During a survey of termites in Wisconsin, one colony was found from a different habitat than the remaining populations. This observation led to further genetic testing which resulted in a determination of Reticulitermes tibialis Banks. This is the first record of a termite species other than Reticulitermes flavipes (Kollar) to be established in the state

Arango, Rachel A

English

Valparaiso University

The Great Lakes Entomologist Volume 48 Numbers 1 & 2 - Spring/Summer 2015 Numbers 1 & 2 - Spring/Summer 2015 Article 8 April 2015 First Record of the Arid-Land Termite, Reticulitermes Tibialis Banks, in Wisconsin Rachel A. Arango Follow this and additional works at: https://scholar.valpo.edu/tgle  Part of the Entomology Commons Recommended Citation Arango, Rachel A. 2015. "First Record of the Arid-Land Termite, Reticulitermes Tibialis Banks, in Wisconsin," The Great Lakes Entomologist, vol 48 (1) Available at: https://scholar.valpo.edu/tgle/vol48/iss1/8 This Peer-Review Article is brought to you for free and open access by the Department of Biology at ValpoScholar. It has been accepted for inclusion in The Great Lakes Entomologist by an authorized administrator of ValpoScholar. For more information, please contact a ValpoScholar staff member at scholar@valpo.edu. 98 THE GREAT LAKES ENTOMOLOGIST Vol. 48, Nos. 1 - 2First Record of the Arid-Land Termite,  Reticulitermes tibialis Banks, in WisconsinRachel A. ArangoAbstractDuring a survey of termites in Wisconsin, one colony was found from a different habitat than the remaining populations.  This observation led to fur-ther genetic testing which resulted in a determination of Reticulitermes tibialis Banks.  This is the first record of a termite species other than Reticulitermes flavipes (Kollar) to be established in the state. ____________________Numerous records of the eastern subterranean termite, Reticulitermes flavipes (Kollar), have been recorded from Wisconsin, a species which has long been thought to be the only termite species established in the state.  Outside of an urban environment, colonies of R. flavipes are often found in shady, wooded areas under logs and in decaying trees.  During a survey of Wisconsin termite populations, one site at Hardscrabble Prairie in Hazel Green showed termite activity under logs and rocks in the middle of a sunny prairie with very little shade cover, prompting further evaluation.As morphological identification of termites is often difficult, genetic-based techniques were utilized for species determination to support observed physical characteristics.  Termites were collected 24 July 2013 in cardboard placed under logs in the middle of the prairie.  Deoxyribonucleic acid (DNA) was extracted from three individual worker termites using the Promega Wizard genomic DNA purification kit following the protocol for isolation of genomic DNA from animal tissues.  Termite DNA (~50ng/µl) was amplified using primers TL2-J-3037 (5’-ATGGCAGATTAGTGCAATGG-3’) and TK-N-3785 (5’-GTTTAAGAGACCAG-TACTTG-3’), which are specific for the mitochondrial COII genes as chosen by Lim and Forschler (2012).  Polymerase chain reaction (PCR) was performed in a standard 50 µl reaction in a MJ 225 DNA Engine Tetrad thermocycler. PCR conditions included a denaturation step at 94°C for 1 minute followed by 30 cycles each consisting of 1 minute at 94°C, 1 minute at 51°C for annealing, and 2 minutes at 72°C for extension.  The cycle finished with a post-cycle extension at 72°C for 5 minutes.  PCR products were cleaned using a Promega Wizard SV gel and PCR cleanup system followed by Big Dye sequencing reaction.  Samples were then cleaned by bead cleanup (Agencourt AMPure XP) prior to submission for sequencing in both directions to UW Biotech (Madison, Wisconsin—Biotech-nology Center).  Molecular analyses and resulting sequences were aligned using MEGA: Molecular Evolutionary Genetics Analysis Free Software (version 5) (Tamura et al. 2011) and searched using the basic local alignment search tool (BLAST).  All three samples resulted in a 99% identity match to the arid-land termite, Reticulitermes tibialis Banks (accession number: AY168206).Unlike R. flavipes, R. tibialis is able to withstand more arid conditions, making it less susceptible to desiccation (Pickens 1934).  This explains its pres-ence in the middle of a sunny, Wisconsin prairie.  Since this species has been found only on the south-western most border of the state, it is likely that this population lies on the northern boundary of R. tibialis distribution.  The discovery 1Arango: First Record of the Arid-Land Termite, <i>Reticulitermes TibialisPublished by ValpoScholar, 20152015 THE GREAT LAKES ENTOMOLOGIST 99is not substantially outside of the known distribution of this species, as it has been found as far north as Iowa and Idaho (Austin et al. 2008).AcknowledgmentsI would like to thank Thomas Meyer at the Wisconsin DNR for permitting insect collection at Hardscrabble Prairie and to Jeff Gruber for helping find this termite population.  Thanks also to Joliene Lindholm for help with genetic analy-sis and Brian Forschler for initial morphological examination of the specimens.Literature CitedAustin, J. W., A. L. Szalanski, J. A. McKern, and R. E. Gold.  2008.  Molecular phylogeography of the subterranean termite Reticulitermes tibialis (Isoptera: Rhi-notermitidae).  Journal of Agricultural and Urban Entomology  25: 63-79.Lim, S. Y. and B. T. Forschler.  201.  Reticulitermes nelsonae, a new species of subterra-nean termite (Rhinotermitidae) from the southeastern United States.  Insects  3:62-90.Pickens, A. L.  1934.  The barren-lands subterranean termite, Reticulitermes tibialis, pp. 184-186. In C. A. Kofoid, S. F. Light, A. C. Horner, M. Randall, W. B. Herms, and E. E. Bowe (eds.), Termites and Termite Control, 2nd ed. University of California Press, Berkeley, CA.Tamura, K., D. Peterson, N. Peterson, G. Stecher, M. Nei, and S. Kumar.  2011. MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods.  Molecular Biology and Evolution  28: 2731-2739.2The Great Lakes Entomologist, Vol. 48, No. 1 [2015], Art. 8https://scholar.valpo.edu/tgle/vol48/iss1/8

First Record of the Arid-Land Termite, \u3ci\u3eReticulitermes Tibialis\u3c/i\u3e Banks, in Wisconsin

ValpoScholar

The Great Lakes Entomologist 
Volume 48 
Numbers 1 & 2 - Spring/Summer 2015 Numbers 
1 & 2 - Spring/Summer 2015 
Article 8 
April 2015 
First Record of the Arid-Land Termite, Reticulitermes Tibialis 
Banks, in Wisconsin 
Rachel A. Arango 
Follow this and additional works at: https://scholar.valpo.edu/tgle 
 Part of the Entomology Commons 
Recommended Citation 
Arango, Rachel A. 2015. "First Record of the Arid-Land Termite, Reticulitermes Tibialis Banks, in 
Wisconsin," The Great Lakes Entomologist, vol 48 (1) 
Available at: https://scholar.valpo.edu/tgle/vol48/iss1/8 
This Peer-Review Article is brought to you for free and open access by the Department of Biology at ValpoScholar. 
It has been accepted for inclusion in The Great Lakes Entomologist by an authorized administrator of ValpoScholar. 
For more information, please contact a ValpoScholar staff member at scholar@valpo.edu. 
98 THE GREAT LAKES ENTOMOLOGIST Vol. 48, Nos. 1 - 2
First Record of the Arid-Land Termite,  
Reticulitermes tibialis Banks, in Wisconsin
Rachel A. Arango
Abstract
During a survey of termites in Wisconsin, one colony was found from a 
different habitat than the remaining populations.  This observation led to fur-
ther genetic testing which resulted in a determination of Reticulitermes tibialis 
Banks.  This is the first record of a termite species other than Reticulitermes 
flavipes (Kollar) to be established in the state.
 
____________________
Numerous records of the eastern subterranean termite, Reticulitermes 
flavipes (Kollar), have been recorded from Wisconsin, a species which has long 
been thought to be the only termite species established in the state.  Outside of 
an urban environment, colonies of R. flavipes are often found in shady, wooded 
areas under logs and in decaying trees.  During a survey of Wisconsin termite 
populations, one site at Hardscrabble Prairie in Hazel Green showed termite 
activity under logs and rocks in the middle of a sunny prairie with very little 
shade cover, prompting further evaluation.
As morphological identification of termites is often difficult, genetic-based 
techniques were utilized for species determination to support observed physical 
characteristics.  Termites were collected 24 July 2013 in cardboard placed under 
logs in the middle of the prairie.  Deoxyribonucleic acid (DNA) was extracted 
from three individual worker termites using the Promega Wizard genomic DNA 
purification kit following the protocol for isolation of genomic DNA from animal 
tissues.  Termite DNA (~50ng/µl) was amplified using primers TL2-J-3037 
(5’-ATGGCAGATTAGTGCAATGG-3’) and TK-N-3785 (5’-GTTTAAGAGACCAG-
TACTTG-3’), which are specific for the mitochondrial COII genes as chosen by 
Lim and Forschler (2012).  Polymerase chain reaction (PCR) was performed in 
a standard 50 µl reaction in a MJ 225 DNA Engine Tetrad thermocycler. PCR 
conditions included a denaturation step at 94°C for 1 minute followed by 30 
cycles each consisting of 1 minute at 94°C, 1 minute at 51°C for annealing, and 
2 minutes at 72°C for extension.  The cycle finished with a post-cycle extension 
at 72°C for 5 minutes.  PCR products were cleaned using a Promega Wizard SV 
gel and PCR cleanup system followed by Big Dye sequencing reaction.  Samples 
were then cleaned by bead cleanup (Agencourt AMPure XP) prior to submission 
for sequencing in both directions to UW Biotech (Madison, Wisconsin—Biotech-
nology Center).  Molecular analyses and resulting sequences were aligned using 
MEGA: Molecular Evolutionary Genetics Analysis Free Software (version 5) 
(Tamura et al. 2011) and searched using the basic local alignment search tool 
(BLAST).  All three samples resulted in a 99% identity match to the arid-land 
termite, Reticulitermes tibialis Banks (accession number: AY168206).
Unlike R. flavipes, R. tibialis is able to withstand more arid conditions, 
making it less susceptible to desiccation (Pickens 1934).  This explains its pres-
ence in the middle of a sunny, Wisconsin prairie.  Since this species has been 
found only on the south-western most border of the state, it is likely that this 
population lies on the northern boundary of R. tibialis distribution.  The discovery 
1
Arango: First Record of the Arid-Land Termite, <i>Reticulitermes Tibialis
Published by ValpoScholar, 2015
2015 THE GREAT LAKES ENTOMOLOGIST 99
is not substantially outside of the known distribution of this species, as it has 
been found as far north as Iowa and Idaho (Austin et al. 2008).
Acknowledgments
I would like to thank Thomas Meyer at the Wisconsin DNR for permitting 
insect collection at Hardscrabble Prairie and to Jeff Gruber for helping find this 
termite population.  Thanks also to Joliene Lindholm for help with genetic analy-
sis and Brian Forschler for initial morphological examination of the specimens.
Literature Cited
Austin, J. W., A. L. Szalanski, J. A. McKern, and R. E. Gold.  2008.  Molecular 
phylogeography of the subterranean termite Reticulitermes tibialis (Isoptera: Rhi-
notermitidae).  Journal of Agricultural and Urban Entomology  25: 63-79.
Lim, S. Y. and B. T. Forschler.  201.  Reticulitermes nelsonae, a new species of subterra-
nean termite (Rhinotermitidae) from the southeastern United States.  Insects  3:62-90.
Pickens, A. L.  1934.  The barren-lands subterranean termite, Reticulitermes tibialis, 
pp. 184-186. In C. A. Kofoid, S. F. Light, A. C. Horner, M. Randall, W. B. Herms, and 
E. E. Bowe (eds.), Termites and Termite Control, 2nd ed. University of California 
Press, Berkeley, CA.
Tamura, K., D. Peterson, N. Peterson, G. Stecher, M. Nei, and S. Kumar.  2011. 
MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, 
evolutionary distance, and maximum parsimony methods.  Molecular Biology and 
Evolution  28: 2731-2739.
2
The Great Lakes Entomologist, Vol. 48, No. 1 [2015], Art. 8
https://scholar.valpo.edu/tgle/vol48/iss1/8


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First Record of the Arid-Land Termite, \u3ci\u3eReticulitermes Tibialis\u3c/i\u3e Banks, in Wisconsin

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