First Record of Aedes stimulans from Louisiana Author(s): Charlie B. Sither, Bruce A. Harrison, Brittania Bintz, Mark R. Wilson, Jung Kim, Monty Graham, Kevin A. Caillouët, Michael L. Hutchinson and Brian D. Byrd Source: Journal of the American Mosquito Control Association, 30(4):305-308. Published By: The American Mosquito Control Association DOI: http://dx.doi.org/10.2987/14-6429R.1 URL: http://www.bioone.org/doi/full/10.2987/14-6429R.1
BioOne (www.bioone.org) is a nonprofit, online aggregation of core research in the biological, ecological, and environmental sciences. BioOne provides a sustainable online platform for over 170 journals and books published by nonprofit societies, associations, museums, institutions, and presses. Your use of this PDF, the BioOne Web site, and all posted and associated content indicates your acceptance of BioOne’s Terms of Use, available at www.bioone.org/page/terms_of_use. Usage of BioOne content is strictly limited to personal, educational, and non-commercial use. Commercial inquiries or rights and permissions requests should be directed to the individual publisher as copyright holder.
BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers, academic institutions, research libraries, and research funders in the common goal of maximizing access to critical research.
Journal of the American Mosquito Control Association, 30(4):305–308, 2014 Copyright E 2014 by The American Mosquito Control Association, Inc.
SCIENTIFIC NOTE FIRST RECORD OF AEDES STIMULANS FROM LOUISIANA CHARLIE B. SITHER,1 BRUCE A. HARRISON,1 BRITTANIA BINTZ,2 MARK R. WILSON,2 JUNG KIM,3 ¨ T,5 MICHAEL L. HUTCHINSON6 AND BRIAN D. BYRD1,7 MONTY GRAHAM,4 KEVIN A. CAILLOUE ABSTRACT. The first confirmed collection of Aedes stimulans in Louisiana was made in St. Tammany Parish, LA. A single adult female was collected by a large-bore aspirator in March 2011, and identified by microscopic and molecular methods. Notes are provided on the morphology, location, habitat, and potential mosquito associates that may be found with Ae. stimulans. KEY WORDS
Aedes (Ochlerotatus) stimulans, Louisiana, Culicidae
We report a new state record of the mosquito species Aedes stimulans (Walker) from St. Tammany Parish, LA. Aedes stimulans, or Ochlerotatus (Woodius) stimulans of Reinert et al. (2009), is a univoltine late winter–early spring species commonly found in the northern Midwest and northeastern USA, and extending south to northern Virginia and west into Oklahoma (Darsie and Ward 2005). However, Dyar (1920) described the collection of Ae. stimulans reared from 2 larval specimens collected by J. A. Le Prince in Electric Mills, Kemper County, Mississippi, approximately 350 km from our collection site in Louisiana. Goddard and Harrison (2005) confirmed the identification of the Mississippi specimens and the state record after borrowing and examining the lectotype and an associated male genitalia preparation located in the National Museum of Natural History (NMNH), Smithsonian Institution. Their findings were in accord with published reports by other authors (Carpenter and LaCasse 1955, Stone and Knight 1956, King et al. 1960, Harden et al. 1967, Knight and Stone 1977, Darsie and Ward 2005). A single adult female of Ae. stimulans was aspirated on March 3, 2011, with the use of a
1 Vector-Borne Infectious Disease Laboratory, Western Carolina University, Cullowhee, NC 28723. 2 Forensic Sciences Program, Western Carolina University, Cullowhee, NC 28723. 3 North Carolina Department of Agriculture & Consumer Services, Structural Pest Control and Pesticide Division, 1631 Mail Service Center, Raleigh, NC 27699. 4 Center for Rapid Product Realization, Western Carolina University, Cullowhee, NC 28723. 5 St. Tammany Parish Mosquito Abatement Program, 62512 Airport Road, Building 23, Slidell, LA 70460. 6 Pennsylvania Department of Environmental Protection, PO Box 1467, Harrisburg, PA 17105. 7 To whom correspondence should be addressed.
large-bore Nasci aspirator (Nasci 1981), in the Big Branch Marsh National Wildlife Refuge, which is managed by the US Fish and Wildlife Service. Mosquito collections were performed under the General Special Use Permit 43558-11-8 in 2011 in St. Tammany Parish, LA. The bottomland hardwood forest in which this mosquito was collected is 2.6 km distant from the shorefront of Lake Pontchartrain. Freshwater marsh and pine flatwood or savannah habitat with occasional bottomland hardwood hammocks dominate the immediate surroundings of the site. During late winter and early spring, numerous shaded and sun-exposed rain-filled puddles and larger vernal pools can be found at this site producing abundant floodwater mosquito species including Aedes vexans (Meigen), Aedes sticticus (Meigen), Aedes canadensis canadensis (Theobald), and Psorophora ferox (von Humboldt). The Louisiana specimen keyed to Ae. stimulans with the use of Darsie and Ward (2005) and Darsie and Hutchinson (2009). To ensure that this specimen was Ae. stimulans and not Aedes excrucians (Walker) or Aedes fitchii (Felt and Young), 2 species commonly confused with Ae. stimulans, a morphological examination was conducted of all 3 species. Photomicrographs of the Louisiana specimen (Fig. 1A–1C) were obtained by using a Nikon SMZ-U stereomicroscope with a 10:1 zoom ratio and EO Plan 1.53 objective and 103 ocular lenses and a phototubemounted Canon EOS 40D. An electron photomicrograph (Fig. 1D) of the foretarsal claws was obtained with the use of an Aspex Scanning Electron Microscope (Aspex Personal SEM, FEI, Hillsboro, OR). Several characters were used to separate Ae. stimulans from Ae. excrucians, including: 1) the shape of the foretarsal claw, which matches the Ae. stimulans photomicrograph in McDaniel and Webb (1974), 2) white scales on the anterodorsal corner of the mesokatepisternum, 3) black scales and setae on the pedicel, and 4) the distribution of the mixed dark and pale scales on the wing veins. Two characters
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Fig. 1. Photomicrographs of the Louisiana Aedes stimulans (LA2011). Morphological characters useful for identifying Ae. stimulans include (A) pale scales on the postprocoxal membrane and on the anterodorsal part of the mesokatepisternum, (B) broad basal pale bands on abdominal terga, (C) pale bands on the hindtarsomeres, and (D) the shape of the foretarsal claw.
that effectively separate Ae. stimulans from Ae. fitchii are 1) the distribution of the mixed dark and pale scales on the wing veins, and 2) black scales and setae on the pedicel. Although our morphological examination of the specimen confirmed the identity as Ae. stimulans, a DNA ‘‘BarCoding’’ approach was pursued to corroborate the identification. The DNA was extracted from a single leg of the Ae. stimulans specimen collected from Louisiana, and from known link-reared Ae. stimulans and Ae. fitchii specimens from Pennsylvania. The DNA was obtained with the use of the QiagenH DNA Investigator kit (Valencia, CA) cellulose materials protocol and polymerase chain reaction (PCR) amplified for the mitochondrial DNA cytochrome oxidase subunit 1 (COI) region with the use of HCO and LCO invertebrate primers and amplification methods as described by Folmer et al. (1994). Reaction mixtures were amplified on a VeritiH thermal cycler (Applied BiosystemsH, Foster City, CA). Resulting PCR products were purified with the use of ExoSAP-ITH (Affymetrix, Santa Clara, CA) then cycle sequenced with the
use of the BigDyeH Terminator v3.1 kit (Applied BiosystemsH, Foster City, CA) with a 36-cm capillary array, and POP-7TM polymer (Life Technologies, Grand Island, NY). The resulting electropherograms were analyzed using CodonCode Aligner (CodonCode Corporation, Centerville, MA) and the subsequent sequence was submitted to GenBank (Accession Number KJ867209). The pinned specimen will be deposited into the National Mosquito Collection, NMNH, Smithsonian Institution, Washington, DC. Sequence and phylogenetic analyses (Fig. 2) on the resulting COI DNA bar codes confirmed the initial morphological identification. The evolutionary history was inferred with the use of neighbor-joining, maximum-likelihood, and maximum-parsimony methods with MEGA 6 (Tamura et al. 2013) and resulted in phylograms with similar topologies. The Louisiana Ae. stimulans (LA 2011) was grouped within the Ae. stimulans clade for each of these models. The analysis involved 15 additional nucleotide sequences (5 Ae. stimulans, 5 Ae. fitchii, 4 Ae. exrucians, and Ae. triseriatus (Say) as the outgroup).
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307
Fig. 2. Phylogenetic analysis of the cytochrome oxidase subunit 1 region (659 base pairs) corroborates the identity of the LA 2011 Aedes stimulans. The phylogram was inferred by maximum-likelihood analysis and the tree is drawn to scale with branch lengths measured in number of substitutions per site. Bootstrap values greater than 90% (1,000 replications) are shown above the corresponding branches.
Prospective sampling for additional adult Ae. stimulans specimens was conducted weekly from December 30, 2013 to March 17, 2014 at the original collection site and at 3 other sites in St. Tammany Parish. Adult mosquito collections were performed by 10-min aspirations of vegetation and other natural resting areas near larval aquatic habitat with the use of a large-bore Nasci aspirator. Larval sampling was conducted by standard larval dipping techniques (O’Malley 1989) of each aquatic habitat present at the site of original specimen collection on 4 occasions including February 17 and 27 and March 10 and 17, 2014. No additional Ae. stimulans specimens were detected by adult or larval sampling in 2013 and 2014. The Louisiana Mosquito Control Association revised its Mosquito Control Training Manual in 2007, and this publication contains identification keys (Fox 2007) for selected mosquitoes in the state. Because these keys are commonly used by mosquito control professionals in Louisiana, the following information is provided to aid in the identification of Ae. stimulans within the state. Using the Louisiana key for adult females (Fox 2007), Ae. stimulans easily proceeds through couplets 1, 2, 3, 6, and ends at couplet 8, where Ae. albopictus (Skuse) and Ae. vexans are separated. Aedes stimulans is easily separated from Ae. albopictus and Ae. vexans by 1) these 2 species lack scales on the postprocoxal membrane; 2) both have leg-banding patterns different from Ae. stimulans; i.e., the former species has long basal white bands on hindtarsomeres 1–4 and 5 is entirely white, and the latter species has very narrow pale bands on the tarsi, and Ae. stimulans has large pale bands, but hind tar-
somere 5 is not entirely pale; and 3) wing scale differences, i.e., Ae. albopictus with dark wing scales and Ae. vexans with dark wing scales except for a few pale scales on the base of the costa and radius. On Ae. stimulans the mixed dark and pale wing scales are primarily concentrated on the anterior wing veins (C, SC, R1, R2+3, R2, and R3), and the remaining posterior veins are primarily dark scaled with few or no pale scales. When keying Ae. stimulans larvae in Fox (2007), a specimen would proceed through couplets 1, 10, 13, 17, and end at couplet 18, where Ae. c. canadensis and Ae. thibaulti Dyar and Knab, are separated. Head setae characters on Ae. stimulans larvae, i.e., 5,6-C single or bifid, are very different from the multibranched setae 5,6-C on Ae. c. canadensis and Ae. thibaulti. Seta 1-A on the antenna of Ae. stimulans is tiny, short, and branched, whereas that seta on Ae. c. canadensis is large, long, branched, and reaches the tip of the antenna. An active role by Ae. stimulans in the transmission of Louisiana zoonotic viruses that affect humans is unknown and unlikely. However in northern states this species has been found infected with Jamestown Canyon (JC) virus (Boromisa and Grimstad 1986) and Snowshoe hare virus (Iversen et al. 1969). Aedes stimulans is an efficient laboratory vector of JC virus (Heard et al. 1991) and West Nile virus has been detected in some collections (Centers for Disease Control and Prevention [CDC] 2014). The new record of Ae. stimulans in Louisiana supports the Dyar (1920) record of this species in Mississippi. Thus, Ae. stimulans has now been confirmed from 2 contiguous Gulf Coast states. However, our subsequent collection efforts
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yielded no additional Ae. stimulans and it remains unknown if local populations of this species exist in Gulf Coast states. We thank 2 anonymous reviewers whose comments and suggestions improved an earlier draft of this manuscript. This project was supported, in part, by funding from the Western Carolina University (WCU) Forensic Science program and the WCU College of Health and Human Sciences. REFERENCES CITED Boromisa RD, Grimstad PR. 1986. Virus–vector–host relationships of Aedes stimulans and Jamestown Canyon virus in a northern Indiana enzootic focus. Am J Trop Med Hyg 35:1285–1295. Carpenter SJ, LaCasse WJ. 1955. Mosquitoes of North America (north of Mexico). Berkeley, CA: Univ. of California Press. CDC [Centers for Disease Control and Prevention]. 2014. For vector control professionals [Internet] [accessed 15 May, 2014]. Available from: http:// www.cdc.gov/westnile/vectorcontrol/. Darsie RF, Hutchinson ML. 2009. The mosquitoes of Pennsylvania. Danville, PA: Pennsylvania Vector Control Association Technical Bulletin 2009-001. Darsie RF, Ward RA. 2005. Identification and geographical distribution of the mosquitoes of North America, north of Mexico. Gainesville, FL: Univ. Press of Florida. Dyar HG. 1920. The American Aedes of the stimulans group (Diptera, Culicidae). Insec Inscit Menst 8: 106–120. Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R. 1994. DNA primers for the amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotechnol 3:294–299. Fox M. 2007. Keys to the medically important mosquitoes of Louisiana. In: Mosquito control training manual. Baton Rouge, LA: Louisiana Mosquito Control Association. p 86–150.
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Goddard J, Harrison BA. 2005. New, recent, and questionable mosquito records from Mississippi. J Am Mosq Control Assoc 21:10–14. Harden FW, Hepburn HR, Ethridge BJ. 1967. A history of mosquitoes and mosquito-borne diseases in Mississippi. Mosq News 27:60–66. Heard PB, Zhang MB, Grimstad PR. 1991. Laboratory transmission of Jamestown Canyon and Snowshoe hare viruses (Bunyaviridae: California Serogroup) by several species of mosquitoes. J Am Mosq Control Assoc 7:94–102. Iversen J, Hanson RP, Papadopoulos O, Morris CV, DeFoliart GR. 1969. Isolation of viruses of the California encephalitis virus group from Boreal Aedes mosquitoes. Am J Trop Med Hyg 18:735–742. King WL, Bradley GH, Smith CN, McDuffie WC. 1960. A handbook of the mosquitoes of the southeastern United States. Washington, DC: Department of Agriculture. Knight KL, Stone A. 1977. A catalog of the mosquitoes of the world (Diptera: Culicidae). 2nd ed. Thomas Say Foundation No. 6. Annapolis, MD: Entomological Society of America. 611 p. McDaniel IN, Webb DR. 1974. Identification of females of the Aedes stimulans group including notes on larval characters and attempts at hybridization. Ann Entomol Soc Am 67:915–918. Nasci RS. 1981. A lightweight battery-powered aspirator for collecting resting mosquitoes in the field. Mosq News 41:808–812. O’Malley CM. 1989. Guidelines for larval surveillance. In: Proceedings of the Seventy-Sixth Annual Meeting of the New Jersey Mosquito Control Association. Lindenwold, NJ: New Jersey Mosquito Control Association. p 44–45. Reinert JF, Harbach RE, Kitching IJ. 2009. Phylogeny and classification of tribe Aedini (Diptera: Culicidae). Zool J Linn Soc 157:700–794. Stone A, Knight KL. 1956. Type specimens of mosquitoes in the United States National Museum. J Wash Acad Sci 46:213–228. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. 2013. MEGA6: Molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729.
BioOne (www.bioone.org) is a nonprofit, online aggregation of core research in the biological, ecological, and environmental sciences. BioOne provides a sustainable online platform for over 170 journals and books published by nonprofit societies, associations, museums, institutions, and presses. Your use of this PDF, the BioOne Web site, and all posted and associated content indicates your acceptance of BioOne’s Terms of Use, available at www.bioone.org/page/terms_of_use. Usage of BioOne content is strictly limited to personal, educational, and non-commercial use. Commercial inquiries or rights and permissions requests should be directed to the individual publisher as copyright holder.
BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers, academic institutions, research libraries, and research funders in the common goal of maximizing access to critical research.
Journal of the American Mosquito Control Association, 30(4):305–308, 2014 Copyright E 2014 by The American Mosquito Control Association, Inc.
SCIENTIFIC NOTE FIRST RECORD OF AEDES STIMULANS FROM LOUISIANA CHARLIE B. SITHER,1 BRUCE A. HARRISON,1 BRITTANIA BINTZ,2 MARK R. WILSON,2 JUNG KIM,3 ¨ T,5 MICHAEL L. HUTCHINSON6 AND BRIAN D. BYRD1,7 MONTY GRAHAM,4 KEVIN A. CAILLOUE ABSTRACT. The first confirmed collection of Aedes stimulans in Louisiana was made in St. Tammany Parish, LA. A single adult female was collected by a large-bore aspirator in March 2011, and identified by microscopic and molecular methods. Notes are provided on the morphology, location, habitat, and potential mosquito associates that may be found with Ae. stimulans. KEY WORDS
Aedes (Ochlerotatus) stimulans, Louisiana, Culicidae
We report a new state record of the mosquito species Aedes stimulans (Walker) from St. Tammany Parish, LA. Aedes stimulans, or Ochlerotatus (Woodius) stimulans of Reinert et al. (2009), is a univoltine late winter–early spring species commonly found in the northern Midwest and northeastern USA, and extending south to northern Virginia and west into Oklahoma (Darsie and Ward 2005). However, Dyar (1920) described the collection of Ae. stimulans reared from 2 larval specimens collected by J. A. Le Prince in Electric Mills, Kemper County, Mississippi, approximately 350 km from our collection site in Louisiana. Goddard and Harrison (2005) confirmed the identification of the Mississippi specimens and the state record after borrowing and examining the lectotype and an associated male genitalia preparation located in the National Museum of Natural History (NMNH), Smithsonian Institution. Their findings were in accord with published reports by other authors (Carpenter and LaCasse 1955, Stone and Knight 1956, King et al. 1960, Harden et al. 1967, Knight and Stone 1977, Darsie and Ward 2005). A single adult female of Ae. stimulans was aspirated on March 3, 2011, with the use of a
1 Vector-Borne Infectious Disease Laboratory, Western Carolina University, Cullowhee, NC 28723. 2 Forensic Sciences Program, Western Carolina University, Cullowhee, NC 28723. 3 North Carolina Department of Agriculture & Consumer Services, Structural Pest Control and Pesticide Division, 1631 Mail Service Center, Raleigh, NC 27699. 4 Center for Rapid Product Realization, Western Carolina University, Cullowhee, NC 28723. 5 St. Tammany Parish Mosquito Abatement Program, 62512 Airport Road, Building 23, Slidell, LA 70460. 6 Pennsylvania Department of Environmental Protection, PO Box 1467, Harrisburg, PA 17105. 7 To whom correspondence should be addressed.
large-bore Nasci aspirator (Nasci 1981), in the Big Branch Marsh National Wildlife Refuge, which is managed by the US Fish and Wildlife Service. Mosquito collections were performed under the General Special Use Permit 43558-11-8 in 2011 in St. Tammany Parish, LA. The bottomland hardwood forest in which this mosquito was collected is 2.6 km distant from the shorefront of Lake Pontchartrain. Freshwater marsh and pine flatwood or savannah habitat with occasional bottomland hardwood hammocks dominate the immediate surroundings of the site. During late winter and early spring, numerous shaded and sun-exposed rain-filled puddles and larger vernal pools can be found at this site producing abundant floodwater mosquito species including Aedes vexans (Meigen), Aedes sticticus (Meigen), Aedes canadensis canadensis (Theobald), and Psorophora ferox (von Humboldt). The Louisiana specimen keyed to Ae. stimulans with the use of Darsie and Ward (2005) and Darsie and Hutchinson (2009). To ensure that this specimen was Ae. stimulans and not Aedes excrucians (Walker) or Aedes fitchii (Felt and Young), 2 species commonly confused with Ae. stimulans, a morphological examination was conducted of all 3 species. Photomicrographs of the Louisiana specimen (Fig. 1A–1C) were obtained by using a Nikon SMZ-U stereomicroscope with a 10:1 zoom ratio and EO Plan 1.53 objective and 103 ocular lenses and a phototubemounted Canon EOS 40D. An electron photomicrograph (Fig. 1D) of the foretarsal claws was obtained with the use of an Aspex Scanning Electron Microscope (Aspex Personal SEM, FEI, Hillsboro, OR). Several characters were used to separate Ae. stimulans from Ae. excrucians, including: 1) the shape of the foretarsal claw, which matches the Ae. stimulans photomicrograph in McDaniel and Webb (1974), 2) white scales on the anterodorsal corner of the mesokatepisternum, 3) black scales and setae on the pedicel, and 4) the distribution of the mixed dark and pale scales on the wing veins. Two characters
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Fig. 1. Photomicrographs of the Louisiana Aedes stimulans (LA2011). Morphological characters useful for identifying Ae. stimulans include (A) pale scales on the postprocoxal membrane and on the anterodorsal part of the mesokatepisternum, (B) broad basal pale bands on abdominal terga, (C) pale bands on the hindtarsomeres, and (D) the shape of the foretarsal claw.
that effectively separate Ae. stimulans from Ae. fitchii are 1) the distribution of the mixed dark and pale scales on the wing veins, and 2) black scales and setae on the pedicel. Although our morphological examination of the specimen confirmed the identity as Ae. stimulans, a DNA ‘‘BarCoding’’ approach was pursued to corroborate the identification. The DNA was extracted from a single leg of the Ae. stimulans specimen collected from Louisiana, and from known link-reared Ae. stimulans and Ae. fitchii specimens from Pennsylvania. The DNA was obtained with the use of the QiagenH DNA Investigator kit (Valencia, CA) cellulose materials protocol and polymerase chain reaction (PCR) amplified for the mitochondrial DNA cytochrome oxidase subunit 1 (COI) region with the use of HCO and LCO invertebrate primers and amplification methods as described by Folmer et al. (1994). Reaction mixtures were amplified on a VeritiH thermal cycler (Applied BiosystemsH, Foster City, CA). Resulting PCR products were purified with the use of ExoSAP-ITH (Affymetrix, Santa Clara, CA) then cycle sequenced with the
use of the BigDyeH Terminator v3.1 kit (Applied BiosystemsH, Foster City, CA) with a 36-cm capillary array, and POP-7TM polymer (Life Technologies, Grand Island, NY). The resulting electropherograms were analyzed using CodonCode Aligner (CodonCode Corporation, Centerville, MA) and the subsequent sequence was submitted to GenBank (Accession Number KJ867209). The pinned specimen will be deposited into the National Mosquito Collection, NMNH, Smithsonian Institution, Washington, DC. Sequence and phylogenetic analyses (Fig. 2) on the resulting COI DNA bar codes confirmed the initial morphological identification. The evolutionary history was inferred with the use of neighbor-joining, maximum-likelihood, and maximum-parsimony methods with MEGA 6 (Tamura et al. 2013) and resulted in phylograms with similar topologies. The Louisiana Ae. stimulans (LA 2011) was grouped within the Ae. stimulans clade for each of these models. The analysis involved 15 additional nucleotide sequences (5 Ae. stimulans, 5 Ae. fitchii, 4 Ae. exrucians, and Ae. triseriatus (Say) as the outgroup).
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Fig. 2. Phylogenetic analysis of the cytochrome oxidase subunit 1 region (659 base pairs) corroborates the identity of the LA 2011 Aedes stimulans. The phylogram was inferred by maximum-likelihood analysis and the tree is drawn to scale with branch lengths measured in number of substitutions per site. Bootstrap values greater than 90% (1,000 replications) are shown above the corresponding branches.
Prospective sampling for additional adult Ae. stimulans specimens was conducted weekly from December 30, 2013 to March 17, 2014 at the original collection site and at 3 other sites in St. Tammany Parish. Adult mosquito collections were performed by 10-min aspirations of vegetation and other natural resting areas near larval aquatic habitat with the use of a large-bore Nasci aspirator. Larval sampling was conducted by standard larval dipping techniques (O’Malley 1989) of each aquatic habitat present at the site of original specimen collection on 4 occasions including February 17 and 27 and March 10 and 17, 2014. No additional Ae. stimulans specimens were detected by adult or larval sampling in 2013 and 2014. The Louisiana Mosquito Control Association revised its Mosquito Control Training Manual in 2007, and this publication contains identification keys (Fox 2007) for selected mosquitoes in the state. Because these keys are commonly used by mosquito control professionals in Louisiana, the following information is provided to aid in the identification of Ae. stimulans within the state. Using the Louisiana key for adult females (Fox 2007), Ae. stimulans easily proceeds through couplets 1, 2, 3, 6, and ends at couplet 8, where Ae. albopictus (Skuse) and Ae. vexans are separated. Aedes stimulans is easily separated from Ae. albopictus and Ae. vexans by 1) these 2 species lack scales on the postprocoxal membrane; 2) both have leg-banding patterns different from Ae. stimulans; i.e., the former species has long basal white bands on hindtarsomeres 1–4 and 5 is entirely white, and the latter species has very narrow pale bands on the tarsi, and Ae. stimulans has large pale bands, but hind tar-
somere 5 is not entirely pale; and 3) wing scale differences, i.e., Ae. albopictus with dark wing scales and Ae. vexans with dark wing scales except for a few pale scales on the base of the costa and radius. On Ae. stimulans the mixed dark and pale wing scales are primarily concentrated on the anterior wing veins (C, SC, R1, R2+3, R2, and R3), and the remaining posterior veins are primarily dark scaled with few or no pale scales. When keying Ae. stimulans larvae in Fox (2007), a specimen would proceed through couplets 1, 10, 13, 17, and end at couplet 18, where Ae. c. canadensis and Ae. thibaulti Dyar and Knab, are separated. Head setae characters on Ae. stimulans larvae, i.e., 5,6-C single or bifid, are very different from the multibranched setae 5,6-C on Ae. c. canadensis and Ae. thibaulti. Seta 1-A on the antenna of Ae. stimulans is tiny, short, and branched, whereas that seta on Ae. c. canadensis is large, long, branched, and reaches the tip of the antenna. An active role by Ae. stimulans in the transmission of Louisiana zoonotic viruses that affect humans is unknown and unlikely. However in northern states this species has been found infected with Jamestown Canyon (JC) virus (Boromisa and Grimstad 1986) and Snowshoe hare virus (Iversen et al. 1969). Aedes stimulans is an efficient laboratory vector of JC virus (Heard et al. 1991) and West Nile virus has been detected in some collections (Centers for Disease Control and Prevention [CDC] 2014). The new record of Ae. stimulans in Louisiana supports the Dyar (1920) record of this species in Mississippi. Thus, Ae. stimulans has now been confirmed from 2 contiguous Gulf Coast states. However, our subsequent collection efforts
308
JOURNAL OF THE AMERICAN MOSQUITO CONTROL ASSOCIATION
yielded no additional Ae. stimulans and it remains unknown if local populations of this species exist in Gulf Coast states. We thank 2 anonymous reviewers whose comments and suggestions improved an earlier draft of this manuscript. This project was supported, in part, by funding from the Western Carolina University (WCU) Forensic Science program and the WCU College of Health and Human Sciences. REFERENCES CITED Boromisa RD, Grimstad PR. 1986. Virus–vector–host relationships of Aedes stimulans and Jamestown Canyon virus in a northern Indiana enzootic focus. Am J Trop Med Hyg 35:1285–1295. Carpenter SJ, LaCasse WJ. 1955. Mosquitoes of North America (north of Mexico). Berkeley, CA: Univ. of California Press. CDC [Centers for Disease Control and Prevention]. 2014. For vector control professionals [Internet] [accessed 15 May, 2014]. Available from: http:// www.cdc.gov/westnile/vectorcontrol/. Darsie RF, Hutchinson ML. 2009. The mosquitoes of Pennsylvania. Danville, PA: Pennsylvania Vector Control Association Technical Bulletin 2009-001. Darsie RF, Ward RA. 2005. Identification and geographical distribution of the mosquitoes of North America, north of Mexico. Gainesville, FL: Univ. Press of Florida. Dyar HG. 1920. The American Aedes of the stimulans group (Diptera, Culicidae). Insec Inscit Menst 8: 106–120. Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R. 1994. DNA primers for the amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotechnol 3:294–299. Fox M. 2007. Keys to the medically important mosquitoes of Louisiana. In: Mosquito control training manual. Baton Rouge, LA: Louisiana Mosquito Control Association. p 86–150.
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Goddard J, Harrison BA. 2005. New, recent, and questionable mosquito records from Mississippi. J Am Mosq Control Assoc 21:10–14. Harden FW, Hepburn HR, Ethridge BJ. 1967. A history of mosquitoes and mosquito-borne diseases in Mississippi. Mosq News 27:60–66. Heard PB, Zhang MB, Grimstad PR. 1991. Laboratory transmission of Jamestown Canyon and Snowshoe hare viruses (Bunyaviridae: California Serogroup) by several species of mosquitoes. J Am Mosq Control Assoc 7:94–102. Iversen J, Hanson RP, Papadopoulos O, Morris CV, DeFoliart GR. 1969. Isolation of viruses of the California encephalitis virus group from Boreal Aedes mosquitoes. Am J Trop Med Hyg 18:735–742. King WL, Bradley GH, Smith CN, McDuffie WC. 1960. A handbook of the mosquitoes of the southeastern United States. Washington, DC: Department of Agriculture. Knight KL, Stone A. 1977. A catalog of the mosquitoes of the world (Diptera: Culicidae). 2nd ed. Thomas Say Foundation No. 6. Annapolis, MD: Entomological Society of America. 611 p. McDaniel IN, Webb DR. 1974. Identification of females of the Aedes stimulans group including notes on larval characters and attempts at hybridization. Ann Entomol Soc Am 67:915–918. Nasci RS. 1981. A lightweight battery-powered aspirator for collecting resting mosquitoes in the field. Mosq News 41:808–812. O’Malley CM. 1989. Guidelines for larval surveillance. In: Proceedings of the Seventy-Sixth Annual Meeting of the New Jersey Mosquito Control Association. Lindenwold, NJ: New Jersey Mosquito Control Association. p 44–45. Reinert JF, Harbach RE, Kitching IJ. 2009. Phylogeny and classification of tribe Aedini (Diptera: Culicidae). Zool J Linn Soc 157:700–794. Stone A, Knight KL. 1956. Type specimens of mosquitoes in the United States National Museum. J Wash Acad Sci 46:213–228. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. 2013. MEGA6: Molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729.
