By Ryan Harrison (Forsyth County)

The Biting Times 52nd and 53rd Annual Conferences Over the next two years (2017 to 2018) we will be doing things a little different. This year Newsletter for the North Carolina Mosquito & Vector Control Association March 2017 we will have a 1.5-day conference in Greenville, NC on July 11-12 (see registration and preliminary program, pages 20-21). In 2018, we will have a joint conference with the Mid- Atlantic Mosquito Control Association (MAMCA) in Carolina Beach (February 12-14, 2018), and then another summer one-day conference in 2018 (location and date TBD). Adult Trapping in Forsyth County, NC. By Ryan Harrison (Forsyth County) One of the coolest aspects of my job here at Forsyth County is the fact that our program consists of a whole lot of adult mosquito trapping. Trapping and testing mosquitoes has become a huge part of my job. It is always one of the best and more rewarding parts of the many duties I have. In my 20 years here, we have streamlined our surveillance efforts and our division runs traps almost three times a week if not more. This has become a great way to see what s really going on in the area. Our tools for adult mosquito collection consist of 8 CDC/CO 2 traps, 10 gravid traps, and new for us in the 2016 season, 4 BG sentinels traps. Each type of trap is designed to catch mosquitoes in different circumstances so by having a variety of traps you can build up quite a bit of data very quickly. Here is a brief description of each type of trap and how Forsyth County uses them. The CDC/ C0 2 light traps are great for getting an idea of the different species that are attracted to carbon dioxide. This type of trap will allow you to get a quick assessment of species looking for a blood meal. (continued on page 5). Inside this issue: 2017 Conference Page 1, 20-21 Adult Trapping Page 1, 5-6 President s Message Page 2 An Intern s Blog Pages 4, 18-19 News Items Pages 2-3 Culex coronator Page 7 Sponsors Pages 11-12 New NC Entos Page 10 Bruce s Corner Pages 14-17 Guess that skeeter! Michael Reiskind, 2017 I m a native North American mosquito, but I often get mistaken for my Old World, invasive congener. We both can transmit West Nile virus to birds. In NC, I come out early in the season, and use natural and artificial larval habitats. Finally, when I am at my freshest, you can see two golden spots on my scutum. What am I? Answer, page 9.

Page 2 Message from the Editor and President Surely, we will look back at 2016 and think of it as the year of Zika. The real question is what will 2017 look like? Will we continue to be threatened by Zika, or will another Aedes driven virus come to the fore, like dengue? Will there be something brand new, like Japanese encephalitis, or the ever-threatening Rift Valley fever? Or will our native arboviruses, like La Crosse encephalitis, coming storming back in reaction to the wildfires in the mountains? Finally, will Lyme disease take hold in the northwest of the state that borders Virginia, where there has been a major outbreak in the last two years? Of course, the answer to all of these questions is I don t know. How disheartening an answer is that? So maybe, a better one is it doesn t matter. Not because of apathy, but because of resilience and flexibility. It doesn t matter, because we have the knowledge and technical ability to address any of these public health threats by attacking the vector and preventing people becoming infected. Now, more than ever, we need to keep people aware of all preventable arthropod-borne threats to public health. We run the risk of a Zika hangover, in which we get accused of crying wolf. We need to emphasize the increasing spread of not just exotic viruses, like Zika and chikungunya, but the sustained threat of dengue and our native La Crosse and eastern equine encephalitis viruses. And we cannot ignore the creeping threat of tickborne pathogens, which are preventable through personal protection. We are the front-line for these threats, and public awareness and education are key to maintaining support for our profession. Don t forget to register and attend our conference this year. Our schedule is a little different this year (page 1, 20-21), but I look forward to seeing each of you there. Check our website: www.ncmvca.org for registration and other information. April is Tick and Mosquito Disease Awareness Month in North Carolina The North Carolina Division of Public Health, Communicable Disease Branch is gearing up for Tick and Mosquito Awareness month in April. The Fight the Bite campaign will be returning to classrooms throughout North Carolina and includes a poster contest for students grades K-12 that promotes tick and mosquito borne illness prevention. State Public Health entomologists will present a lecture titled, Ticks and mosquitoes suck! How you can prevent vector-borne diseases this summer at the Daily Planet Café in the North Carolina Museum of Natural Sciences. Additional lectures will be provided at the annual NC Communicable Disease conference, the Chatham County tick-borne infections meeting, and the NC Association of Local Health Directors conference. Contact Dr. Alexis Barbarin (see Page 10) for more information.

Page 3 AMCA s Washington Day By Joe Andrews, Legislative Chair of NCMVCA I will be attending the AMCA Washington Day again this year. My goal in this message is to coax a few of you to join me at the event. We would work in unison to visit as many of our NC legislators as possible to instill in them the need to reinvest in science based Vector Management. If all goes right, our new and capable Medical Entomologist, Michael Doyle (page 10) will be joining us. The time is nigh to register for the event and to most assuredly apply for the stipends to help offset your expenses. Day one meeting is chock full of information about the processes and updates about policy at the federal level; something I have found invaluable in the past. Then, the following morning, it s off to the Hill! These are exciting and crucial times with the evolution of a young and strong university partnership with WCU, ECU and NC State along with the dedication of DHHS to reestablish a sustainable surveillance driven public health vector management infrastructure. Now, I challenge you to step up and do your part working to make a change for Public Health back here in North Carolina. Here are some facts provided me by our AMCA coordinator Dennis Salmen: 1) Parking is $48/day if overnight and $20/day if you're gone by days' end. 2) This hotel is about 3 short blocks from Foggy-Bottom Metro station. 3) You don't have to be an AMCA member to attend...it s FREE to all. 4) You do have to be a member to apply for the Central Life Science stipends. 5) The stipend submission deadline is only MAR 23rd. 6) INFORMATIONAL conference call session scheduled for MAR 17, 2PM for new or interested folks. See AM- CA conference webpage to view info (http://www.mosquito.org/washington-conference-info-conf-call ). I am going to Amtrak up there, and would love company! (Joe s contact: joea@allprovector.com; 919-343-8440) The Biting Times, March 2017 Editor-in-chief: Dr. Michael Reiskind Associate Editor: Chrystal Swinger Contributors: Ryan Harrison Jeff Brown Bruce Harrison Shannon Garrick Joe Andrews Photos: Matt Bertone Meredith Spence Chrystal Swinger Biogents.com JWHock.com Bruce Harrison Culex restuans ID: B. Harrison (thanks!) Photo: Chrystal Swinger

Page 4 An Interns Point of View: Excerpts from a Blog By Shannon Garrick (Catawba College), edited by C. Swinger Internship Week 1: Vector Control technician (6/17/2016) It is hard to believe I have been at my internship for a week now. I found out about this opportunity from another source back in late April and I am glad that I took advantage of it. The ironic thing is that I am planning on working with my professor/mentor this coming school year on her mosquito research. I had shadowed and been a part of her research in the past, but I took a break from it for about a year. Partly because I wasn t sure what I wanted to do and was a bit apprehensive of the people doing the research. BUT I had the experience, interest, and knowledge that helped my application stand out. This internship is a grant based one due to the Zika virus concern and its connection to cases of microcephaly. We are doing mosquito collection and ID ing. Essentially, we are looking for Aedes aegypti - which is the species responsible for causing Zika virus, along with other mosquito-borne diseases. (Other species cause disease as well). The place I work at is very interesting. Interning here will give me a feel for public health and if I want to pursue that avenue after graduation. They have a medical and dental clinic (to help those who can t otherwise afford medical care), WIC, international travel health, environmental health, and wellness programs including exercise, cooking, and others. So far, the environmental health seems very interesting to me. The department I work for is very nice the head of the department is laid back and very understanding, The person I work under reminds me a lot of my entomology professor (who I had for my environmental science courses) and of course, my mentor. She can identify mosquito species really fast!!! I work with two other guys who are very nice and will be doing mainly collections and ID ing. So far, I tried ID ing while waiting the other day and have shadowed and helped with collection setting out traps and retrieving them, I hope to become decent at ID ing and bring some of what I learn this summer to my Independent Research this coming school year. Internship end of week 3 (7/2/16) Hard to believe it is July 1st! The summer is half way over and my birthday has come and gone My internship is going well. Basically, we are learning more in-depth about mosquitoes both identification and anatomy/physiology of them, along with collection methods and surveying/checking out complaints. (continued on page 18)

Page 5 (from page 1). A CDC light trap works by using light and carbon dioxide to attract the mosquitoes; these insects are then pulled into a small container (figure 1). CDC light traps are great for catching large numbers of Aedes, Anopheles, Psorophora and Culex. To give you an idea of the efficiency of a CDC light trap, in 2015, I was able to capture 24 of our 36 species (in Forsyth) in one trap. We regularly retrieve traps with several thousand mosquitoes; in fact one of the main problems we run into using the CDC traps is that we catch way too many mosquitoes to identify quickly. Often light traps are not the best choice for disease surveillance unless you can target mosquitoes that have been around for a while. This season we caught a total 31 species in our CO 2 /CDC light traps for a total of 4,221 mosquitoes. The majority of these specimens were: Aedes vexans, Psorophora ferox, Figure 2. Gravid trap. Photo: JW Hock and company. Aedes sticticus, and Coquillettidia perturbans. We also caught a few of less common species for Forsyth County as well; Aedes hendersoni, Aedes fulvus pallens, and also Uranotaenia sapphirina. Gravid traps are an excellent tool for collecting container breeding species of mosquitoes that have already taken a blood meal and are developing eggs. Gravid mosquitoes are more likely to have been exposed to virus. These traps are used all around Forsyth County to capture the mosquitoes that we submit to the state for virus testing. The traps come in several different styles, but they all function on the same principles. A gravid trap attracts mosquitoes by using water mixed with organic materials (Figure 2). You can adjust or change the contents of the water infusion to accommodate the species of mosquitoes you are targeting. Forsyth County uses two types of infusions to attract mosquitoes. I make a hay infusion by taking a handful of hay (some people use rabbit food pellets) that is collected from a barn stall and let it soak in a 5 gallon bucket of water with a lid on it. After about a week you get a mixture of water that has really foul odor, we jokingly call this the special tea. This particular hay infusion is really good at attracting gravid Culex. If we want to attract more Aedes, we can make a separate tea using oak leaves instead of hay. In Forsyth we have found that by placing the gravid traps in shaded areas of the older parts of our municipalities, we can really increase the number of Culex we catch. Be aware that this type of trap catches significantly less mosquitoes than the CDC light trap, however it s much easier to keep up with the data collection, identification and you are also more likely to get virus positive pools. The gravid trap does not catch as many species as the light traps do. (continued, next page). Figure 1. CDC light trap. Photo: M. Spence

(from previous page) You tend to catch more Ae. albopictus, Ae. japonicus, Ae. triseriatus, Cx. pipiens complex, and Cx. restuans. Occasionally you can trap other species in gravid traps but primarily you stick to these container breeding species. It s important to note these 5 container species are our primary species implicated in the arbovirus cycles in NC. If you plan on doing disease surveillance the gravid trap is the way to go. Figure 3. BG-Sentinel trap. Photo: Biogents.com. This season we were able to capture 430 mosquitoes in our gravid traps, 246 of these were gravid Cx. pipiens complex, followed by Ae. albopictus with 134 specimen, and several other species. Usually in Forsyth we catch hundreds of Ae. japonicus in our gravid traps, but this year we only caught 9! It appears as though 2016 was not a great year for Ae. japonicus. Last, but not least, are BG sentinel traps we acquired with our Zika funding. The BG trap is primarily targeting Aedes species such as: Ae. albopictus and Ae. aegypti, however, you can catch other species as well. This is a trap that attracts the mosquitoes with its shape, size, it sits on the ground and uses a proprietary attractant called BG lure (Figure 3) You can also use octanol, CO 2 or really any other attractant out there. Octanol and BG lure at the same time gave us the best results. In our first season using this trap I was astonished at how well it caught Ae.albopictus. This season we caught 4,625 mosquitoes in our Sentinels, 4,610 of these mosquitoes were Aedes albopictus! We even caught 1,880 females in one afternoon! This trap instantly became an integral part to our Zika response. When these travel related Zika cases were called into our EPI teams, we would have a BG trap onsite within a few hours to conduct surveillance and testing. I cannot wait to deploy the BG sentinel this coming season with other attractants like dry ice (CO 2 ). There are many options for collecting female mosquitoes. If you are considering using traps, it would be best to pinpoint your goals in surveillance and use the trap that best fits your needs. Ask for help from programs and academics around your area. Most of us in the field love to share the little details that took us years to figure out. Any previous knowledge is a plus. Perhaps a program in your area already has information that you could use. The most important thing to remember is trapping is a fun and creative way to gain valuable information about the mosquitoes that are found in your area. There is nothing like picking up traps when there are billions of mosquitoes around you! What an experience! One of the best ways to practice integrated mosquito management is developing a program that has a foundation of science and surveillance. After all, if you don t know what the bugs are and when the bugs are flying, you can t knock them out of the air! When I first started at Forsyth County in 1997, surveillance data came from 10-15 sites around the county. Most of these data had been collected by Parker Whitt and Dr Bruce Harrison with PHPM, so the data we had was thorough and consistent. It really gave me a great starting point to work with. (Contact Ryan: harrisrl@forsyth.cc, phone: 336-703-3170). Page 6

Page 7 The Seasonal Distribution of Culex coronator in South East North Carolina in 2016, an Operational Perspective By Jeff Brown (Brunswick Co.), Avian White (ECU), Rick Hickman (Brunswick Co.), Marie Hemmen (New Hanover Co.), Justin Bunn (ECU) Several investigators have described collections of Cx. coronator Dyer and Knab since it was first described by Dyar and Knab in1906. Debboun et al. (2005) collected specimens from May to August in Louisiana in 2004. Goddard et al. (2006) collected specimens from August and October in 2005 in Mississippi. McNelly et al. (2007), described post-hurricane Katrina collections ranging from September to November 2005 in Alabama. Smith et al. (2006), reported 17 separate collections ranging from August to November in 2005 in Florida. Moulis et al. (2008), described collections in Georgia and South Carolina during October through December 2007. Culex coronator was first collected in Brunswick and New Hanover Counties in southeastern NC in September 2008. The second collection of Cx. coronator was October 2012 in New Hanover County (Harrison et al. 2016). In surveillance conducted during weeks 28-47 of 2013, three trap stations in New Hanover County produced 4 separate collections totaling 37 Cx. coronator, suggesting this introduced species may be here to stay. Finally, in 2015, two more Cx. coronator were collected in New Hanover County at their South Wilmington station on week 48 of 2015. This brings us to 2016 where things get interesting with Cx. coronator populations. During the last half of the 2016 mosquito season, multiple collections of Cx. coronator were collected in Brunswick, Columbus and New Hanover counties in southeastern N C between August 15-21 (Week 33) and December 19 th (week 51) 2016 (Figure 1). The weekly collections in New Hanover and Brunswick Counties were part of each program s ongoing mosquito monitoring activities. New Hanover County deploys eight miniature light traps with baited dry ice which are set Monday evenings and collected Tuesday mornings. Brunswick County uses three New Jersey traps baited with only light which are collected every morning. Both surveillance systems captured the weekly seasonal distribution of Cx. coronator in 2016. (continued next page) Figure 1. Total Culex coronator collected all trap types and frequencies. BRU: Brunswick County collections; NHC: New Hanover County collections, Col: Columbus County collections.

Page 8 Figure 2. Each point represents a Cx. coronator collection location in southeastern NC in 2016. (from previous page) Hurricane Matthew made landfall in the region on October 8, 2016. On October 10, 2016 (week 41) post disaster baseline traps were placed at six locations across Brunswick County. All six trap sites collected Cx. coronator. As part of the N C Department of Public Health emergency response, graduate students from East Carolina University in Greenville, NC were recruited and deployed to set CO 2 baited miniature light traps across Columbus County to document the post-disaster emergence of mosquito populations on October 27, 2016 (week 43). Six of the eight traps set in Columbus County collected 88 Cx. coronator specimens, hence expanding the range of Cx. coronator in southeastern NC to include Columbus County. During the 2016 season, 857 Cx. coronator were collected across 26 locations in southeastern NC (Figures 1, 2). The ongoing surveillance programs and the historical weather data in our counties provides the opportunity to examine seasonal distribution of Cx. coronator. By considering the weekly rainfall and temperature data with the 2016 seasonal distribution of Cx. coronator, we can assess whether environmental variables may have driven this year s increase of Cx. coronator populations. The North Myrtle Beach South Carolina (2017) National Oceanic and Atmospheric Administration (NOAA) weather station provides weather data for the southern end of Brunswick County, while the Wilmington North Carolina (2017) NOAA weather station collects similar information and represents the New Hanover County to the north. (continued next page) Figure 3a (top). Wilmington, NC NOAA weather data. 3b (bottom) North Myrtle Beach, NC NOAA Weather data.

Page 9 (from previous page) Using both data sets, we can see the changes and similarities in the environmental conditions across a distance of approximately 90 miles. Figures 3a and 3b show an above average temperature range for both weather stations for weeks 26 through 52 of 2016. The rainfall amounts at both stations for the last half of 2016 should be considered. The North Myrtle Beach weather station had three precipitation events from week 34 to 41 that deposited a combined total of 24.88 inches of precipitation in 2016. The Wilmington weather station displayed the same three events over the same time frame but only produced 20.60 inches of rain. The weather data for 2015 and 2016 suggests the above average rainfall and temperatures at the end of the 2015 season may have provided the opportunity for successful overwintering of Cx. coronator. The two consecutive years with above average temperatures and rainfall totals in the late fall likely led to the increased activity in Cx. coronator observed in the fall of 2016. Our historical surveillance data has allowed us to have a better operational understanding of the environmental conditions required to produce increased Cx. coronator populations. This, coupled with a better idea of where to look for Cx. coronator larvae on the high landscapes, presents an operational opportunity for larval collections of Cx. coronator in the future. We anticipate Cx. coronator populations to follow the same seasonal distribution as the resident populations of Cx. nigripalpus. We typically see adult Cx. nigripalpus activity ranging from weeks 34 to 48 during the season in southeastern NC. Finally, New Hanover County set CO 2 baited light traps on February 20, 2017 and collected 2 Cx. coronator at their South Wilmington site along the Cape Fear River. This is the first winter collection of Cx. coronator we are aware of in NC. (contact Jeff Brown: Jeff.Brown@brunsiwckcountync.gov) Acknowledgements We would like to express our sincere thanks to Dr. Stephanie Richards for the opportunity to be a part of this wonderful project, working with such a superb group of people. Her insight, supplies and knowledge of vectors was an outstanding asset to us. (references on next page) (from page 1). Answer to the Skeeter quiz: Culex restuans! Other facts about Cx. restuans. It is a vector, likely mostly between birds, of West Nile virus, St. Louis encephalitis virus, and other viruses. It is found from the Atlantic to the Pacific Oceans, and down into Central America. The adults look like Cx. pipiens/quinquefasciatus, as do the larvae, but there are differences in setae.

Page 10 (from previous page) References Debboun M, Kuhr DD, Rueda LM, Pewcor JE. 2005. First Record of Culex (Culex) coronator in Louisiana. USA. J Am Mosq Control Assoc 21:455-457. Dyer HG, Knab F. 1906. The larva of mosquitoes classified as independent organisms. J NY Ent Soc 14:169-230. Goddard J, Varnado WC, Harrison BA. 2006 Notes on the ecology of Culex coronator Dyer and Knab, in Mississippi. J Am Mosq Control Assoc 22:622-625. Harrison, B.A., Byrd,B.D., Sither,C.B. and P.B. Whitt. 2016. The Mosquitoes of the Mid-Atlantic Region: An Identification Guide. Mosquito and Vector-borne Infectious Diseases Laboratory Publication 2016-1. Western Carolina University, Cullowhee, NC, pg173. McNelly JR, Smith M, Micher-Stevens KM, Harrison BA. 2007. First records of Culex coronator from Alabama. J Am Mosq Control Assoc 23:473-475. Moulis RA, Russell JD, Lewandowski Jr. HB, Thompson PS, Heusel JL 2008.. Culex coronator in Coastal Georgia and South Carolina. J Am Mosq Control Assoc 24(4):588-590. National Oceanic and Atmospheric Administration. 2017National Weather Forecast Office Wilmington NC. http:// w2.weather.gov/climate/index.php?wfo=ilm. Smith JP, Walsh JD, Cope EH, Tennant RA Jr, Kozak JA III, Darsie RF Jr. 2006. Culex coronator Dyar and Knab, a new Florida Species Record. J Am Mosq Control Assoc 22:330-332. Introducing your Two New State Medical Entomologists Alexis M. Barbarin (alexis.barbarin@dhhs.nc.gov) is a Public Health Entomologist in the State of North Carolina Division of Public Health, Communicable Disease Branch. She began her career in Entomology researching bed bugs at Penn State University, where she received a Ph.D. in Entomology and a master of education in Agricultural and Extension Education. As a postdoctoral researcher at North Carolina State University, Dr. Barbarin investigated the dispersal patterns of bed bugs using molecular tools like microsatellite markers, coupled with the use of human social network analysis to uncover how human relationships mediate bed bug dispersal. In her current position, Dr. Barbarin hopes to use her entomological knowledge to alleviate the tick-borne disease burden of residents in the state of North Carolina. Michael Doyle (michael.doyle@dhhs.nc.gov) started his work life knee deep in water as an Environmental Consultant doing wetland delineations, then as a Regulatory Biologist for the Army Corps of Engineers. His mosquito control experience on a two-man crew with Colorado Mosquito Control, Inc., eventually running large city and county integrated mosquito control programs. After graduate school at Colorado State University (M.S. Entomology) he served as an Entomologist at the CDC, studying vector biology. This was followed by becoming the Executive Director of the Florida Keys Mosquito Control District, supervising a $10m/ year operation. Mike is now proud to join the NCDHHS as a State Public Health Entomologist, specializing in mosquito disease issues. His primary roles are to: assist counties with mosquito surveillance/control improvement; serve as a central resource vector information; and represent the State in the NCMVCA, AMCA and other organizations.

Page 11 2017 Sustaining Members Steve Molnar 877-881-6105 smolnar@myadapco.com Joe Andrews 888-603-1008 joea@allprovector.com Peter Connelly 706-338-4737 PeterC@amvac-chemical.com Kurt Vandock 919-549-2226 kurt.vandock@bayer.com Joe Strickhouser 630-894-2000 jstrickhouser@clarke.com Jeff Hottenstein 703-498-9362 jhottenstein@clarke.com

Page 12 2017 Sustaining Members Tommy Bowen 980-875-5422 Tommy.Bowen@duke-energy.com Joey Osbourne 828-308-2181 joey@themosquitoauthority.com Jim Andrews 910-547-8070 Jim.Andrews@valent.com Official NCMVCA Address: NCMVCA (c/o Dr. Stephanie Richards) 3403 Carol Belk Building 300 Curry Court Greenville, NC 27858

Page 13 2017 Executive Committee and Committee Chairs Executive Committee (Board Members) President Michael Reiskind mhreiski@ncsu.edu 919-515-0719 Vice-President Stephanie Richards richardss@ecu.edu 252-328-2526 Secretary-Treasurer Mark Phelps mark.phelps@pittcountync.gov 252-902-3216 Past President James Gardner james.gardner@pittcountync.gov 252-902-3210 Chairs of Standing Committees Award and Nominations Parker Whitt parker.whitt@ncagr.gov 336-462-6084 Constitution and By-Laws Michael Doyle michael.doyle@dhhs.nc.gov 919-546-1637 Finance Mark Phelps richardss@ecu.edu 252-328-2526 Legislative Joe Andrews joea@allprovector.com 919-343-8440 Membership and Communications Michael Reiskind mhreiski@ncsu.edu 919-515-0719 Program Stephanie Richards richardss@edu.ecu 252-328-2526 Member-At-Large Abram Young abram.young@brunswickcountync.gov 910-616-2475 Member-At-Large (Industry Rep) Joe Strickhouser jstrickhouser@clarke.com 704-333-2523 2017 Committee Members Award and Nominations Joe Andrews joea@allprovector.com Constitution and Bylaws Mark Phelps mark.phelps@pittcountync.gov Constitution and Bylaws Dennis Salmen Dennis_salmen@yahoo.com Finance Stephanie Richards richardss@ecu.edu Finance Michael Reiskind mhreiski@ncsu.edu Legislative Abram Young abram.young@brunswickcountync.gov Membership and Communications Michael Reiskind mhreiski@ncsu.edu Membership and Communications Chrystal Swinger clwsinger@cabarrushealth.org Membership and Communications Stephanie Richards (Website) richardss@ecu.edu Membership and Communications Joe Strickhouser (Industry/Vendor Support) jstrickhouser@clarke.com Program Stephanie Richards richardss@ecu.edu Program Brian Byrd (Student Competition) bdbyrd@wcu.edu Program Joe Strickhouser (Industry/Vendor Support) jstrickhouser@clarke.com Parliamentarian Michael Doyle michael.doyle@dhhs.nc.gov Auditor Ray Silverthorne rsilverthorne@cravencountync.gov Do you want to be more involved with the NCMVCA as a member on a committee? Speak to any of the above individuals, or contact the new officers at the end of the conference to see how you can help your organization continue to grow and promote vector control in North Carolina.

Page 14 Bruce s Corner There be Skeeters in Those Mountains By Bruce A. Harrison Many years ago in the late 1940s, my dad drove me from south Georgia to Spruce Pine, NC, to visit an aunt, and I remember the huge mountains, forests, and I marveled at the trout in the stream behind her house. Actually, I caught 4-5 small trout using worms and crickets for bait and they were my first taste of rainbow trout. That short trip rang the bell for my love of the mountains of North Carolina. This was enhanced further during the summer of 1958 when I was a student in Wildlife and Fisheries at N.C. State College and worked as a fishery biologist helper for 13 weeks camped out on seven of the beautiful reservoirs in the mountains. I was in love (and still am) with nature during that period (and girls), but I vividly remember that there were mosquitoes coming out of those forests late in the afternoon and at night that did not mind the stench of fish on me and would bite anything that wasn t covered by clothing or a blanket. Coming from the flatlands, swamps, rivers and creeks of southern Georgia where mosquitoes can be top dog outdoors I had never contemplated finding mosquitoes in a mountainous region. However, I suspect that there are a significant number of people from the coastal plain and piedmont of North Carolina today that have not considered mosquitoes being in or being of importance in the mountains. In fact, I m sure of this because a friend told me that a legislative representative from a mountainous county that was being interviewed in 2010 commented You mean there are still mosquitoes in the mountains? This occurred just seven years ago when efforts were being made to save the state elimination of the Public Health Pest Management Section of the NCDENR. When I returned to NC in March, 1993 to work in Public Health Pest Management it was fortuitous that I was assigned as the medical entomologist for 47 of the most-western counties in the state. This opened the door for me to repeatedly travel to the mountains. When Parker Whitt was hired in 1995 to work with me that doubled our efforts. We obtained permits to work in the state parks and when we had time, other than responding to complaints about ticks, flies, fleas, head lice, bed bugs, midges, rats, mice, bees, wasps, ants, delusions of parasitosis, post-hurricane surveillance and mosquito pooling work, etc., we made mosquito collections in our counties as often as possible. These trips increased dramatically after West Nile virus arrived in NC in 2000 and TVA contracted us with annual funding assistance to collect mosquitoes for WNv testing in mountainous counties with drainage into the French Broad River system that flows into Tennessee. Thus, we quickly built up collections and records for mosquito species that we found in the mountains. We also discovered that the few roads, predominately forested mountains, and our inability to get into certain areas made mosquito surveillance difficult. But like other North Carolinians the people of the mountains are friendly, provided you are courteous and respectful of their property. (continued next page)

Page 15 (from previous page) Unfortunately, there are many counties in the mountains that currently need considerable additional mosquito collecting to determine their species. Regardless, we were surprised at the mosquito diversity and abundance that we found in state parks, in the valleys, in city parks, in nurseries, on farms, and in small communities. Now that I have established the fact that mosquitoes exist in the mountains, I m going to provide proof of this. For a few nay sayers, it is true that our NC forefathers and families left the coastal plain in the summers to go live in the piedmont and mountains to avoid getting malaria. However, that was not because there were no mosquitoes in the mountains and piedmont, it was because the malaria vector, Anopheles quadrimaculatus, was not as abundant in the mountains and piedmont as in the coastal plain. If you wonder into the right place there are plenty of different species in abundance in the mountains. And, they are no longer confined to the countryside and forests. Currently, there are 10 genera and 39 species of mosquitoes that occur in the 24 mountain counties. This represents all 10 of the genera found in the state and 58 % of the 67 species now recognized in NC. Aedes albopictus and Aedes j. japonicus are the only species recognized from all 24 counties, and Aedes triseriatus is recognized from 22 counties, but almost certainly in all 24 counties (Figure 1). The two former species are Asian invasive species, with Ae. albopictus first found in NC in 1987 and Ae. j. japonicus in 2003. Until the arrival of those two invading species a native species, Ae. triseriatus, was the only recognized vector of La Crosse encephalitis virus in the mountain counties. Now, all three of these species are recognized as vectors of La Crosse encephalitis virus to humans and this disease occurs primarily in children. All three species can be very abundant in the growing cities in the western counties, and the rampant increase in summer human populations in the mountains dramatically increases human exposure to these species. Although politically overshadowed by West Nile virus, in the last decade La Crosse encephalitis virus has been responsible on an annual basis for more mosquito-borne human disease in North Carolina than eastern equine encephalomyelitis, West Nile, and Cache Valley viruses combined. Of the most recent series of arbovirus introductions, i.e. dengue, chikungunya and Zika, only the last virus is likely to strongly impact human health in NC and that will primarily be through travel related infected embryos in women in NC that contracted the virus elsewhere or through sexual contact in NC. Another interesting tidbit about vector mosquitoes in the mountains is Eugene Powell finding Ae. aegypti in used tires in Swain County in the late 1980s, and in 1994 during a used tire survey Eugene and I again found Ae. aegypti in the same area. Later in the early 2000s, Dr. Will Reeves collected a larva and reared a single Ae. aegypti female from a gutter on the Park Headquarters in the Great Smokey Mountain National Park in Swain County. (continued, next page)

THE BITING TIMES Page 16 (from previous page) Other commonly found species in the 24 mountain counties include Anopheles punctipennis (22/24), Culex restuans (20/24), Culex territans (19/24), Aedes atropalpus (18/24), Culex pipiens complex (17/24), Aedes vexans (14/24) and Culex salinarius (14/24). Nearly all of the 14 remaining species have been found in <11 counties, which clearly indicates the need for further surveillance. Regarding other records of interest, our malaria vector, Anopheles quadrimaculatus, was collected as females in Henderson and McDowell counties, and females of An. crucians s.l. were collected in 10 counties. Seven of the eight Psorophora species found in NC occur in the mountains, and Ps. horrida and Ps. mathesoni were typically found in Figure 1. Three most abundant vectors of La Crosse encephalitis in Western North Carolina. riverine flood waters in adjacent woodlands. Other females collected following woodland riverine flooding include Ae. cinereus (8 counties), Ae. sticticus (7 counties) and Ae. trivittatus (10 counties). Gene Powell also collected a female of Psorophora cyanescens in Buncombe County. Females of Culiseta melanura were collected in Alleghany, Buncombe, Henderson, McDowell and Rutherford counties. Females of Culex nigripalpus were collected in Henderson, Jackson, Macon, McDowell, Rutherford and Transylvania counties. Females of Ae. dupreei were collected in Buncombe, Cherokee, Henderson, Macon and Rutherford counties. Females of Ae. infirmatus were collected in Buncombe, Henderson, Rutherford and Transylvania counties. Females of Anopheles barberi were collected in Buncombe, Haywood, Jackson, Polk, Swain and Transylvania counties. One female of Ae. atlanticus was collected in Buncombe County, two females of Ae. tormentor were collected in Jackson and Henderson counties, and five specimens combined under Ae. atlanticus/tormentor were collected in Cherokee, Henderson, Macon, McDowell and Rutherford counties before the females of these two species could be differentiated. Wyeomyia smithii were collected as larvae in Jackson and Macon counties. (continued next page).

(from previous page). Although I have not addressed all 39 species, I think you now get the picture. Mosquitoes have run me out of Jackson Park in Henderson County, beside the Hiawassee River in Cherokee County, in Nurseries in Henderson and McDowell counties and in many yards in Figure 2. The kind of artificial containers that harbor larval Ae. albopictus, Ae. japonicus, and Ae. triseriatus. those changes above. (contact Bruce: skeeterdoc@gmail.com) towns and communities (particularly old sections) in the mountains. Since humans are responsible for and driving most of the mosquito problems in the mountains I think it is safe to say that things will get worse with the rapidly growing human populations across the mountains, more developments creating mosquito friendly habitats, climate change and probable new invasive viruses arriving in the U.S. Thus, more concentrated mosquito surveillance efforts are needed to keep track of burgeoning mosquito populations that will walk hand-in-hand with Acknowledgements. Although I cannot remember all of the individuals that helped accrue the above data, several clearly stand out and deserve thanks. First and foremost, Parker Whitt deserves special recognition for 17+ years of working with me on countless collection trips. His hard work greatly enhanced our knowledge of the mosquitoes in the mountains and western piedmont. Dr. Nolan Newton infrequently trapped mosquitoes near his home in Rutherford County and most of our records from that county came from his efforts. Dr. Brian Byrd and Charlie Sither have greatly assisted our knowledge of the mosquitoes in Haywood and Jackson counties and also in filling in county records for Ae. j. japonicus in five other counties in the mountains. Eugene Powell worked with Dr. Charles Apperson and his graduate students in Swain and Jackson counties primarily before I arrived, but he always sent specimens for confirmation if he felt they were of interest and a number of those specimens were very important additions to our knowledge. Page 17 Student Scholarships Our NCMVCA/MAMCA (Mid-Atlantic Mosquito Control Association) meeting is a ways off (February 12-14, Carolina Beach, NC), but we will have student scholarships available ($250). Students must send Brian Byrd (bdbyrd@wcu.edu) their presentation, along with an abstract by December 15, 2017. Please contact Dr. Byrd with further questions.

Page 18 (from page 4). For collections, we use several types of traps which include a light trap with CO 2 (dry ice), light trap with a lure (bait), and sentinel trap (large black trap with an inverted collection bag). Each of these traps appear to attract different amounts of mosquitoes (and other insects) and species of mosquitoes. We also use dippers, which is a method used to gather larvae samples to ID or send off to Western Carolina for further use. ID ing has turned out not to be as bad as I had originally thought. We were taught by looking backwards for original characteristics (using the manual) and told which specific features to look for. I definitely enjoyed how C.S has passion when it comes to teaching us the characteristics of the different species. She becomes very animated and uses sound effects when explaining it. The other aspect (public health) is going to check out the sites where complaints are coming from the first one we went to was at the Purdue Chicken Plant and I only went on the first visit where we went all the way around wood lines. We also saw someone with a large tent set up that was very elaborate in the middle of the woods. The other visit was a neighborhood with mixture of trailers and modular homes. This was an interesting adventure to say the least I ended up on the side with some questionable houses. Tuesday, me and T left notes about checking the properties later in the week and Thursday all of us followed up by looking for larvae, using the dipping method to collect any if found, and letting the residents know what we had found and what they needed to. Some were quick to tell us who had mosquitoes in their yard or that they didn t have any. We have also contributed data and on Wednesday we were introduced to 5 department heads and C.S. presented what we are doing and what we have found thus far (I was glad not to have to get up and present). Definitely looking forward to what the rest of the summer has in store!!!! The final leg of summer (7/13/2016) So hard to believe that summer break is almost over. I love my internship - I have learned so much this summer and am looking forward to continuing with this particular topic (mosquito - vectors) for independent study this coming school year. We have been setting up traps and gathering ovitraps in different areas all over the county the last few weeks. Recently, we went to old permanent sites to deliver notices about checking for mosquito larvae. These places ranged from Kannapolis, Harrisburg, and Concord. We had 4 or 5 places that were non-existent from 10 years ago, since C.S last visited these sites (funding for monitoring abroviruses/other vector borne diseases was cut back in 2008 and they started funding again because of the Zika virus threat). (continued on next page).

Page 19 (continued from previous page). We have also gone to check on complaints. Yesterday, we followed-up (with CS) on a complaint near CHA, and found a surprising breeding ground on the premise with mosquitoes. The person lived next to someone who put old cans of soda in a plastic bin filled with water from the recent rains, Yet - when the larvae samples were dead - due to someone putting pertherin (not sure on spelling) or another similar insecticide in the bucket to prevent other insects. The other complaint we went to had breeding all over - including a storm pipe, water on a trailer, and some miscellaneous buckets filled with water. It was also interesting to learn how to control mosquito breeding - which they use 3 types (I only remember 2 off the top of my head). MMF - is a layer of molecules that inhibit growth of mosquito larvae (Larvae stage is the stage before the mosquito is fully grown) but isn't harmful to other insects. C.S had a bottle of this (looks like cleaner) but has told us that they stopped making this particular kind of insecticide. The other control method is using gambusia - which are fish that eat mosquitoes (I am not sure if it is larvae or adult). (Disclaimer: we don't do the control methods as interns). Some Thoughts About my Final Year of Undergrad (8/3/2016) The internship at CHA working with mosquitoes has taught me so much in just 2 months. First off, I was planning on doing independent research in upcoming semesters but couldn't seem to find an area to focus on with mosquitoes and wasn't confident enough in my own abilities. The research that takes place on campus is both field work (setting out/picking up the traps, taking larval samples) and molecular work (looking for certain disease RNA in the mosquitoes DNA). This internship taught me to more about field work (as well as public health) and ID'ing. Also - it helped me realized that I am meant to do some type of research. Second, the people at CHA and establishing new connections (in terms of references). I was impressed when I first was oriented to CHA how friendly everyone was and how welcoming the HR department made me feel. I learned early on my mentor's (Dr. H) mentor worked with my boss with mosquito identification and public health, even after the funding was cut for vector borne disease control. And speaking of my boss - she has been a great person to work for and learn under. She is very animated and passionate about mosquitoes - and educating others. (contact Chrystal Swinger: Chrystal.Swinger@cabarrushealth.org). Editor s Note: Shannon will be graduating from Catawba College this June with a B.A. in Biology and a minor in psychology, environmental science and chemistry. She will be moving to Connecticut immediately after and will pursue a graduate degree in vector-borne disease, specifically tick related.

Page 20 2017 NCMVCA Mosquito Workshop Registration July 11-12, 2017 Pitt County Agricultural Extension Center 403 Government Circle, Greenville, NC 27834 Registration is free for NCMVCA Members who register before June 19, 2017. This workshop includes the annual business meeting of the NCMVCA. If you have questions about your 2017 membership status, please check with Stephanie Richards (richardss@ecu.edu). Confirmation of your registration will be emailed to you upon receipt. You may pick up your name badge and other materials at the registration desk at the workshop. Day 1 Registration (mosquito surveillance/control) Received before June 19, 2017 After June 19, 2017 (Late) 10:00 a.m. 5:00 p.m. Member Free $20 Non-Member (includes regular membership [$20] for 2017) $20 $20 Attending FREE lunch on Day 1 (please circle one)? YES NO Please explain any dietary restrictions Day 2 Registration (mosquito larval identification) Received before June 19, 2017* After June 19, 2017 (Late)* 8:00 a.m. 12:00 p.m. Member $50 $70 Non-Member (includes NCMVCA membership [$20] for 2017) $70 $90 *Note that the number of seats in the Day 2 larval identification course is limited due to availability of microscopes and specimens. NCMVCA membership fee for 2018 (optional) $20 $20 Vendor Additional Sponsorship (optional) $ Total: $ Name: Email Address: Phone Number: Employer: Mailing Address: Method of Payment: Check (payable to NCMVCA) Invoice required (emailed to address given) Credit card (see below) Card number: Code on back of card: Expiration date: Billing zip code: If you are a 2017 NCMVCA member and plan to attend only Day 1 of the workshop (free), please still send the registration form to Stephanie Richards (see below) for planning purposes. Mail Completed Registration Form and Payment (if applicable) to: Stephanie Richards (NCMVCA) 3403 Carol Belk Building, 300 Curry Court Greenville, NC 27858 Email: richardss@ecu.edu ; Phone: 252-328-2526

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Page 22 Order Form The Mosquitoes of the Mid-Atlantic Region: An Identification Guide Bruce Harrison, Brian Byrd, Charles Sither, and Parker Whitt This 201 page spiral-bound 8.5 X 11 inch guide includes dichotomous keys to the adult females and fourth instar larvae for 8 states (DE, GA, NC, MD, PA, SC, VA, and WV) with more than 585 novel Adobe Illustrator figures. Printed on 100 lb gloss paper, this guide includes sections such as: 1) Taxonomic Interpretations, 2) State Records, 3) Basic Morphology, 4) The Acquisition of Characters to Separate Larval Instars, 5) Extensive Notes, 6) Couplet Sequences, 7) Illustration Index, 8) How to Use a Dichotomous Key, 9) Glossary, and others. The keys were thoughtfully reviewed by experts from the Smithsonian Institute, North American Mosquito Control Districts, and Academia. Item Quantity Price per key Total Mosquito key $30 Grand total: Name: Email Address: Phone Number: Employer: Mailing Address (where key will be mailed): Method of Payment: Check (payable to NCMVCA) Invoice required (emailed to address given) Credit card (see below) Card number: Code on back of card: Expiration date: Billing zip code: Email or Mail Completed Form and Payment To: NCMVCA c/o Stephanie Richards 3403 Carol Belk Building, 300 Curry Court Greenville, NC 27858 richardss@ecu.edu NCMVCA: North Carolina Mosquito and Vector Control Association www.ncmvca.org