Rapid Differentiation of Vector Species: Development of Microsatellite Markers for Population Genetics of Biting Midges and a Potential Tool for Species Identification of Culicoides Sonorensis

Abstract Background: Proper vector surveillance relies on the ability to identify species of interest accurately and efficiently, though this can be difficult in groups containing cryptic species. Culicoides is a genus of small biting flies responsible for the transmission of numerous pathogens to a multitude of vertebrates. Regarding pathogen transmission, the C. variipennis species complex is of particular interest in North America. Of the six species within this group, only C. sonorensis is a proven vector of bluetongue virus and epizootic hemorrhagic disease virus. Unfortunately, subtle morphological differences, cryptic species, and mitonuclear discordance make species identification in the C. variipennis complex challenging. Recently, a SNP analysis enabled discrimination between the species of this group; however, this demanding approach is not practical for vector surveillance. Methods: The aim of the current study was to develop a reliable and affordable way of differentiating the species within the C. variipennis complex, especially C. sonorensis. Twenty-five putative microsatellite markers were identified using the C. sonorensis genome and tested for amplification within five species of the C. variipennis complex. Machine learning was then used to determine which markers best explain the genetic differentiation between species. This led to the development of a subset of four and seven markers which were also tested for species differentiation.Results: A total of 21 microsatellite markers were successfully amplified in the species tested. Clustering analyses of all of these markers recover the same species-level identification as the previous SNP data. Additionally, the subset of seven markers was equally capable of accurately differentiating the members of the C. variipennis complex as the 21 microsatellite markers. Finally, one microsatellite marker (C508) was found to be species-specific, only amplifying in the vector species C. sonorensis among the samples tested. Conclusions: These microsatellites provide an affordable way in which to differentiate the species of the C. variipennis complex and could lead to a better understanding of the species dynamics within this group. Additionally, after further testing, marker C508 may allow for the identification of C. sonorensis with a single-tube assay, potentially providing a powerful new tool for vector surveillance in North America.

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Benchmarking DNA barcodes: an assessment using available primate sequences

Genome ◽

10.1139/g06-025 ◽

2006 ◽

Vol 49 (7) ◽

pp. 851-854 ◽

Cited By ~ 48

Author(s):

Mehrdad Hajibabaei ◽

Gregory AC Singer ◽

Donal A Hickey

Keyword(s):

New Species ◽

Cryptic Species ◽

Dna Barcoding ◽

Species Identification ◽

Dna Sequences ◽

Phylogenetic Relationships ◽

Primate Species ◽

Dna Barcodes ◽

Global Biodiversity ◽

Efficient Sequence

DNA barcoding has been recently promoted as a method for both assigning specimens to known species and for discovering new and cryptic species. Here we test both the potential and the limitations of DNA barcodes by analysing a group of well-studied organisms—the primates. Our results show that DNA barcodes provide enough information to efficiently identify and delineate primate species, but that they cannot reliably uncover many of the deeper phylogenetic relationships. Our conclusion is that these short DNA sequences do not contain enough information to build reliable molecular phylogenies or define new species, but that they can provide efficient sequence tags for assigning unknown specimens to known species. As such, DNA barcoding provides enormous potential for use in global biodiversity studies.Key words: DNA barcoding, species identification, primate, biodiversity.

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Geographic range in Chthamalus along the west coast of North America

Journal of the Marine Biological Association of the United Kingdom ◽

10.1017/s0025315405011227h ◽

2005 ◽

Vol 85 (2) ◽

pp. 327-331 ◽

Cited By ~ 17

Author(s):

J.P. Wares ◽

A.E. Castañeda

Keyword(s):

North America ◽

Cryptic Species ◽

Genetic Markers ◽

West Coast ◽

Environmental Gradients ◽

Geographic Range ◽

The West ◽

Barnacle Species ◽

Range Overlap

Identification of the range boundaries and microgeographic distribution of cryptic species is greatly facilitated by the use of genetic markers. Here we characterize the geographic range overlap between two cryptic species, Chthamalus fissus and C. dalli, and show that as with other barnacle species, their distribution and abundance is probably dictated more by microhabitat characteristics and the presence of conspecifics than broader environmental gradients. We also show that C. dalli appears to be panmictic across the studied range.

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Evaluation of A Baculovirus-Expressed VP2 Subunit Vaccine for the Protection of White-Tailed Deer (Odocoileus virginianus) from Epizootic Hemorrhagic Disease

Vaccines ◽

10.3390/vaccines8010059 ◽

2020 ◽

Vol 8 (1) ◽

pp. 59

Author(s):

Sun Young Sunwoo ◽

Leela E. Noronha ◽

Igor Morozov ◽

Jessie D. Trujillo ◽

In Joong Kim ◽

...

Keyword(s):

North America ◽

Neutralizing Antibodies ◽

Subunit Vaccine ◽

Rna Virus ◽

Expression System ◽

Clinical Disease ◽

Vascular Lesions ◽

Hemorrhagic Disease ◽

Epizootic Hemorrhagic Disease ◽

Homologous Virus

Epizootic hemorrhagic disease virus (EHDV) is an arthropod-transmitted RNA virus and the causative agent of epizootic hemorrhagic disease (EHD) in wild and domestic ruminants. In North America, white-tailed deer (WTD) experience the highest EHD-related morbidity and mortality, although clinical disease is reported in cattle during severe epizootics. No commercially licensed EHDV vaccine is available in North America. The objective of this study was to develop and evaluate a subunit vaccine candidate to control EHD in WTD. Recombinant VP2 (rVP2) outer capsid proteins of EHDV serotypes 2 (EHDV-2) and 6 (EHDV-6) were produced in a baculovirus-expression system. Mice and cattle vaccinated with EHDV-2 or EHDV-6 rVP2 produced homologous virus-neutralizing antibodies. In an immunogenicity/efficacy study, captive-bred WTD received 2 doses of EHDV-2 rVP2 or sham vaccine, then were challenged with wild-type EHDV-2 at 30 d post vaccination. None of the rVP2-vaccinated deer developed clinical disease, no viral RNA was detected in their blood or tissues (liver, lung, spleen, kidney), and no EHDV-induced lesions were observed. Sham-vaccinated deer developed clinical disease with viremia and typical EHD vascular lesions. Here, we demonstrate a rVP2 subunit vaccine that can provide protective immunity from EHDV infection and which may serve as an effective tool in preventing clinical EHD and reducing virus transmission.

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Multivariate discrimination among cryptic species of the mite genus Chaetodactylus (Acari: Chaetodactylidae) associated with bees of the genus Lithurgus (Hymenoptera: Megachilidae) in North America

Experimental and Applied Acarology ◽

10.1023/b:appa.0000032927.78170.c1 ◽

2004 ◽

Vol 33 (3) ◽

pp. 157-182 ◽

Cited By ~ 8

Author(s):

Pavel B. Klimov ◽

Barry M. OConnor

Keyword(s):

North America ◽

Cryptic Species ◽

Mite Genus

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Molecular evidence of allodiploidy in F1 gametophytic blades from a cross between Neopyropia yezoensis and a cryptic species of the Neopyropia yezoensis complex (Bangiales, Rhodophyta) by the use of microsatellite markers

Aquaculture Reports ◽

10.1016/j.aqrep.2020.100489 ◽

2020 ◽

Vol 18 ◽

pp. 100489

Author(s):

Kyosuke Niwa ◽

Fumina Sano ◽

Takashi Sakamoto

Keyword(s):

Microsatellite Markers ◽

Cryptic Species ◽

Molecular Evidence

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Psilolechia clavulifera, a Lichen Species New to Canada

The Canadian Field-Naturalist ◽

10.22621/cfn.v124i2.1058 ◽

2010 ◽

Vol 124 (2) ◽

pp. 181 ◽

Cited By ~ 2

Author(s):

Chris Lewis

Keyword(s):

North America ◽

Cryptic Species ◽

First Record ◽

Lichen Species

Psilolechia clavulifera is reported as new to Canada, where it was been found growing under a shaded rock overhang near Temagami, Ontario. This is the first record of the species for Canada. This occurrence represents only the fourth documented record of the species for North America since it was first discovered in 1939 by J. Lowe. This specimen was found growing with other rarely collected cryptic species found in unique microhabitats: Psilolechia lucida, Protothelenella corrosa, and Microcalicium arenarium.

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Three Colopterus Beetle Species Carry the Oak Wilt Fungus to Fresh Wounds on Red Oak in Missouri

Plant Disease ◽

10.1094/pdis-92-2-0270 ◽

2008 ◽

Vol 92 (2) ◽

pp. 270-275 ◽

Cited By ~ 14

Author(s):

Maya Hayslett ◽

Jennifer Juzwik ◽

Bruce Moltzan

Keyword(s):

Abundant Species ◽

Pathogen Transmission ◽

Red Oak ◽

Beetle Species ◽

Vector Species ◽

Oak Wilt ◽

Ceratocystis Fagacearum ◽

Oak Trees ◽

The Family ◽

Nitidulid Beetles

Beetles in the family Nitidulidae can transmit the oak wilt fungus, Ceratocystis fagacearum, to fresh wounds on healthy oak trees, leading to infection and disease development. Historically, nitidulid beetles have not been considered important vectors of the pathogen in Missouri. Studies were conducted in the spring of 2005 and 2006 to determine frequencies of nitidulid beetle species contaminated with C. fagacearum visiting fresh wounds on red oak trees in central Missouri. Colopterus truncatus, C. niger, and C. semitectus were the most abundant species collected from fresh wounds and the only species found to be contaminated with Ceratocystis fagacearum. Of 230 beetles assayed for C. fagacearum, 23 yielded the fungus. Contamination frequencies were higher for beetles collected in April than May; no beetles collected in June were contaminated. We hypothesize that Colopterus truncatus, C. niger, and C. semitectus are principal nitidulid beetle vector species in Missouri during spring. The risk for pathogen transmission by these beetles appears to be greatest in April and least in June.

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Transmission and Epidemiology of Bluetongue and Epizootic Hemorrhagic Disease in North America: Current Perspectives, Research Gaps, and Future Directions

Vector-Borne and Zoonotic Diseases ◽

10.1089/vbz.2014.1703 ◽

2015 ◽

Vol 15 (6) ◽

pp. 348-363 ◽

Cited By ~ 60

Author(s):

Mark G. Ruder ◽

Timothy J. Lysyk ◽

David E. Stallknecht ◽

Lane D. Foil ◽

Donna J. Johnson ◽

...

Keyword(s):

North America ◽

Hemorrhagic Disease ◽

Research Gaps ◽

Future Directions ◽

Epizootic Hemorrhagic Disease

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Geographic Distribution of Cryptic Species of Plasmopara viticola Causing Downy Mildew on Wild and Cultivated Grape in Eastern North America

Phytopathology ◽

10.1094/phyto-08-13-0225-r ◽

2014 ◽

Vol 104 (7) ◽

pp. 692-701 ◽

Cited By ~ 22

Author(s):

Mélanie Rouxel ◽

Pere Mestre ◽

Anton Baudoin ◽

Odile Carisse ◽

Laurent Delière ◽

...

Keyword(s):

North America ◽

Cryptic Species ◽

Downy Mildew ◽

Geographic Distribution ◽

Host Species ◽

Large Scale ◽

Plasmopara Viticola ◽

Eastern North America ◽

Grape Downy Mildew ◽

Parthenocissus Quinquefolia

The putative center of origin of Plasmopara viticola, the causal agent of grape downy mildew, is eastern North America, where it has been described on several members of the family Vitaceae (e.g., Vitis spp., Parthenocissus spp., and Ampelopsis spp.). We have completed the first large-scale sampling of P. viticola isolates across a range of wild and cultivated host species distributed throughout the above region. Sequencing results of four partial genes indicated the presence of a new P. viticola species on Vitis vulpina in Virginia, adding to the four cryptic species of P. viticola recently recorded. The phylogenetic analysis also indicated that the P. viticola species found on Parthenocissus quinquefolia in North America is identical to Plasmopara muralis in Europe. The geographic distribution and host range of five pathogen species was determined through analysis of the internal transcribed spacer polymorphism of 896 isolates of P. viticola. Among three P. viticola species found on cultivated grape, one was restricted to Vitis interspecific hybrids within the northern part of eastern North America. A second species was recovered from V. vinifera and V. labrusca, and was distributed across most of the sampled region. A third species, although less abundant, was distributed across a larger geographical range, including the southern part of eastern North America. P. viticola clade aestivalis predominated (83% of isolates) in vineyards of the European winegrape V. vinifera within the sampled area, indicating that a single pathogen species may represent the primary threat to the European host species within eastern North America.

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