Quote- "Seven spirochetal strains have been successfully recovered from mosquitoes and black flies: BR149 (Culex pipiens), BR151 (Cx. pipiens), BR173 (Cx. pipiens), BR177 (Cx. pipiens), BR193 (Aedes cinereus), BR208 (Cx. pipiens), and BR231 (Simulium noelleri).
The strains have been adapted to laboratory conditions (BSK-H Complete medium). Their preliminary determination based on 16S rRNA gene sequencing has shown that they differ from the Lyme disease spirochete Borrelia burgdorferi sensu lato as well as other members of the Order Spirochaetales indicating novel bacterial species in the Family Spirochaetaceae." Source
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The first study below was done in CT in 1988. The rest of the studies are from other countries, with the most recent ones on top of the list.
https://www.ncbi.nlm.nih.gov/pubmed/3170711
https://www.ncbi.nlm.nih.gov/pubmed/26631488
https://www.ncbi.nlm.nih.gov/pubmed/20618647
https://www.ncbi.nlm.nih.gov/pubmed/20047261
https://www.ncbi.nlm.nih.gov/pubmed/18274258
https://www.ncbi.nlm.nih.gov/pubmed/17199258
https://www.ncbi.nlm.nih.gov/pubmed/17044261
https://www.ncbi.nlm.nih.gov/pubmed/15236506
https://www.ncbi.nlm.nih.gov/pubmed/12498597
https://www.ncbi.nlm.nih.gov/pubmed/12088398
https://www.ncbi.nlm.nih.gov/pubmed/10885843
https://www.ncbi.nlm.nih.gov/pubmed/9513946
https://www.ncbi.nlm.nih.gov/pubmed/7711309
https://www.ncbi.nlm.nih.gov/pubmed/1983455
No Borrelia in Arkansas Spiders
https://www.ncbi.nlm.nih.gov/pubmed/10321947
Abstracts
Ticks Tick Borne Dis. 2016 Mar;7(2):256-63. doi: 10.1016/j.ttbdis.2015.10.018. Epub 2015 Nov 12.
Occurrence of Borrelia burgdorferi s.l. in different genera of mosquitoes (Culicidae) in Central Europe.
Melaun C1, Zotzmann S1, Santaella VG1, Werblow A1, Zumkowski-Xylander H2, Kraiczy P3, Klimpel S4.
Abstract
Lyme disease or Lyme borreliosis is a vector-borne infectious disease caused by spirochetes of the Borreliaburgdorferi sensu lato complex. Some stages of the borrelial transmission cycle in ticks (transstadial, feeding and co-feeding) can potentially occur also in insects, particularly in mosquitoes. In the present study, adult as well as larval mosquitoes were collected at 42 different geographical locations throughout Germany.
This is the first study, in which German mosquitoes were analyzed for the presence of Borrelia spp.
Targeting two specific borrelial genes, flaB and ospA encoding for the subunit B of flagellin and the outer surface protein A, the results show that DNA of Borrelia afzelii, Borrelia bavariensis and Borrelia garinii could be detected in ten Culicidae species comprising four distinct genera (Aedes, Culiseta, Culex, and Ochlerotatus).
Positive samples also include adult specimens raised in the laboratory from wild-caught larvae indicating that transstadial and/or transovarial transmission might occur within a given mosquito population.
Copyright © 2015 Elsevier GmbH. All rights reserved.
KEYWORDS:
Borrelia burgdorferi complex; Culicidae; Lyme borreliosis; Mosquitoes; Spirochetes
PMID:
26631488
DOI:
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Microbiol Res. 2014 May-Jun;169(5-6):348-52. doi: 10.1016/j.micres.2013.10.002. Epub 2013 Oct 19.
Selected phenotypic features of BR91, a unique spirochaetal strain isolated from the Culex pipiens mosquito.
Šikutová S1, Bunková L2, Krejčí E3, Halouzka J1, Sanogo YO4, Rudolf I5.
1
Institute of Vertebrate Biology, v.v.i., Academy of Sciences of the Czech Republic, Květná 8, 603 65 Brno, Czech Republic.
2
Tomas Bata University, Faculty of Technology, Department of Environmental Protection and Engineering, náměstí T. G. Masaryka 275, 762 72 Zlin, Czech Republic.
3
Institute of Public Health, Department of Bacteriology, Partyzánské náměstí 7, 702 00 Ostrava, Czech Republic.
4
University of Illinois at Urbana-Champaign, WM Keck Centre for Comparative & Functional Genomics, Urbana, IL 61801, USA.
5
Institute of Vertebrate Biology, v.v.i., Academy of Sciences of the Czech Republic, Květná 8, 603 65 Brno, Czech Republic; Masaryk University, Department of Experimental Biology, Section of Microbiology, Kotlářská 2, 611 37 Brno, Czech Republic. Electronic address: rudolf@ivb.cz.
Abstract
Growth temperature range, resistance to selective antibiotics, activities of 23 enzymes, protein fingerprints and fatty acids composition of the spirochaetal strain BR91, isolated from the Culex pipiens mosquito, were tested. The spirochaetes were grown in BSK-H Complete liquid medium. The optimal in vitro growth temperature of the strain was 33 °C. Strain BR91 was sensitive to trimethoprim, nalidixic acid, 5-fluorouracil, and tolerated phosphomycin. The strain produced acid and alkaline phosphatase, esterase (C4), esterase-lipase (C8), leucine arylamidase, naphthol-AS-BI-phosphohydrolase and α-fucosidase. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) assay revealed several major proteins in the size range of 13-16 kDa, 22-30 kDa and 37-131 kDa. Fatty acid methyl ester (FAME) analysis showed that C₁₄:₀, C₁₆:₀, C₁₈:₁ ω9c and summed feature 5 (C₁₈:₂ ω6,9c and/or C₁₈:₀ anteiso) are major fatty acids. This study highlights certain phenotypic differences between strain BR91 and the Lyme disease spirochaete Borrelia burgdorferi, and supports the hypothesis that strain BR91 represents a unique taxonomical entity in a system of spirochaetal species.
Copyright © 2013 Elsevier GmbH. All rights reserved.
KEYWORDS:
Culex pipiens; Enzyme activity; FAME; SDS-PAGE; Spirochaete
PMID:
24239193
DOI:
[Indexed for MEDLINE]
Free full text
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J Vector Ecol. 2010 Jun;35(1):50-5.
Novel spirochetes isolated from mosquitoes and black flies in the Czech Republic.
Sikutová S, Halouzka J, Mendel J, Knoz J, Rudolf I.
Institute of Vertebrate Biology, v.v.i., Academy of Sciences of the Czech Republic, Kvetná 8, CZ-603 65 Brno, Czech Republic.
Abstract
During the years 1999-2002, a total of 4,898 individuals of 26 species of hematophagous insects (4,149 mosquitoes, 583 black flies, and 166 tabanid flies) was examined for the presence of spirochetes using dark-field microscopy. There was an overall recovery of spirochetes from the midguts of Culicidae and Simuliidae of 23.5% and 11.4%, respectively. Spirochetes were not detected in Tabanidae.
Seven spirochetal strains have been successfully recovered from mosquitoes and black flies: BR149 (Culex pipiens), BR151 (Cx. pipiens), BR173 (Cx. pipiens), BR177 (Cx. pipiens), BR193 (Aedes cinereus), BR208 (Cx. pipiens), and BR231 (Simulium noelleri).
The strains have been adapted to laboratory conditions (BSK-H Complete medium). Their preliminary determination based on 16S rRNA gene sequencing has shown that they differ from the Lyme disease spirochete Borrelia burgdorferi sensu lato as well as other members of the Order Spirochaetales indicating novel bacterial species in the Family Spirochaetaceae.
PMID: 20618647
***
Ann Agric Environ Med. 2008 Dec;15(2):225-30.
Ribotyping and whole-cell protein analysis of spirochetes isolated from arthropods in the Czech Republic.
Bunková L, Svec P, Halouzka J, Rudolf I, Nemec M.
Department of Lipids, Tensides and Cosmetics Technology, Faculty of Technology, Tomas Bata University-Zlin, Namesti T. G. Masaryka 275, Zlin, Czech Republic. bunkova@ft.utb.cz
Abstract
In the presented work, six Borrelia burgdorferi sensu lato isolates were recovered from Ixodes ricinus ticks and one strain of spirochete with uncertain taxonomic position isolated from Culex pipiens mosquito collected in the Valtice area (South Moravia, Czech Republic), as well as representative type strains, were further characterized by ribotyping and whole-cell protein fingerprinting.
The ribotype patterns obtained with HindII and EcoRV ribotype patterns showed good correlations with the recently proposed genotypes and grouped each genospecies in a well-separated cluster. The whole-cell protein profiles of genospecies were more heterogeneous than the ribotype patterns. The non-identified spirochetal strain was clearly separated from the Borrelia isolates in both methods thus proving their good capacity to characterize Borrelia burgdorferi sensu lato strains.
PMID: 19061259 [PubMed - indexed for MEDLINE]Free Article
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Folia Biol (Krakow). 2007;55(3-4):143-6.
Ticks and mosquitoes as vectors of Borrelia burgdorferi s. l. in the forested areas of Szczecin.
Kosik-Bogacka DI, Kuźna-Grygiel W, Jaborowska M.
Department of Biology and Medical Parasitology, Pomeranian Medical University, Powstancow Wielkopolskich Av. 72, 70-111 Szczecin, Poland. kodan@sci.pam.szczecin.pl
Abstract
The aim of the study was to determine the infection level of adult forms and larvae of ticks and mosquitoes with Borrelia burgdorferi in the forested areas of Szczecin. A total of 1699 ticks Ixodes ricinus, including 1422 nymphs, 277 adult forms and 2862 mosquito females representing the genera Aedes (89.6%) and Culex (10.4%) were collected between the years 2004 and 2005.
A further 3746 larvae and 1596 pupae of Culex pipiens pipiens were colleted from water bodies. Borrelia burgdorferi s. l. was detected in the arthropods by the method of indirect immunofluorescence assay (IFA). A positive immunological reaction was detected in 16.6% of the adult forms and in 16.5% of the nymphs of Ixodes ricinus.
Spirochetes were also detected in 1.7% of mosquito females, 3.2% of larvae and in 1.6% of pupae of Culex pipiens pipiens. The results of the present study confirm that contact with ticks constitutes the main risk of contracting Lyme disease, although mosquitoes play a role as vectors as well.
PMID: 18274258 [PubMed - indexed for MEDLINE]
***
Infect Genet Evol. 2009 May;9(3):308-13. Epub 2008 Jan 3.
Genomic resources for invertebrate vectors of human pathogens, and the role of VectorBase.
Megy K, Hammond M, Lawson D, Bruggner RV, Birney E, Collins FH.
EMBL, European Bioinformatics Institute, Wellcome Trust Genome Campus, Hinxton CB10 1SD, UK. kmegy@ebi.ac.uk
Abstract
High-throughput genome sequencing techniques have now reached vector biology with an emphasis on those species that are vectors of human pathogens. The first mosquito to be sequenced was Anopheles gambiae, the vector for Plasmodium parasites that cause malaria.
Further mosquitoes have followed: Aedes aegypti (yellow fever and dengue fever vector) and Culex pipiens (lymphatic filariasis and West Nile fever). Species that are currently in sequencing include the body louse Pediculus humanus (Typhus vector), the triatomine Rhodnius prolixus (Chagas disease vector) and the tick Ixodes scapularis (Lyme disease vector).
The motivations for sequencing vector genomes are to further understand vector biology, with an eye on developing new control strategies (for example novel chemical attractants or repellents) or understanding the limitations of current strategies (for example the mechanism of insecticide resistance); to analyse the mechanisms driving their evolution; and to perform an exhaustive analysis of the gene repertory. The proliferation of genomic data creates the need for efficient and accessible storage.
We present VectorBase, a genomic resource centre that is both involved in the annotation of vector genomes and act as a portal for access to the genomic information (http://www.vectorbase.org).
PMID: 18262474 [PubMed - indexed for MEDLINE]PMCID: PMC2720521Free PMC Article
***
Ann Agric Environ Med. 2006;13(2):345-8.
Isolation of Borrelia afzelii from overwintering Culex pipiens biotype molestus mosquitoes.
Zákovská A, Capková L, Serý O, Halouzka J, Dendis M.
Department of Comparative Animal Physiology and General Zoology, Faculty of Science, Masaryk University, Kotlarska 2, CZ-611 37 Brno, Czech Republic. alenazak@sci.muni.cz
Abstract
A total of 662 samples (winter period: 469; summer period: 193 specimens) of female mosquitoes of the genus Culex, Aedes and Anopheles were collected during the period March 2000-April 2001 from the locality of Vysoke Myto (Eastern Bohemia, Czech Republic). They were examined by dark field microscopy for the presence of spirochetes.
The motile spirochetes were observed in 4.2 % of all species of investigated mosquitoes. One spirochetal strain out of the 8 isolation attempts (BRZ14) was obtained (cultivation rate was 12.5 %) and the spirochetal strain was then successfully cultivated and identified using PCR for the presence of Borrelia burgdorferi s.l., and subsequently with the RFLP as genomospecies Borrelia afzelii.
This strain was derived from overwintering Culex (Culex) pipiens biotype molestus female mosquitoe. This is apparently one of the sporadic cases of the occurrence of pathogenic borreliae in haematophagous arthropods, other than Ixodes ricinus complex ticks.
PMID: 17199258 [PubMed - indexed for MEDLINE]Free Article
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Folia Biol (Krakow). 2006;54(1-2):55-9.
Borrelia burgdorferi sensu lato infection in mosquitoes from Szczecin area.
Kosik-Bogacka DI, Kuźna-Grygiel W, Górnik K.
Chair and Department of Biology and Medical Parasitology, Pomeranian Medical University, Powstańców Wielkopolskich 72, 70-111 Szczecin, Poland. kodan@sci.pam.szczecin.pl
Abstract
The aim of the study was to determine the level of infection in mosquitoes with spirochetes Borrelia burgdorferi sensu lato in the woody areas of Szczecin. The mosquitoes were collected from May to September 2003.
The spirochetes, Borrelia burgdorferi s. l., present in mosquitoes were detected in mosquitoes with indirect immunofluorescence assay (IFA) using rabbit anti-Borrelia burgdorferi antibodies and goat anti-rabbit IgG marked with fluorescein isocyanate (FITC).
A total of 1557 females and 58 males were collected. They represented the genera Aedes (63%) and Culex (37%). The infection level of the mosquitoes from the area studied amounted to 1.7%. The results of the present study confirm the potential of these arthropods to spread Lyme borreliosis.
PMID: 17044261 [PubMed - indexed for MEDLINE]
***
Cell Biochem Funct. 2007 Mar-Apr;25(2):185-8.
Seroprevalence study of Tick-borne encephalitis, Borrelia burgdorferi, Dengue and Toscana virus in Turin Province.
Pugliese A, Beltramo T, Torre D.
Department of Medical and Surgical Sciences, Section of Clinical Microbiology of Turin University, Amedeo di Savoia Hospital, Turin, Italy. agostino.pugliese@unito.it
Abstract
Tick borne encephalitis virus (TBEV) is present in some European countries and it is transmitted by a tick bite. Ixodes ricinus is the main vector of the infection in Italy, where fortunately clinical neurological manifestations, typical of the more serious phase of the disease, are very rarely observed.
This behaviour is different from other endemic Euroasiatic areas where numerous cases of encephalitis are described. However TBE transmission has not been widely investigated in Italy and available epidemiological data have been obtained only by studies performed in Central and Northern Regions of the country.
In addition seroepidemiological researches were made prevalently on subjects at high risk of tick bite, such as hunters or forest guards from Trentin and Central Italy. No precise information about TBE virus diffusion was available in the Piedmont before our investigations.
We found that hunters and wild boar breeders seem to be particularly exposed to the risk of TBE virus infection in Turin Province and in particular in the Susa valley, although no neurological involvement was observed in our population.
In particular a seroprevalence of about 5% was detected by the use of purified antigens ELISA test, amongst the subjects at high risk of tick bite. Moreover low risk individuals showed a seroprevalence of below 2%. In addition a parallel seroepidemiological study was performed in Turin Province for Borrelia burgdorferi, the aetiological agent of Lyme disease, also transmitted by tick bite (e.g. Ixodes ricinus), for Dengue and Toscana (TOS) arboviruses, respectively transmitted by Aedes mosquitoes and phlebotomes.
Data reported here demonstrate only a sporadic presence in our population of antibodies against Borrelia and Dengue infection. Moreover using an ELISA test performed with viral purified nucleoprotein, we reported a total percentage of about 3% of subjects positive for TOSV. Copyright 2005 John Wiley & Sons, Ltd.
PMID: 16312014 [PubMed - indexed for MEDLINE]
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Ann Agric Environ Med. 2004;11(1):105-8.
The prevalence of spirochete Borrelia burgdorferi sensu lato in ticks Ixodes ricinus and mosquitoes Aedes spp. within a selected recreational area in the city of Szczecin.
Kosik-Bogacka D, Kuźna-Grygiel W, Bukowska K.
Department of Biology and Medical Parasitology, Pomeranian Medical University, 70-111 Szczecin, Al. Powstánców Wielkopolskich 72, Poland. kodan@sci.pam.szczecin.pl
Abstract
The aim of this study was to determine the prevalence of spirochete Borrelia burgdorferi s.l. in ticks Ixodes ricinus and mosquitoes Aedes spp. within the Bukowa Forest, collected between 2000 and 2001. The study covered 215 ticks (193 nymphs and 22 adults) and 947 mosquitoes female of the genus Aedes.
Spirochetes of Borrelia burgdorferi s.l. were detected in the arthropods studied with the method of indirect immunofluorescence assay (IFA). Positive readings of the immunological reaction were stated in 17.7 % of the collected nymphs and adult forms of Ixodes ricinus, and in 0.8 % of mosquito females of the genus Aedes.
The number of B. burgdorferi observed in a view field (400 x) of microscopic preparations of all infected mosquitoes and about 10 % of the infected ticks, ranged from 1-10. This number in 50 % of the nymphs was from 11-50 spirochetes.
View fields of the preparations of the other 50 % of nymphs and adult forms featured more then 50 spirochetes.
The observed low values of the prevalence and infection intensity of female mosquitoes Aedes spp. compared to ticks suggest that the former do not pose a serious epidemiological threat in the spreading of Lyme disease.
PMID: 15236506 [PubMed - indexed for MEDLINE]Free Article
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Ann Agric Environ Med. 2009;16(2):273-6.
What is the percentage of pathogenic borreliae in spirochaetal findings of mosquito larvae?
Nejedla P1, Norek A, Vostal K, Zakovska A.
Abstract
In this study, larvae (1,179 ex.) of mosquito genera Culex were examined for the presence of spirochaetes by Dark Field Microscopy (DFM) at the locality of Blansko (Czech Republic) in of 2004-2008. DFM spirochaete positive samples (25.4 %) were investigated by nested PCR; only 4 samples were positive for the presence of Borrelia burgdorferi sensu lato, which is 0.3 % of the total examined samples. We can conclude that only a low percentage of pathogenic borreliae are presented in mosquito larvae, while the spirochaete of undefined genera infect larvae in high amounts.
PMID:
20047261
[Indexed for MEDLINE]
Free full text
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Ann Agric Environ Med. 2002;9(2):257-9.
Positive findings of Borrelia burgdorferi in Culex (Culex) pipiens pipiens larvae in the surrounding of Brno city determined by the PCR method.
Zákovská A, Nejedla P, Holíková A, Dendis M.
Department of Comparative Animal Physiology and General Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic. alenazak@sci.muni.cz
Abstract
After first finding Borrelia in the midgut of imago mosquitoes, we concentrated on the presence of Borrelia in mosquito development stages--larvae of the third or fourth instar.
In the summer season in the years of 2000-2001 a total of 439 Culex (Culex) pipiens pipiens larvae were collected from a barrel of rainwater in the Obrany holiday area of Brno city (East Moravia, Czech Republic).
The larvae midgut was observed under dark-field microscopy. Ten DFM positive samples (2.28%) were further analysed using the single-tube nested PCR method for the presence of flagellum DNA sequence specific for Borrelia burgdorferi sensu lato, of which 5 were positive.
Borrelian positivity of Culex (C.) pipiens pipiens larvae was 1.14%. One spirochete isolated strain in BSK-H medium was obtained. PCR detection for borrelian DNA of the isolated strain was negative. From these results we can conclude that a low percentage Borreliae can be also found in mosquito larvae and are likely to survive into imago stage.
PMID: 12498597 [PubMed - indexed for MEDLINE]Free Article
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Ann Agric Environ Med. 2002;9(1):55-7.
Detection of Borrelia burgdorferi sensu lato in mosquitoes (Culicidae) in recreational areas of the city of Szczecin.
Kosik-Bogacka D, Bukowska K, Kuźna-Grygiel W.
Department of Biology and Medical Parasitology, Pomeranian Medical University, 70-111 Szczecin, al. Powstancow Wielkopolskich 72, Poland. kodan@sci.pam.szczecin.pl
Abstract
The mosquitoes were caught within the period of their highest activity, i.e. June August 2001, in recreational areas of the city of Szczecin. Spirochetes, Borrelia burgdorferi sensu lato were detected in mosquitoes with the aid of the method of indirect immunofluorescence (IFA) using rabbit anti-Borrelia burgdorferi antibodies and goat anti-rabbit IgG marked with fluorescein isothiocyanate (FITC). A total of 639 mosquito females representing genera Aëdes (99.1%) and Culex (0.9%) were collected.
The mean value of the infection rate of mosquitoes from the area studied was 1.25%. The highest infection rate was recorded in June (3.2%), while the lowest -- in July (0.6%). All mosquitoes infected with Borrelia burgdorferi spirochetes belonged to the genus Aëdes. The results of the present study confirm a potential role of those arthropods in epidemiology of Lyme borreliosis
PMID: 12088398 [PubMed - indexed for MEDLINE]Free Article