INSTALLATION PEST MANAGEMENT CONSULTATION, IOWA ARMY AMMUNITION PLANT, MIDDLETOWN, IOWA, 2-5 DECEMBER 1988 DEPARTMENT OF THE ARMY U.S. Army Environmental Hygiene Agency Aberdeen Proving Ground, Maryland 21010-6422 [Seal of Department of Defense, United States of America] REPLY TO ATTENTION OF: HSHB-MR-EMG INSTALLATION PEST MANAGEMENT CONSULTATION NO. 16-44-0520-90 IOWA ARMY AMMUNITION PLANT MIDDLETOWN, IOWA 2-5 DECEMBER 1988 1. REFERENCES. a. AR 40-5, 30 August 1986, Preventive Medicine. b. AR 420-76, 3 June 1986, Pest Management. c. Message, HQDA, ATTN: DASD-PSP-D, 191800Z June 1987, subject: Lyme Disease Surveillance and Prevention. 2. AUTHORITY. Memorandum, HSC, HSCL-P, 10 February 1988, subject: Lyme disease on Army Installations. 3. PURPOSE. a. To characterize the spatial distribution and relative frequency of the deer tick, Ixodes dammini, on the Iowa Army Ammunition Plant (IAAP), as determined by infestations on white-tailed deer, Odocoileus virginianus. b. To determine the prevalence of infection of indigenous tick species with the Lyme disease agent, Borrelia burgdorferi. c. To assess the prevalence of specific antibody to B. burgdorferi infection in the white-tailed deer population. d. To gather information on possible human cases of Lyme disease acquired at IAAP. e. To present information concerning the epidemiology and symptomatology of this idsease to installation medical and civilian personnel, and to discuss protective measures to reduce the risk of exposure to the vector species. f. To assess the potential risk of exposure of personnel who train, work or recreate on IAAP to the tick vector(s) of Lyme disease. 4. GENERAL. a. Personnel Contacted. See Appendix A for a list of personnel contacted and/or briefed. b. Survey Conduct. The survey was conducted by CPT George W. Korch, Entomologist, Entomological Sciences Division (ESD), U.S. Army Environmental Hygiene Agency (USAEHA), and by Dr. Darrell Hale, Entomologist, U.S. Army Environmental Hygiene Agency - West (USAEHA-W), Fitzsimons Army Medical Center, Aurora, Colorado. c. Background. (1) The IAAP is located 11 miles west of the Mississippi River in Des Moines County, Iowa. It measures approximately 30 square miles in area and is comprised of rolling terrain with interspersed hardwood, deciduous forest, and open pasture land which is leased for agriculture. Most administrative and industrial operations are located on the northern half of the installation. The southern half is more wooded, being deeply cut with streams draining to the Skunk River along the southern boundary. A map of the installation is provided as Appendix B. (2) The IAAP is a government-owned, contractor-operated site, managed by Mason & Hanger-Silas Mason Co., Inc (M&HSM). In addition to the civilian workers employed by this contractor, the Department of Army (DA) maintains an oversight office staffed by DA civilian employees and two permanent party military personnel, including the installation commander and the executive officer. (3) Other military personnel who routinely use the installation for training purposes include the 224th Engineering Battalion (Bn), Burlington, Iowa; the University of Iowa ROTC program; the 410th Infantry Bn, Ottumwa, Iowa; and the 134th Medical Co, Wahington, Iowa. An estimated 1,200-1,600 military personnel per year train on the installation during 5,000 - 8,000 man days. The installation is also used by the local Boy Scouts and by area sportsmen, hunters, and fishermen. (4) One human case of Lyme disease was reportedly acquired on IAAP by a National guardsman during training exercises in April 1986 [sic], prompting concern for the risk of additional human exposure to this disease on the installation. (5) Lyme disease is a bacterial disease transmitted to man and animals via the bite of an infected tick. A synopsis of this disease is presented in Appendix C. One of the three major U.S. foci of human disease occurs in adjacent Minnesota. Human disease was first reported in Iowa around 1985-86, and the principal vector for the disease in the upper midwestern U.S., Ixodes dammini, was first found in three eastern counties in Iowa around 1986. It is likely that additional human involvement with Lyme disease in Iowa may occur as the enzootic focus expands from neighboring Minnesota. 5. FINDINGS AND DISCUSSION. a. LD Surveillance, Occurrence, and Practices to Date. (1) The one human case of Lyme disease reportedly acquired at IAAP occurred in a member of Company A, 224th Engineering Battalion, Iowa Army National Guard, who was likely infected while training at IAAP on 25-26 April 1987. The patient was a 22-year old male, from Washington, Iowa, who recalled finding five immature ticks attached to his body following the field exercise. This individual subsequently developed symptoms of Lyme disease which required hospitalization and prolonged antibiotic therapy. (2) The medical authority on IAAP, Dr. Saar, suggested that there may have been two possible additional cases of Lyme disease acquired on IAAP, although no clinical or serological evidence was presented to substantiate either case. One individual was a 5-year old male, who had accompanied his father onto the installation on 12 September 1987. He experienced numerous tick bites, which resulted in a reaction suggestive of a typical immunologic response to the tick salivary components; however, no other symptoms suggestive of Lyme disease were reported. The second instance involved a foreman at one of the industrial yards, although no specifics of this infection were given. Therefore, only one human case of Lyme disease can be ascribed to IAAP at this time. (3) The recognized tick vector of Lyme disease, Ixodes dammini, has been found in studies performed on IAAP by non-military personnel but in very low numbers. These studies were performed by Dr. Nixon Wilson, Professor at the University of Northern Iowa, Cedar Falls, who collected ticks from deer during the 1986 and 1987 hunting seasons and provided no cost consultation and tick identification services to the installation from December 1986 to mid-July 1988. A compilation of the species of ticks collected and identified by Dr. Wilson, as well as results of selective screening for the etiologic agent in these tick specimens, is given in Appendix D. (4) No B. burgdorferi infected I. dammini were found by Dr. Wilson at IAAP; however, three spirochete infected winter ticks, Dermacentor albipictus, were removed from deer during the 1987 hunting season. The assay was performed by Dr. Vincent Marshall, Animal Vaccine Laboratory, Council Bluffs, Iowa, and the spirochetes were presumed to be Borrelia burgdorferi, although the tick species from which the spirochetes were recovered is not generally associated with Lyme disease epizootiology. (5) The collection and dissemination of Lyme disease information was recently instituted at IAAP, due to its newly identified status on the installation. Mr. Buren, of the M&HSM security office, has compiled news reports, scientific articles and other information on the disease. Dr. Wilson provided 1,000 copies of a Lyme disease summary to members of the installation (see Appendix E); however, this did not include a discussion of personal protective measures. b. Deer Management Practices. (1) The deer management program at IAAP is commendable. The current size of the deer herd on IAAP is estimated at around 1,100-1,600 according to Mr. Bob Arbige, Chief, Operations Review Division. The deer hunting program has succeeded in lowering this population over 4 years from an estimated herd size of 3,600 deer. (2) Proper deer herd management is essential to the reduction of Lyme disease transmission. These animals serve as the principal hosts for the adult stage of the deer tick, thus for the reproductive potential of the tick population. Deer herd statistics are collected annually by the IAAP (see Appendix F). The long range goal is to maintain the deer herd population around 1,000. One potential difficulty presented to the management program, however, is the lack of exclusion barriers to entry by deer onto the installation from the surrounding area, especially along the southern boundary of the installation. c. Lyme Disease Briefings. (1) The survey officer presented a 1-hour seminar on Lyme disease symptoms, ecology, epidemiology, and personal protective measures to management personnel from the Safety Office, Security Office, Commander's Office, and Operations Review Division. A similar presentation was delivered separately to the entire medical staff. (2) Over 300 hunters were briefed on Lyme disease during a safety meeting regularly scheduled the evening before the opening of gun season. Attendance at this meeting was mandatory for participation in the installation's deer hunting program. The survey officer requested hunters' cooperation in collecting blood samples and ticks at this time. The subsequent level of participation by the hunters in the sampling efforts was greatly enhanced by this presentation, with nearly 60 percent of the hunters providing ticks and/or blood samples from their respective kills. d. Tick and Serum Collection Activities. (1) Three hundred blood/tick collection kits (see Appendix G for kit contents) were distributed to hunters as they arrived at 0530 hours to select hunting areas. Kits not used on the first day were returned to the survey team and redistributed the following day. (2) Approximately 400 deer were killed by hunters during the 2 days of the survey. Hunter completed data sheets were collected on 177 animals, and these data are compiled by age, sex and location in Appendix H (Table H-1). Hunters submitted blood samples from 168 deer at check-in, yielding a 56 percent return rate. We examined 310 deer for evidence of tick infestation during routine weighing and check-in procedures. Each deer was examined for 5 to 10 minutes according to the USAEHA Lyme disease surveillance protocol, concentrating search efforts on the ears, head, and neck. (3) A total of 294 ticks from 120 (39 percent) of the deer were collected and later identified at the Agency. Only 1 adult and 1 nymphal I. dammini were found on infested deer (see Appendix H, Table H-2), representing 0.6 percent of the total tick sample. The vast majority of ticks (99.1 percent) were Dermacentor albipictus, the winter tick. One Amblyomma americanum nymph was also found. The low level of I. damini infestation seen on IAAP deer corroborates previous tick survey statistics compiled by Dr. Wilson with 0.4 percent of the 1986 tick sample removed from deer, and none of the 1987 sample, represented by this species (see Appendix D). (4) An average of 0.9 ticks/deer (all species included) were found. No significant differences were seen in mean infestation level or prevalence of infestation between male and female deer. Regression analysis of the infestation level for all ticks observed on deer by weight of the deer specimen revealed a significant positive regression coefficient (p less than 0.05) indicating greater numbers of ticks on larger deer. The highest level of tick infestation seen on deer (Appendix H, Table H-3) occurred in hunting area 5 (3.05 ticks/deer) and the lowest level observed was from area 2 (0.91 ticks/deer). e. Analysis of Ticks and Serum Samples for Evidence of B. burgdorferi Infection. (1) Ticks were screened at the Agency for infection with Borrelia burgdorferi by immunofluorescent antibody (IFA) assay (see Appendix G for methods). A total of 2 I. dammini, 1 Amblyomma americanum and 195 Dermacentor albipictus were examined by IFA. No spirochetes were seen in any of these samples. (2) Ninety-six Dermacentor albipictus ticks were sent to the Connecticut Agricultural Research Station, New Haven, Connecticut, for isolation attempts for spirochetes. Tissues from 44 ticks surviving transport were individually placed into duplicate tubes Barbour-Stonner- Kelley (BSK) medium, with and without the antibiotic rifampin. No spirochetes were isolated from any of these ticks (Appendix H, Table H-4). (3) A total of 163 deer sera were screened for specific antibody to the Lyme disease spirochete using by [sic] IFA according to methods presented in Appendix G. Overall, 4 percent of the deer sera indicated positive antibody titer to B. burgdorferi (Appendix H, Table H-5). The available data for these positive animals are broken down according to mass, sex and location of the deer (Appendix H, Table H-6). Of cases with available data, all seropositive animals were females ranging between 45 and 55 kg in mass, which indicates infection in relatively young animals. There was no apparent clustering of seropositive animals in particular hunting areas. (4) The only evidence that Lyme disease spirochetes may be extant on IAAP derives from the occurrence of one confirmed human case of disease which is believed to have been acquired on the installation and evidence of infection in deer according to finding of positive antibody in this population. The discovery of positive serology in 4 percent of deer indicate that the spirochete is likely being transmitted within this population, albeit at a low rate. While there is no well-defined association between the serum antibody prevalence rate in white-tailed deer and prevalence of human disease, the finding does suggest that human exposure to the spiorchete will occur. (5) The results also suggest that the infestation level of the vector species on deer is fairly low at this time. Survey data from previous years also suggest a low vector density. Deer survey studies performed in areas in the country where Lyme disease is endemic typically yield higher I. dammini infestation levels on white-tailed deer. Carey et al. (1980) reported that 65 percent of deer were infested with a mean level of 25.7 adult deer ticks per deer in Connecticut.* --------- *Carey A.B., W.L. Krinsky, and A.J. Main. 1980. Ixodes dammini (Acari:Ixodidae) and associated ixodid ticks in south-central Connecticut, USA. J. Med. Entomol., 17(1):89-99. (6) There are several possible reasons for the low number of I. dammini observed during this survey i.e., the tick species was not active at this time in this area; infestations were missed; or the tick is just becoming established in this geographic area. The accumulated evidence strongly suggests the latter explanation. The deer tick is likely being introduced into the region from the established Minnesota focus to the northeast, possibly through bird migration. Further surveys would be required to evaluate whether deer ticks are increasing in number on IAAP. (7) The proper conditions for endemic Lyme disease exist at IAAP. The necessary components to support an endemic focus for the deer tick, and hence for Lyme disease include, but are not limited to, a well-established deer herd, wooded habitat, and presence of the white-footed mouse, Peromyscus leucopus. The first two factors are present, and it is very likely that the mouse is present as well, since the installation falls within the geographic range of the species and has a suitable habitat. (8) The observation made by Dr. Wilson of a Borrelis sp. in the winter tick population suggests that infection may be "spilling over" into an atypical vector species. Similar observations have been noted on Long Island, New York, and in Maryland, with American dog ticks, Dermacentor variabilis, or lone star ticks, Amblyomma americanum, showing Borrelia burgdorferi infections as a result of feeding on the same mammalian host as the deer tick. Isolation and characterization of the spirochetes found in the winter ticks from IAAP is required, however, to definitively establish whether the observed bacteria were in fact B. burgdorferi. Our attempts to isolate B. burgdorferi in the IAAP winter tick specimens were unsuccessful; therefore, we are unable to substantiate this observation. These findings do not indicate that the spirochete is absent from IAAP, but more likely, that the prevalence of infected ticks is currently low. 6. CONCLUSIONS. Lyme disease appears to be in a hypo-endemic status at this time but may be increasing in this geographic region. The possibility of increasing deer tick and spirochete activity on IAAP necessitates further surveillance effort for the tick vectors. It is also necessary to educate individuals at risk of exposure to the tick about Lyme disease. 7. RECOMMENDATIONS. a. To ensure good pest management practice, the following recommendations are made: (1) Select a Lyme disease coordinator to oversee training of installation personnel concerning Lyme disease prevention, and to coordinate with this Agency or other appropriate institutions for further Lyme disease surveillance. (2) Conduct an annual safety briefing(s) for installation personnel concerning Lyme disease prevention, including use of personal protective measures. (3) Provide Lyme disease information and personal protective measures via a pamphlet or information sheet to unaffiliated persons or groups using the installation for recreational purposes. (4) Inform DA military units using IAAP for training purposes of the possible threat due to Lyme disease, and provide appropriate Lyme disease information to these units coordinated through the 224th Engineering Bn. (5) Monitor changes in the population of the tick vector(s) of Lyme disease and of the infection status of Ixodes dammini by providing ticks from at least one spring sampling period, using drag cloth collection, to this Agency. (6) Continue the commendable deer management program to achieve further reduction of the deer herd. b. To ensure regulatory compliance, the following recommendation is made: Report human cases of Lyme disease acquired on IAAP to the Office of the Army Surgeon General in accordance with AR 40-5, para 3-1. [signature of Alfred L. Hoch] for GEORGE W. KORCH, JR. CPT,MS Entomologist Entomological Sciences Division APPROVED: [signature] ALFRED L. HOCH MAJ, MS Chief, Pest Management Branch Entomological Sciences Division APPENDIX A PERSONNEL CONTACTED 1. LTC Jack Conway, Commander, IAAP. 2. Mr. Robert Haines, Safety Office, IAAP. 3. Mr. Allan Buren, Security Training Officer, Security Office, M&HSM Company, Burlington, Iowa. 4. Dr. Saar, Medical Officer, Dispensary, M&HSM Company, Burlington, Iowa. 5. SSG Ross, Training NCO, 224th Engineering Bn, Iowa National Guard, Burlington, Iowa. 6. Dr. Nixon Wilson, Department of Biology, University of Northern Iowa, Cedar Falls, Iowa. 7. Mr. Richard Herman, Security Officer, Safety Office, IAAP. 8. Mr. Bob Arbige, Chief, Operations Review Division, IAAP. 9. Mr. Ron White, MACOM Entomologist, Installation and Service Activities, AMC. 10. LTC Benjamin McLaurin, Medical Entomologist, Surgeon's Office, U.S. Army Material Command. 11. Dr. John F. Anderson, Director, Connecticut Agricultural Experiment Station, New Haven, Connecticut. APPENDIX B [hunting and fishing map - omitted] APPENDIX C GENERAL LYME DISEASE INFORMATION AND PERSONAL PROTECTIVE MEASURES 1. Lyme disease is a multisystemic bacterial disease which is transmitted to man generally via the bite of an infected tick. The disease progresses from an early stage involving a rash, malaise, fever and several other general symptoms to a secondary stage characterized by neurologic, or cardiologic abnormalities. Intermittent or transient arthritis and chronic debilitating neurologic disturbance are typically seen in the third stage of illness and can last months to years in duration. Not all patients present the entire range of symptoms. Many individuals apparently become asymptomatic, although it is not clear whether this necessarily indicates elimination of the causative organism from the patient. Serological findings are used to corrobarate clinical evidence of disease but should not be relied upon as diagnostic of Lyme disease. Early antibiotic therapy is effective in reducing likelihood of further symptoms, although failures to alleviate symptoms with antibiotic therapy have been reported. Tetracycline and its derivatives are the current drugs of choice in treatment. 2. The spirochete resides in the midgut of ticks, primarily of the genus Ixodes. The organism is transmitted to a mammal (or avian) host via the tick's salivary secretions during ingestion of a blood meal. The white- footed mouse is a particularly important host for the subadult tick stages (larvae and nymphs), and participates in the natural transmission cycle of the spirochete. Infected nymphal (i.e. second stage) ticks pass the infection to white-footed mice early in the spring. These infected woodland rodents then transmit the bacterium to the new generation of ticks (larvae) which feed in the late summer. Deer support the adult tick population and provide an opportunity for mating and dispersal of gravid females. Deer probably do not play an important role as far as transmitting infection to the ticks. 3. The deer tick is most likely to be encountered in wooded settings, seeking hosts while waiting on low-lying vegetation. The nymph measures only 1.2 mm in length and is dark brown to black in color, making it difficult to see on military field uniforms. Frequent inspection of the garments for ticks while in the field is necessary to prevent later tick attachment. Attached ticks must be removed as soon as possible, using tweezers to grasp the tick at the skin surface and gently pull away from the skin. 4. Use of repellents such as DEET provide only limited protection against tick attachment. The clothing-based permethrin repellents administered as a 0.5-percent aerosol spray provide a very high level of repellency (due to toxic effect against ticks); however, are not available for use under current law in the State of Iowa. Use of permethrin spray would be possible only after State application for a local use permit under section 24 (c) of the Federal Insecticide Fungicide and Rodenticide Act (FIFRA), or under national or military registration for this particular application route. APPENDIX D PREVIOUS SURVEILLANCE ACTIVITIES FOR DEER TICKS ON IAAP Previous collections of ticks were performed on IAAP by Dr. Nixon Wilson, Biology Department, University of Northern Iowa. TABLE D-1. A COMPENIUM OF THE TICKS COLLECTED BY DR. N. WILSON DURING DEER HUNTING SEASONS ON IAAP ---------------------------------------------------------------- Number of Number of Number of Deer Deer ticks Survey dates Examined Infested Tick species collected ---------------------------------------------------------------- 6-7 Dec 86 182 19 D. albipictus 220 I. dammini 1 5-6 Dec 87 NA(1) 43 D. albipictus 205 (1) Data not available ---------------------------------------------------------------- TABLE D-2. COMPILATION OF ALL TICKS COLLECTED BY DR. N. WILSON FROM IAAP DURING THE PERIOD FROM APRIL 1987 UNTIL JULY 1988 TICK SPECIES ---------------------------------------------------------------------- Ixodes Amblyomma Dermacentor Dermacentor STAGE dammini americanum variabilis albipictus ---------------------------------------------------------------------- Adult 2 234 1437 446 Nymph 0 231 1 10 Larva 0 1 0 2 ---------------------------------------------------------------------- APPENDIX E LYME DISEASE: AN EMERGING PUBLIC HEALTH PROBLEM IN IOWA Lyme disease, first recognized in 1975 in Connecticut, is initially characterized by a slowly expanding red rash and flu-like symptoms. This frequently is followed weeks to months later by recurrent inflammatory joint arthritis. Neurologic or cardiac abnormalities also may be manifested. The majority of patients with Lyme disease have reported tick exposures. The recently discovered causative organism is a bacterium which is passed to humans when the tick bites. Lyme disease is known to occur in three distinct geographical areas: 1) the northeast where it was first discovered and where most of the cases have been reported; 2) the midwest (Minnesota and Wisconsin) where an increasing number of cases have been reported; and 3) the west where it recently has been reported. Additional cases are being reported from other parts of the country and it is now known from over 30 states. Ixodes dammini is the tick species incriminated in transmission of the disease in the northeast and midwest while another species is infected in the west. It is suspected still another species is the vector in the south. All of these species are related and all feed readily on man. The adults and immature stages of I. dammini prefer deer and mice, respectively, although they can be found on a variety of other domestic and wild animals. There have been several cases of Lyme disease reported in Iowa but the primary vector, I. dammini, was unknown from the state. Research was initiated in the spring of 1986 in an attempt to find this tick and gather information on other ticks occurring in Iowa. The first specimen of I. dammini was found on a dog in Fayette Co., in May 1986. Since then 8 other specimens have been found on dogs and deer in Allamakee, Des Moines, and Fayette Cos. The adult ticks occur on these hosts during the cooler months of the year, whereas the immature stages are active in the spring and summer. Research on tick-host relationships and distribution in Iowa is continuing. You can help by sending me any ticks you find. They can be preserved in rubbing alcohol in a small pill bottle or vial and sent along with information on the host, locality collected (nearest town), date, and collector. All specimens received will be acknowledged. Thank you. Nixon Wilson Biology Department University of Northern Iowa Cedar Falls, IA 50614-0421 (319) 273-2010 APPENDIX F DEER POPULATION CENSUS ON IOWA ARMY AMMUNITION PLANT 2/11/87 IOWA ARMY AMMUNITION PLANT WINTER AERIAL DEER SURVEYS YEAR DATE COUNT COMMENTS ------- ------ ----- --------------------- 1972-73 18 DEC 187 80% OF TIMBER COVERED 1973-74 NONE 0 1974-75 FEB 183 40% OF TIMBER COVERED 1975-76 FEB 211 80% OF TIMBER COVERED 1976-77 NONE 0 1977-78 24 JAN 729 100% OF TIMBER COVERED 16 APR 370 1978-79 6 MAR 665* NOT GOOD COUNT 1979-80 19 FEB 697 3 IN SNOW COVER 1980-81 NONE 0 1981-82 NONE 0 1982-83 22 MAR 979 1 IN SNOW-MARGINAL 1983-84 24 JAN 1598 2-4 IN SNOW COVER 1984-85 29 JAN 1917 4-8 IN SNOW COVER(IDEAL) 1985-86 12 FEB 1240 3-4 IN SNOW COVER(EXCELLENT) 1986-87 7 APR 1086 NO SNOW, FOLIAGE, POOR CONDITION 1987-88 11 FEB 1101 4-8 IN SNOW COVER(IDEAL) * COUNT VARIED FROM 570 TO 665 2/11/87 IOWA AMMUNITION PLANT BOW TO GUN HARVEST RATIO YEAR BOW GUN BUCKS % BUCKS DOE % DOE TOTAL ---- --- --- ----- ------- --- ----- ----- 1976* 10 97 73 68.2% 34 31.8% 107 1977 63 62 77 60.6% 50 39.4% 127 1978 67 112 119 66.5% 60 33.5% 179 1979 129 115 161 66.0% 83 34.0% 244 1980 138 133 208 71.5% 83 28.5% 291 1981 134 176 199 64.2% 111 35.8% 310 1982 138 210 236 67.8% 112 32.2% 348 1983 178 225 281 69.7% 122 30.3% 403 1984 241 532 430 55.6% 343 44.4% 773 1985 345 *296 726 39.4% 1117 60.6% 1843 1986 498 *296 709 39.5% 1085 60.5% 1794 1987 254 910 474 40.7% 690 59.3% 1164 * PLANT STRIKE CURTAILED BOW HUNTING ACTIVITY 11/2/88 IOWA ARMY AMMUNITION PLANT DEER HARVEST STATISTICS (1977-1987) YEAR BUCKS DOES TOTAL %BUCKS %DOES ---- ----- ---- ----- ------ ----- 1961 9 10 19 47.4% 52.6%* 1962 10 9 19 52.6% 47.4%* 1963 32 33 65 49.2% 50.8%* 1964 33 32 65 50.8% 49.2%* 1965 78 79 157 49.7% 50.3%* 1966 129 128 247 52.2% 51.8%* 1967 60 59 119 50.4% 49.6%* 1968 57 56 113 50.4% 49.6%* 1969 66 65 131 50.4% 49.6%* 1970 48 60 108 44.4% 55.6% 1971 65 73 138 47.1% 52.9% 1972 48 75 123 39.0% 61.0% 1973 88 50 138 63.8% 36.2% 1974 106 51 157 67.5% 32.5% 1975 100 57 157 63.7% 36.3% 1976 73 34 107 68.2% 31.8% 1977 77 50 127 60.6% 39.4% 1978 133 46 179 74.3% 25.7% 1979 161 82 243 66.3% 33.7% 1980 208 83 291 71.5% 28.5% 1981 199 111 310 64.2% 35.8% 1982 236 112 348 67.8% 32.2% 1983 281 122 403 69.7% 30.3% 1984 430 343 773 55.6% 44.4% BOW 1985 258 287 545 47.3% 52.7% GUN #1 1985 210 333 543 38.7% 61.3% 1985 TOTAL GUN #2 1985 165 318 483 34.2% 65.8% 1843 GUN #3 1985 93 179 272 34.2% 65.8% ==== BOW 1986 234 264 498 47.0% 53.0% GUN #1 1986 194 257 481 40.3% 59.7% GUN #2 1986 190 357 547 34.7% 65.3% 1986 TOTAL GUN #3 1986 77 122 199 38.7% 61.3% 1794 GUN #4 1986 9 38 47 19.1% 80.9% ==== GUN #5 1986 5 17 22 22.7% 77.3% BOW 1987 122 132 254 48.0% 52.0% GUN #1 1987 176 276 452 38.9% 61.1% 1987 TOTAL GUN #2 1987 143 225 368 38.9% 61.1% 1164 GUN #3 1987 33 57 90 36.7% 63.3% ==== ------------------------------------------------- TOTALS 4636 4712 9348 49.6% 50.4% ==== ==== ==== ==== ==== *BUCK/DOE RATIO IS AN ESTIMATE APPENDIX G LYME DISEASE DETECTION METHODS AND EQUIPMENT 1. DEER HUNTER TICK/BLOOD COLLECTION KITS. Kits provided to hunters for collection of samples each contained a 7 milliliter (ml) sterile Vacutainer (tm) tube; one single piece, molded plastic pipet; one paper cleaning wipe; Lyme disease health information sheet; specific instructions for specimen collection; data recording sheet; and a screw-capped, plastic vial for tick specimens all enclosed in a plastic bag. Each kit was assigned a unique identification number, and this number was printed on the blood tube, tick vial, data sheet and outer surface of the kit. 2. IMMUNOFLUORESCENT ANTIBODY ASSAY METHODS. a. Infection status was determined by dissecting the midgut from the tick in phosphate buffered saline (PBS); triturating the tissues, and air drying on 10 well Teflon [Registered]-coated spot slides (Cel-line, Inc.) These slides were fixed in -20 degrees C for 7 minutes and stored at -70 degrees C until assayed. B. burgdorferi were observed using an IFA technique. The primary antibody used to detect the spirochete was a monoclonal antibody directed against the outer surface A protein (H 5332), the secondary detection antibody was Fluorescein isothiocyanate (FITC) conjugated Goat antimouse IgG (Heavy and Light chain, Kirkegaard & Perry Labs, Inc.). b. An immunofluorescent antibody test was used to determine the prevalence of infection with Borrelia burgdorferi in deer. Sera were initially screened for antibody response at a 1:32 dilution in PBS, and all samples yielding positive reaction were further titrated by fourfold dilution to 1:512. Serum samples were overlain in replicate on the B. burgdorferi coated antigen spot slides (Cel-line, Inc.) for 1 hour at room temperature in a humidified chamber. Slides were washed twice with PBS and a detector antibody, FITC conjugated Goat antideer IgG (Heavy and Light chain, Kirkegaard & Perry Labs, Inc.), was added to each well of the spot slides. Following a 30-minute incubation at room temperature, slides were washed twice with PBS, cover-slipped. A known positive deer serum control (provided courtesy of Dr. Edward Bosler, New York State Department of Health) was used as a quality control. Slides were read under ultraviolet light at 400 x magnification and response was graded as either strong (+), weak (+/-) or negative. Final antibody titer category for a sample was assigned based on the highest dilution yielding at least one strong response. Samples yielding weak responses on both replicates were assigned a titer 1/2 that of the corresponding dilution. Sera demonstrating antibody reactivity at titers greater than or equal to 1:256 dilution were considered indicative of B. burgdorferi infection. ----------- [Registered]Teflon is a registered trademark of E. I. Dupont de Nemours and Co., Inc., Wilmington, Delaware. Use of trademarked names does not imply endorsement by the U.S. Army but is intended only to assist in identification of a specific product. APPENDIX H LABORATORY FINDINGS TABLE H-1. SEX, FIELD DRESSED WEIGHT RANGE AND SITE OF CAPTURE OF DEER REMOVED FROM THE IAAP DURING 3-4 DECEMBER 1988 GUN SEASON. DIFFERENCES BETWEEN COLUMN TOTALS FOR THE VARIOUS CATEGORIES REFLECT DISCREPANCIES IN REPORTING BY HUNTERS. --------------------------------------------------------------------------- SEX WEIGHT RANGE (kgs) -------------- --------------------------------------------- SITE Male Female 0-23 24-45 46-68 69-91 Total --------------------------------------------------------------------------- 1 17 37 4 29 23 2 58 2 10 32 7 16 20 2 45 4 8 24 2 18 14 0 34 5 7 13 1 8 10 1 20 6 10 12 3 9 11 0 23 Total 52 118 17 70 78 5 177 --------------------------------------------------------------------------- TABLE H-2. SUMMARY OF TICKS COLLECTED FROM DEER ON IAAP, 3-4 DECEMBER 1988, BY SPECIES AND STAGE. TICK SPECIES ---------------------------------------- Ixodes Dermacentor Amblyomma STAGE dammini albipictus americanum Total --------------------------------------------------------------------------- Adult 1 243 1 245 Nymph 1 49 1 50 Total 2 291 1 294 --------------------------------------------------------------------------- TABLE H-3. MEAN TICK INFESTATION LEVEL OF DEER FROM IAAP, 3-4 DECEMBER 1988, ACCORDING TO HUNTING AREA. HUNTING SITE ------------------------------------- 1 2 4 5 6 ----------------------------------------------------------- Total ticks 93 41 42 61 41 Average ticks/deer 1.60 0.91 1.23 3.05 1.78 ----------------------------------------------------------- TABLE H-4. RESULTS OF ISOLATION ATTEMPTS FOR BORRELIA BURGDORFERI FROM DERMACENTOR ALBIPICTUS COLLECTED FROM DEER ON IAAP, 3-4 DECEMBER 1988. ALL ISOLATION ATTEMPTS WERE CONDUCTED THROUGH THE KIND COOPERATION OF THE CONNECTICUT AGRICULTURAL RESEARCH STATION, NEW HAVEN, CONNECTICUT. ------------------------------------------------------------ NUMBER Stage ---------------------------------------------- of Ticks Cultures Cultures Cultures Tick Tested Positive Negative Contaminated ------------------------------------------------------------ Male 23 0 17 6 Female 20 0 18 2 Nymph 1 0 1 0 ------------------------------------------------------------ TABLE H-5. RESULTS OF IMMUNOFLUORESCENT ANTIBODY ASSAY OF DEER SERUM SAMPLES FOR ANTIBODY TO BORRELIA BURGDORFERI. DEER SERA WERE COLLECTED ON THE IAAP, 3-4 DECEMBER 1988. A SERUM TITERS GREATER THAN OR EQUAL TO 1:256 IS CONSIDERED A POSITIVE INDICATION OF INFECTION (INDICATED BY BOLDFACE). ANTIBODY NUMBER TITER TESTED % OF TOTAL --------------------------------------------- greater than 1:32 110 67 1:32 24 15 1:64 10 6 1:128 12 7 1:256 4 2 1:512 3 2 --------------------------------------------------------------------------- TABLE H-6. CHARACTERISTICS OF DEER YIELDING SEROPOSITIVE ANTIBODY RESPONSE (GREATER THAN OR EQUAL TO 1:256) TO BORRELIA BURGDORFERI. WEIGHTS ARE GIVEN IN KILOGRAMS, N/A INDICATES UNAVAILABLE DATA. --------------------------------------------------------------------------- SAMPLE HUNTING NUMBER WEIGHT SEX AREA ------------------------------------------- 314 N/A N/A N/A 317 55.5 Female 1 337 45.5 Female 6 340 51.4 Female 5 544 N/A Female 2 557 N/A Female 2 590 N/A N/A N/A APPENDIX I TECHNICAL ASSISTANCE 1. Technical advice or consultation regarding issues concerning pest management may be obtained from your Major Command Pest Management Consultant, Mr. Ron White, at the following address and telephone number: Commander USAMC Installations and Services Activity ATTN: AMXEN-U Rock Island, IL 61299-7190 AUTOVON: 793-4067 COMMERCIAL: (309) 782-4062 2. Technical advice concerning the findings and recommendations forwarded in this report may be obtained from Entomological Sciences Division, USAEHA, Aberdeen Proving Ground, Maryland at the following address and telephone number: Commander U.S. Army Environmental Hygiene Agency ATTN: HSHB-MR-EMG APG, MD 21010-5422 ATUOVON: 584-3613/3015 COMMERCIAL: (301) 671-3613/3015 3. Technical information regarding Lyme disease surveillance may be acquired either from ESD (address provided in para 2) or from USAEHA-W at the following address and phone number: Commander U.S. Army Environmental Hygiene Activity Direct Support Activity ATTN: HSHB-AW-P Fitzsimons Army Medical Center, Aurora, CO 80045-5001 AUTOVON: 943-8090 COMMERCIAL: (303) 361-8090 4. Medical reporting of human cases of Lyme disease must be transmitted via Report of Selected Condition (MED 16(R4)) through the Preventive Medicine Services, MEDDAC, Fort Leonard Wood, Missouri, at the following address and telephone number: Commander U.S. Army Medical Department Activity ATTN: HSXP-PM Fort Leonard Wood, MO 65473-5700 AUTOVON: 581-9471 COMMERCIAL: (314) 368-9471 5. Requests for USAEHA services should be directed through command channels of the requesting activity to the address listed in para 2 above. 6. An information copy of any request for USAEHA services should be sent to: Commander U.S. Army Health Services Command ATTN: HSCL-P Fort Sam Houston, TX 78234-6000