The Acetylcholinesterase Gene Ace: a Diagnostic Marker for the Pipiens and Quinquefasciatus Forms of the Culex Pipiens Complex

Total Page:16

File Type:pdf, Size:1020Kb

The Acetylcholinesterase Gene Ace: a Diagnostic Marker for the Pipiens and Quinquefasciatus Forms of the Culex Pipiens Complex Journal oJ'the American Mosquito Control Association, 14(4):39O_396, l99g Copyright O 1998 by the American Mosquito Control Association, Inc. THE ACETYLCHOLINESTERASE GENE ACE: A DIAGNOSTIC MARKER FOR THE PIPIENS AND QUINQUEFASCIATUS FORMS OF THE CULEX PIPIENS COMPLEX DENIS BOURGUET'2 DINA FONSECA,S GwENAEI- vouRcn,, MARIE-pIERRE DUBOIS,I FABRICE CHANDRE,4 CARLO SEVERINIs AND MICHEL RAYMONDI,6 ABSTRACT' The taxonomy of the Culex pipiens complex remains a controversial issue in mosquito sysrem- atics. Based on morphologic characters, 2 allopatric taxa are recognized, namely pipiens (including "molestus") Cx. the form in temperate areas and Cx. quinquefasciatus in tropical areas. Here we report on variability at the nucleotide level of an acetylcholinesterase gene in several strains and natural populations of this species complex. Few polymorphisms were found in coding regions within a subspecies but many polymorphismi were observed between subspecies in noncoding regions. We describe a method based on a restriction enzyme polymorphism in polymerase chain reaction-amplified DNA, in which the presence or absence of one restriction site diicrim- inates Cx. pipiens, Cx. quinquefasciatus, and their hybrids. This technique reliably discriminates mosquitoes from more than 30 worldwide strains or populations. Polymerase chain reaction amplification of specific alleles may also be a useful tool for characterizing specific alleles of each sibling taxon. KEY WORDS Acetylcholinesterase gene, Culex pipiens complex, diagnostic markeq sibling species, Cule-r torrentium, Culex pipiens "molestus" INTRODUCTION feed on mammals (mammophily). In contrast, fe- males from epigeous The mosquito Culex pipiens represents a species habitats require a blood meal produce complex that is incompletely understood (see Har- to their first batch of eggs (anautogeny), bach et al. [1985] for a review). Based on morpho- are unable to mate in confined spaces, such as in logic characters, 3 types have thus far been de- laboratory conditions (eurygamy), hibernate during scribed: Culex quinquefasciatus Say (Sirivanakarn the winter (heterodynamy), and have a propensity "molestus" and White 1978), Culex pipiens For- to feed on birds (ornithophily). The same associa- skil (Harbach et al. 1984), and Culex pipiens Lin- tion between physiologic traits and habitat types is naeus (Harbach et al. 1985). The last 2 types are observed in northern Europe and in North Ameri- sympatric and are considered by some authors to can and Australian regions with cold winters (Rou- be ecotypes of the same form (Roubaud 1933; Mat- baud 1933, Marshall and Stanley 1937, Spielman tingly 1951; Pasteur 19771,Barr l98l; Chevillon et 1964, Miles 1976). "molesras") al. 1995a,1998; Vinogradova et al. 1996; Eritja, Culex pipiens (including Cx. p. is 1998), as they are mainly distinguished by ecolog- largely a temperate form, whereas Cx. quinquefas- "mo- ical and physiologic characteristics. Culex p. ciatus is cosmotropical (Mattingly et al. 1951, Barr lestus" breeds in underground urban habitats (hy- 1957). Cx. quinquefascialus is homodynamous, pogeous habitats such as cellars, sanitary spaces stenogamous, and anautogenous. Extensive areas of under buildings, and septic tanks), and Cx. pipiens overlap and hybridization exist in the Middle and breeds in rural open-air habitats (epigeous habitats Far East, North and South America, Australia, and such as brooks, rivers, swamps, ditches, or nny ar- Africa (Barr 1982; Urbanelli et al. 1995, 1997). The tificial open-air collection of water). Females from main morphologic differences between Cx. pipiens hypogeous habitats do not require a blood meal to and Cx. quinquefasciatus are found in the male produce their first batch of eggs (autogeny), are genitalia, and can be quantified using the DV/D ra- able to mate in confined spaces (stenogamy), do not tio (Sundararaman 1949), where DV is the distance hibernate (homodynamy), and have a tendency to from the tip of the ventral arm of the phallosome to its intersection with the dorsal arm and D is the I Institut des Sciences de I'Evolution (UMR 5554), La- distance between the tips of the dorsal arms of the boratoire Gdn6tique et Environnement, Universitd Mont- phallosome. Values of DV/D below 0.2 characteize pellier II, Place Eugdne Bataillon, 34 090 Montpellier, France. Cx. pipiens, whereas values above O.4 characteize 2 Present address: Station de Recherche de Lutte Bio- Cx. quinquefasciatus. Although this ratio has prov- logique, INRA La Minidre, 78285 Guyancourt, France. en to be reliable outside hybrid zones by several 3 Molecular Genetics Laboratory-DZR, National Zoo- authors (Mattingly et al. 1951, Ban 1957), its use logical Park, Smithsonian Institution, Washington, DC, is restricted to adult males. More recently, bio- 20008. chemical and molecular techniques have been used 4 ORSTOM, Laboratoire de Lutte contre les Insectes to find diagnostic markers (Miller et al. 1996, Sev- Nuisibles, BP 5045, 34032 Montpellier, France. 5 Laboratorio di Parassitologia, Istituto Superiore di erini et al. 1996, Crabtree et al. 1997). Recently, Sanitd, Viale Regina Elena, 299, OOl6l Rome, Italy. part of an acetylcholinesterase gene, referred to as 6 To whom correspondence should be addressed. Ace, was cloned for a Cx. pipiens strain (Malcolm 390 Deceranen1998 DIAGNosTIc Mnnren tN C. prplalrs Cotr,tplex 391 et al. 1998), thus offering a new opportunity to the PCR primers. For the DC3, Hilo, McCandless, compare Cx. pipiens and Cx. quinquefasciatus at and Macapi populations, the PCR conditions were the genomic level. Here we report partial sequences identical to those described above but reagents of the Ace locus for different collections of the Cx. from ABVPerkin Elmer (Norwalk, CT) and an MJ pipiens complex. Variation in this region clearly ResearchPeltier thermocycler (MJ Research,Inc., discriminates Cx. pipiens from Cx. quinquefascia- Waterton, MA) were used instead. The PCR prod- /zs. Based on these sequences we propose and test ucts were purifled with a QlAquick PCR purifica- a restriction enzyme pattern as a diagnostic marker tion kit (Qiagen, Valencia, CA). One microliter of for the 2 subspecies. clean DNA was cycle sequencedusing AmpliTaq DNA FS polymerase and dye-labeled terminators (PE Biosystems, Foster City, CA), and was exam- MATERIALS AND METHODS ined on an automated sequencer(ABI/Perkin El- Mosquitoes: Origins and references of the strains mer). and populations used in this study are given in Ta- Restriction fragment length polymorphism ble 1. Mosquitoes from populations or strains close (RFLP) analysis: At least 3 mosquitoesfrom each to putative hybrid zones (Mattingly et al. 1951) strain or population were analyzed except for Fort such as BED (South Africa), Killcare (Australia), Knox and Simpson where RFLP analyseswere per- DC3 (Washington DC, USA), and BEIJING (Chi- formed on genomic DNA of up to 10Omosquitoes. na) were classified as Cx. pipiens or Cr. quinque- Single mosquito genomic DNAs were obtained fol- fasciatus by means of DV/D ratios of male genitalia lowing Qiao and Raymond (1995). The 700-bp (Ban 1957). Females of the strain S-LAB (Cx. fragment of the Ace gene was amplified as de- quinquefasciafus) were crossed with males of 2 dif- scribed above. Aliquots of l0 pl of each amplifi- ferent Cx. pipiens strains from southern France to cation were digested with the ScaI restriction en- obtain hybrid individuals that were referred to as zyme and loaded onto a 1.5-2Vo(w/v) agarosegel MSE-FI and RSV respectively (Table 1). Mosqui- with tris borate EDTA (TBE) buffer. toes from 2 populations of Culex torrentium Mar- tini (see Thble 1) were also used for comparison. Polymerase chnin reaction (PCR) amplification RESULTS AND DISCUSSION and sequencing.' For the MSE, BRUGES A, Praias, Ace polymorphism S-LAB, SUPERCAR, MRES, and BEUING strains, genomic DNA extraction of up to l0O mosquitoes At least 363 nucleotide sites (44 in exon 2, 158 was perfonned as described by Raymond et al. in intron 2, arrd 161 in exon 3) have been se- (1989). The DNA from the DC3. Hilo. and Mc- quenced at the Ace locus for several strains and Candless strains was extracted from individual populations from various geographic areas (China, mosquitoes using a standard phenol-chloroform Hawaii, Brazil, Ivory Coast, Cuba, and California protocol (Sambrook et al. 1989). A 7Oo-base pair for Cx. quinquefasciatusartd France, Belgium, Por- (bp) fragment (which encompassed part of exon 2, tugal, and Washington,DC, for Cx. pipiens). Yari- intron 2, and part of exon 3, see Fig. 1 and Malcolm able nucleotidesare shown in Fig. 2. Variable sites et al. [1998]) of the Ace gene was amplified using are mainly located in intron 2 and substitutions in the oligonucleotide primers F 1457 (5'-GAGGA- the exons did not change the inferred amino-acid GATGTGGAATCCCAA-3') and B 1246 (5'- sequence.This indicates that Ace is probably not a TGGAGCCTCCTCTTCACGGC-3') (Eurogentec, pseudogene,although its exact function remains Seraing, Belgium). Amplifications were performed unknown (Malcolm et al. 1998).The polymorphism in a 50-pl volume containing 75 mM Tfis-HCl (pH zrmongstrains of the same subspeciesis low and 9.O), 20 mM (NH4),SO', O.l7o (w/v) Tlveen 20, Taq er'rorsmay not be excluded. In contrast, we 1.25 mM MgClr, 250 pM of each deoxynucleoside found many differences (37 variable sites out of triphosphate (dNTP), IOO ng of each primer, 10- 7lO sequenced)between the Ace sequencesof Cx. 100 ng of DNA, and 2.5 units of Taq polymerase pipiens and those of Cx. quinquefasciatus.The Ace (Eurogentec). The tubes were then quickly trans- gene of the Cx. pipiens complex is characterizedby ferred to the thermal cycler (Thermocycler Croco- the presenceof 10 introns (Malcolm et al. 1998). dile II, Appligene, Illkirch, France). After 5 min at With the exception of intron 4, these introns are 93oC, reactions were cycled 35 times through the very large, resembling more the structure of the following temperature profile: 93oC for 1 min, 52oC Drosophila melanogasterAce Meigen gene (Four- for 1 min, and 72"C for 90 sec.
Recommended publications
  • Mosquitoes (Diptera: Culicidae) in the Dark—Highlighting the Importance of Genetically Identifying Mosquito Populations in Subterranean Environments of Central Europe
    pathogens Article Mosquitoes (Diptera: Culicidae) in the Dark—Highlighting the Importance of Genetically Identifying Mosquito Populations in Subterranean Environments of Central Europe Carina Zittra 1 , Simon Vitecek 2,3 , Joana Teixeira 4, Dieter Weber 4 , Bernadette Schindelegger 2, Francis Schaffner 5 and Alexander M. Weigand 4,* 1 Unit Limnology, Department of Functional and Evolutionary Ecology, University of Vienna, 1090 Vienna, Austria; [email protected] 2 WasserCluster Lunz—Biologische Station, 3293 Lunz am See, Austria; [email protected] (S.V.); [email protected] (B.S.) 3 Institute of Hydrobiology and Aquatic Ecosystem Management, University of Natural Resources and Life Sciences, Vienna, Gregor-Mendel-Strasse 33, 1180 Vienna, Austria 4 Zoology Department, Musée National d’Histoire Naturelle de Luxembourg (MNHNL), 2160 Luxembourg, Luxembourg; [email protected] (J.T.); [email protected] (D.W.) 5 Francis Schaffner Consultancy, 4125 Riehen, Switzerland; [email protected] * Correspondence: [email protected]; Tel.: +352-462-240-212 Abstract: The common house mosquito, Culex pipiens s. l. is part of the morphologically hardly or non-distinguishable Culex pipiens complex. Upcoming molecular methods allowed us to identify Citation: Zittra, C.; Vitecek, S.; members of mosquito populations that are characterized by differences in behavior, physiology, host Teixeira, J.; Weber, D.; Schindelegger, and habitat preferences and thereof resulting in varying pathogen load and vector potential to deal B.; Schaffner, F.; Weigand, A.M. with. In the last years, urban and surrounding periurban areas were of special interest due to the Mosquitoes (Diptera: Culicidae) in higher transmission risk of pathogens of medical and veterinary importance.
    [Show full text]
  • Copyright © and Moral Rights for This Thesis Are Retained by the Author And/Or Other Copyright Owners
    Copyright © and Moral Rights for this thesis are retained by the author and/or other copyright owners. A copy can be downloaded for personal non-commercial research or study, without prior permission or charge. This thesis cannot be reproduced or quoted extensively from without first obtaining permission in writing from the copyright holder/s. The content must not be changed in any way or sold commercially in any format or medium without the formal permission of the copyright holders. When referring to this work, full bibliographic details including the author, title, awarding institution and date of the thesis must be given e.g. AUTHOR (year of submission) "Full thesis title", Canterbury Christ Church University, name of the University School or Department, PhD Thesis. Renita Danabalan PhD Ecology Mosquitoes of southern England and northern Wales: Identification, Ecology and Host selection. Table of Contents: Acknowledgements pages 1 Abstract pages 2 Chapter1: General Introduction Pages 3-26 1.1 History of Mosquito Systematics pages 4-11 1.1.1 Internal Systematics of the Subfamily Anophelinae pages 7-8 1.1.2 Internal Systematics of the Subfamily Culicinae pages 8-11 1.2 British Mosquitoes pages 12-20 1.2.1 Species List and Feeding Preferences pages 12-13 1.2.2 Distribution of British Mosquitoes pages 14-15 1.2.2.1 Distribution of the subfamily Culicinae in UK pages 14 1.2.2.2. Distribution of the genus Anopheles in UK pages 15 1.2.3 British Mosquito Species Complexes pages 15-20 1.2.3.1 The Anopheles maculipennis Species Complex pages
    [Show full text]
  • Clearing up Culex Confusion
    Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology 1185 Clearing up Culex Confusion A Basis for Virus Vector Discrimination in Europe JENNY C. HESSON ACTA UNIVERSITATIS UPSALIENSIS ISSN 1651-6214 ISBN 978-91-554-9044-7 UPPSALA urn:nbn:se:uu:diva-232726 2014 Dissertation presented at Uppsala University to be publicly examined in Zootissalen, Villavägen 9, 2 tr, Uppsala, Friday, 7 November 2014 at 10:00 for the degree of Doctor of Philosophy. The examination will be conducted in English. Faculty examiner: Professor Laura D Kramer (Wadsworth Center, New York State Department of Health, USA). Abstract Hesson, J. C. 2014. Clearing up Culex Confusion. A Basis for Virus Vector Discrimination in Europe. Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology 1185. 56 pp. Uppsala: Acta Universitatis Upsaliensis. ISBN 978-91-554-9044-7. Mosquito species of the Culex genus are the enzootic vectors for several bird-associated viruses that cause disease in humans. In Europe, these viruses include Sindbis (SINV), West Nile and Usutu viruses. The morphologically similar females of Cx. torrentium and Cx. pipiens are potential vectors of these viruses, but difficulties in correctly identifying the mosquito species have caused confusion regarding their respective distribution, abundance, ecology, and consequently their importance as vectors. Species-specific knowledge from correctly identified field material is however of crucial importance since previous research shows that the relatively unknown Cx. torrentium is a far more efficient SINV vector than the widely recognized Cx. pipiens. The latter is involved in the transmission of several other viruses, but its potential importance for SINV transmission is debated.
    [Show full text]
  • Polymorphism of Mitochondrial COI and Nuclear Ribosomal ITS2 in the Culex Pipiens Complex and in Culex Torrentium (Diptera: Culicidae)
    © Comparative Cytogenetics, 2010 . Vol. 4, No. 2, P. 161-174. ISSN 1993-0771 (Print), ISSN 1993-078X (Online) Polymorphism of mitochondrial COI and nuclear ribosomal ITS2 in the Culex pipiens complex and in Culex torrentium (Diptera: Culicidae) E.V. Shaikevich, I.A. Zakharov N.I. Vavilov Institute of General Genetics, 119991 Moscow, Russia E-mails: [email protected], [email protected] Abstract. Polymorphism of the mtDNA gene COI encoding cytochrome C oxidase subunit I was studied in the mosquitoes Culex pipiens Linnaeus, 1758 and C. torren- tium Martini, 1925 from sixteen locations in Russia and in three laboratory strains of subtropical subspecies of the C. pipiens complex. Representatives of this complex are characterized by a high ecological plasticity and there are signifi cant ecophysiologi- cal differences between its morphologically similar members. The full-size DNA se- quence of the gene COI spans 1548 bp and has a total A+T content of 70.2 %. The TAA is a terminating codon in all studied representatives of the C. pipiens complex and C. torrentium. 64 variable nucleotide sites (4 %) were found, fi fteen haplotypes were detected, and two heteroplasmic specimens of C. torrentium were recorded. COI haplotype diversity was low in Wolbachia–infected populations of the C. pipiens complex. Monomorphic haplotypes were found in C. p. quinquefasciatus and C. p. pipiens f. molestus. Three haplotypes were detected for the C. p. pipiens, but these haplotypes were not population-specifi c. On the other hand, each of the ten studied Wolbachia-uninfected C. torrentium individuals from three different populations had unique mitochondrial haplotypes.
    [Show full text]
  • Single Mosquito Metatranscriptomics Identifies Vectors, Emerging Pathogens and Reservoirs in One Assay
    TOOLS AND RESOURCES Single mosquito metatranscriptomics identifies vectors, emerging pathogens and reservoirs in one assay Joshua Batson1†, Gytis Dudas2†, Eric Haas-Stapleton3†, Amy L Kistler1†*, Lucy M Li1†, Phoenix Logan1†, Kalani Ratnasiri4†, Hanna Retallack5† 1Chan Zuckerberg Biohub, San Francisco, United States; 2Gothenburg Global Biodiversity Centre, Gothenburg, Sweden; 3Alameda County Mosquito Abatement District, Hayward, United States; 4Program in Immunology, Stanford University School of Medicine, Stanford, United States; 5Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, United States Abstract Mosquitoes are major infectious disease-carrying vectors. Assessment of current and future risks associated with the mosquito population requires knowledge of the full repertoire of pathogens they carry, including novel viruses, as well as their blood meal sources. Unbiased metatranscriptomic sequencing of individual mosquitoes offers a straightforward, rapid, and quantitative means to acquire this information. Here, we profile 148 diverse wild-caught mosquitoes collected in California and detect sequences from eukaryotes, prokaryotes, 24 known and 46 novel viral species. Importantly, sequencing individuals greatly enhanced the value of the biological information obtained. It allowed us to (a) speciate host mosquito, (b) compute the prevalence of each microbe and recognize a high frequency of viral co-infections, (c) associate animal pathogens with specific blood meal sources, and (d) apply simple co-occurrence methods to recover previously undetected components of highly prevalent segmented viruses. In the context *For correspondence: of emerging diseases, where knowledge about vectors, pathogens, and reservoirs is lacking, the [email protected] approaches described here can provide actionable information for public health surveillance and †These authors contributed intervention decisions.
    [Show full text]
  • Meta-Transcriptomic Comparison of the RNA Viromes of the Mosquito Vectors Culex Pipiens and Culex Torrentium in Northern Europe
    bioRxiv preprint doi: https://doi.org/10.1101/725788; this version posted August 5, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Meta-transcriptomic comparison of the RNA viromes of the mosquito 2 vectors Culex pipiens and Culex torrentium in northern Europe 3 4 5 John H.-O. Pettersson1,2,3,*, Mang Shi2, John-Sebastian Eden2,4, Edward C. Holmes2 6 and Jenny C. Hesson1 7 8 9 1Department of Medical Biochemistry and Microbiology/Zoonosis Science Center, Uppsala 10 University, Sweden. 11 2Marie Bashir Institute for Infectious Diseases and Biosecurity, Charles Perkins Centre, 12 School of Life and Environmental Sciences and Sydney Medical School, the University of 13 Sydney, Sydney, New South Wales 2006, Australia. 14 3Public Health Agency of Sweden, Nobels väg 18, SE-171 82 Solna, Sweden. 15 4Centre for Virus Research, The Westmead Institute for Medical Research, Sydney, Australia. 16 17 18 *Corresponding author: [email protected] 19 20 Word count abstract: 247 21 22 Word count importance: 132 23 24 Word count main text: 4113 25 26 1 bioRxiv preprint doi: https://doi.org/10.1101/725788; this version posted August 5, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
    [Show full text]
  • The Role of Culex Pipiens Mosquitoes in Transmission of West Nile Virus in Europe Mosquitoes in Transmission of West Nile Virus in Europe Chantal B.F
    The role of Culex pipiens The role of Culex pipiens mosquitoes in transmission of West Nile virus in Europe mosquitoes in transmission of West Nile virus in Europe Chantal B.F. Vogels Nile virus in Europe Chantal B.F. mosquitoes in transmission of West Chantal B.F. Vogels 2017 Propositions 1. Northern Europe must prepare for West Nile virus transmission. (this thesis) 2. West Nile virus can affect the brain of both its vertebrate host and mosquito vector. (this thesis) 3. Conformism is a poor evolutionary strategy. 4. The individual-based character of the scientific review process makes publishing a lottery. 5. Sufficient sleep is key to a successful career. 6. Benefits of doing sports outweigh the risks of injury. Propositions belonging to the thesis, entitled ‘The role of Culex pipiens mosquitoes in transmission of West Nile virus in Europe’ Chantal B.F. Vogels Wageningen, 8 September 2017 The role of Culex pipiens mosquitoes in transmission of West Nile virus in Europe Chantal B.F. Vogels Thesis committee Promotor Prof. Dr M. Dicke Professor of Entomology Wageningen University & Research Co-promotor Dr C.J.M. Koenraadt Assistant professor, Laboratory of Entomology Wageningen University & Research Other members Prof. Dr R.A.A. van der Vlugt, Wageningen University & Research Dr M.A.H. Braks, National Institute for Public Health and the Environment, Bilthoven Dr L.S. van Overbeek, Wageningen University & Research Dr W.F. de Boer, Wageningen University & Research This research was conducted under the auspices of the C.T. de Wit Graduate School for Production Ecology & Resource Conservation The role of Culex pipiens mosquitoes in transmission of West Nile virus in Europe Chantal B.F.
    [Show full text]
  • Том 15. Вып. 2 Vol. 15. No. 2
    РОССИЙСКАЯ АКАДЕМИЯ НАУК Южный научный центр RUSSIAN ACADEMY OF SCIENCES Southern Scientific Centre CAUCASIAN ENTOMOLOGICAL BULLETIN Том 15. Вып. 2 Vol. 15. No. 2 Ростов-на-Дону 2019 © “Кавказский энтомологический бюллетень” составление, редактирование compiling, editing На титуле оригинальная фотография С. Маршалла (Stephen Marshall) Argyrochlamys marshalli Grichanov, 2010 Адрес для переписки: Максим Витальевич Набоженко [email protected] E-mail for correspondence: Dr Maxim Nabozhenko [email protected] Русская электронная версия журнала – http://www.ssc-ras.ru/ru/journal/kavkazskii_yntomologicheskii_byulleten/ English online version – http://www.ssc-ras.ru/en/journal/caucasian_entomological_bulletin/ Издание осуществляется при поддержке Южного научного центра Российской академии наук (Ростов-на-Дону) ­e journal is published by Southern Scientific Centre of the Russian Academy of Sciences under a Creative Commons Attribution- NonCommercial 4.0 International License Журнал индексируется в eLibrary.ru, ­omson Reuters (Zoological Record, Biological Abstracts, BIOSIS Previews, Russian Science Index Citation), ZooBank, DOAJ, Crossref ­e journal is indexed/referenced in eLibrary.ru, ­omson Reuters (Zoological Record, Biological Abstracts, BIOSIS Previews, Russian Science Index Citation), ZooBank, DOAJ, Crossref Техническое редактирование и компьютерная верстка номера – С.В. и М.В. Набоженко; корректура – С.В. Набоженко Кавказский энтомологический бюллетень 15(2): 233–235 © Caucasian Entomological Bulletin 2019 Synaphosus shirin Ovtsharenko, Levi et Platnik, 1994 (Gnaphosidae) и Holocnemus pluchei (Scopoli, 1763) (Pholcidae) – два новых вида пауков (Aranei) в фауне Кавказа Synaphosus shirin Ovtsharenko, Levi et Platnik, 1994 (Gnaphosidae) and Holocnemus pluchei (Scopoli, 1763) (Pholcidae) – two new species of spiders (Aranei) in the fauna of the Caucasus © А.В. Пономарёв1, Н.Ю. Снеговая2, В.Ю. Шматко1 © A.V. Ponomarev1, N.Yu. Snegovaya2, V.Yu.
    [Show full text]
  • Downloaded from NCBI on Mar 27, 2019.) 636 Default Parameters Were Used, Except the E-Value Cutoff Was Set to 1E-2
    bioRxiv preprint doi: https://doi.org/10.1101/2020.02.10.942854; this version posted February 13, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. bioRxiv PREPRINT 1 Single MOSQUITO 2 METATRANSCRIPTOMICS RECOVERS 3 MOSQUITO species, BLOOD MEAL 4 SOURces, AND MICROBIAL CARgo, 5 INCLUDING VIRAL DARK MATTER 1† 3† 2† 1*† 6 Joshua Batson , Gytis Dudas , Eric Haas-Stapleton , Amy L. Kistler , Lucy M. 1† 1† 1,4† 5† 7 Li , Phoenix Logan , Kalani Ratnasiri , Hanna Retallack *For CORRespondence: Chan ZuckERBERG Biohub, 499 ILLINOIS St, San FrANCISCO CA 94158; Alameda County [email protected] (AK) 8 1 2 Mosquito Abatement District, 23187 Connecticut St., Hayward, CA 94545; 3GothenburG † 9 These AUTHORS CONTRIBUTED EQUALLY Global Biodiversity Centre, Carl SkOTTSBERGS GATA 22B, 413 19, Gothenburg, Sweden; TO THIS WORK 10 PrOGRAM IN IMMUNOLOGY, StanforD University School OF Medicine, StanforD CA 94305; 11 4 Department OF Biochemistry AND Biophysics, University OF California San Francisco, San 12 5 FrANCISCO CA 94158 13 14 15 AbstrACT Mosquitoes ARE A DISEASE VECTOR WITH A COMPLEX ECOLOGY INVOLVING INTERACTIONS BETWEEN 16 TRANSMISSIBLE pathogens, ENDOGENOUS MICRobiota, AND HUMAN AND ANIMAL BLOOD MEAL SOURces. 17 Unbiased METATRANSCRIPTOMIC SEQUENCING OF INDIVIDUAL MOSQUITOES OffERS A STRAIGHTFORWARD AND 18 RAPID WAY TO CHARACTERIZE THESE dynamics. Here, WE PROfiLE 148 DIVERSE wild-caught MOSQUITOES 19 COLLECTED IN California, DETECTING SEQUENCES FROM eukaryotes, PRokaryotes, AND OVER 70 KNOWN AND 20 NOVEL VIRAL species.
    [Show full text]
  • Single Mosquito Metatranscriptomics Identifies Vectors, Emerging Pathogens and Reservoirs in One Assay
    bioRxiv preprint doi: https://doi.org/10.1101/2020.02.10.942854; this version posted December 21, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. Single mosquito metatranscriptomics identifies vectors, emerging pathogens and reservoirs in one assay Joshua Batson, Gytis Dudas, Eric Haas-Stapleton, Amy L. Kistler, Lucy M. Li, Phoenix Logan, Kalani Ratnasiri, Hanna Retallack Abstract Mosquitoes are major infectious disease-carrying vectors. Assessment of current and future risks associated with the mosquito population requires knowledge of the full repertoire of pathogens they carry, including novel viruses, as well as their blood meal sources. Unbiased metatranscriptomic sequencing of individual mosquitoes offers a straightforward, rapid and quantitative means to acquire this information. Here, we profile 148 diverse wild-caught mosquitoes collected in California and detect sequences from eukaryotes, prokaryotes, 24 known and 46 novel viral species. Importantly, sequencing individuals greatly enhanced the value of the biological information obtained. It allowed us to a) speciate host mosquito, b) compute the prevalence of each microbe and recognize a high frequency of viral co-infections, c) associate animal pathogens with specific blood meal sources, and d) apply simple co-occurrence methods to recover previously undetected components of highly prevalent segmented viruses. In the context of emerging diseases, where knowledge about vectors, pathogens, and reservoirs is lacking, the approaches described here can provide actionable information for public health surveillance and intervention decisions.
    [Show full text]
  • Establishment of Culex Modestus in Belgium and a Glance Into the Virome of Belgian
    bioRxiv preprint doi: https://doi.org/10.1101/2020.11.27.401372; this version posted February 22, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Establishment of Culex modestus in Belgium and a glance into the virome of Belgian 2 mosquito species 3 4 Lanjiao Wanga*, Ana Lucia Rosales Rosasa*, Lander De Coninck b, Chenyan Shi b, Johanna 5 Bouckaerta, Jelle Matthijnssens b, Leen Delanga# 6 aKU Leuven Department of Microbiology, Immunology and Transplantation, Rega Institute for Medical Research, Laboratory of Virology and Chemotherapy, Leuven, Belgium. bLaboratory of Viral Metagenomics, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium. 7 Running Head: Establishment of Culex modestus in Belgium 8 *Lanjiao Wang and Ana Lucia Rosales Rosas contributed equally to this work. # Address correspondence to Prof. Leen Delang, [email protected] 9 10 Abstract: 11 Culex modestus mosquitoes are known transmission vectors of West Nile virus and Usutu 12 virus. Their presence has been reported across several European countries, including only 13 one larva confirmed in Belgium in 2018. Mosquitoes were collected in the city of Leuven 14 and surroundings in the summer of 2019 and 2020. Species identification was performed 15 based on morphological features and partial sequences of the mitochondrial cytochrome 16 oxidase subunit 1 (COI) gene. The 107 mosquitoes collected in 2019 belonged to eight 17 mosquito species: Cx. pipiens (24.3%), Cx.
    [Show full text]
  • Culex Pipiens Biotype Molestus and Culex Torrentium Are Vector‑Competent for Usutu Virus Cora M
    Holicki et al. Parasites Vectors (2020) 13:625 https://doi.org/10.1186/s13071-020-04532-1 Parasites & Vectors RESEARCH Open Access German Culex pipiens biotype molestus and Culex torrentium are vector-competent for Usutu virus Cora M. Holicki1, Dorothee E. Scheuch1, Ute Ziegler1, Julia Lettow2,3, Helge Kampen2, Doreen Werner4 and Martin H. Groschup1* Abstract Background: Usutu virus (USUV) is a rapidly spreading zoonotic arbovirus (arthropod-borne virus) and a consider- able threat to the global avifauna and in isolated cases to human health. It is maintained in an enzootic cycle involv- ing ornithophilic mosquitoes as vectors and birds as reservoir hosts. Despite massive die-ofs in wild bird populations and the detection of severe neurological symptoms in infected humans, little is known about which mosquito species are involved in the propagation of USUV. Methods: In the present study, the vector competence of a German (i.e. “Central European”) and a Serbian (i.e. “Southern European”) Culex pipiens biotype molestus laboratory colony was experimentally evaluated. For comparative purposes, Culex torrentium, a frequent species in Northern Europe, and Aedes aegypti, a primarily tropical species, were also tested. Adult female mosquitoes were exposed to bovine blood spiked with USUV Africa 2 and subsequently incubated at 25 °C. After 2 to 3 weeks saliva was collected from each individual mosquito to assess the ability of a mosquito species to transmit USUV. Results: Culex pipiens biotype molestus mosquitoes originating from Germany and the Republic of Serbia and Cx. tor- rentium mosquitoes from Germany proved competent for USUV, as indicated by harboring viable virus in their saliva 21 days post infection.
    [Show full text]