Chewing lice of Bearded Reedling (Panurus biarmicus) and diversity of louse-host associations of birds in reed beds in Slovakia.

A total of 1,621 wild birds representing 34 species were examined for chewing lice in reed beds in southwestern Slovakia during the pre-breeding migration 2008-2009 and 2016-2019. A total of 377 (23.3%) birds representing 15 species were parasitized by 26 species of chewing lice of 12 genera. Dominant genera were Penenirmus (with dominance 32.6%) and Menacanthus (29.4%), followed by Brueelia (12.6%), Acronirmus (10.8%), Philopterus (7.7%), and Myrsidea (4.2%). We evaluated 33 host-louse associations including both 1) host-generalist, parasitizing more than one host species and host-specific lice, occurring only on a single host species, and 2) lice species with large range geographic distribution, reported across the range of the distribution of their hosts and lice species with only occasional records from a limited area within the range of their hosts. The Bearded Reedling, Panurus biarmicus (Linnaeus, 1758), was parasitized by two species of chewing lice, Menacanthus brelihi Balát, 1981 and Penenirmus visendus (Zlotorzycka, 1964), with conspicuously different prevalences (5.6% vs. 58.2%, respectively; n = 251). New material enabled us to redescribe both species of lice: the first one is resurrected from previous synonymy as a valid species. A fragment of the mitochondrial cytochrome oxidase I gene was sequenced from these two species in order to assess their relative phylogenetic position within their genera. Our study demonstrates the importance of an adequate identification of parasites, especially on rarely examined and endangered hosts.

Title: Mallophages de la Panure à moustaches (Panurus biarmicus) et diversité des associations mallophages-hôtes des oiseaux dans les roselières en Slovaquie. Abstract: Au total, 1 621 oiseaux sauvages représentant 34 espèces ont été examinés à la recherche de mallophages dans les roselières du sud-ouest de la Slovaquie au cours de la migration de pré-reproduction 2008­2009 et 2016­2019. Parmi ceux-ci, 377 oiseaux (23,3 %), représentant 15 espèces, étaient parasités par 26 espèces de mallophages de 12 genres. Les genres dominants étaient Penenirmus (avec une dominance de 32,6 %) et Menacanthus (29,4 %), suivis de Brueelia (12,6 %), Acronirmus (10,8 %), Philopterus (7,7 %) et Myrsidea (4,2 %). Nous avons évalué 33 associations mallophage-hôte comprenant à la fois 1) des espèces de mallophages généralistes, parasitant plus d'une espèce hôte, et des mallophages spécifiques, présents uniquement sur une seule espèce hôte et 2) des espèces de mallophages ayant une large répartition géographique, signalées à travers l'étendue de la répartition de leurs hôtes, et des espèces de mallophages avec seulement des observations occasionnelles dans une zone limitée à l'intérieur de l'aire de répartition de leurs hôtes. La Panure à moustaches, Panurus biarmicus (Linnaeus, 1758), était parasitée par deux espèces de mallophages, Menacanthus brelihi Balát, 1981 et Penenirmus visendus (Zlotorzycka, 1964), avec des prévalences nettement différentes (respectivement 5,6 % et 58,2 %, n = 251). Du nouveau matériel nous a permis de redécrire les deux espèces de mallophages, la première étant ressuscitée de la synonymie précédente en tant qu'espèce valide. Un fragment du gène mitochondrial de la cytochrome oxydase I a été séquencé à partir de ces deux espèces afin d'évaluer leur position phylogénétique relative au sein de leurs genres. Notre étude démontre l'importance d'une identification adéquate des parasites, en particulier sur les hôtes rarement examinés et menacés.

Chewing lice of passerine birds in reed beds in Slovakia, with a special focus on Panurus biarmicus.

A total of 1185 passerine birds representing five species were examined for chewing lice in reed beds in southwestern Slovakia in spring (April) 2008, 2009 and 2016. Additional collecting focused only on chewing lice from Panurus biarmicus (Linnaeus, 1758) (Passeriformes: Panuridae) was carried out in spring (April), summer (July) and autumn (October) 2019. A total of 283 (24%) birds were parasitized by 10 species of chewing lice of four genera: Penenirmus, Menacanthus, Philopterus, and Brueelia. Most birds showed only very light (1-10 lice/host; 74%) to light infestations (11-20 lice/host; 16%). The authors found significantly higher prevalences and mean abundances of chewing lice on residents/short-distance migrants, that is, P. biarmicus, Acrocephalus melanopogon (Temminck, 1823) (Passeriformes: Acrocephalidae), than on long-distance migratory birds, that is, Acrocephalus scirpaceus (Hermann, 1804), Acrocephalus schoenobaenus (Linnaeus, 1758) (Passeriformes: Acrocephalidae), Locustella luscinioides (Savi, 1824) (Passeriformes: Locustellidae). No significant difference was found in the total mean intensity of chewing lice between these two groups of birds. Ischnoceran lice were more prevalent and abundant than amblyceran lice on residents and short-distance migrants, whereas the opposite was found on bird species that migrate long distances. A total of 146 (58%, n = 251) P. biarmicus were parasitized by 1490 chewing lice. Males of P. biarmicus showed higher prevalence and mean abundance than females with gradually descending values of prevalence, mean abundance and mean intensity from spring to autumn. The knowledge of the occurrence and population dynamics of lice on wild passerine birds can be useful in endangered species conservation programs and can also be applied to captive passerine birds, which may be analogous to resident birds in this sense.

Dermacentor reticulatus - a tick on its way from glacial refugia to a panmictic Eurasian population.

The ornate dog tick (Dermacentor reticulatus) shows a recently expanding geographic distribution. Knowledge on its intraspecific variability, population structure, rate of genetic diversity and divergence, including its evolution and geographic distribution, is crucial to understand its dispersal capacity. All such information would help to evaluate the potential risk of future spread of associated pathogens of medical and veterinary concern. A set of 865 D. reticulatus ticks was collected from 65 localities across 21 countries, from Portugal in the west to Kazakhstan and southern Russia in the east. Cluster analyses of 16 microsatellite loci were combined with nuclear (ITS2, 18S) and mitochondrial (12S, 16S, COI) sequence data to uncover the ticks' population structures and geographical patterns. Approximate Bayesian computation was applied to model evolutionary relationships among the found clusters. Low variability and a weak phylogenetic signal showing an east-west cline were detected both for mitochondrial and nuclear sequence markers. Microsatellite analyses revealed three genetic clusters, where the eastern and western cluster gradient was supplemented by a third, northern cluster. Alternative scenarios could explain such a tripartite population structure by independent formation of clusters in separate refugia, limited gene flow connected with isolation by distance causing a "bipolar pattern", and the northern cluster deriving from admixture between the eastern and western populations. The best supported demographic scenario of this tick species indicates that the northern cluster derived from admixture between the eastern and western populations 441 (median) to 224 (mode) generations ago, suggesting a possible link with the end of the Little Ice Age in Europe.

Genomic analysis of qnr-harbouring IncX plasmids and their transferability within different hosts under induced stress.

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.

Combining Nuclear and Mitochondrial Loci Provides Phylogenetic Information in the Philopterus Complex of Lice (Psocodea: Ischnocera: Philopteridae).

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.

New species and additional data on the chewing louse genus Myrsidea (Phthiraptera: Menoponidae) from wild Neotropical Passeriformes (Aves).

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.

Extensive Genetic Commonality among Wildlife, Wastewater, Community, and Nosocomial Isolates of Escherichia coli Sequence Type 131 (H30R1 and H30Rx Subclones) That Carry blaCTX-M-27 or blaCTX-M-15.

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.

Molecular characterization of plasmid-mediated AmpC beta-lactamase- and extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae among corvids (Corvus brachyrhynchos and Corvus corax) roosting in Canada.

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.

Variability of species of Babesia Starcovici, 1893 in three sympatric ticks (Ixodes ricinus, Dermacentor reticulatus and Haemaphysalis concinna) at the edge of Pannonia in the Czech Republic and Slovakia.

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.

Quinolone-resistant Escherichia coli in Poultry Farming.

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.

Plasmid-mediated resistance to cephalosporins and quinolones in Escherichia coli from American crows in the USA.

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.

Complete Nucleotide Sequences of Two VIM-1-Encoding Plasmids from Klebsiella pneumoniae and Leclercia adecarboxylata Isolates of Czech Origin.

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 IncFIIY-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.

'Candidatus Rickettsia mendelii', a novel basal group rickettsia detected in Ixodes ricinus ticks in the Czech Republic.

A novel rickettsial sequence in the citrate synthase gltA gene indicating a novel Rickettsia species has been detected in 7 out of 4524 Ixodes ricinus ticks examined within several surveys performed in the Czech Republic from 2005 to 2009. This new Candidatus Rickettsia sp. sequence has been found in 2 nymphs feeding on wild birds (Luscinia megarhynchos and Erithacus rubecula), in a male tick from vegetation, and 4 ticks feeding on a dog (3 males, 1 female tick). Portions of the ompA, ompB, sca4, and htrA genes were not amplifiable in these samples. A maximum likelihood tree of rickettsiae based on comparisons of partial amino acid sequences of citrate synthase and nucleotide sequences of 16S rDNA genes and phylogenetic analysis revealed a basal position of the novel species in the proximity of R. bellii and R. canadensis. The novel species has been named 'Candidatus Rickettsia mendelii' after the founder of genetics, Gregor Mendel.

Characteristics of Quinolone Resistance in Escherichia coli Isolates from Humans, Animals, and the Environment in the Czech Republic.

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.

Spotted fever group rickettsiae detected in immature stages of ticks parasitizing on Iberian endemic lizard Lacerta schreiberi Bedriaga, 1878.

Spotted fever rickettsioses are tick-borne diseases of growing public health concern. The prevalence of rickettsia-infected ticks and their ability to parasitize humans significantly influence the risk of human infection. Altogether 466 Ixodes ricinus ticks (428 nymphs and 38 larvae) collected from 73 Lacerta schreiberi lizards were examined by PCR targeting the citrate synthetase gene gltA for the presence of Rickettsia spp. Rickettsial DNA was detected in 47% of nymphs and 31.6% of larvae. They were subsequently subjected to a second PCR reaction using primers derived from the outer membrane protein rOmpA encoding gene (ompA) to detect spotted fever group rickettsiae (SFG). This analysis shows that 41.4% of nymphs and 7.9% of larvae collected from the lizards contain DNA of SFG rickettsiae. Sequencing of 43 randomly selected samples revealed two different haplotypes, both closely related to R. monacensis (39 and 4 samples, respectively). The remaining ompA negative Rickettsia spp. samples were determined to be R. helvetica based on sequencing of ompB and gltA fragments. Our results indicate that the role of Iberian endemic lizard L. schreiberi and its ectoparasites in the ecology and epidemiology of zoonotic SFG rickettsioses may be appreciable.

Plasmid-mediated resistance to cephalosporins and fluoroquinolones in various Escherichia coli sequence types isolated from rooks wintering in Europe.

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.

Low rates of antimicrobial-resistant Enterobacteriaceae in wildlife in Taï National Park, Côte d'Ivoire, surrounded by villages with high prevalence of multiresistant ESBL-producing Escherichia coli in people and domestic animals.

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.

Chewing lice (Phthiraptera: Amblycera and Ischnocera) from wild birds in southern Vietnam, with descriptions of two new species.

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.

Hidden threat of tortoise ticks: high prevalence of Crimean-Congo haemorrhagic fever virus in ticks Hyalomma aegyptium in the Middle East.

It is the first time that Crimean-Congo haemorrhagic fever virus (CCHFV), causing potentially lethal disease of humans, has been reported from the Middle East region and from the tortoise tick Hyalomma aegyptium from a tortoise host, whose epidemiological significance may have remained almost completely overlooked so far. We used RT-PCR to screen for 245 ticks collected from 38 Testudo graeca tortoise individuals. Results of our genetic screening provide unambiguous evidence of occurrence of CCHFV in this region and host, suggesting a potentially important role of H. aegyptium in CCHF epidemiology.

Antimicrobial-resistant Enterobacteriaceae from humans and wildlife in Dzanga-Sangha Protected Area, Central African Republic.

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 ß-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 blaCTX-M-15 and qepA.