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American crow ( Corvus brachyrhynchos) faeces were tested for Escherichia coli with plasmid-mediated quinolone resistance (PMQR), extended-spectrum beta-lactamases (ESBL) and AmpC beta-lactamases. A total of 590 faecal samples were collected at four roosting sites in the USA and cultivated on selective media. Pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST) were performed to assess clonality. Transferability of resistance genes was studied using conjugation and transformation bioassays. In total, 78 (13%, n = 590) cefotaxime-resistant isolates were obtained, of which 66 and 12 displayed AmpC and ESBL phenotypes, respectively. Fifty-four AmpC-producing isolates carried blaCMY-2. Isolates producing ESBLs contained genes blaCTX-M-27 (5 isolates), blaCTX-M-15 (4), blaCTX-M-14 (2) and blaCTX-M-1 (1). Ninety isolates (15%, n = 590) with reduced susceptibility to ciprofloxacin were obtained, among which 14 harboured PMQR genes aac(6′)-Ib-cr (4 isolates), qnrB19 (3), qnrS1 (2), qnrA1 (2), qnrB2 (1), qnrB6 (1) and qnrD3 (1). High genetic diversity was revealed by PFGE and MLST. Epidemiologically important E. coli clones (e.g., ST131, ST405) were identified. Plasmids carrying blaCMY-2 were assigned predominantly to IncA/C (8 plasmids), IncI1/ST23 (5) and IncI1/ST12 (3). The study demonstrates a widespread occurrence of E. coli with ESBL, AmpC and PMQR genes associated with clinically important multidrug-resistant clones and epidemic plasmids, in American crows. [ABSTRACT FROM AUTHOR]

Escherichia coli is a common commensal bacterial species of humans and animals that may become a troublesome pathogen causing serious diseases. The aim of this study was to characterize the quinolone resistance phenotypes and genotypes in E. coli isolates of different origin from one area of the Czech Republic. E. coli isolates were obtained from hospitalized patients and outpatients, chicken farms, retailed turkeys, rooks wintering in the area, and wastewaters. Susceptibility of the isolates grown on the MacConkey agar with ciprofloxacin (0.05 mg/L) to 23 antimicrobial agents was determined. The presence of plasmid-mediated quinolone resistance (PMQR) and ESBL genes was tested by PCR and sequencing. Specific mutations in gyrA, gyrB, parC, and parE were also examined. Multilocus sequence typing and pulsed-field gel electrophoresis were performed to assess the clonal relationship. In total, 1050 E. coli isolates were obtained, including 303 isolates from humans, 156 from chickens, 105 from turkeys, 114 from the rooks, and 372 from wastewater samples. PMQR genes were detected in 262 (25%) isolates. The highest occurrence was observed in isolates from retailed turkey (49% of the isolates were positive) and inpatients (32%). The qnrS1 gene was the most common PMQR determinant identified in 146 (56%) followed by aac(6')-Ib-cr in 77 (29%), qnrB19 in 41 (16%), and qnrB1 in 9 (3%) isolates. All isolates with high level of ciprofloxacin resistance (>32 mg/L) carried double or triple mutations in gyrA combined with single or double mutations in parC. The most frequently identified substitutions were Ser(83)Leu; Asp(87)Asn in GyrA, together with Ser(80)Ile, or Glu(84)Val in ParC. Majority of these isolates showed resistance to beta-lactams and multiresistance phenotype was found in 95% isolates. Forty-eight different sequence types among 144 isolates analyzed were found, including five major clones ST131 (26), ST355 (19), ST48 (13), ST95 (10), and ST10 (5). No isolates sharing 100% relatedness and originating from different areas were identified. In conclusion, our study identified PMQR genes in E. coli isolates in all areas studied, including highly virulent multiresistant clones such as ST131 producing CTX-M-15 beta-lactamases. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance genes can be found in all ecosystems, including those where antibiotic selective pressure has never been exerted. We investigated resistance genes in a collection of faecal samples of wildlife (non-human primates, mice), people and domestic animals (dogs, cats) in Côte d’Ivoire; in the chimpanzee research area of Taï National Park (TNP) and adjacent villages. Single bacteria isolates were collected from antibiotic-containing agar plates and subjected to molecular analysis to detect Enterobacteriaceae isolates with plasmid-mediated genes of extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated quinolone resistance (PMQR). While the prevalence of ESBL-producing E. coli in the villages was 27% in people (n = 77) and 32% in dogs (n = 38), no ESBL-producer was found in wildlife of TNP (n = 75). PMQR genes, mainly represented by qnrS1, were also present in human- and dog-originating isolates from the villages (36% and 42% in people and dogs, respectively), but no qnrS has been found in the park. In TNP, different variants of qnrB were detected in Citrobacter freundii isolates originating non-human primates and mice. In conclusion, ESBL and PMQR genes frequently found in humans and domestic animals in the villages were rather exceptional in wildlife living in the protected area. Although people enter the park, the strict biosecurity levels they are obliged to follow probably impede transmission of bacteria between them and wildlife. [ABSTRACT FROM AUTHOR]

Commensal Escherichia coli isolates from healthy zoo animals kept in Ostrava Zoological Garden, Czech Republic, were investigated to evaluate the dissemination of extended-spectrum beta-lactamase ( ESBL) and plasmid-mediated quinolone resistance ( PMQR) genes. A total of 160 faecal samples of various animal species were inoculated onto Mac Conkey agar with cefotaxime (2 mg L−1) or ciprofloxacin (0.05 mg L−1) to obtain ESBL- or PMQR-positive E. coli isolates. Clonality of E. coli isolates was investigated by multilocus sequence typing and pulsed-field gel electrophoresis. Plasmids carrying ESBL or PMQR genes were typed by PCR-based replicon typing, plasmid multilocus sequence typing and restriction fragment length polymorphism. Forty-nine (71%, n = 69) cefotaxime-resistant and 15 (16%, n = 94) ciprofloxacin-resistant E. coli isolates harboured ESBL or PMQR genes. Isolates were assigned to 18 sequence types ( ST) and 20 clusters according to their macrorestriction patterns by pulsed-field gel electrophoresis. The genes bla CTX-M-1 and qnrS1 were detected on highly related IncI1 plasmids assigned to clonal complex 3 ( ST3, ST38) and on non-related IncN plasmids of ST1 and ST3, respectively. The gene qnrS1 was located on related IncX1 plasmids. Dissemination of antibiotic resistance is associated with spreading of particular E. coli clones and plasmids of specific incompatibility groups among various animal species. [ABSTRACT FROM AUTHOR]

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 bla CTX-M-1 (n = 3), bla CTX-M-14 (n = 2), bla CTX-M-32 (n = 2) and bla CTX-M-124 (n = 1) were detected in eight E. coli isolates, whereas bla SHV-2 (2) was found in two K. pneumoniae. E. coli isolates contained bla CMY-2 (n = 83) and bla CMY-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. [ABSTRACT FROM AUTHOR]

Extended-spectrum-beta-lactamase (ESBL)-producing, AmpC beta-lactamase-producing, and plasmid-mediated quinolone resistance (PMQR) gene-positive strains of Escherichia coli were investigated in wintering rooks (Corvus frugilegus) from eight European countries. Fecal samples (n = 1,073) from rooks wintering in the Czech Republic, France, Germany, Italy, Poland, Serbia, Spain, and Switzerland were examined. Resistant isolates obtained from selective cultivation were screened for ESBL, AmpC, and PMQR genes by PCR and sequencing. Pulsed-field gel electrophoresis and multilocus sequence typing were performed to reveal their clonal relatedness. In total, from the 1,073 samples, 152 (14%) cefotaxime-resistant E. coli isolates and 355 (33%) E. coli isolates with reduced susceptibility to ciprofloxacin were found. Eighty-two (54%) of these cefotaxime-resistant E. coli isolates carried the following ESBL genes: blaCTX-M-1 (n = 39 isolates), blaCTX-M-15 (n = 25), blaCTX-M-24 (n = 4), blaTEM-52 (n = 4), blaCTX-M-14 (n = 2), blaCTX-M-55 (n = 2), blaSHV-12 (n = 2), blaCTX-M-8 (n = 1), blaCTX-M-25 (n = 1), blaCTX-M-28 (n = 1), and an unspecified gene (n = 1). Forty-seven (31%) cefotaxime-resistant E. coli isolates carried the blaCMY-2 AmpC beta-lactamase gene. Sixty-two (17%) of the E. coli isolates with reduced susceptibility to ciprofloxacin were positive for the PMQR genes qnrS1 (n = 54), qnrB19 (n = 4), qnrS1 and qnrB19 (n = 2), qnrS2 (n = 1), and aac(6')-Ib-cr (n = 1). Eleven isolates from the Czech Republic (n = 8) and Serbia (n = 3) were identified to be CTX-M-15-producing E. coli clone B2-O25b-ST131 isolates. Ninety-one different sequence types (STs) among 191 ESBL-producing, AmpC-producing, and PMQR gene-positive E. coli isolates were determined, with ST58 (n = 15), ST10 (n = 14), and ST131 (n = 12) predominating. The widespread occurrence of highly diverse ESBL- and AmpC-producing and PMQR gene-positive E. coli isolates, including the clinically important multiresistant ST69, ST95, ST117, ST131, and ST405 clones, was demonstrated in rooks wintering in various European countries. [ABSTRACT FROM AUTHOR]

Abstract: Antimicrobial resistance is a worldwide concern of public health. Unfortunately, resistant bacteria are spreading to all ecosystems, including the strictly protected ones. We investigated antimicrobial resistance in gastrointestinal Enterobacteriaceae of wild mammals and people living within Dzangha-Sangha Protected Areas, Central African Republic, with an emphasis on extended-spectrum β-lactamase (ESBL) and plasmid-mediated quinolone resistance (PMQR) genes. We compare resistance genes found in microbiota of humans, gorillas habituated and unhabituated to humans and other wildlife. In gorillas, we additionally investigate the presence of ESBL resistant isolates after treatment by ceftiofur. We found a considerable prevalence of multiresistant Enterobacteriaceae isolates with ESBL and PMQR genes in humans (10% and 31%, respectively). Among wildlife the most significant findings were CTX-M-15-producing Klebsiella pneumoniae in a habituated gorilla and a multiresistant Escherichia coli isolate with gene qepA in an unhabituated gorilla. Other isolates from wildlife were mostly represented by qnrB-harboring Citrobacter spp. The relatedness of resistant E. coli was investigated in a PFGE-based dendrogram; isolates from gorillas showed less than 80% similarity to each other and less than 80% similarity to human isolates. No ESBL-producing isolates were found in animals treated by ceftiofur. Although we did not detect any bacterial clone common to wildlife and humans, we detected an intersection in the spectrum of resistance genes found in humans and gorillas, represented by bla CTX-M-15 and qepA. [Copyright &y& Elsevier]

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., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

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 bla CTX-M-15 H 30Rx (60%) and bla CTX-M-27 H 30R1 (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 bla CTX-M-15 H 30Rx 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 bla CTX-M region. In contrast, the bla CTX-M-27 H 30R1 isolates typically exhibited virotype C (86%), an F1:A2:B20 replicon (76%), and a conserved IS 26 -ΔIS Ecp1-bla CTX-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., (Copyright © 2018 American Society for Microbiology.)

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.

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.

Two multidrug resistance (MDR) plasmids, carrying the VIM-1-encoding integron In110, were characterized. Plasmid pLec-476cz (311,758 bp), from a Leclercia adecarboxylata isolate, consisted of an IncHI1 backbone, a MDR region, and two accessory elements. Plasmid pKpn-431cz (142,876 bp), from a sequence type 323 (ST323) Klebsiella pneumoniae isolate, comprised IncFII Y -derived and pKPN3-like sequences and a mosaic region. A 40,400-bp sequence of pKpn-431cz was identical to the MDR region of pLec-476cz, indicating the en bloc acquisition of the VIM-1-encoding region from one plasmid by the other., (Copyright © 2017 American Society for Microbiology.)

A total of 239 individuals of 50 bird species were examined for chewing lice (Insecta: Phthiraptera) in southern Vietnam. Fifty-six birds of 20 species were parasitised by 15 species of lice belonging to 10 genera from two suborders, Amblycera: Menacanthus, Meromenopon, Myrsidea, and Ischnocera: Alcedoecus, Brueelia, Cuculicola, Meropoecus, Penenirmus, Philopteroides and Philopterus. Thirteen louse samples from Passeriformes were identified to genus only because they contain inadequate material. A total of 29 host-louse associations were found, of which nine are new, including: (1) two new species of the genus Brueelia, which are described and named in this paper: Brueelia binhchauensis from Megalaima lineata (Vieillot, 1816) (Piciformes: Megalaimidae), and Brueelia malacocincla from Malacocincla abbotti Blyth, 1845 (Passeriformes: Pellorneidae); (2) first records of lice from Cyornis hainanus (Ogilvie-Grant, 1900); and (3) the first record of Myrsidea claytoni Hellenthal & Price, 2003 from Cymbirhynchus macrorhynchos (Gmelin, 1788) (Passeriformes: Eurylaimidae), here regarded as a case of natural host-switching. A portion of the mitochondrial cytochrome oxidase I (COI) gene for some species of chewing lice was sequenced in order to assess their genetic divergences.

This study concerned the occurrence of fecal bacteria with plasmid-mediated quinolone resistance (PMQR) genes in rooks (Corvus frugilegus, medium-sized corvid birds) wintering in continental Europe during winter 2010/2011. Samples of fresh rook feces were taken by cotton swabs at nine roosting places in eight European countries. Samples were transported to one laboratory and placed in buffered peptone water (BPW). The samples from BPW were enriched and subcultivated onto MacConkey agar (MCA) supplemented with ciprofloxacin (0.06 mg/L) to isolate fluoroquinolone-resistant bacteria. DNA was isolated from smears of bacterial colonies growing on MCA and tested by PCR for PMQR genes aac(6')-Ib, qepA, qnrA, qnrB, qnrC, qnrD, qnrS, and oqxAB. All the PCR products were further analyzed by sequencing. Ciprofloxacin-resistant bacteria were isolated from 37% (392 positive/1,073 examined) of samples. Frequencies of samples with ciprofloxacin-resistant isolates ranged significantly from 3% to 92% in different countries. The qnrS1 gene was found in 154 samples and qnrS2 in 2 samples. The gene aac(6')-Ib-cr was found in 16 samples. Thirteen samples were positive for qnrB genes in variants qnrB6 (one sample), qnrB18 (one), qnrB19 (one), qnrB29 (one), and qnrB49 (new variant) (one). Both the qnrD and oqxAB genes were detected in six samples. The genes qnrA, qnrC, and qepA were not found. Wintering omnivorous rooks in Europe were commonly colonized by bacteria supposedly Enterobacteriaceae with PMQR genes. Rooks may disseminate these epidemiologically important bacteria over long distances and pose a risk for environmental contamination.