CTX-M15-producing Escherichia coli clone B2-O25b-ST131 and Klebsiella spp. isolates in municipal wastewater treatment plant effluents

Objectives: The global occurrence of antibiotic resistance genes in bacteria in water environments is an increasing concern. Treated wastewater was sampled daily over a 45 day period from the outflow of a municipal wastewater treatment plant in Brno, Czech Republic, and examined for extended-spectrum b-lactamase (ESBL)-producing bacteria. Methods: Water samples were cultivated on MacConkey agar with cefotaxime (2 mg/L) and individual colonies were examined for ESBL production. Phenotypic ESBL-positive bacteria identified as Escherichia coli or Klebsiella spp. were tested for the presence of antibiotic resistance genes, the virulence gene afa/dra and the bla CTX-M upstream region. Genetic relatedness was analysed by PFGE, multilocus sequence typing and plasmid analysis. Results: A total of 68 ESBL-producing Enterobacteriaceae isolates were detected in 34 out of 45 wastewater samples. ESBL-producing isolates included 26 E. coli isolates, 4 Klebsiella pneumoniae isolates and 1 Klebsiella oxytoca isolate. The pandemic and multiresistant B2-O25b-ST131 clone was predominant, being detected among 19 E. coli isolates, and 17 of the B2-O25b-ST131 isolates were positive for the FIA replicon and the afa/dra operon and had an IS26 element flanking bla CTX-M-15 . Seventeen of the B2-O25b-ST131 isolates showed closely related PFGE profiles (defined by 84% band similarity) and belonged to identical clonal groups. Conclusions: The results highlight the inadequacy of the treatment process in removing multiresistant bacteria from municipal wastewater and point to a risk of transmission of clinically important multiresistant strains, such as the pandemic ST131 clone, to the environment. This is the first study demonstrating the pandemic ST131 clone in wastewater

Host dispersal shapes the population structure of a tick-borne bacterial pathogen

Birds are hosts for several zoonotic pathogens. Because of their high mobility, especially of longdistance migrants, birds can disperse these pathogens, affecting their distribution and phylogeography. We focused on Borrelia burgdorferi sensu lato, which includes the causative agents of Lyme borreliosis, as an example for tick-borne pathogens, to address the role of birds as propagation hosts of zoonotic agents at a large geographical scale. We collected ticks from passerine birds in 11 European countries. B. burgdorferi s.l. prevalence in Ixodes spp. was 37% and increased with latitude. The fieldfare Turdus pilaris and the blackbird T. merula carried ticks with the highest Borrelia prevalence (92 and 58%, respectively), whereas robin Erithacus rubecula ticks were the least infected (3.8%). Borrelia garinii was the most prevalent genospecies (61%), followed by B. valaisiana (24%), B. afzelii (9%), B. turdi (5%) and B. lusitaniae (0.5%). A novel Borrelia genospecies "Candidatus Borrelia aligera" was also detected. Multilocus sequence typing (MLST) analysis of B. garinii isolates together with the global collection of B. garinii genotypes obtained from the Borrelia MLST public database revealed that: (a) there was little overlap among genotypes from different continents, (b) there was no geographical structuring within Europe, and (c) there was no evident association pattern detectable among B. garinii genotypes from ticks feeding on birds, questing ticks or human isolates. These findings strengthen the hypothesis that the population structure and evolutionary biology of tick-borne pathogens are shaped by their host associations and the movement patterns of these hosts.Peer reviewe

Combined Analysis of Variation in Core, Accessory and Regulatory Genome Regions Provides a Super-Resolution View into the Evolution of Bacterial Populations

The use of whole-genome phylogenetic analysis has revolutionized our understanding of the evolution and spread of many important bacterial pathogens due to the high resolution view it provides. However, the majority of such analyses do not consider the potential role of accessory genes when inferring evolutionary trajectories. Moreover, the recently discovered importance of the switching of gene regulatory elements suggests that an exhaustive analysis, combining information from core and accessory genes with regulatory elements could provide unparalleled detail of the evolution of a bacterial population. Here we demonstrate this principle by applying it to a worldwide multi-host sample of the important pathogenic E. coli lineage ST131. Our approach reveals the existence of multiple circulating subtypes of the major drug-resistant clade of ST131 and provides the first ever population level evidence of core genome substitutions in gene regulatory regions associated with the acquisition and maintenance of different accessory genome elements.Peer reviewe

Mites of the genus Neharpyrhynchus Fain (Acariformes, Harpirhynchidae) from Neotropical birds

Three new species of parasitic mites of the genus Neharpyrhynchus Fain (Acariformes, Harpirhynchidae) are described from Neotropical birds, N. chlorospingus sp. n. from Chlorospingus pileatus (Passeriformes, Emberizidae) from Costa Rica, N. mironovi sp. n. from Dacnys cayana (Passeriformes, Thraupidae) and N. tangara sp. n. from Tangara cayana (Thraupidae) both from Brazil. Neharpyrhynchus trochilinus (Fain) is recorded from 3 new hosts of the family Trochilidae (Apodiformes), Panterpe insignis and Eugenes fulgens from Costa Rica, and Amazilia lactea from Brazil. Emended diagnosis of the genus and a key to species are provided; all records of Neharpyrhynchus species are summarized

Chewing lice of genus Ricinus (Phthiraptera, Ricinidae) deposited at the Zoological Institute of the Russian Academy of Sciences, Saint Petersburg, Russia, with description of a new species

We revised a collection of chewing lice deposited at the Zoological Institute of the Russian Academy of Sciences, Saint Petersburg, Russia. We studied 60 slides with 107 specimens of 10 species of the genus Ricinus (De Geer, 1778). The collection includes lectotype specimens of Ricinus ivanovi Blagoveshtchensky, 1951 and of Ricinus tugarinovi Blagoveshtchensky, 1951. We registered Ricinus elongatus Olfers, 1816 ex Turdus ruficollis, R. ivanovi ex Leucosticte tephrocotis and Ricinus serratus (Durrant, 1906) ex Calandrella acutirostris and Calandrella cheleensis which were not included in Price’s world checklist. New records for Russia are R. elongatus ex Turdus ruficollis; Ricinus fringillae De Geer, 1778 ex Emberiza aureola, Emberiza leucocephalos, Emberiza rustica, Passer montanus and Prunella modularis; Ricinus rubeculae De Geer, 1778 ex Erithacus rubecula and Luscinia svecica; Ricinus serratus (Durrant, 1906) ex Alauda arvensis. New records for Kyrgyzstan are R. fringillae ex E. leucocephalos and ex Fringilla coelebs. A new record for Tajikistan is R. serratus ex Calandrella acutirostris. The new species Ricinus vaderi Valan n. sp. is described with Calandra lark, Melanocorypha calandra; from Azerbaijan, as a type host

Ricinus dalgleishi

Ricinus dalgleishi Nelson, 1972 (Figs 1–6, 19, Table 1) Ricinus dalgleishi Nelson, 1972: 82, plate 22, figs 1–5. Ricinus dalgleishi Nelson, 1972; Price et al. 2003: 247. Type host. Geothlypis trichas (Linnaeus, 1766) —Common yellowthroat (Passeriformes: Parulidae) Type locality. State College, Mississippi, United States of America. Diagnosis. This species belongs to the marginatus species-group (Nelson 1972) by having lunar nodi, no ovoid sclerite evident, and mandibles without finger-like extension. By having 4 setae on the paramere apices, R. dalgleishi is distinguished from Ricinus marginatus (Children, 1836), Ricinus pallens (Kellogg, 1899), and Ricinus emersoni Nelson, 1972, which have 3 setae. Further, it differs from Ricinus dendroicae Nelson, 1972, Ricinus picturatus (Carriker, 1902) and Ricinus frenatus (Burmeister, 1838) in having setae a6, and from Ricinus flavicans Carriker, 1964, Ricinus seiuri Nelson, 1972 and Ricinus polioptilus Carriker, 1964 in having temple apices hooked outside. Description. Head subconical, with straight lateral margins. Frons narrowly convex; lateral margins convergent in the front and rounded; not continuous with head lateral margin. Temple apices acute, hooked outside. Occipital margin nearly straight. Eyes protruded. Transverse carina present; nearly straight. Lunar nodi present; tentorial nodi present; and lunar nodi nearly equal to tentorial. Mandibles monomorphic; blades long, thin and sharp needle-like; tips not notched; galea not evident; basal lobe without finger-like process. Maxillary plates sickle shaped (sausage-like); pigment pattern absent; palpi geniculate; not reaching the margin of head. Ovoid sclerite not evident. Gula with 2 pairs of setae; top of gular plate sclerite truncate; medial part with concavity; posterior projections present; directing posteriorly. Anterior margin of labium concave. The cf series composed of 10 setae; df series present; f1 evidently longer than f2; a1 far shorter than m4; with two associated sensilla; a3 absent; a4 present; a6 present; m1–m3 equal; m2 off the marginal carinae; m4 evidently longer than pa. Labium with 13 pairs of setae. Mental setae positioned laterally to each other. Preantennal setae strongly spinose. Setae along the antennal lappets 12–13. Three pilose, subequal postocular setae. Thorax. Prothorax hexagonal; anterior margin distinctly concave; lateral margins slightly notched; posterior margin concave. Posterolateral angles of prothorax slightly pointed. Prosternal sclerites thin, parallel, not joined. Anterior margin of sternal plate rounded, without lateral depression; posterior angles acute. L3 present; L6 present; L5 nearly equal to L4 and L6; L9 evidently shorter than L7 and L8; c4 present; c1 twice or more longer than c2; c2 pilose; c3 twice or more longer than c4; w series composed of 6 setae; anterior setae unequal. Long sternal setae situated medially on pterothorax 3; q series composed of 2 spines; q2 strongly spinose; q2 not shorter than w series; b1 evidently shorter than b2. Coxa I with 2 tactile setae; femur I with 2 tactile setae; femur II and III with present tactile setae. Abbreviations: a, dorsal setae on temples; ant. n., antennal nodi; b, dorsal setae on posterior margin; c, 4 pairs of dorsal setae on pterothorax; d, dorsal setae on head; dps, dorsal pleural setae; L, lateral prothoracic setae; lun. n., lunar nodi; m, dorsoventral setae on marginal carinae; max. palp., maxillary palpi; max., maxillary setae; ment., mental setae; pa, paraantennal setae; pm, paramental setae; po, postocular setae; pr, dorsal prothoracic setae; preant., preantennal setae; prst. pl., prosternal plate; prst. s., prosternal plate setae; ps. s., postspiracular setae; q, ventral and submarginal setae on pterothorax; sc, sternal central setae; sen., sensilla; sl, sternal lateral setae; spir., spiracle; st. s., sternal setae; t, dorso-lateral setae on temples; tent. n., tentorial nodi; term. s., terminal setae of tergite IX; VI–VIII, tergites; vps, ventral pleural setae; w, series of lateral setae on pterothorax. Scale bars are in mm. Note: Complete anal fringes are drawn as in Nelson (1972). Abdomen. Lateral margin of abdomen nearly straight. Segment IX nearly equal to VIII. Pleural nodi on segment IX present, relatively wide, margins smooth; lateral part of abdominal pleurites lightly pigmented. Second ventral pleurite with 3 setae; II vps 1–3 large spinose; III vps 1–2 small spinose; III vps 3 small spine; IV vps 1 large spine; IV vps 2 long pilose; IV vps 3 large spine; V vps 1 small spine; V vps 3 small pilose; VI vps 1,3 small pilose; VII vps 1,3 small pilose; VIII vps 1 small pilose; VIII vps 3 moderately long pilose. Two pairs of setae on tergite VIII. Sternolateral setae equal to sternocentral on II–VI sternites. Vulva with 4 setae. Abdominal segment IX with 2 long terminal setae. Male genitalia. Symmetrical, basal plate narrow anteriorly and lateral margin slightly concave. Parameres triangular, with blunt endings. Mesosomal plate pigmented, lightly pointed and without medial extension. Four setae on the distal tips of the parameres. Dimensions. Female (n = 8). Head length 0.66–0.67; width 0.59–0.60; head index 110–113. Labrum width 0.28–0.29. Prothoracic length 0.31–0.32. width 0.52–0.53, ratio 0.59–0.60. Distance between prosternal setae 0.063–0.072. Abdominal width 0.82–0.85. Total length 3.20–3.35. Ratio of total length and abdomen width 3.90– 3.94. Male (n = 6). Head length 0.60–0.62; head width 0.54–0.55; head index 111–113. Labrum width 0.24–0.25. Prothoracic length 0.29–0.31, width 0.46–0.47, ratio 0.63–0.66. Distance between prosternal setae 0.058–0.065. Abdominal width 0.73–0.74. Total length 2.95–3.00. Ratio of total length and abdomen width 4.04–4.05. Length of male genitalia 0.41–0.43. Width of mesosomal plate 0.160–0.168. Material examined. Paratypes. Ex Geothlypis trichas: 2♀, Pearlington, Mississippi, U.S.A., 3 Jun. 1910, G.G. Rohwer, slide Bish-29874 (Lot 40-14138); 1♀, Damarest, New Jersey, U.S.A., 20 May 1926, B.S. Bowdish; 5♀, Elmhurst, New York, U.S.A., 9 May 1932, M.V. Beals, slide Bish-19646; 2♀, Groton, Massachusetts, U.S.A., 21 May 1933, W.P. Wharton, slide Bish-21187; 1♀, New London, North Carolina, U.S.A., 4 Apr. 1945, R.C. Simpson. [Note: all these paratypes are held in the USNM]. Non types. Ex Helmitheros vermivorum (Gmelin, 1789) —Worm-eating warbler (Passeriformes: Parulidae): 2♂, 2♀, Utila Island, Islas de la Bahía, Honduras (16°06' N, 86°54' W), 23 Aug. 2014, I. Literak, slides UT22a–b [also 3♂, 2♀, 6N in alcohol]; 1♀, same collecting data as previous sample, 26 Aug. 2014, slide UT65; 1♂, 1♀, 2N, Marshall’s Pen, Mandeville, Jamaica, Feb. 1981, R. Dalgleish, slide 814 (USNM); 2♀ same collecting data as previous sample, 24–30 Dec. 1982 (USNM). Remarks. Ricinus dalgleishi was described by Nelson (1972) from females only. Our comparison of female paratypes of this species against females from Helmitheros vermivorum has shown that those from H. vermivorum are only slightly smaller, but without significant morphological differences. Therefore, we regard samples from both hosts as belonging to the same species. Considering that we have examined adult lice of both sexes and nymphs from two host individuals of H. vermivorum collected in two different localities, and that these specimens share the same morphological and morphometric characteristics, we confirm H. vermivorum as a natural and regular host for R. dalgleishi and establish it as new host-parasite association. Also, we record and describe the male of R. dalgleishi for the first time.Published as part of Valan, Miroslav, Sychra, Oldrich & Literak, Ivan, 2016, Redescriptions and new host records of chewing lice of the genus Ricinus (Phthiraptera: Ricinidae) from the Neotropical Region, pp. 179-189 in Zootaxa 4154 (2) on pages 180-182, DOI: 10.11646/zootaxa.4154.2.5, http://zenodo.org/record/26649

Ricinus vireoensis

Ricinus vireoensis Nelson, 1972 (Fig. 21, Table 1) Ricinus vireoensis Nelson, 1972: 95, plate 33, figs 1–6. Ricinus vireoensis Nelson, 1972; Price et al. 2003: 251. Type host. Vireo griseus (Boddaert, 1783) —White-eyed vireo (Passeriformes: Vireonidae) Type locality. Leon County, Florida, United States of America. Material examined. Ex Vireo pallens Salvin, 1863 —Mangrove vireo (Passeriformes: Vireonidae): 2♀, Utila Island, Islas de la Bahía, Honduras (16°06' N, 86°54' W) 27 Aug. 2014, I. Literak, slide UT87 [also 2♀, 4N in alcohol]. Ex Vireo olivaceus (Linnaeus, 1766) Red-eyed vireo (Passeriformes: Vireonidae): 3♀, Utila Island, Islas de la Bahía, Honduras (16°06' N, 86°54' W), 24 Aug. 2014, I. Literak, slide UT43 [also 7N in alcohol]. Remarks. Vireo pallens is an additional host species for R. vireoensis (see Price et al. 2003: 251), bringing the total number of known hosts for this louse species to eight. Bird family Bird species Country P A % Louse species ♀ ♂ N Totals Parulidae Helmitheros vermivorum Honduras 2/9 22.2 Ricinus dalgleishi * 5 5 6 1 6 Tyrannidae Corythopis delalandi Paraguay 1/10 10.0 Ricinus sp.* 2 - 2 4 Fringillidae Euphonia laniirostris Costa Rica 2/26 7.7 Ricinus tanagraephilus 14 1 9 24 Vireonidae Vireo pallens Honduras 1/9 11.1 Ricinus vireoensis * 4 - 4 8 Vireo olivaceus Honduras 1/12 8.3 Ricinus vireoensis 3 - 7 1 0Published as part of Valan, Miroslav, Sychra, Oldrich & Literak, Ivan, 2016, Redescriptions and new host records of chewing lice of the genus Ricinus (Phthiraptera: Ricinidae) from the Neotropical Region, pp. 179-189 in Zootaxa 4154 (2) on page 188, DOI: 10.11646/zootaxa.4154.2.5, http://zenodo.org/record/26649

Ricinus tanagraephilus Eichler 1956

Ricinus tanagraephilus Eichler, 1956 (Figs 12–18, 22–23, Table 1) Ricinus tanagraephilus Eichler, 1956: 133. Ricinus tanagraephilus Eichler, 1956; Nelson 1972: 111. Species inquirendae. Ricinus tanagraephilus Eichler, 1956; Price et al. 2003: 251. Type host. Euphonia laniirostris hypoxantha von Berlepsch & Taczanowski, 1884 —Thick-billed euphonia (Passeriformes: Fringillidae) Type locality. Peru. Diagnosis. Ricinus tanagraephilus belongs to the subangulatus species-group (Nelson 1972) by having lunar nodi, monomorphic mandibles, evident ovoid sclerite with pit-like depressions, and pattern on terminal tergite of female iIIi x iIIi, but can be distinguished from Ricinus subangulatus (Carriker, 1903), Ricinus complicatus Carriker, 1964, Ricinus ramphoceli Nelson, 1972 and Ricinus volatiniae Nelson, 1972 by having setae a6 and 13 labial setae. Also, it differs from R. vireoensis and Ricinus wolfi Nelson, 1972 by having 3 setae on the parameres. Further, R. tanagraephilus differs from Ricinus subhastatus (Durrant, 1906) by having setae a6, and inner setae on pleurites VI–VIII small and pilose. Description. Head subconical, with straight lateral margins. Frons broadly convex; lateral margins divergent, not continuous with head lateral margin. Temple apices acute, hooked outside. Occipital margin nearly straight. Eyes not evident or slightly raised. Transverse carina present, convex. Lunar nodi present; tentorial nodi present, and lunar nodi nearly equal to tentorial. Mandibles monomorphic, blades long, thin and sharp needle-like, with tips notched; galea evident; basal lobe without finger-like process. Maxillary plates sausage-like; pigment pattern present; palpi geniculate, reaching the head margin. Ovoid sclerite evident, ornamentation deeply pitted. Gula with 2–3 pairs of setae; top of gular plate sclerite truncate; medial part without concavity; posterior projections present, directing posteriorly. Anterior margin of labium concave. The cf series composed of 10 setae; df series present; f1 evidently longer than f2; a1 far shorter than m4; with two associated sensilla; a3 absent; a4 present; a6 present; m1–m3 equal; m2 off the marginal carinae; m4 evidently longer than pa. Labium with 13 pairs of setae. Mental setae positioned laterally to each other. Mental setae shorter than maxillary. Preantennal setae strongly spinose. Setae along the antennal lappets 11–13. Two spinose and subequal postocular setae. Thorax. Prothorax hexagonal; anterior margin distinctly concave; lateral margins slightly notched; posterior margin concave. Posterolateral angles of prothorax evidently rounded. Prosternal sclerites thin, parallel, not joined. Anterior margin of sternal plate concave, without lateral depression; posterior angles straight. L3 present; L6 present; L5 larger than L4 and L6; L9 evidently shorter than L7 and L8; c4 present; c1 and c2 nearly equal; c2 spinose; c3 and c4 nearly equal; w series composed of 6 setae; anterior setae unequal. Long sternal setae situated medially on pterothorax 1; q series composed of 2 spines; q2 strongly spinose; q2 not shorter than w series; b1 evidently shorter than b2. Coxa I with 2 tactile setae; femur I with 2 tactile setae; femur II and III with present tactile setae. Abdomen. Lateral margin of abdomen nearly straight. Segment IX nearly equal to VIII. Pleural nodi on segment IX present; relatively wide; margins smooth; lateral part of abdominal pleurites lightly pigmented. Second ventral pleurite with 3 setae; II vps 1–3 large spinose; III vps 1–2 small spinose; III vps 3 small spine; IV vps 1 large spine; IV vps 2 long pilose; IV vps 3 large spine; V vps 1 small spine; V vps 3 small pilose; VI vps 1, 3 small pilose; VII vps 1, 3 small pilose; VIII vps 1 small pilose; VIII vps 3 small to medium pilose. Two pairs of setae on tergite VIII. Sternolateral setae not equal to sternocentral on sternites II–VI. Vulva with 11 setae. Abdominal segment IX with 2 long terminal setae. Male genitalia symmetrical; basal plate not narrow anteriorly; lateral margin slightly concave. Parameres triangular, with blunt endings. Mesosomal plate pigmented, lightly pointed and without medial extension. Three setae on the distal tips of the parameres. Dimensions. Female (n = 15). Head length 0.75–0.77; width 0.61–0.63; head index 120–124. Labrum width 0.30–0.32. Prothoracic length 0.37–0.38; width 0.58–0.59; ratio 0.63–0.66. Distance between prosternal setae 0.059–0.062. Abdominal width 1.01–1.05. Total length 3.70–3.78. Ratio of total length and abdomen width 3.60– 3.66. Male (n = 1). Head length 0.70; head width 0.59; head index 119. Labrum width 0.29. Prothoracic length 0.34; width 0.53; ratio 0.64. Distance between prosternal setae 0.062. Abdominal width 0.84. Total length 3.325. Ratio of total length and abdomen width 3.96. Length of male genitalia 0.48; width of mesosomal plate 0.188. Material examined. Holotype ♀, Perú, date unknown, von Koepcke Coll., slide WEC3066a (ZMHU). Non-types. Ex Euphonia laniirostris d'Orbigny & Lafresnaye, 1837: 1 ♂, 4♀, La Amistad Lodge, Las Tablas, Costa Rica (8°54’ N, 82°47’ W; 1300 m a.s.l.) 18–19 Aug. 2010, I. Literak, slides LT15a–b, LT65 [also 10♀, 9N in alcohol]. Remarks. Eichler’s (1956) brief description of R. tanagraephilus was based on one female only. Carriker (1964: 50) subsequently redescribed this species, but Nelson (1972: 111) examined the specimen from Euphonia laniirostris studied by Carriker (1964) and recognized it as a distorted specimen of R. marginatus. For that reason, Nelson (1972) designated R. tanagraephilus as a species inquirendae. We also examined Carriker’s specimen and we are able to confirm Nelson’s (1972) identification. Although our samples originated from a different country than the type locality of R. tanagraephilus and, therefore, also from a different subspecies of E. laniirostris than the type host, our comparison against the holotype of this louse species allowed us to confirm that this is a distinct and valid species of Ricinus. Also, we record and describe the male of R. tanagraephilus for the first time.Published as part of Valan, Miroslav, Sychra, Oldrich & Literak, Ivan, 2016, Redescriptions and new host records of chewing lice of the genus Ricinus (Phthiraptera: Ricinidae) from the Neotropical Region, pp. 179-189 in Zootaxa 4154 (2) on pages 184-187, DOI: 10.11646/zootaxa.4154.2.5, http://zenodo.org/record/26649