Study Proposal Effects of chronic wasting disease on the fecundity on female mule deer (from birth to weaning) Jessie R. Dulberger Graduate Degree Program in Ecology Colorado State University Dr. N. Thompson Hobbs Natural Resource Ecology Laboratory Colorado State University Dr. Michael W. Miller Colorado Division of Wildlife Graduate Degree Program in Ecology Colorado State University Dr. Michael F. Antolin Professor and Assistant Chair Department of Biology Colorado State University Project Summary The goal of this study is to understand how much chronic wasting disease affects fecundity on female mule deer. The study will be conducted in a non-hunted natural area in the foothills of Boulder, Colorado. Our objectives of this study are to compare fecundity (i.e., potential reproductive capacity) rates for chronic wasting disease infected and non-infected female deer during the first month of birth and during weaning in the 3 month of birth. In order to meet these goals we will observe forty-two radio-collared does and assess fawn presence or absence for three summers after parturition (in June) and during weaning (in August). Twice a week each animal will be monitored for mortality. Initial results from the pilot study suggest that the fecundity of chronic wasting disease infected does is much lower than non-infected does. This study is unique because the mule deer population is not affected by hunting. Introduction Chronic wasting disease (CWD) is a natural, infectious and fatal prion disease that negatively affects members of the deer family, mainly Mule Deer (Odocoileus hemionus), Elk (Cervus elaphus nelsoni), and White-Tailed Deer (Odocoileus virginianus (Miller et al. 2005). CWD causes neural degeneration and ultimately death (Williams et al. 2002, Hobbs et al. 2002). Wildlife managers in many states with wildlife recreation based economies are concerned about the possible devastating effect of chronic wasting disease on the state’s deer populations. Population models suggest that fawn survival is key to maintaining deer population dynamics (Jensen et al. 1999). We hypothesize that chronic wasting disease does have a significant affect on fecundity in female mule deer. Objectives Important factors that affect the stability of a healthy deer population is the fecundity of does and fawn recruitment (Pojar and Bowden 2004). Therefore it is important to study how CWD affects both the fecundity and fawn recruitment up to weaning, in a freeranging mule deer population. The specific objectives of this study are to: (1) Compare the presence or absence of fawns during the neonatal period with infected vs. uninfected does. (2) Compare fawn survival of infected vs. uninfected does until weaning (16 weeks after birth. (3) Compare presence or absence of fawns of captive infected and non-infected female mule deer fecundity (Colorado Division of Wildlife unpublished data). Background In the last 39 years, CWD has become a threat to states with wildlife recreationbased economies and thus a concern to both the public and wildlife managers throughout the country (Miller 2002, Williams et al. 2002, and Hobbs et al. 2001). Currently CWD is not a direct threat to humans. There have been no cases of human prion disease associated with CWD (World Health Organization 2000, Belay et al. 2001), but a recent study has shown that CWD specific prions ( PrPres) are in skeletal muscles. This raises the possibly that through consumption of infected cervid meat, dietary exposure to prions could occur (Angers et al. 2006). The disease is thought to be transmitted indirectly to deer horizontally via prions in feces (Miller et al. 2004), but a recent study with captive deer has shown that fawns orally given blood, saliva and chronic wasting disease infected brain tissue, tested positive for the chronic wasting disease prions within 12 months of age (Mathiason et al. 2006). Models show that CWD epidemics drive populations to extinction, specifically when prevalence rates reach 5% larger (Miller et al. 2000). The reduction of cervid populations across North America may harm the balance within many ecosystems (Hobbs et al. 1996). Culling deer herds in CWD hotspot areas had been the practiced management technique, until recently. The Colorado Division of Wildlife has halted the practice of culling, because the prevalence of CWD in deer populations has not been reduced over the past 5 years (Colorado Division of Wildlife, unpublished report). Further studies are needed to understand how CWD spreads, how to stop it from spreading, and how it affects deer populations. Declining mule deer densities have caused wildlife researchers to focus on studying fawn survival and recruitment (Pojar and Bowden 2004). Population models for deer suggest long-term population trends are most sensitive to the survival of fawns, (Jensen et al. 1999). Andelt et al. 2004 showed that mule deer fawn:doe ratios in December had declined by 0.015 fawns/doe/yr from 1972 through 1995 in Colorado. Many hypotheses have been offered to explain fawn:doe declines including: failure to breed, decreased fecundity, resorption/abortion of fetuses, low summer and fall fawn survival, habitat deterioration, increased predation, drought, hunting, disease or a combination of these factors (Unsworth et al. 1999, Andelt et al. 2004, Pojar and Bowden 2004). Andelt et al, 2004 concluded that failure to breed or maintain pregnancy through at least early January or February was not the cause of low fawn:doe ratios observed in Colorado, since 90-93% of deer in the study were pregnant. Predation, specifically coyotes (Canis latrans) has been questioned as a significant mortality factor for fawn numbers (Jensen, 1999), but it has since been shown that coyotes (Canis latrans) and mountain lions (Felis concolor) appear to kill fawns in very poor condition (Bishop, 2005). Mortality rate of fawns were not directly related to population levels of coyotes (Hamlin, 1984). Pojar et al. found that the highest cause of fawn mortality was sickness/starvation, and second were coyotes. Another study showed that poor doe condition was the primary cause of early fawn loss (Wood 1962). We hypothesis that chronic wasting disease negatively affects does fecundity in two possible ways. We have observations, from the main Boulder mule deer study, that does with chronic wasting disease exhibit non-normal behavior, (not staying as close with the herd and decreased alertness), that could be linked to bad parenting skills. We also hypothesis that chronic wasting disease negatively affects the body condition of does. Yoakam 1965 observed that poor body condition of does led to fawn/s being born weak or handicapped, and then death within their first week. This study will evaluate the effects of CWD on the fecundity of adult does in the foothills of Boulder, Colorado. An ongoing mule deer survival study with the City of Boulder’s Open Space and Mountain Parks and the Colorado Division of Wildlife has thus far shown a relatively high prevalence rate (50% in bucks and 30 % in does). When animals were retested a year later 25% of the does that were not infected had contracted chronic wasting disease (unpublished data). Models forecast population declines once CWD prevalence exceeds about 5% (Miller 2000). Therefore the deer population along the Front Range in the City of Boulder is in jeopardy, and will be a good example of how CWD affects a non hunted free-ranging mule deer population. The City of Boulder Open Space and Mountain Parks (OSMP) has had several mule deer population studies using the Lincoln-Peterson estimator. From 1983 to 1990 there was an increase in the mule deer population from 783 to 1,238 in a 17 square mile area (Kelsey, 1990). The study area for this past study was twice as large as the current study and it ranged North of El dorado Springs Drive to just south of Lee Hill Road. The 2005 markresight population count of mule deer averaged 270 using the Bowden estimator. The study area ranged North of El dorado Springs Dr. and South of Baseline Rd. Even though the 2005 study is half the size of the study done in 1983-1990, if doubled it would only average around 540, which is the lowest City of Boulder population seen in many years. The Pilot Study Summer 2006 From June to September in 2006, a pilot study was conducted. The power calculation performed from this initial data set showed that we will need twelve infected and equal numbered non-infected does to detect a difference of 0.9 in fecundity rates. Figures 2,3 and 4 demonstrate that sample sizes of seven and ten were too small, while fifteen is not needed. The methods during the pilot were not as rigorous as the methods will be for the following two summers, due to lack of time. There were seven matched pairs of does in the study for the summer of 2006. Extra non-infected does were also observed for presence or absence of fawns, mainly in August/September, (please see figure 1.). Using Likelihood and Akaike’s information criterion corrected for small sample bias (AICc) for model selection, we were able to detect a difference using a two parameter comparison. The two parameter model had more support in the data than the one parameter model. The limiting factor for this study is the sample size of chronic wasting disease infected does, since only one out of three female deer sampled tested positive for CWD. Number does with Number total does Fawns Observed CWD Positive 3 7 23 32 CWD Negative Figure 1. Pilot study data from summer 2006. total # of does (7) 1.2 1 wr 0.8 0.6 0.4 0.2 0 0 1 2 3 4 5 6 1 parameter 2 parameters # of does w ith faw ns Figure 2. One versus two parameters for seven does. Total # of does 10 1.2 1 wr 0.8 0.6 0.4 0.2 0 0 1 2 3 # of does with fawns 4 5 1 parmater 2 parmaters Figure 3. One versus two parameters for ten does. 6 Total # of does 12 1.2 1 wr 0.8 0.6 0.4 0.2 0 0 1 2 3 4 5 # of does with fawns 6 1 parameter 2 parameters Figure 4. One versus two parameters for twelve does. Total # of does 15 1.2 1 wr 0.8 0.6 0.4 0.2 0 0 1 2 3 4 # of does with fawns 5 6 1 paramter 2 paramters Figure 5. One versus two parameters for fifteen does. Material and Methods Study Area: The study area will focus on low-elevation mule deer winter range. The study area in Boulder boundaries are as follows: Baselines Rd on the north, Broadway Rd and the east, El dorado Springs Dr. on the south and the Flatiron Mountain range to the west ( See figure 7.). Figure 7. Study area in Boulder, Colorado Methods 200 meters Figure 8. Fawn search method. Approach: Adult does have been previously marked with unique numbered ear tags and collars with mortality-sensing radio transmitters. In May each doe’s summer fawning locations will be located. Twelve CWD infected and twelve non-infected does will be paired as of June 1st or last date seen in May, on the basis of infected or not infected with chronic wasting disease, age, location and genotype. Additional non-infected does will also be monitored the same as the paired individuals, time permitting. The location and body condition of each doe has been determined at least once a month, since their original capture date. They are also monitored twice weekly for survival status. When an animal dies it is found via the mortality emitting radio transmitter and telemetry. The cause of death is recorded and samples are taken to retest for chronic wasting disease, and if the cause is unknown the body is submitted for a necropsy to the Colorado Division of Wildlife laboratory. Beginning June 1st, each doe will be observed 1x/week for two hours a day to determine the presence or absence of fawn/s. During these observations does will be located, and monitored from a distance far enough to avoid alerting them, but close enough to identify the radio-collar number and udder development, (approximately 40 m) (Shively et al. 2005). If fawns are not seen during the initial two hour observation, then a fawn search will be performed for an additional thirty to forty minutes. The search will consist of marking the original location of the doe upon first approach and then searching out to 200 meters in a circle around that original point (see figure 8). If searches are unsuccessful, but the doe remains in the same vicinity, a follow-up searches will made 1-2 days later, time permitting. As fawns are located, the technician will slowly leave the area, as to not stress the fawn. When fawns and does are seen together, fawn less than 20 meters away will be considered associated with the doe (Schwede et al, 1994). Once the presence of fawns is established, does will be observed twice a month until weaning when fawns are 16 weeks of age, or the end of September. At each observation the body condition, location of the doe and fawn, presence or absence of fawn/s and vegetation type will be recorded. Since there are forty-two deer to observe, deer will be observed at different times to utilize all daylight hours. There will be two technicians working on this project, therefore eighty hours a week of observations. There will be two time shifts, to cover sunrise and sunset, (6AM-2PM and 12PM-8PM). Does that show behavioral and physical characteristics, such as moving alone, udder development and sunken flanks, are prime candidates for having fawns (Pojar and Bowden 2004, Huegel et al. 1985). During the first eight weeks after birth mothers associate very little with fawns and periods of contact are brief, occurring predominantly around sunrise and sunset (Schede et al. 2006). It is also crucial to detect presence or absence of fawns after in during the neonatal period because in June the probability of survival is lowest, 0.33 in June compared to 0.9 in September (Raganella-Pelliccioni, 2006). Mothers also keep the shortest distances during the first two weeks (Schwede et al. 1994). During this time does can also be approached closely before flushing (Huegel et al. 1985). Huegal et al. 1985 found that does always remained in the general vicinity of their fawns while the crew searched. Analysis Conditional logistic regression will be used to evaluate the relative strength of models predicting female fecundity. Several factors that will contribute to these models are: location, genotype, and age and CWD status. We will create a candidate set of models via a priori knowledge and use likelihood inference and AICc to select the model that has the most support in the data. We will also similarly analyze a Colorado Division of Wildlife data set of captive does and their fecundity rates. There are several years of fecundity data with specific birth dates and death dates of fawns, their genotypes, whether they are CWD infected or not, and how long they have had the disease. Budget June 2006-September 2006 Research Assistantship ($/month * 4 monthes) (first field season)…$(OSMP) Field Equipment (scope, digital camera, logbook, binoculars, miscellaneous)………..$2,000.00 Tom’s Salary ? Mike’s Salary ? 1st Field Season Subtotal……………………………………………………………….$2,000.00 + Ten Percent…………………………………………………………………………..$200.00 1st Field Season total…………………………………………………………………..$2200.00 October 2006-September 2007 Research Assistantship ($1500/month * 12 months + fringe) (first field season)……..$14,400.00 Tuition (two semesters)………………………………………………………………...$3,073.98 Field Assistant (2 monthes at $1760/month+fringe)..………………………………….$3,520.00 Materials and Supplies…………………………………………………………………$300.00 Travel and Conferences……………… ………………………………………………$500.00 Tom’s Salary ? Mike’s Salary ? Year 1 Subtotal……………………………………………………………………….$21,793.98 + Ten Percent…………………………………………………………………………$2,179.39 Year 1 Total…………………………………………………………………………..$23,973.37 October 2007-September 2008 Research Assistantship ($1500/month * 12 months + fringe) (first field season)……..$14,400.00 Tuition (two semesters)………………………………………………………………...$4,810.90 Mileage (,000 miles/week at 36 cents/mile...…………………………………………..$? Field Assistant (2 monthes @$11/hr 40hr/wk +fringe)………………………..………$3,520.00 Materials and Supplies………………………………………………………………….$300.00 Tom and Mike’s Salary ? Year 2 Subtotal…………………………………………………………………………$23030.90 + Ten Percent…………………………………………………………………………..$2,303.09 Year 2 Total……………………………………………………………………………$25333.99 October 2008-September 2009 Research Assistantship ($1500/month * 12 months + fringe) (first field season)……..$14,400.00 Materials and Supplies…………… ………………………………………………….$300.00 Tom and Mike’s Salary ? Year 3 Subtotal……………………………………………………………………….$14,700.00 + Ten Percent…………………………………………………………………………$1,470.00 Year 3 Total…………………………………………………………………………..$16170.00 3 ½ YEAR PROJECT TOTAL……………………………………………………….$63,904.27 Budget for Jessie Dulberger for 2 ½ years. 3 field seasons, 5 semesters of classes, 1 semester to write thesis. Literature Cited: Andelt,William F., Thamas M. Pojar, and Laurie W. Johnson, 2004. Long-term trend in mule deer pregnancy and fetal rates in Colorado. Journal of Wildlife Management: Vol. 68, No. 3, pp. 542-549. Angers, Rachel C., Shawn R. Browning, Tanya S. Seward, Christina J. Sigurdson, Michael W. Miller, Edward A. Hoover and Glenn C. Telling, 2006. Prions in skeletal muscles of deer with Chronic Wasting Disease. Sciencexpress: www.sciencexpress.org, pp. 1-3. Belay, Ermias D., Ryan A. Maddox, Elizabeth S. Williams, Michael W. Miller, Pierluigi Gambetti, and Laurence B. Schonberger, 2004. Chronic wasting disease and potential transmission to humans. 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