evaluated trees derived from these genes individually as well as from concatenated sequences. All trees show 20 clearly demarcated taxa (i.e., putative species) divided into five well-supported clades. Percent sequence divergence between putative species (~5-30%) for the COI gene tended to be much higher than those for the nuclear genes (~1-15%), as expected. In cases where species are described, the lineages identified based on molecular divergence correspond to morphologically defined species. In some cases, species that are host generalists exhibit additional underlying genetic variation and such cases need to be explored by further future taxonomic revisions of the Philopterus Complex.
- MeSH
- buněčné jádro MeSH
- fylogeneze * MeSH
- genetické markery MeSH
- hmyzí proteiny analýza MeSH
- Ischnocera klasifikace genetika MeSH
- mitochondriální proteiny analýza MeSH
- respirační komplex IV analýza MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Publikační typ
- abstrakt z konference MeSH
This study evaluated the carriage of AmpC and extended-spectrum beta-lactamase (ESBL) genes and associated plasmids in faecal bacteria of Canadian corvids. Faecal samples from 449 birds in five roosting sites across Canada were analyzed using selective media, screening for AmpC and ESBL genes by PCR, and sequencing. Genomic relatedness was determined by PFGE and MLST. Plasmid mobility was studied by conjugation and transformation experiments, followed by plasmid typing. In total, 96 (21%, n = 449) cefotaxime-resistant Escherichia coli and three (0.7%) Klebsiella pneumoniae isolates were identified. ESBL genes blaCTX-M-1 (n = 3), blaCTX-M-14 (n = 2), blaCTX-M-32 (n = 2) and blaCTX-M-124 (n = 1) were detected in eight E. coli isolates, whereas blaSHV-2 (2) was found in two K. pneumoniae. E. coli isolates contained blaCMY-2 (n = 83) and blaCMY-42 (n = 1). The high genetic diversity of the isolates and presence of clinically important E. coli ST69 (n = 1), ST117 (n = 7) and ST131 (n = 1) was revealed. AmpC genes were predominantly carried by plasmids of incompatibility groups I1 (45 plasmids), A/C (10) and K (7). The plasmid IncI1/ST12 was most common and found in diverse E. coli STs in all sites. Highly diverse E. coli isolates containing AmpC and ESBL genes, including clinically important clones and emerging plasmids, are in circulation throughout Canadian wildlife.
- MeSH
- bakteriální proteiny genetika MeSH
- beta-laktamasy genetika MeSH
- Escherichia coli klasifikace enzymologie genetika izolace a purifikace MeSH
- feces mikrobiologie MeSH
- Klebsiella pneumoniae enzymologie genetika izolace a purifikace MeSH
- multilokusová sekvenční typizace MeSH
- plazmidy genetika MeSH
- vrány mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Kanada MeSH
Escherichia coli sequence type 131 (ST131) is currently one of the leading causes of multidrug-resistant extraintestinal infections globally. Here, we analyzed the phenotypic and genotypic characteristics of 169 ST131 isolates from various sources (wildlife, wastewater, companion animals, community, and hospitals) to determine whether wildlife and the environment share similar strains with humans, supporting transmission of ST131 between different ecological niches. Susceptibility to 32 antimicrobials was tested by disc diffusion and broth microdilution. Antibiotic resistance genes, integrons, plasmid replicons, 52 virulence genes, and fimH-based subtypes were detected by PCR and DNA sequencing. Genomic relatedness was determined by pulsed-field gel electrophoresis (PFGE). The genetic context and plasmid versus chromosomal location of extended-spectrum beta-lactamase and AmpC beta-lactamase genes was determined by PCR and probe hybridization, respectively. The 169 ST131 study isolates segregated predominantly into blaCTX-M-15H30Rx (60%) and blaCTX-M-27H30R1 (25%) subclones. Within each subclone, isolates from different source groups were categorized into distinct PFGE clusters; genotypic characteristics were fairly well conserved within each major PFGE cluster. Irrespective of source, the blaCTX-M-15H30Rx isolates typically exhibited virotype A (89%), an F2:A1:B- replicon (84%), and a 1.7-kb class 1 integron (92%) and had diverse structures upstream of the blaCTX-M region. In contrast, the blaCTX-M-27H30R1 isolates typically exhibited virotype C (86%), an F1:A2:B20 replicon (76%), and a conserved IS26-ΔISEcp1-blaCTX-M-like structure. Despite considerable overall genetic diversity, our data demonstrate significant commonality between E. coli ST131 isolates from diverse environments, supporting transmission between different sources, including humans, environment, and wildlife.
- MeSH
- antibakteriální látky farmakologie MeSH
- beta-laktamasy genetika MeSH
- Escherichia coli účinky léků genetika MeSH
- infekce vyvolané Escherichia coli genetika mikrobiologie MeSH
- infekce získané v komunitě genetika mikrobiologie MeSH
- lidé MeSH
- mnohočetná bakteriální léková rezistence genetika MeSH
- plazmidy genetika MeSH
- proteiny z Escherichia coli genetika MeSH
- pulzní gelová elektroforéza MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
Twenty-four species of chewing lice of the genus Myrsidea Waterston, 1915 (Phthiraptera: Menoponidae) from Neotropical Suboscines (Passeriformes: Formicariidae, Furnariidae, Pipridae, Thamnophilidae, Tityridae, Tyrannidae) are recorded and discussed. They include: eight new species which are described and illustrated (Myrsidea capeki new species ex Chiroxiphia caudata; Myrsidea leptopogoni new species ex Leptopogon superciliaris; Myrsidea leucophthalmi new species ex Automolus leucophthalmus; Myrsidea pachyramphi new species ex Pachyramphus polychopterus; Myrsidea philydori new species ex Philydor rufum; Myrsidea pyriglenae new species ex Pyriglena leucoptera; Myrsidea scleruri new species ex Sclerurus scansor and Myrsidea zuzanae new species ex Furnarius rufus), as well as nine previously known species with additional data on intraspecific morphological variability, host associations and geographical distribution (Myrsidea barbati Price, Hellenthal Dalgleish, 2005; Myrsidea dalgleishi Valim, Price Johnson, 2011; Myrsidea flaviventris Price, Hellenthal Dalgleish, 2005; Myrsidea klimesi Sychra, 2006; Myrsidea meyi Valim, Price Johnson, 2011; Myrsidea oleaginei Price, Hellenthal Dalgleish, 2005; Myrsidea olivacei Price, Hellenthal Dalgleish, 2005; Myrsidea pitangi Price, Hellenthal Dalgleish, 2005 and Myrsidea spellmani Price, Johnson Dalgleish, 2008b). Seven further species are recorded at genus level only due to lack of adequate material. A 379 bp portion of the mitochondrial cytochrome oxidase I (COI) gene was sequenced from seven species in order to assess relative genetic divergences among Myrsidea populations.
- MeSH
- Amblycera MeSH
- Ischnocera * MeSH
- nemoci ptáků MeSH
- Passeriformes * MeSH
- Phthiraptera MeSH
- zavšivení MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Publikační typ
- abstrakt z konference MeSH
Increasing bacterial resistance to quinolone antibiotics is apparent in both humans and animals. For humans, a potential source of resistant bacteria may be animals or their products entering the human food chain, for example poultry. Between July 2013 and September 2014, samples were collected and analyzed in the Moravian regions of the Czech Republic to isolate the bacterium Escherichia coli. As a result, 212 E. coli isolates were obtained comprising 126 environmental isolates from poultry houses and 86 isolates from cloacal swabs from market-weight turkeys. Subsequently, the E. coli isolates were tested for susceptibility to selected antibiotics. Resistance of the poultry isolates to quinolones ranged from 53% to 73%. Additionally, the presence of plasmid-mediated resistance genes was studied. The genes were confirmed in 58% of the tested strains. The data on resistance of isolates from poultry were compared with results of resistance tests in human isolates obtained in the same regions. The high levels of resistance determined by both phenotyping and genotyping methods and reported in the present study confirm the fact that the use of fluoroquinolones in poultry should be closely monitored.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence MeSH
- chinolony farmakologie MeSH
- drůbež mikrobiologie MeSH
- Escherichia coli účinky léků izolace a purifikace MeSH
- zemědělství MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
The distribution, variability and host specificity of species of Babesia Starcovici, 1893 were studied in questing ticks collected on the northwestern edge of the Pannonian Basin in the south-easternmost part of the Czech Republic and in western Slovakia. The area is characterised by relatively natural floodplain habitats and the sympatric occurrence of three tick species possessing wide host spectra, namely Ixodes ricinus (Linnaeus), Dermacentor reticulatus (Fabricius) and Haemaphysalis concinna Koch. Analysis was carried out on 1,408 I. ricinus, 2,999 D. reticulatus and 150 H. concinna altogether, collected from 59 localities. We documented the presence of Babesia spp. not only in I. ricinus but also in H. concinna in the Czech Republic. Two isolates from I. ricinus were classified as B. venatorum Herwaldt, Cacciò, Gherlinzoni, Aspöck, Slemenda, Piccaluga, Martinelli, Edelhofer, Hollenstein, Poletti, Pampiglione, Löschenberger, Tura et Pieniazek, 2003 (formerly determined as Babesia sp. EU1), which is a zoonotic parasite and can cause human babesiosis. The rest of our amplicons were very similar to B. canis (Piana et Galli-Valerio, 1895), which is usually transmitted by D. reticulatus. Despite the huge amount of examined samples, all D. reticulatus ticks were Babesia-free. Due to this finding, we did not consider our obtained isolates to be B. canis, but other closely related species possessing a similar sequence of the studied portion of 18S rDNA. Although this genetic marker is most frequently used in PCR-based diagnostic methods of babesias, its low variability compromises its reliability in studies based only on this marker.
- MeSH
- arachnida jako vektory parazitologie MeSH
- Babesia klasifikace genetika izolace a purifikace MeSH
- babezióza parazitologie přenos MeSH
- Dermacentor parazitologie MeSH
- hostitelská specificita MeSH
- infestace klíšťaty parazitologie MeSH
- Ixodidae parazitologie MeSH
- klíště parazitologie MeSH
- lidé MeSH
- RNA ribozomální 18S genetika MeSH
- zoonózy MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Slovenská republika MeSH
