MULTIPLE PATERNITY IN WHITE-TAILED DEER (ODOCOILEUS VIRGINIANUS) REVEALED BY DNA MICROSATELLITES

Journal of Mammalogy, 8():88 89, 00 MULTIPLE PATERNITY IN WHITE-TAILED DEER (ODOCOILEUS VIRGINIANUS) REVEALED BY DNA MICROSATELLITES RANDY W. DEYOUNG,* STEPHEN DEMARAIS, ROBERT A. GONZALES, RODNEY L. HONEYCUTT, AND KENNETH L. GEE Department of Wildlife and Fisheries, Box 9690, Mississippi State, MS 976 (RWD, SD) Samuel Roberts Noble Foundation, P.O. Box 80, Ardmore, OK 70 (RAG, KLG) Department of Wildlife and Fisheries Sciences, Texas A&M University, College Station, TX 778 (RLH) Multiple paternity in single litters (siring of offspring by male) has been documented in several taxa with different mating systems. However, information on occurrence of multiple paternity in ungulates is lacking. We used 9 DNA microsatellite markers to assign parentage in litters of captive white-tailed deer (Odocoileus virginianus) born in 6 pens with multiple males during 997 999. We detected multiple paternity in 7 of 7 litters with offspring, occurring in of 6 pens: of triplet litters and 5 of twin litters. This is the first reported evidence of multiple paternity for single ungulate litters, which indicates that some aspects of ungulate reproductive ecology are not well understood. The occurrence of multiple paternity in litters of free-ranging ungulates would have implications for ungulate mating systems and reproductive strategies. Sex-linked microsatellite markers may provide a promising method for investigating female promiscuity in free-ranging ungulate populations where litter size is typically offspring. Key words: female promiscuity, microsatellites, multiple paternity, Odocoileus virginianus, parentage, paternity, reproductive ecology, white-tailed deer Many hypotheses pertaining to mammalian reproductive ecology are testable only using genetic techniques. The advent of new molecular markers and molecular techniques has enabled researchers to both establish parentage and estimate relationships among individuals with a high degree of confidence (Fleischer 996; Marshall et al. 998; Strassmann et al. 996). Parentage assignment enables estimation of reproductive success, social structure, kinship, and fitness (Neff et al. 000; Petrie and Kempenaers 998). Genetic methods are providing important breakthroughs in the study of animal mating systems. For instance, several genetic studies have revealed patterns of male mating success that are quite different from * Correspondent: rwd@msstate.edu those derived from behavioral observations (Amos et al. 99; Coltman et al. 999; Pemberton et al. 99). Genetic techniques have also verified multiple paternity, the fathering of individuals within a single litter by male, in birds (Petrie and Kempenaers 998) and several species of mammals, including deer mice (Peromyscus maniculatus Birdsall and Nash 97), common shrews (Sorex araneus Tegelstrom et al. 99), black bears (Ursus americanus Schenk and Kovacs 995), and grizzly bears (U. arctos Craighead et al. 995). These recent findings indicate that reproductive strategies in mammals, especially as they relate to individual fitness and overall mating success, differ between males and females. A genetic dissection of mating patterns is requisite for study of the evolution of mating systems (Fleischer 996). Fur- 88

August 00 DEYOUNG ET AL. MULTIPLE PATERNITY IN DEER 885 thermore, male reproductive success and female strategies may change depending on sex ratio, resource availability, etc., and a knowledge of how these parameters influence mating systems is important for estimation of effective population size and studies of basic population structure (Sugg and Chesser 99). Although instances of multiple sires for a single litter have been documented in mammalian species with litter sizes of, verification of multiple paternity for polygynous and promiscuous species of mammals that typically have offspring per litter (e.g., many ungulates) is more difficult, especially when sampling of potential sires is incomplete (Xia 999). As indicated for red deer (Cervus elaphus Pemberton et al. 99) and gray seals (Halichoerus grypus Ambs et al. 999), evidence of dominance does not always equate to mating success of males. Therefore, in ungulate species where twins and triplets are produced with a reasonable frequency, multiple paternity might occur. The white-tailed deer (Odocoileus virginianus) is a likely candidate for evidence of multiple paternity in ungulates. Twinning is common in white-tailed deer, and studies of captive individuals indicate that females sometimes mate with male during the -h estrous period (Verme 965). We used a panel of DNA microsatellite loci to investigate the possibility of multiple paternity in a group of captive white-tailed deer. MATERIALS AND METHODS We conducted our investigation using captive white-tailed deer in the Mississippi State University Captive Animal Facility. The facility contains 6 pens ranging from 0. to. ha in size. Each year, deer were assigned to different pens according to needs of the facility and research conducted. Different groupings of males and females were allowed to breed each year, and surplus males were housed in male-only pens. Deer density did not exceed 0/ha. There were 6 breeding pens that contained males during 997 999 (Table ). The sole pen in 997 contained 6 males (ages 8.5, 6.5,.5,.5,.5, and.5 years) and adult females. There were pens in 998, of which contained males (ages.5,.5, and.5 years) and 7 adult females (998a); the other (998b) contained males (ages.5 years) and adult females. Three pens were available in 999: 999a ( males, ages 5.5 and.5 years, and adult females); 999b (three.5-year males, 6 adult females); 999c ( males, ages 5.5,.5, and.5 years, and adult females). Adult deer were manually restrained or sedated in October each year with a Telazol xylazine mixture composed of. mg/kg Telazol (Fort Dodge Animal Health, Fort Dodge, Iowa) plus. mg/kg xylazine (Phoenix Scientific, St. Joseph, Missouri), antagonized with 0.5 mg/ kg yohimbine (Abbott Laboratories, North Chicago, Illinois) delivered by cartridge-fired dart (Pneu-Dart Incorporated, Williamsport, Pennsylvania) or a pole syringe. Fawns were manually restrained within days postpartum, and birth dates were recorded. Blood ( 5 ml) was obtained by venipuncture from all individuals and stored at C in vacuum tubes containing ethylene diamine tetraacetic acid (EDTA; Vacutainer, Becton-Dickson and Company, Franklin Lakes, New Jersey). We isolated DNA from whole blood through either phenol chloroform extraction or a commercial kit (Puregene DNA isolation kit; Gentra Systems Incorporated, Minneapolis, Minnesota). For the extraction protocol, we mixed 0.5 ml of the blood EDTA mixture with sucrose Triton X buffer (0. M sucrose, 0.0 M Tris, ph 7.6, 0.005 M MgCl, % Triton X-00) and centrifuged (,000 g for 0 min) for collection of white cells. This procedure was repeated times. White blood cells were then placed in 500 l of Laird s buffer (Laird et al. 99) and digested overnight with Proteinase K (0 mg/ml) at 7 C. We extracted DNA by adding sample to an equal volume of phenol chloroform isoamyl alcohol mixture and precipitating with one-twentieth volume of 5 M NaCl and volumes of 95% ethanol, followed by centrifugation (,000 g for 5 min). Samples were vacuum-dried and redissolved in double-distilled H O or Tris EDTA. DNA concentration was determined using a fluorometer and DNA standards. We determined the genotype of each deer on the basis of 9 microsatellite loci from the - locus cervid microsatellite panel described by

886 JOURNAL OF MAMMALOGY Vol. 8, No. TABLE. Pen setup and birth records for 6 breeding pens of Odocoileus virginianus with males maintained at the Mississippi State University Captive Animal Facility during 997 999. Pen number Number of adults Male Females Number of litters a Multiple paternity 997 6 i () ii () iii () iv () 998a 7 0 i () ii () iii () iv () v () vi () vii () viii () b ix () x () 998b 0 i () ii () iii () iv () 999a 8 i () ii () iii () iv () v () c vi () vii () viii () 999b 6 5 i () ii () iii () iv () v () 999c 0 0 i () ii () iii () iv () v () vi () vii () viii () ix () x () Estimated conception date November December 7 December 7 December November 5 November 5 December December 8 December 5 December 5 December 0 December 6 January 0 January December December 7 December January December December 6 December 6 December 0 December December 9 January January 9 November November 0 December 0 December December 0 November November December 7 December December December December December February 6 February a Roman numerals indicate birth order, numbers in parentheses indicate numbers of fawns. b Paternity of of the fawns in this litter not assigned because of a low-quality DNA sample. c Sired by a male from an adjacent pen after an interior fence was damaged. Females in concurrent estrous 6 5 5 5 5

August 00 DEYOUNG ET AL. MULTIPLE PATERNITY IN DEER 887 Anderson et al. (00). A full description of the panel and reaction conditions is contained therein. We omitted the OCAM and BM0 loci because of many untyped individuals, leaving 9 for evaluation. In brief, fragments were amplified via polymerase chain reaction using fluorescent-tagged primers in single or multiplexed reactions on a PE Gene Amp 9600 thermocycler (Applied Biosystems Incorporated, Foster City, California). For each individual, the products from polymerase chain reactions were mixed together ( l of each reaction), and l of this mixture was added to a denaturing mixture of size standard and formamide (Genescan rox 500; Applied Biosystems). The reaction product-denaturing mixes were loaded on an ABI Prism 0 Genetic Analyzer (Applied Biosystems) for separation and detection. DNA fragments were quantified and analyzed with GeneScan software, and alleles were assigned using Genotyper software (both Applied Biosystems). We assigned parentage for all fawns using the software CERVUS.0 (Marshall et al. 998). The software also provides estimates of mean allelic diversity (A), expected multilocus heterozygosity (H), mean polymorphism information content (PIC), and exclusion probabilities with and without known parents. This procedure demonstrated confidence in assigning paternity consistent with simulated values (Slate et al. 000) and has also been used to assign paternity in other terrestrial vertebrates (Zamudio and Sinervo 000). Multiple paternity was inferred when paternity in litters with offspring assigned to a single female was shared by different males. We used estimated conception dates to examine qualitative trends in paternity assignment. Because reported gestation periods for whitetailed deer are variable (range: 87 days, X 00 days, SD days Adams 960; Haugen 959; Haugen and Davenport 950; Verme 965), we estimated fawn conception dates by subtracting 00 days from the birth date. We constructed intervals of days from the estimated conception dates and compared litters born in the same pen to determine if multiple paternity was influenced by the presence of concurrently estrous female. RESULTS The marker panel was highly variable in this group of deer (A 9.9, H 0.7, PIC 0.69). Exclusion probabilities were 0.99, which enabled us to assign parentage with 95% confidence. Females maintained in the 6 multiple-sire breeding pens produced a total of litters ( single, twin, and triplet Table ). The genotypic database included these 7 fawns and their potential parents, for a total of 9 genotypes. We identified 7 instances of multiple paternity in the 7 compound litters (those with offspring), occurring in of 6 pens. Five of the 7 multiply sired litters were conceived in the probable presence of concurrently estrous female (median Table ). We assigned multiple paternity for of litters born in the 997 pen. Of the 6 fawns, were assigned to the 6.5-year-old male and were assigned to of the.5- year-old males; these males shared paternity of a twin litter. Two of 0 litters born in pen 998a were assigned to multiple sires. One twin litter shared paternity between the.5- and.5-year-old males, and another twin litter shared paternity between the.5- and.5-year-old males. The paternity of fawn was undetermined because of a lowquality DNA sample. There were multiple paternity assignments in two 999 breeding pens. Two of 8 litters born in pen 999a had sire. The males in this pen shared paternity for twin litter and the triplet litter. Two of 5 litters were assigned to sire in pen 999b. The pen was intended to be occupied by three.5-year-old males and 6 adult females. Ten additional males were temporarily housed in this pen for days during late November 998. Only of the intended.5-year-old males successfully reproduced, siring 5 fawns, including shared paternity of a twin and triplet litter with an 8.5-year-old male. A different 8.5-year-old male and a.5-year-old male each sired a single fawn. There were no instances of multiple paternity in of the 6 breeding pens. A single male in pen 998b sired all litters. Ten litters were born in pen 999c, where 5 of

888 JOURNAL OF MAMMALOGY Vol. 8, No. 6 fawns were sired by the.5-year-old male and fawn was sired by the 5.5-yearold male. DISCUSSION Multiple paternity occurred in about 5% of compound litters, though the actual incidence of copulation with multiple males could have been greater because not all copulations may have resulted in conception. This suggests that copulation with multiple males is common in captivity under these conditions. Multiple paternity occurred predominantly in the presence of concurrently estrous female. We attempted to be conservative in our estimates of possible concurrent estrus by using a consensus of reported gestation periods and standard deviations. All 7 dams of the multiply sired litters could have been in concurrent estrus with at least other female if the window of possible overlap were increased to 5 days (Table ). These observations indicate that a single male cannot continuously monopolize several estrous females in a pen. Therefore, competition for mates probably influenced the occurrence of multiple paternity. Examination of chronology of paternity revealed situations in addition to multiple paternity where competition for mates was apparent. For example, 6 of the 0 litters produced in the 999c pen were compound and median number of females concurrently in estrus was.5. Though there were no multiply sired litters born in the 999c pen, the only fawn sired by the 5.5-year-old male was a single birth occurring when there were an estimated concurrently estrous females and immediately before a large cluster of 5 6 concurrently estrous females (pen 999c, litter iii Table ). A similar situation occurred in pen 999a when a male from an adjacent pen gained access for 5 days after damage to an interior fence. This male sired a litter during a period intermediate to the multiple paternity assignments when there were concurrent estrous females (pen 999a, litter v Table ). There were no multiple paternity assignments in of the 6 breeding pens. However, the uneven occurrence of multiple paternity in the captive-born litters cannot be attributed to any single factor. The 998b pen produced only litters ( of which were single births), limiting the opportunity to detect multiple mating. Sex ratio in the pens varied from 0.8 to.5 males : female but did not show a clear association with multiple paternity. Individual differences in male aggressiveness coupled with high deer density in a small area may facilitate monopoly of females by a single aggressive male in some situations. It is unlikely that a single male could simultaneously monopolize female outside of captive conditions. This is because the breeding system of white-tailed deer in free-ranging populations involves formation of a tending bond, where a male guards, tends, and courts an individual estrous female (Hirth 977). The reproductive behavior of captive white-tailed deer suggests that estrous females will copulate only within a -h period (Haugen 959; Haugen and Davenport 950; Verme 965) but may copulate with male during this - h estrus (Verme 965). Therefore, multiple paternity is possible in free-ranging populations of white-tailed deer. The occurrence and rate of multiple paternity in a free-ranging deer population probably depends upon population demographics. Male reproductive success in cervids is influenced by male social rank (Clutton-Brock et al. 98; McElligot and Hayden 000). McCullough (979) suggested that males probably do not achieve the social status required to breed until they are physically mature, after.5 years of age. In populations with a balanced sex ratio and diverse age structure, male white-tailed deer are thought to maintain a strict dominance hierarchy in which physically mature males dominate immature males. A small number of dominant males probably possess breed-

August 00 DEYOUNG ET AL. MULTIPLE PATERNITY IN DEER 889 ing rights (Marchinton and Hirth 98) and may displace subordinate males that are tending females. Subordinate males are thought to achieve reproductive success only through surreptitious fertilization or kleptogamy. However, in populations that lack physically mature males, yearling males may breed without establishing a formal social hierarchy (Ozoga and Verme 985), thereby increasing the likelihood that an individual male will sire offspring. Differences in numbers of potential mates contribute to variability of reproductive success of individual males (Wade 979; Wade and Arnold 980) and probably affect frequency of multiple paternity. Population sex ratio and male age structure can influence length of breeding season in white-tailed deer (Gruver et al. 98). A female-skewed sex ratio coupled with a preponderance of young males may result in longer breeding seasons for white-tailed deer (Ozoga and Verme 985). Higher female : male ratios may provide more opportunities for breeding success for younger or less dominant males, as suggested for gray seals (Ambs et al. 999). Komers et al. (997) reported that male male interactions decreased and male female interactions increased with increasing female : male ratios in captive fallow deer (Dama dama). Thus, higher female : male ratios may decrease the frequency of multiple paternity in freeranging populations by reducing competition for mates and decreasing the opportunity for females to mate with male. It is possible that frequency of multiple paternity in deer populations is influenced by geographic variation in breeding season length. Length of breeding season in whitetailed deer is more strictly defined in northern climates because of the extreme environmental conditions (Demarais et al. 000). A concise breeding period is favored in northern climates because optimal fawn nutrition and growth are necessary for overwinter survival. White-tailed deer populations exhibit more variation in breeding date in the less-demanding southern climates (S. Demarais, in litt.; Richter and Labisky 985). Frequency of multiple paternity may increase with longer breeding seasons because there would be fewer females in concurrent estrous during any portion of the breeding season, increasing competition for estrous females. Multiple paternity could occur by displacement of a tending male, but it may also be influenced by female choice. If females select mates on the basis of fitness benefits to their offspring, then there is greater reward in mating with multiple males when male genetic quality is variable (Petrie et al. 998). Frequency of multiple paternity increases as population genetic diversity increases both between species and between populations of birds (Petrie et al. 998). White-tailed deer are the most polymorphic large grazing mammals (Breashears et al. 988; Smith et al. 98) and would be expected to show multiple paternity in free-ranging populations under this hypothesis. However, the role of female selection in cervid reproduction is unclear. Bubenik (98) suggests that female cervids may selectively breed with large-antlered males. Examination of fluctuating asymmetry (proposed as an index of phenotypic quality) in moose (Alces alces) antlers reveals that asymmetry is less in largeantlered males, which presumably do most of the breeding (Bowyer et al. 00). A study of genetic variability in the major histocompatibility complex in white-tailed deer suggests that antler development and body size may provide an advertisement of heritable male quality (Ditchkoff et al. 00). Females may act on these cues to select mates. Nevertheless, Clutton-Brock et al. (98) found no evidence of mate selection in red deer, with the exception that females avoid copulating with young males. Several other North American ungulates produce compound litters and exhibit mating systems that could result in multiple paternity in single litters. Mule and blacktailed deer (O. hemionus) commonly produce twin litters and have a polygynous,

890 JOURNAL OF MAMMALOGY Vol. 8, No. tending-bond breeding system (Kie and Czech 000) similar to that of white-tailed deer. Moose may produce twin litters and exhibit both a harem breeding system, in tundra-dwelling moose, and a pair breeding system, in forest-dwelling moose (Franzmann 000). Twinning is common in pronghorn (Antilocapra americana), which exhibit both harem-type and territorial mating systems, depending on spatial dispersion and population density (Yoakum and O Gara 000). The occurrence of multiple paternity in deer has interesting implications for our understanding of ungulate mating strategies. Further research is needed to clarify the role and prevalence of female promiscuity in the reproductive ecology of deer. The frequency of promiscuous female mating may differ both among species of the same genera (Foltz 98; Xia and Millar 99) and among populations of the same species (Boonstra et al. 99; Sheridan and Tamarin 986). Therefore, extrapolation of these results to free-ranging populations must be critically evaluated, especially regarding population sex ratio, density, male age structure, and breeding date. Sex-linked microsatellite markers may be the most promising method for investigating multiple paternity in free-ranging populations. ACKNOWLEDGMENTS R. Dawkins, S. McKinney, M. Harrison, and B. Taylor maintained the Mississippi State University Captive Facility and assisted with sampling. We thank A. Harris and J. Anderson for assistance with DNA extraction and amplification, and anonymous reviewers for helpful editorial comments. R. DeYoung was supported by a fellowship from the Rob and Bessie Welder Wildlife Foundation. The Mississippi Department of Wildlife, Fisheries, and Parks provided funding through Federal Aid in Wildlife Restoration funds (Project W-8-7, Study 5). This manuscript is contribution WF70 of the Mississippi State University Forest and Wildlife Research Center and 577 of the Rob and Bessie Welder Wildlife Foundation. Research was conducted under Mississippi State University Institutional Animal Care and Use Protocol 98-0. LITERATURE CITED ADAMS, W. H., JR. 960. Population ecology of whitetailed deer in northeastern Alabama. Ecology : 706 75. AMBS, S. M., D. J. BONESS,W.D.BOWEN,E.A.PERRY, AND R. C. FLEISCHER. 999. Proximate factors associated with high levels of extraconsort fertilization in polygynous grey seals. Animal Behaviour 58: 57 55. AMOS, B., S. TWISS, P. POMEROY, AND S. ANDERSON. 99. Male mating success and paternity in the grey seal, Halichoerus grypus: a study using DNA fingerprinting. Proceedings of the Royal Society of London Series B 5:99 07. ANDERSON, J. D., ET AL. 00. Development of microsatellite DNA markers for the automated genetic characterization of white-tailed deer populations. Journal of Wildlife Management 66:67 7. BIRDSALL, D. A., AND D. NASH. 97. Occurrence of successful multiple insemination of females in natural populations of deer mice (Peromyscus maniculatus). Evolution 7:06 0. BOONSTRA, R., X. XIA, AND L. PAVONE. 99. Mating system of Microtus pennsylvanicus. Behavioral Ecology :8 89. BOWYER, R. T., K. M. STEWART, J.G.KIE, AND W. C. GASAWAY. 00. Fluctuating asymmetry in antlers of Alaskan moose: size matters. Journal of Mammalogy 8:8 8. BREASHEARS, D. D., M. H. SMITH, E. G. COTHRAN, AND P. E. JOHNS. 988. Genetic variability in white-tailed deer. Heredity 60:9 6. BUBENIK, A. B. 98. The behavioral aspects of antlerogenesis. Pp. 89 9 in Antler development in Cervidae (R. D. Brown, ed.). Caesar Kleberg Wildlife Research Institute, Kingsville, Texas. CLUTTON-BROCK, T. H., F. E. GUINNESS, AND S. D. AL- BON. 98. Red deer: behavior and ecology of two sexes. University of Chicago Press, Chicago, Illinois. COLTMAN, D. W., D. R. BANCROFT, A. ROBERTSON, J. A. SMITH, T. H. CLUTTON-BROCK, AND J. M. PEM- BERTON. 999. Male reproductive success in a promiscuous mammal: behavioral estimates compared with genetic paternity. Molecular Ecology 8:99 09. CRAIGHEAD, L., D. PAETKAU, H. V. REYNOLDS, E. R. VYSE, AND C. STROBECK. 995. Microsatellite analysis of paternity and reproduction in Arctic grizzly bears. Journal of Heredity 86:55 6. DEMARAIS, S., K. V. MILLER, AND H. A. JACOBSON. 000. White-tailed deer. Pp. 60 68 in Ecology and management of large mammals in North America (S. Demarais and P. R. Krausman, eds.). Prentice- Hall, Inc., Upper Saddle River, New Jersey. DITCHKOFF, S. S., R. L. LOCHMILLER, R. E. MASTERS, S. R. HOOFER, AND R. A. VAN DEN BUSSCHE. 00. Major histocompatibility complex-associated variation in secondary sexual traits of white-tailed deer (Odocolieus virginianus): evidence for good-genes advertisement. Evolution 55:66 65.

August 00 DEYOUNG ET AL. MULTIPLE PATERNITY IN DEER 89 FLEISCHER, R. C. 996. Application of molecular methods to the assessment of genetic mating systems in vertebrates. Pp. 6 in Molecular zoology: advances, strategies, and protocols (J. D. Ferraris and S. R. Palumbi, eds.). Wiley-Liss, New York. FOLTZ, D. W. 98. Genetic evidence for long-term monogamy in a small rodent, Peromyscus polionotus. American Naturalist 7:665 675. FRANZMANN, A. W. 000. Moose. Pp. 578 600 in Ecology and management of large mammals in North America (S. Demarais and P. R. Krausman, eds.). Prentice-Hall, Inc., Upper Saddle River, New Jersey. GRUVER, B. J., D. C. GUYNN, JR., AND H. A. JACOBSON. 98. Simulated effects of harvest strategy on reproduction in white-tailed deer. Journal of Wildlife Management 8:55 5. HAUGEN, A. O. 959. Breeding records of captive white-tailed deer in Alabama. Journal of Mammalogy 0:08. HAUGEN, A. O., AND L. A. DAVENPORT. 950. Breeding records of white-tailed deer in the Upper Peninsula of Michigan. Journal of Wildlife Management : 90 95. HIRTH, D. H. 977. Social behavior of white-tailed deer in relation to habitat. Wildlife Monographs 5: 55. KIE, J. G., AND B. CZECH. 000. Mule and black-tailed deer. Pp. 69 657 in Ecology and management of large mammals in North America (S. Demarais and P. R. Krausman, eds.). Prentice-Hall, Inc., Upper Saddle River, New Jersey. KOMERS, P. E., C. PELABON, AND D. STENSTROM. 997. Age at first reproduction in male fallow deer: agespecific versus dominance-specific behaviors. Behavioral Ecology 8:56 6. LAIRD, P. W., A. ZIJDERVELD, K. LINDERS, M. A. RUD- NICKI, R. JAENISCH, AND A. BERNS. 99. Simplified mammalian DNA isolation procedure. Nucleic Acids Research 9:9. MARCHINTON, R. L., AND D. H. HIRTH. 98. Behavior. Pp. 9 68 in White-tailed deer: ecology and management (L. K. Halls, ed.). Stackpole Books, Harrisburg, Pennsylvania. MARSHALL, T. C., J. SLATE, L.E.B.KRUUK, AND J. M. PEMBERTON. 998. Statistical confidence for likelihood-based paternity inference in natural populations. Molecular Ecology 7:69 655. MCCULLOUGH, D. R. 979. The George Reserve deer herd: population ecology of a K-selected species. University of Michigan Press, Ann Arbor. MCELLIGOT, A. G., AND T. J. HAYDEN. 000. Lifetime mating success, sexual selection, and life history of fallow bucks (Dama dama). Behavioral Ecology and Sociobiology 8:0 0. NEFF, B. D., J. REPKA, AND M. R. GROSS. 000. Parentage analysis with incomplete sampling of candidate parents and offspring. Molecular Ecology 9: 55 58. OZOGA, J. J., AND L. J. VERME. 985. Comparative breeding behavior and performance of yearling vs. prime-age white-tailed bucks. Journal of Wildlife Management 9:6 7. PEMBERTON, J. M., S. D. ALBON, F.E.GUINNESS, T.H. CLUTTON-BROCK, AND G. A. DOVER. 99. Behavioral estimates of male mating success tested by DNA fingerprinting in a polygynous mammal. Behavioral Ecology and Sociobiology :66 75. PETRIE, M., C. DOUMS, AND A. P. MOLLER. 998. The degree of extra-pair paternity increases with genetic variability. Proceedings of the National Academy of Sciences of the United States of America 95:990 995. PETRIE, M., AND B. KEMPENAERS. 998. Extra-pair paternity in birds: explaining variation between species and populations. Trends in Ecology and Evolution :5 58. RICHTER, A. R., AND R. F. LABISKY. 985. Reproductive dynamics among disjunct white-tailed deer herds in Florida. Journal of Wildlife Management 9:96 97. SCHENK, A., AND K. M. KOVACS. 995. Multiple mating between black bears revealed by DNA fingerprinting. Animal Behaviour 50:8 90. SHERIDAN, M., AND R. H. TAMARIN. 986. Space use, longevity, and reproductive success in meadow voles. Behavioral Ecology and Sociobiology : 575 578. SLATE, J., T. C. MARSHALL, AND J. M. PEMBERTON. 000. A retrospective assessment of the accuracy of the paternity inference program CERVUS. Molecular Ecology 9:80 808. SMITH, M. H., R. BACCUS, H.O.HILLESTAD, AND M. N. MANLOVE. 98. Population genetics of the white-tailed deer. Pp. 9 8 in White-tailed deer ecology and management (L. K. Halls, ed.). Stackpole Books, Harrisburg, Pennsylvania. STRASSMANN, J. E., C. R. SOLIS, J.M.PETERS, AND D. C. QUELLER. 996. Strategies for finding and using highly polymorphic DNA microsatellite loci for studies of genetic relatedness and pedigrees. Pp. 6 80 in Molecular zoology: advances, strategies, and protocols (J. D. Ferraris and S. R. Palumbi, eds.). Wiley-Liss, New York. SUGG, D. W., AND R. K. CHESSER. 99. Effective population sizes with multiple paternity. Genetics 7: 7 55. TEGELSTROM, J., J. SEARLE, J. BROOKFIELD, AND S. MERCER. 99. Multiple paternity in common wild shrews (Sorex araneus) is confirmed by DNA fingerprinting. Heredity 66:7 79. VERME, L. J. 965. Reproductive studies on penned white-tailed deer. Journal of Wildlife Management 9:7 79. WADE, M. J. 979. Sexual selection and variance in reproductive success. American Naturalist :7 76. WADE, M. J., AND S. J. ARNOLD. 980. The intensity of sexual selection in relation to male sexual behaviour, female choice, and sperm precedence. Animal Behaviour 8:6 6. XIA, X. 999. Estimating the frequency of promiscuous females by using molecular data. Pp. 6 5 in The theory, methods, and application of molecular ecology (X. G. Zhu, M. Sun, and K. Le, eds.). China Higher Education Press and Springer-Verlag, Beijing, China. XIA, X., AND J. S. MILLAR. 99. Genetic evidence of promiscuity in Peromyscus leucopus. Behavioral Ecology and Sociobiology 5:7 78. YOAKUM, J. D., AND B. W. O GARA. 000. Pronghorn.

89 JOURNAL OF MAMMALOGY Vol. 8, No. Pp. 559 577 in Ecology and management of large mammals in North America (S. Demarais and P. R. Krausman, eds.). Prentice-Hall, Inc., Upper Saddle River, New Jersey. ZAMUDIO, K. R., AND B. SINERVO. 000. Polygyny, mate-guarding, and posthumous fertilization as alternative male mating strategies. Proceedings of the National Academy of Sciences 97:7. Submitted June 00. Accepted 0 November 00. Associate Editor was Meredith J. Hamilton.