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Pathogenesis of Mycobacterium bovis infection in American bison.

Eighteen 12-month-old bison were randomly placed in each of 3 groups (6 animals/group): group-1 bison were exposed to live Mycobacterium bovis, group-2 bison were inoculated with killed M bovis in oil, and group-3 bison were noninfected controls. Six, 6-month-old, tuberculin test-negative calves were placed (pen contact) with group-1 bison 30 days after exposure to M bovis. Tuberculin skin test responses (caudal fold and/or comparative cervical) were detected in all bison in groups 1 and 2 at 2, 4, 6, 10, and 12 months. Tuberculin skin test responses were observed in 2 of 6 calves at 9 and 11 months after pen contact with M bovis-exposed bison (group 1). Statistically significant lymphocyte blastogenic responses to M bovis purified protein derivative were detected in group-1 bison exposed to live M bovis at 2 months after exposure (P less than 0.025). Significant ELISA reactions were detected in sera of bison at 2 months after exposure to killed M bovis in oil (P less than 0.005) and in bison 2 months after exposure to live M bovis (P less than 0.01). Significant tuberculin skin responses, ELISA reactions, or lymphocyte blastogenic responses to M bovis purified protein derivative were not observed in the 6 control bison. Grossly visible tuberculous lesions were observed in lymph nodes and/or lung collected at necropsy in 4 of 6 bison at 12 months after exposure to live M bovis. Microscopic granulomas compatible with tuberculosis were detected in 5 of 6 bison; M bovis was isolated from tissues of each of the 6 bison.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Safety of revaccination of pregnant bison with Brucella abortus strain RB51.

From December 1998 through February 1999, a study was conducted in a Brucella-infected bison herd to evaluate the safety of booster vaccination of adult bison (Bison bison) with 6 x 10(9) colony forming units (CFU) of Brucella abortus strain RB51 (SRB51) that had previously been vaccinated as yearlings with 1 x 10(10) CFU of SRB51. Abortions or other adverse effects were not observed after SRB51 booster vaccination. At 10 wk after adult vaccination, pregnant and nonpregnant bison (n = 65) were randomly selected for bacteriologic sampling of targeted maternal tissues during abattoir processing. Fetal tissues were also sampled in pregnant bison. The SR351 recovered from tissue samples of eight of 48 pregnant bison and none of 17 nonpregnant bison. In three of the eight culture-positive bison, SRB51 was recovered from fetal tissues. In three additional bison, one pregnant and two nonpregnant, B. abortus biovar 1 field strain was recovered from internal iliac or supramammary lymphatic tissues. Results of this study suggest the possibility that the SRB51 vaccine can be safely used to booster vaccinate pregnant bison in a Brucella-infected bison herd. Our data also reaffirms the potential for B. abortus field strains to persist in bison until attainment of reproductive age, despite extensive use of vaccination and serologic testing.

Abortion, Veterinary↗

Pathology of brucellosis in bison from Yellowstone National Park.

Between February 1995 and June 1999, specimens from seven aborted bison (Bison bison) fetuses or stillborn calves and their placentas, two additional placentas, three dead neonates, one 2-wk-old calf, and 35 juvenile and adult female bison from Yellowstone National Park (USA) were submitted for bacteriologic and histopathologic examination. One adult animal with a retained placenta had recently aborted. Serum samples from the 35 juvenile and adult bison were tested for Brucella spp. antibodies. Twenty-six bison, including the cow with the retained placenta, were seropositive, one was suspect, and eight were seronegative. Brucella abortus biovar 1 was isolated from three aborted fetuses and associated placentas, an additional placenta, the 2-wk-old calf, and 11 of the seropositive female bison including the animal that had recently aborted. Brucella abortus biovar 2 was isolated from one additional seropositive adult female bison. Brucella abortus was recovered from numerous tissue sites from the aborted fetuses, placentas and 2-wk-old calf. In the juvenile and adult bison, the organism was more frequently isolated from supramammary (83%), retropharyngeal (67%), and iliac (58%) lymph nodes than from other tissues cultured. Cultures from the seronegative and suspect bison were negative for B. abortus. Lesions in the B. abortus-infected, aborted placentas and fetuses consisted of necropurulent placentitis and mild bronchointerstitial pneumonia. The infected 2-wk-old calf had bronchointerstitial pneumonia, focal splenic infarction, and purulent nephritis. The recently-aborting bison cow had purulent endometritis and necropurulent placentitis. Immunohistochemical staining of tissues from the culture-positive aborted fetuses, placentas, 2-wk-old calf, and recently-aborting cow disclosed large numbers of B. abortus in placental trophoblasts and exudate, and fetal and calf lung. A similar study with the same tissue collection and culture protocol was done using six seropositive cattle from a B. abortus-infected herd in July and August, 1997. Results of the bison and cattle studies were similar.

Abortion, Veterinary↗

Genetic variation within and relatedness among wood and plains bison populations.

There are two recognized subspecies of bison, wood (Bison bison athabascae) and plains (Bison bison bison) bison. The establishment of most bison populations from a small number of individuals has raised concerns about their genetic variation. To this end, 11 bison populations were surveyed with 11 microsatellite loci in order to calculate genetic variation and genetic distances. Mean number of alleles ranged between 3.18 at Antelope Island State Park (Utah) and 6.55 at Wood Buffalo National Park (Alberta and Northwest Territories). Mean heterozygosity ranged from 0.295 at Antelope Island State Park to 0.669 at Custer State Park (South Dakota). The amount of genetic variability present in the bison populations as measured by mean number of alleles and overall probability of identity was found to correlate with the number of founders for all sampled populations. The G-test for heterogeneity revealed some evidence for the existence of subpopulations at Wood Buffalo National Park, however very small genetic distances between these subpopulations suggest that nuclear material from the plains bison introduced into Wood Buffalo National Park has diffused throughout the park. Genetic distances between the sampled populations were generally larger between than within the two bison subspecies.

Alleles↗

The pathogenicity of Brucella suis biovar 4 for bison.

The pathogenicity of Brucella suis biovar 4 for bison (Bison bison) was evaluated by inoculation of 2.1 x 10(7) colony forming units (CFU) in 0.1 ml saline into the conjunctival sac of six pregnant cows. Six pregnant bison were inoculated with 1.27 x 10(7) CFU of Brucella abortus strain 2308 as a positive control. Bison were inoculated on 23 January 1992, and observed until calving or abortion after which they were euthanized, and necropsied. Bacteriological and histological examinations were conducted on lymph nodes, reproductive tract, mammary gland, and internal organs. Terminal serum samples from calves and cows were evaluated by card, rivanol precipitation, standard tube agglutination, cold complement fixation tube, indirect bison conjugated enzyme linked immunosorbent assay (ELISA), competitive ELISA, and particle-concentration fluorescence immunoassay. No clinical signs of brucellosis were seen in bison inoculated with B. suis biovar 4, and infection was found only in lymph nodes of two animals. There was no evidence of metastasis of this organism to the mammary gland or the reproductive tract. There were no detectable levels of antibodies to Brucella spp. in terminal blood samples taken from B. suis biovar 4-challenged bison. Brucella abortus was isolated from several tissues in all control bison. All B. abortus-challenged animals developed uterine infection and five developed mammary gland infection. Reproductive disease resulted in abortions in five B. abortus-challenged bison and neonatal death in the remaining calf. Brucella suis biovar 4 does not appear to be pathogenic for bison.

Abortion, Veterinary↗

Conservation of germ plasm from bison infected with Brucella abortus.

Reproductive procedures for cattle were adapted to American bison (Bison bison) to evaluate the potential preservation of germ plasm from bison infected with Brucella abortus without transmission of the pathogen to the recipient or offspring. Two of four experimentally inoculated bison bulls excreted B. abortus in the semen. Four healthy calves were produced from non-infected, un-vaccinated bison cows by natural breeding with a bison bull excreting B. abortus in the semen. There was no seroconversion of the cows or their calves. Two culture negative bison calves were produced by superovulation of infected bison donor cows followed by artificial insemination and embryo transfer without transmitting B. abortus to recipient cows or calves. These limited data indicate that embryo manipulatory procedures and natural breeding in bison may facilitate preservation of valuable germ plasm from infected bison while reducing the risk of transmission of B. abortus to recipients and progeny.

Animals↗

Evaluation of serologic responses, lymphocyte proliferative responses, and clearance from lymphatic organs after vaccination of bison with Brucella abortus strain RB51.

OBJECTIVE: To evaluate clearance of the vaccine strain, immunologic responses, and potential shedding of Brucella abortus strain RB51 organisms after vaccination of bison calves. ANIMALS: Fourteen 7-month-old female bison calves. PROCEDURE: 10 bison calves were vaccinated SC with 1.22 x 10(10) colony-forming units of B abortus strain RB51. Four bison calves were vaccinated SC with 0.15M NaCl solution. Rectal, vaginal, nasal, and ocular swab specimens were obtained to evaluate potential shedding by vaccinated bison. The superficial cervical lymph node was biopsied to evaluate clearance of the vaccine strain. Lymphocyte proliferative responses to strain RB51 bacteria were evaluated in lymph node cells obtained from biopsy specimens and also in peripheral blood mononuclear cells. RESULTS: Strain RB51 was recovered from superficial cervical lymph nodes of vaccinates examined 6, 12, and 18 weeks after vaccination (4/4, 3/4, and 1/4, respectively) but not in vaccinates examined at 24 weeks (0/3) after vaccination or nonvaccinates examined at all sample collection times (n = 1 bison/sample period). Serologic, immunologic, and bacterial culture techniques failed to reveal shedding of strain RB51 by vaccinates or infection of nonvaccinated bison. Lymphocyte proliferative responses were evident in lymph node cells and blood mononuclear cells from strain RB51-vaccinated bison beginning 12 weeks after vaccination. CONCLUSION: Strain RB51 was cleared from bison by 18 to 24 weeks after vaccination. Bison vaccinated with strain RB51 did not shed the vaccine strain to nonvaccinated bison housed in close proximity. Strain RB51 did not induce antibody responses in bison that would interfere with brucellosis surveillance tests, but did stimulate cell-mediated immunity.

Agglutination Tests↗

Effects of ration, season and animal handling on composition of bison and cattle blood.

Composition of ration and season of sampling markedly affected the composition of blood in six tamed bison (Bison bison) steers and eight Hereford cattle (Bos taurus) steers. Observed values extended reported ranges for albumin, phosphorus and blood urea nitrogen (BUN) in bison serum. There were several differences between species in blood composition. In particular, erythrocytic and BUN values were higher in bison than in cattle. Overall mean values for bison and cattle receiving experimental rations were, respectively: BUN, 17.1 mg/dl and 14.1 mg/dl; hemoglobin, 17.8 g/dl and 13.3 g/dl; packed cell volume (PCV), 47.6% and 35.6%; red blood cells, 9.3 x 10(6)/mm3 and 8.2 x 10(6)/mm3; mean corpuscular volume (MCV), 51.3 mean 3; mean corpuscular hemoglobin, 18.9 pg and 16.1 pg. The significant changes in blood composition associated with changes in ration composition support the use of blood composition as an index of nutritional status. There were no sex-specific differences in blood of 20 bison from Elk Island National Park and 34 bison from Wood Buffalo National Park, Alberta. Alkaline phosphatase (ALP) level was higher in juvenile than in adult bison. Impoundment of wild bison for 24 hr was accompanied by a decrease in BUN and an increase in PCV. Wild bison that were killed during handling had significantly higher blood levels of ALP, glutamate oxaloacetate transaminase, MCV and phosphorus.

Animal Feed↗

Serum biochemical and hematologic values of normal and Mycobacterium bovis-infected American bison.

Hematologic and serum biochemical tests were used to monitor the health of 3 groups of bison in an experimental study of tuberculosis. Bison were randomly assigned to Mycobacterium bovis-infected, M. bovis-sensitized, and uninfected control groups. Hematologic measurements included total and differential leukocyte counts, hemoglobin (Hb), packed cell volume (PCV), fibrinogen, and plasma proteins. Biochemical tests included serum urea nitrogen, creatinine, aspartate amino transferase, sorbitol dehydrogenase, serum calcium, and serum phosphorus. There were no significant differences (P = 0.05) in any test values between groups of bison. The bison data were combined and compared to similar data of cattle. The mean values for PCV and Hb were higher than values (PCV 24-46%, Hb 8-15 g/dl) for cattle. Mycobacterium bovis-infected bison had a slight increase in the number of blood monocytes and lymphocytes when compared to the uninfected bison but were within the normal ranges for bison and cattle. Other hematologic parameters were within normal ranges reported for cattle. Creatinine levels in all bison were above the normal range (1.0-1.5 mg/dl) for cattle. Phosphorus levels for M. bovis-infected and M. bovis-sensitized bison exceeded the normal range (5.6-8.0 mg/dl) reported for cattle. The level for uninfected bison was near the upper limit of normal for cattle. Mean values for other serum biochemical tests were within the normal ranges reported for cattle.

Animals↗

Recent studies on the characterization of Anaplasma marginale isolated from North American bison.

Anaplasma marginale (Rickettsiales: Anaplasmataceae), a tick-borne pathogen of cattle, is endemic in tropical and subtropical regions of the world. Many geographic isolates of A. marginale occur worldwide that have been identified by major surface protein (MSP) 1a, which varies in sequence and molecular weight owing to different numbers of tandem 28-29 amino acid repeats. Although serologic tests have identified American bison, Bison bison, as being infected with A. marginale, the present studies were undertaken to confirm A. marginale infection in bison, to characterize bison isolates, and to compare the phylogenetic relationship of the bison isolates with other A. marginale isolates from North America. Nine A. marginale isolates derived from Canadian bison possessed identical msp4 sequences with one characteristic silent nucleotide change. The sequence of MSP1a was determined for one Canadian and two U.S. bison isolates of A. marginale, and these isolates contained 4 and 5 tandem repeats, respectively. One U.S. bison isolate tested for infectivity proved to be infective for cattle and transmitted by Dermacentor variabilis ticks. the results of this study demonstrated that these A. marginale isolates obtained from bison were similar to ones derived from naturally infected cattle.

Anaplasma marginale↗

Behavioral responses of bison and elk in Yellowstone to snowmobiles and snow coaches.

Managers of public lands are charged with protecting some of our most important natural resources and ecosystems, while providing for their use and enjoyment by visitors. Almost one million visitors entered Yellowstone National Park by motorized means on snowmobiles (87%) or snow coaches (13%) during 1992-2003. Most vehicles toured the central portion of the park where bison (Bison bison) and elk (Cervus elaphus) concentrate in geothermal areas. We sampled >6500 interactions between groups of these species and groups of snowmobiles and snow coaches (collectively, OSV, over-snow vehicles) during five winters (1999-2000, 2002-2004). Multinomial logits models were used to identify conditions leading to behavioral responses. Elk responded three times as often (52%) as bison (19%) during interactions with groups of snowmobiles and snow coaches due to increased vigilance responses (elk, 44%; bison, 10%). However, the frequency of higher-intensity movement responses by bison and elk were similar (6-7% travel, 1-2% flight, <1% defense) and relatively low compared to other studies of ungulates and snowmobile disturbance. The likelihood of active responses by bison and elk increased significantly if animals were on or near roads, groups were smaller, or humans approached. The likelihood of an active response by bison decreased within winters having the largest visitation, suggesting some habituation to snowmobiles and snow coaches. There was no evidence that snowmobile use during the past 35 years affected the population dynamics or demography of bison or elk. Thus, we suggest that regulations restricting levels and travel routes of over-snow vehicles (OSVs) were effective at reducing disturbances to bison and elk below a level that would cause measurable fitness effects. We recommend park managers consider maintaining OSV traffic levels at or below those observed during our study. Regardless, differing interpretations of the behavioral and physiological response data will continue to exist because of the diverse values and beliefs of the many constituencies of Yellowstone.

Animals↗

Duration of bluetongue viremia in experimentally infected American bison.

Six yearling American bison (Bison bison bison) bulls and one yearling ewe (Ovis aries) were inoculated intradermally and subcutaneously with 2 x 10(5) plaque forming units (pfu) of bluetongue (BT) virus serotype 11. Two uninoculated yearling bison bulls served as negative controls. Blood samples were collected for serology and virus isolation on 0, 4, 7, 11 and 14 days post-inoculation (dpi) and every 2 wk thereafter to 127 dpi. Every 4 wk a new ewe was inoculated with a pooled sample of whole blood from the six infected bison, and each sheep was monitored for 28 days for clinical signs of BT and seroconversion. Bluetongue viremia was detected in all six inoculated bison starting at 4 to 28 dpi and was no longer detectable from 42 dpi onward. Pooled blood samples collected at 28, 56, 84 and 112 dpi from the six infected bison were not infectious for sheep. The six infected bison seroconverted by 11 to 28 dpi on a competitive enzyme-linked immunosorbent assay and by 28 dpi on the serum neutralization test, and all remained seropositive thereafter. No clinical signs or lesions attributable to BT were observed in the infected bison or controls. There was evidence that a small amount of epizootic hemorrhagic disease virus type 2 had been present in the BT virus inoculum; reasons are given for concluding that this did not affect the results of the BT study.

Animals↗

R51163 as a sedative for handling and transporting plains bison and wapiti.

Forty captive wapiti (Cervus elaphus) and thirty-two bison (Bison bison bison) were tested in April and October 1988, respectively, for their response to the sedative R51163. Treatment animals were injected with either 0.1, 0.2, or 0.3 mg of R51163/kg and then observed for 72 hr. Behavior was significantly altered by the drug. Hyperactive, aggressive, and milling behavior was characteristic of treated wapiti and they were extremely dangerous and reared when hind quarters were touched. Although treated plains bison displayed some milling behavior, they were generally more calm than wapiti. There was a marked difference between sexes in plains bison for all behavioral categories. Male bison were more ataxic, often observed in sternal or lateral recumbency, less conscious, and were slower to respond than females or controls. Respiratory rate increased in treated wapiti and plains bison, and heart rates of treated wapiti increased. Because of the powerful sedative effect on large, male bison, R51163 may be useful for handling unmanageable or dangerous animals and warrants further studies.

Aggression↗

Biosafety and antibody responses of adult bison bulls after vaccination with Brucella abortus strain RB51.

OBJECTIVE: To evaluate clearance, antibody responses, potential shedding, and histologic lesions in reproductive tissues of adult bison bulls after vaccination with Brucella abortus strain RB51. ANIMALS: 61 two- and 3-year-old bison bulls. PROCEDURE: 12 bison bulls were vaccinated s.c. with B abortus strain RB51, 3 were inoculated s.c. with 0.15 M NaCl, and antibody responses were evaluated. Various specimens were obtained to evaluate bacterial shedding. Four vaccinates and 1 control were necropsied 10, 20, and 30 weeks after vaccination. In a separate experiment, bison bulls were vaccinated s.c. with 0.15 M NaCl, or by hand or ballistically with strain RB51. Antibody responses were monitored 6 weeks after vaccination and during necropsy 13 weeks after vaccination. Tissue specimens obtained during necropsy from both studies were evaluated bacteriologically and histologically. RESULTS: Strain RB51 was recovered at various times from semen of 3 of 12 vaccinated bison bulls in experiment 1. During necropsy, strain RB51 was recovered 10 and 20, but not 30, weeks after vaccination. In experiment 2, strain RB51 was recovered from lymphoid tissues of hand- and ballistic-vaccinated bison bulls during necropsy. In both experiments, microscopic lesions in testes, epididymis, and seminal vesicles were minimal and did not differ between strain RB51-vaccinated and saline-inoculated bison bulls. CONCLUSIONS AND CLINICAL RELEVANCE: Strain RB51 does not induce relevant inflammatory lesions in reproductive tissues of adult bison bulls. Shedding of strain RB51 in semen may be transient in some bison bulls; however, the importance of this observation is unknown.

Agglutination Tests↗

Anaplasma marginale infections in American bison: experimental infection and serologic study.

Anaplasma marginale was experimentally transmitted from cattle to bison and back to cattle. Of the 2 splenectomized and 1 intact American bison calves (Bison bison) inoculated with a North Texas A marginale stabilate, 1 splenectomized and 1 intact bison exhibited clinical signs of anaplasmosis. Active parasitemias in these bison were observed along with positive reactions in the rapid card agglutination and complement fixation tests. Blood from the infected bison produced disease in splenectomized bovine calves. Screening tests for anti-Anaplasma antibodies in 178 blood samples collected from adult bison from the National Bison Range, Montana, revealed 1 rapid card agglutination test-positive sample, and 110 negative, 40 suspect, and 28 positive (15.7%) complement fixation test samples.

Agglutination Tests↗

Fecal androgens of bison bulls during the rut.

The influence of sex hormones is a key proximate factor underlying male reproductive behavior in mammals. Effective conservation policies for the remaining purebred plains bison (Bison bison bison) herds require knowledge of the physiology underlying bison reproductive biology. We used fecal steroid analysis to characterize androgen levels in adult bison bulls before, during, and after the rut, and to examine androgen levels of bulls differing in reproductive status, age, and mating success. Fieldwork was carried out at the Fort Niobrara National Wildlife Refuge in north-central Nebraska. All adult bison in the herd were individually known by unique brands. Fecal samples were collected during 2003 from bulls during pre-rut (June), rut (July-August), and post-rut (September), and behavioral observations focused on reproductive status and mating success during the rut. Matched sample data indicated that androgen levels (ng/g feces) of bulls peaked during the rut, doubling from pre-rut to rut and then declining by 75% during post-rut. Dominant bulls that tended (guarded) cows maintained higher androgen levels than bulls that were not tending. There was a positive correlation between bull age (associated with mating success) and androgens, with higher androgen levels in prime-aged bulls compared with younger bulls. Nonetheless, there was no correlation between mating success (measured by number of copulations observed) and androgen level. This suggests that while androgens may provide the proximate motivation to compete for matings, other factors determine the mating success of bison bulls.

Age Factors↗

Efficacy of calfhood vaccination with Brucella abortus strain RB51 in protecting bison against brucellosis.

In the studies reported here, protection induced by calfhood vaccination of bison with 1.2-6.1 x 10(10)CFU of Brucella abortus strain RB51 (SRB51) against a virulent strain of B. abortus was evaluated. Non-vaccinated and SRB51-vaccinated bison were intraconjunctivally challenged during midgestation with 3 x 10(7)CFU of virulent B. abortus strain 2308 (S2308). Maternal and fetal tissues were obtained within 24hour after abortion or parturition. Incidence of abortion was greater (P<0.05) in non-vaccinated as compared to SRB51-vaccinated bison (62% and 15%, respectively), with abortions occurring between 5 and 8 weeks after experimental challenge. Calves from bison vaccinated with SRB51 had a reduced (P<0.05) prevalence of fetal infection with S2308 as compared to calves from non-vaccinated bison (19% and 62%, respectively). Although the ability to recover the 2308 challenge strain from maternal tissues did not differ (P>0.05) between nonvaccinates and vaccinates (100% and 78%, respectively), calfhood vaccination with SRB51 reduced (P<0.05) recovery of S2308 from uterine or mammary gland tissues. In bison which did not abort, S2308 was routinely recovered in low numbers from maternal lymphatic tissues; particularly the parotid, bronchial, supramammary, and mandibular lymph nodes. The RB51 vaccine strain was not recovered at any time from maternal or fetal samples obtained at necropsy. Histological lesions associated with Brucella-induced abortions were suppurative placentitis, fetal broncho-interstitial pneumonia, and fetal histiocytic splenitis. The results of our studies suggest that calfhood vaccination of bison with SRB51 is efficacious in protecting against intramammary, intrauterine, and fetal infection following exposure to a virulent strain of B. abortus during pregnancy. As brucellosis is transmitted horizontally through fluids associated with the birth or abortion of an infected fetus, or vertically to the calf through the ingestion of milk containing B. abortus, our data suggest that calfhood vaccination with SRB51 will be beneficial in preventing transmission of brucellosis in bison.

Abortion, Veterinary↗

Differential introgression of uniparentally inherited markers in bison populations with hybrid ancestries.

Historical hybridization between Bison bison (bison) and Bos taurus (cattle) has been well documented and resulted in cattle mitochondrial DNA (mtDNA) introgression, previously identified in six different bison populations. In order to examine Y chromosome introgression, a microsatellite marker (BYM-1) with non-overlapping allele size distributions in bison and cattle was isolated from a bacterial artificial chromosome (BAC) clone, and was physically assigned to the Y chromosome by fluorescence in situ hybridization. BYM-1 genotypes for a sample of 143 male bison from 10 populations, including all six populations where cattle mtDNA haplotypes were previously identified, indicated that cattle Y chromosome introgression had not occurred in these bison populations. The differential permeability of uniparentally inherited markers to introgression is consistent with observations of sterility among first generation hybrid males and a sexual asymmetry in the direction of hybridization favouring matings between male bison and female cattle.

Animals↗