Red Wolf Research Articles

Mitochondrial DNA extracted from eastern North American wolves killed in the 1800s is not of gray wolf origin

We analysed the mitochondrial DNA (mtDNA) from two historical samples of eastern North American wolves: the last wolf reported to have been killed in northern New York State (ca. 1890s) and a wolf killed in Maine in the 1880s. These wolves represent eastern wolves, presently classified as the gray wolf (Canis lupus) subspecies Canis lupus lycaon, which were present well before the expansion of western coyotes (Canis latrans) into these regions. We show the absence of gray wolf mtDNA in these wolves. They both contain New World mtDNA, supporting previous findings of a North American evolution of the eastern timber wolf (originally classified as Canis lycaon) and red wolf (Canis rufus) independently of the gray wolf, which originated in Eurasia. The presence of a second wolf species in North America has important implications for the conservation and management of wolves. In the upper Great Lakes region, wolves of both species may exist in sympatry or interbreed with each other, which impacts the accuracy of estimates of numbers of wolves of each species within this geographic region. Furthermore, the historical distribution of the eastern timber wolf (C. lycaon), as revealed by these skin samples, has important implications for the reintroduction of wolves into the northeastern U.S. states, such as New York and Maine.

Major histocompatibility complex variation in red wolves: evidence for common ancestry with coyotes and balancing selection.

We examined variation at a class II major histocompatibility complex (MHC) gene (DRB1) in the captive red wolf population and samples of coyotes from Texas and North Carolina. We found 4 alleles in the 48 red wolves, 8 alleles in the 10 coyotes from Texas and 15 alleles in the 29 coyotes from North Carolina. Two of the four alleles found in red wolves, Caru-2 and Caru-4, were found in both the Texas and North Carolina coyote samples. Allele Caru-1, previously found in gray wolves, was also found in the North Carolina sample. The most frequent red wolf allele, Caru-3, was not found in any of the coyote samples. However, an allele found in both the Texas and North Carolina coyote samples is only one nucleotide (one amino acid) different from this red wolf allele. Overall, it appears from examination of this MHC gene that red wolves are more closely related to coyotes than to gray wolves. There were a number of different types of evidence supporting the action of balancing selection in red wolves. Namely, there was: (i) an excess of heterozygotes compared with expectations; (ii) a higher rate of nonsynonymous than synonymous substitution for the functionally important antigen-binding site positions; (iii) an eight times higher average heterozygosity of individual amino acids at the positions identified as part of the antigen-binding site than those not associated with it; (iv) the amino acid divergence of four red wolf alleles was greater than that expected from a simulation of genetic drift; and (v) the distribution of alleles, and the distributions of amino acids at many positions were more even than expected from neutrality. Examination of the level and pattern of linkage disequilibria between pairs of sites suggest that the heterozygosity, substitution and frequencies at individual amino acids are not highly dependent upon each other.

THE ORIGINAL STATUS OF WOLVES IN EASTERN NORTH AMERICA

Assessment was made of all available cranial specimens of wild Canis dating since the Blancan and prior to AD 1918 in the region east of the Great Plains and south of the Prairie Peninsula, Lakes Erie and Ontario, and the St. Lawrence River. The small wolf C. priscolatrans(= C. edwardii) of the early Irvingtonian seems unrelated to the modern red wolf (C. rufus), but gave rise to a lineage including the larger C. armbrusteri and culminating in C. dirus of the late Rancholabrean. A small wolf, possibly a descendant of the Eurasian C. mosbachensis, did not reappear in the east until near the end of the Rancholabrean. At the same time, the coyote (C. latrans) disappeared from the east, not to return until the small wolf was extirpated in the 20th century. Fragmentary remains of the small wolf, dating from around 10,000 and 2,000-200 ybp, show continuity with 14 complete, mostly modern, eastern skulls. Multivariate analysis indicates those 14 represent a well-defined species, C. rufus, distinct from large series of the western gray wolf (C. lupus) and coyote. There is no evidence that the red wolf originated as a hybrid of the latter two species, though early specimens from central Texas suggest it began to interbreed with C. latrans by about 1900. Three long-recognized red wolf subspecies appear valid: C. r. floridanus, Maine to Florida; C. r. gregoryi, south-central United States; and C. r. rufus, central and coastal Texas, southern Louisiana, and probably now represented in the captive/reintroduced populations. The subspecies C. lupus lycaon of southeastern Ontario and southern Quebec is statistically intermediate to C. rufus and western C. lupus, and may have resulted from natural hybridization of those two species. Such could explain how the red and gray wolf differ so sharply where their ranges meet in the west but morphologically approach one another in the east.

Cardiorespiratory effects of medetomidine-butorphanol, medetomidine-butorphanol-diazepam, and medetomidine-butorphanol-ketamine in captive red wolves (Canis rufus).

Safe, effective, and reversible immobilization protocols are essential for the management of free-ranging red wolves (Canis rufus). Combinations using an alpha2-adrenoceptor agonist and ketamine have been shown to be effective for immobilization but are not reversible and can produce severe hypertension and prolonged or rough recoveries. To minimize hypertension and provide reversibility, 24 red wolves were immobilized using three medetomidine-butorphanol (MB) combinations without the use of ketamine in the initial injection. All wolves were administered medetomidine (0.04 mg/kg i.m.) and butorphanol (0.4 mg/kg i.m.). Seven wolves received no other immobilization agents (MB wolves), nine received diazepam (0.2 mg/kg i.v.) at the time they were instrumented (MBD wolves), and eight received ketamine (1 mg/kg i.v.) 30 min after instrumentation (MBK30 wolves). Physiologic parameters were monitored during immobilization. The heart rate was similar among the three groups for the first 30 min, and marked bradycardia was noted in one wolf from each group. Hypertension was observed initially in all three groups but was resolved within 10-30 min. The MBK30 wolves had significant elevations in heart rate and transient hypertension after intravenous ketamine administration. Most wolves had mild to moderate metabolic acidemia. Immobilizing drugs were antagonized in all wolves with atipamezole (0.2 mg/kg i.m.) and naloxone (0.02 mg/kg i.m.). The medetomidine-butorphanol-diazepam wolves were also given flumazenil (0.04 mg/kg i.v.). All wolves were standing within 12 min and were fully recovered within 17 min. Medetomamine-butorphanol and MBD combinations provided effective and reversible immobilization of red wolves without the sustained hypertension associated with the use of alpha2-adrenoceptor agonist-ketamine combinations. Delaying the administration of ketamine reduced its hypertensive effects.

Reproductive endocrine patterns in captive female and male red wolves (Canis rufus) assessed by fecal and serum hormone analysis

AbstractReproductive steroid profiles in female (n=13) and male (n=5) red wolves (Canis rufus) were characterized in fecal samples collected during the breeding season (December—May) and over a 1 year period, respectively. Blood samples from females (n=12) also were collected during the periovulatory period for luteinizing hormone (LH) and steroid analysis. High performance liquid chromatography (HPLC) of fecal extracts determined that estradiol and estrone constituted the major and minor forms, respectively, of fecal estrogen metabolites. Although native progesterone was present, pregnane metabolites predominated as the major forms of fecal progestins. HPLC analysis of fecal extracts from males revealed no native testosterone, but rather the predominance of more polar androgen metabolites. Based on hormone profiles and/or pup production, females were classified as pregnant (n=3), ovulatory‐nonpregnant (n=9), or acyclic (n=3). Longitudinal monitoring of females indicated no pregnancy‐specific differences in concentrations of either fecal progestagen or estrogen metabolites compared to ovulatory‐nonpregnant individuals; however, baseline progestagen concentrations were consistently elevated in acyclic females. There was good correspondence between serum and fecal steroid concentration during the periovulatory period. A rise in serum estrogens preceded the ovulatory LH surge which was then followed by a significant progesterone rise during the luteal phase. In males, changes in fecal androgen metabolite concentrations coincided with photoperiod fluctuations, increasing in late autumn and reaching peak concentrations during mid‐ to late winter just before the start of the breeding season. Collectively, these results serve as a database of ovarian and testicular endocrine events in this species, which can be utilized in population management and application of assisted reproductive technologies. Zoo Biol 21:321–335, 2002. © 2002 Wiley‐Liss, Inc.

α1-Antitrypsin polymorphism and systematics of eastern North American wolves

We used data on the polymorphic status of α1-antitrypsin (α1AT) to study the relationship of Minnesota wolves to the gray wolf (Canis lupus), which was thought to have evolved in Eurasia, and to red wolves (Canis rufus) and coyotes (Canis latrans), which putatively evolved in North America. Recent evidence had indicated that Minnesota wolves might be more closely related to red wolves and coyotes. Samples from wild-caught Minnesota wolves and from captive wolves, at least some of which originated in Alaska and western Canada, were similarly polymorphic for α1AT, whereas coyote and red wolf samples were all monomorphic. Our findings, in conjunction with earlier results, are consistent with the Minnesota wolf being a gray wolf of Eurasian origin or possibly a hybrid between the gray wolf of Eurasian origin and the proposed North American wolf.

Breeding habitat and lair structure of the ringed seal ( Phoca hispida ladogensis ) in northern Lake Ladoga in Russia

The breeding habitat and lair structures of the ringed seal (Phoca hispida ladogensis) were studied by snowmobile expeditions in northern Lake Ladoga, Russia, during 1996 to 1999. Mostly flat ice and no large pack ice existed in northern Lake Ladoga during this study. All birth lairs and 88% of haul-out lairs were located in snowdrifts in the shorelines of islands or islets and 12% of the haul-out lairs occurred in pressured ice ridges. Some large haul-out lairs had been used communally by the seals. A total of 15% of the lairs observed had been attacked or marked by red fox (Vulpes vulpes), wolf (Canis lupus) or unidentified canines. The colour of the lanugo fur of the Ladoga ringed seal pup appeared to be dark grey. We conclude that the northern part of Lake Ladoga is an active breeding area for the ringed seal, which attests conservation needs for this area.

Serum concentrations of vitamin D metabolites, vitamins A and E, and carotenoids in six canid and four ursid species at four zoos

Nutritional status for six captive canid species ( n=34) and four captive ursid species ( n=18) were analyzed. The species analyzed included: African wild dog ( Lycaon pictus), arctic fox ( Alopex lagopus), gray wolf ( Canis lupus), maned wolf ( Chrysocyon brachyurus), Mexican wolf ( Canis lupus baleiyi), red wolf ( Canis rufus), brown bear ( Ursus arctos), polar bear ( Ursus maritimus), spectacled bear ( Tremarctos ornatus), and sun bear ( Ursus malayanus). Diet information was collected for these animals from each participating zoo (Brookfield Zoo, Fort Worth Zoo, Lincoln Park Zoological Gardens, and North Carolina Zoological Park). The nutritional composition of the diet for each species at each institution met probable dietary requirements . Blood samples were collected from each animal and analyzed for vitamin D metabolites 25(OH)D and 1,25(OH) 2D, vitamin A (retinol, retinyl stearate, retinyl palmitate), vitamin E (α-tocopherol and γ-tocopherol) and selected carotenoids. Family differences were found for 25(OH)D, retinol, retinyl stearate, retinyl palmitate and γ-tocopherol. Species differences were found for all detectable measurements. Carotenoids were not detected in any species. The large number of animals contributing to these data, provides a substantial base for comparing the nutritional status of healthy animals and the differences among them.

DNA profiles of the eastern Canadian wolf and the red wolf provide evidence for a common evolutionary history independent of the gray wolf

The origin and taxonomy of the red wolf (Canis rufus) have been the subject of considerable debate and it has been suggested that this taxon was recently formed as a result of hybridization between the coyote and gray wolf. Like the red wolf, the eastern Canadian wolf has been characterized as a small "deer-eating" wolf that hybridizes with coyotes (Canis latrans). While studying the population of eastern Canadian wolves in Algonquin Provincial Park we recognized similarities to the red wolf, based on DNA profiles at 8 microsatellite loci. We examined whether this relationship was due to similar levels of introgressed coyote genetic material by comparing the microsatellite alleles with those of other North American populations of wolves and coyotes. These analyses indicated that it was not coyote genetic material which led to the close genetic affinity between red wolves and eastern Canadian wolves. We then examined the control region of the mitochondrial DNA (mtDNA) and confirmed the presence of coyote sequences in both. However, we also found sequences in both that diverged by 150 000 - 300 000 years from sequences found in coyotes. None of the red wolves or eastern Canadian wolf samples from the 1960s contained gray wolf (Canis lupus) mtDNA sequences. The data are not consistent with the hypothesis that the eastern Canadian wolf is a subspecies of gray wolf as it is presently designated. We suggest that both the red wolf and the eastern Canadian wolf evolved in North America sharing a common lineage with the coyote until 150 000 - 300 000 years ago. We propose that it retain its original species designation, Canis lycaon.

Piecing together the puzzle of carnivore reproduction

Recent advances in feline and canine reproductive studies demonstrate how methodically piecing this information together is beginning to reap rewards for wildlife conservation programs. Non-invasive endocrinology can be used to monitor female reproductive function, time con-specific introductions or AI, and diagnose pregnancy. Sperm morphology characteristics and cell membrane function may be genetically inherited and differ between genetically diverse and inbred species/populations in felids. It is not clear if the same is true for the endangered red wolf. While standards exist for freezing feline and canine sperm, new information using fluorescent staining and zona penetration assays (ZPA) indicates that significant damage can occur during pre-freeze cooling, and may also be related to a species' genetic diversity. Posthumous gamete salvage from genetically valuable animals not only provides a means to study sperm and oocyte physiology but also to assist with genetic management of populations. Using the knowledge gained, IVM/IVF and ICSI have been successful in the domestic cat and AI has resulted in offspring in numerous non-domestic felids. However, understanding the processes of IVM/IVF is still not well understood in canids. New information reveals that sperm and the cumulus cells may be integral to oocyte maturation and that canine epididymal sperm are not capable of undergoing fertilization. The acquisition of knowledge and application of biotechnologies lags behind for non-domestic canid conservation programs.

Brief communication. Polymorphism of α1-antitrypsin in North American species of Canis

alpha 1-Antitrypsin (A1AT) is a major protease inhibitor present in all mammalian sera that have thus far been investigated. A1AT is also highly polymorphic and is therefore a useful genetic marker. Previously reported A1AT polymorphism in domestic dogs consisted of two alleles designated as PiM and PiS which exhibited frequencies of 0.72 and 0.28, respectively, in a group of randomly collected mongrel dogs. North American species of Canis, which included gray wolves (n = 29), Mexican wolves (n = 20), coyotes (n = 24), wolf-dog crosses (n = 9), and red wolves (n = 27) were tested for A1AT polymorphism. A1AT phenotypes were determined by isoelectric focusing, followed by direct immunoblotting using a specific antiserum. A1AT concentrations were determined by radial immunodiffusion. Concentrations of A1AT were similar to those found in domestic dogs (2.26 +/- 0.3, +/- SD mg/ml, +/- SD) and tended to be higher in females than in males, possibly indicating that A1AT may be hormonally influenced in females. Three phenotypic band patterns were observed (M, MS, S). The allele frequencies for domestic dogs and gray wolves were very similar, 0.72 and 0.67 for PiM and 0.28 and 0.33 for PiS, respectively. The Mexican wolves had a significantly lower frequency of PiS = 0.10. Coyotes and red wolves were all found to be monomorphic for the PiS allele and were indistinguishable from each other in that respect.

Survey of necropsy results in captive red wolves (Canis rufus), 1992-1996.

Through the Red Wolf Species Survival Plan, the captive red wolf (Canis rufus) population was developed with the intent of reestablishing wild populations. One part of the plan was a survey for diseases that might occur as a result of population homogeneity or that might impede breeding success and reintroduction. For this survey, complete necropsies and histopathologic analyses were performed on 62 red wolves from 1992 to 1996. Major causes of 22 neonatal deaths were parental trauma, parasitic pneumonia, and septicemia. Common neonatal lesions included pododermatitis and systemic ascariasis. Cardiovascular anomalies and systemic parasitism were found in two juveniles. Causes of death in the 38 adults included conspecific trauma, neoplasia, or gastrointestinal diseases such as necrotizing enteritis, intestinal perforation, and gastric volvulus. Lymphosarcoma represented 50% of the fatal neoplasms. Three adults died from cardiovascular failure or hyperthermia during handling, and several adults were euthanized for suspected genetic diseases. Overall, the captive population had few significant health problems, but population fitness might be improved by continued removal of potentially deleterious genes from the breeding population and by modifying the husbandry of neonates and adults.

Predicting Red Wolf Release Success in the Southeastern United States

Although the red wolf (Canis rufus) was once found throughout the southeastern United States, indiscriminate killing and habitat destruction reduced its range to a small section of coastal Texas and Louisiana. Wolves trapped from 1973 to 1980 were taken to establish a captive breeding program that was used to repatriate 2 mainland and 3 island red wolf populations. We collected data from 320 red wolf releases in these areas and classified each as a success or failure based on survival and reproductive criteria, and whether recaptures were necessary to resolve conflicts with humans. We evaluated the relations between release success and conditions at the release sites, characteristics of released wolves, and release procedures. Although <44% of the variation in release success was explained, model performance based on jackknife tests indicated a 72-80% correct prediction rate for the 4 operational models we developed. The models indicated that success was associated with human influences on the landscape and the level of wolf habituation to humans prior to release. We applied the models to 31 prospective areas for wolf repatriation and calculated an index of release success for each area. Decision-makers can use these models to objectively rank prospective release areas and compare strengths and weaknesses of each.

DNA profiles of the eastern Canadian wolf and the red wolf provide evidence for a common evolutionary history independent of the gray wolf

The origin and taxonomy of the red wolf (Canis rufus) have been the subject of considerable debate and it has been suggested that this taxon was recently formed as a result of hybridization between the coyote and gray wolf. Like the red wolf, the eastern Canadian wolf has been characterized as a small "deer-eating" wolf that hybridizes with coyotes (Canis latrans). While studying the population of eastern Canadian wolves in Algonquin Provincial Park we recognized similarities to the red wolf, based on DNA profiles at 8 microsatellite loci. We examined whether this rela - tionship was due to similar levels of introgressed coyote genetic material by comparing the microsatellite alleles with those of other North American populations of wolves and coyotes. These analyses indicated that it was not coyote ge- netic material which led to the close genetic affinity between red wolves and eastern Canadian wolves. We then exam- ined the control region of the mitochondrial DNA (mtDNA) and confirmed the presence of coyote sequences in both. However, we also found sequences in both that diverged by 150 000 - 300 000 years from sequences found in coyotes. None of the red wolves or eastern Canadian wolf samples from the 1960s contained gray wolf (Canis lupus) mtDNA sequences. The data are not consistent with the hypothesis that the eastern Canadian wolf is a subspecies of gray wolf as it is presently designated. We suggest that both the red wolf and the eastern Canadian wolf evolved in North Amer- ica sharing a common lineage with the coyote until 150 000 - 300 000 years ago. We propose that it retain its original species designation, Canis lycaon. 2166 Resume : Les origines et la taxonomie du Loup roux (Canis rufus) font l'objet d'une controverse importante et une hypothese a ete emise, a savoir qu'il s'agit d'un taxon recent issu de l'hybridation entre le Coyote et le Loup gris. Comme le Loup roux, le Loup de l'est du Canada est decrit comme un petit loup « mangeur de cerfs » qui s'hybride avec le Coyote (Canis latrans). L'etude de la population de Loups de l'est du Canada dans le parc provincial Algonquin nous a permis de reconnaitre des similarites avec le Loup roux d'apres les profils d'AD Na8l ocus microsatellites. Nous avons tente de determiner si cette relation etait due a des degres semblables d'introgression de materiel genetique en comparant les alleles microsatellites avec ceux d'autres populations nord-americaines de loups et de coyotes. Les analyses ont revele que ce n'est pas du materiel genetique de coyote qui a mene a la grande affinite genetique entre le Loup roux et le Loup de l'est du Canada. Nous avons ensuite examine la region de controle de l'ADN mitochondrial (ADNmt) et confirme la presence de sequences du Coyote chez les deux loups. Cependant, nous avons egalement

No Inbreeding Depression Observed in Mexican and Red Wolf Captive Breeding Programs

Abstract: Inbreeding depression is expected to affect populations of outbreeding mammals in inverse proportion to their population size and can affect whether small populations persist or go extinct. We used studbook records to examine the effect of inbreeding upon juvenile viability and litter size in two endangered species that have recently been reintroduced to the wild: the Mexican wolf (Canis lupus baileyi) and the red wolf (C. rufus). We found that neither juvenile viability nor litter size was lowered by inbreeding in either taxon. In fact, both captive breeding programs appear to have less lethal equivalents than the median estimate for mammals. We did find that year of birth was correlated with increasing viability in both taxa. We conclude that there is no evidence that inbreeding depression will prove a major obstacle to the success of either recovery effort.

Genetic evidence for a recent origin by hybridization of red wolves.

Genetic data suggest that red wolves (Canis rufus) resulted from a hybridization between coyotes (C. latrans) and grey wolves (C. lupus). The data of the hybridization, however, is uncertain. According to one hypothesis, the two species came into contact as coyotes increased their geographical range in conjunction with the advance of European settlers and as grey wolves were extirpated from the American south. Alternatively, the red wolves could have originated tens of thousands of years ago as a result of climate and habitat changes that disturbed the ecology of the two parent species. To obtain an upper limit on the date of hybridization that would help to distinguish the two scenarios, we compared microsatellite allele length distributions from red wolves, coyotes and grey wolves. Subject to the assumptions of our analysis, we conclude that the red wolves originated as a result of hybridizations that occurred during the past 12,800 years, and probably during the past 2500 years.

Frequency of the codon 807 mutation in the cGMP phosphodiesterase beta-subunit gene in Irish setters and other dog breeds with hereditary retinal degeneration.

Rod-cone dysplasia 1 (rcd1) in Irish setters is caused by a nonsense mutation in the cGMP phosphodiesterase beta-subunit gene (PDE6B). We examined the frequency of the mutant allele in the Irish setter population and determined if the defect is present in dogs of other breeds which are affected with other inherited photoreceptor diseases. Between 1994 and 1997, samples were obtained from 436 clinically normal Irish setters, a red wolf, and dogs from 23 different breeds. The mutation in codon 807 of PDE6B was detected in genomic DNA by heteroduplex analysis, allele-specific PCR, or restriction enzyme digestion. Of the 436 samples from clinically normal setters, 34 contained the mutation in one of the two PDE6B alleles (carrier rate = 7.8%). In contrast, the same mutation was not found in the red wolf or dogs of other breeds affected with PRA or inherited photoreceptor diseases. The high percentage of tested carriers, however, is not representative of the number of carriers in the population since some dogs tested were closely related and did not represent a random sample of the Irish setter breed.

Booklist and Notes

Booklist and Notes George Brosi Allen, Thomas J. The Butterflies of West Virginia and Their Caterpillars. Pittsburgh: University ofPittsburgh Press, 1997. 388 pages with index, glossary, figures, tables and color plates. Trade paperback. $22.95. One hundred and twenty-eight species of butterflies are displayed brilliantly in color plates and described meticulously in this outstanding guidebook. Background information on the butterfly and butterfly gardening round out the book. The author is a wildlife biologist with the West Virginia Division of Natural Resources. Berry, Wendell. Two More Stories of the Port William Membership. Frarikfort, Kentucky: Gnomon, 1997. 62 pages. Hardback in dust jacket. $15.00. These stories, like those in the three previous coUections and five novels by Wendell Berry, are set in Port WUliam, Kentucky, a fictional rural community which strongly resembles Port Royal, his own home. The residents he writes about form a kind of membership because each relies explicitly upon the land to make a living and a life. The first story, "A Friend of Mine," focuses upon a farmer, Elton Penn, while the second story, "Inheritors," centers around a lawyer who raises cattle. As Wade Hall points out in his review for the Lexington Herald-Leader, "Berry's intention is not merely to paint a nostalgic portrait ofyesteryear. In fact, he offers this lifestyle as a viable alternative to dependence on the supermarket and Wal-Mart." Carnuto, Christopher. Another Country:Journeying Toward the Cherokee Mountains. NewYork: Henry Holt and Company, 1997. 351 pages with notes, sources, and an index. Hardback in dust jacket. $25.00. The central occasion for this book of nature writing is the release of red wolves, long extinct in this area, back into the Smoky Mountains National Park. An implicit question raised in the musings captured here is whether long-extinct peoples could ever repopulate the area and what that would mean. Thus much of die book is an exploration of the region's Cherokee heritage and its meaning. "He limns the good—the George Brosi sells both new and out-of-print books. His address is 123 Walnut Street, Berea, Kentucky 40403. Hisphone: (606)986-3262. 71 Cherokee language and customs, the reintroduction of the red wolf, the improvisational jazz to be found in the veery's song; and the bad—the savagery and sadism ofthe European conquest, cultural dismemberment, environmental degradation, pauperization ofthe land's spirit, the loss of native plants and animals, native ideas and images . . . Carnuto is humble enough, nimble enough, to sojourn successfully in these mythopoeic climes, conjuring a place portrait of swarming, satisfying complexity." —Kirkus Reviews. The author lives in Rockbridge County, Virginia, and is tiie author ofA Fly Fisherman's Blue Ridge. Conley, Robert J. War Woman: A Novel ofthe Real People. New York: St. Martin's Press, 1997. 357 pages with a glossary. Hardback in dust jacket. $25.95. Limited historical records indicate contact in the fifteenth century between the Cherokee and both the Spanish in the Florida peninsula and the English at Jamestown as well as offering some evidence of Spanish efforts to mine gold in Cherokee territory. This novel, centering around a strong female protagonist, provides a speculative fictional link between these prehistorical events. Robert J. Conley, an Oklahoma Cherokee, has made an unprecedented contribution to the literature of his people with his easily accessible writing. This is the eighth novel in this series ofearly Cherokee history and his twenty-fourth book. Fisher, Noel C. War at Every Door: Partisan Politics and Guerrilla Violence in East Tennessee: 1860-1869. Chapel Hill: University of North Carolina Press, 1997. 250 pages with maps, photos, notes, bibliography and index. Hardback in dust jacket. $29.95. "This book represents a major fresh perspective on the many faces of the Civil War, especially the relationship between conventional and partisan conflict ... It is Civil War history as it ought now to be written— across the spectrum ofpolitical, economic, social and military, events and the people who made them." —B. Franklin Cooling, author of Fort Donelson's Legacy. "No book so well demonstrates why we must go beyond traditional drum-and-bugle military history to understand the American Civil War. War at Every Door is destined to become a classic...

Characteristics of fresh and frozen-thawed red wolf (Canis rufus) spermatozoa.

Ejaculates of the red wolf (Canis rufus) were evaluated immediately after collection and freeze-thawing to initiate a reproductive database for this endangered species. Electroejaculates from 13 adult red wolves collected during the breeding season (February-March; n=25; 1-3 collections/male) had a mean volume of 4.7+/-0.7 ml, 146.5+/-25.7 x 10(6) spermatozoa/ml and 71.2% motile spermatozoa. The mean proportion of cells with normal morphology was 73.6+/-3.2% (range, 20.3-93.7%), with 64% of ejaculates (16/25) containing 70-90% normal spermatozoa. The four most predominant abnormalities were a coiled flagellum (8.1%), a bent flagellum (4.7%), a bent midpiece with no cytoplasmic droplet (3.3%;), and a detached head defect (6.4%). After cooling in glycerolated extender, semen was frozen using a pelleting method on dry ice before plunging into liquid nitrogen. Pellets were thawed in phosphate buffered saline and examined for % sperm motility, normal morphology, viability and intact acrosomes. There was a decline (P < 0.05) in sperm motility (approximately 40%) and percentage of normal sperm (11.9%) after freezing, but no change in the proportion of viable cells. After freezing, there was a marked decline (P < 0.05) in the proportion of intact acrosomes from 74.5% to 55.5% which was accompanied by an increased proportion (P < 0.05) of partial acrosomes from 11.9% to 35.8%. These data demonstrate that, although red wolf spermatozoa can survive freeze-thawing using a technique common for domestic dog sperm, the finding of significant acrosome damage reveals (1) likely species specificity in the Canis genus and (2) the need for refining sperm cryopreservation technology for the red wolf.

Cranial and dental abnormalities of the endangered red wolf Canis rufus

Skulls of 3 captive-raised female endangered red wolves Canis rufus Audubon and Bachman, 1851 exhibited severe malocclusion of the jaws. Cranial and dental abnormalities (including crowding of upper toothrows, and an extra tooth behind the lower left M3 in one of the three mandibles) were also evident.