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Brucellosis in the Greater Yellowstone Area (1998)

Chapter: Appendix B: Meeting Agendas and Presentations

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Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
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Appendix B

Bison in the Greater Yellowstone Area Draft Agenda 24-25 July 1997 Room 108, Reid Hall Bozeman, MT

24 July 1997

 

8:45

 

 

a. Opening remarks, introductions

 

Lee Paulson, Project Director

Norman Cheville, Principal Investigator

Dale McCullough, Principal Investigator

 

b. Comments

 

Dan Huff, National Park Service, Department of the Interior

Jack Rhyan, APHIS, U. S. Department of Agriculture

Bob Hillman, GYBIC

Factors in transmission

 

 

c. Serology and infection; epidemiology and pathogenesis

 

Tom Roffe

 

d. Brucella abortus and reproductive tissues

 

Jack Rhyan

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

 

e. Risk of transmission

 

Paul Nicoletti

10:15

Break

10:45

f. Relationship among serology, culture test results, and likelihood of infectiousness

 

Tom Roffe

 

g. Genetic diversity and disease resistance in bison with active disease eradication

 

Joe Templeton

 

h. Molecular biology and Brucella abortus

 

Peter Gogan

12:15

Lunch break

1:30

i. Modelling

 

Mike Miller

 

k. Population changes and distribution

 

Mary Meagher

3:15

Break

3:45

m. Population dynamics, preliminary data

 

Peter Gogan

 

n. Issues in vaccination

 

Fred Enright; Phil Elzer

 

o. Safety and effectiveness of existing vaccines

 

Steve Olsen

5:00

Adjourn

25 June 1997

 

8:45

p. Elk as a reinfection pathway for bison

 

Terry Kreeger

 

q. Outcomes for vaccination program specific to bison given the presence of Brucella abortus in elk and other wildlife

 

Terry Kreeger

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

10:15

Break

10:45

r. Role of vaccine development for bison and elk

 

Can Brucella abortus be eliminated totally from the GYA by development and use of a vaccine?

 

What would be the theoretical tradeoffs between a vaccine-only approach and a vaccination approach combined with a test and slaughter program?

 

Steve Olsen

Fred Enright

Phil Elzer

11:45

Discussion

12:15

Lunch break

1:30

Public comments from interested parties

3:30

Adjourn

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

Brucellosis in the Greater Yellowstone Area Agenda 4 August 1997 National Museum of Wildlife Art Jackson, Wyoming

8:45

Opening remarks, introductions

 

Lee Paulson, Project Director; Norman Cheville, Principal Investigator; Dale McCullough, Principal Investigator

 

Comments

 

Bob Schiller, National Park Service, Department of the Interior; Jack Rhyan, APHIS, U.S. Department of Agriculture; Art Reese, Wyoming Game and Fish Department

9:45

Molecular genetics

 

Betsy Bricker, National Animal Disease Center

10:15

Break

10:45

Experience in Montana

 

Keith Aune, Montana Department of Fish, Wildlife and Parks

 

Issues in transmission

 

Beth Williams, University of Wyoming

 

Brucellosis and wildlife research in Yellowstone and Grand Teton National Parks

 

Wayne Brewster, National Park Service

12:00

Lunch

1:00

Research in elk

 

Terry Kreeger, Wyoming Game and Fish

 

RB51 in elk

 

Phil Elzer, Louisiana State University

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

 

Vaccine applications at feedgrounds; habitat improvement

 

Scott Smith, Wyoming Game and Fish

2:30

Break

3:00

Public comment

5:00

Adjourn

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

EVALUATION OF BRUCELLA ABORTUS VACCINE STRAIN RB51 IN BISON

Philip H. Elzer1 and Donald S. Davis2

1  

2  

Department of Veterinary Science, Louisiana State University Agricultural Center, Baton Rouge, LA 70803 and Department of Veterinary Pathobiology, Texas A&M University, College Station, TX 77843.

Introduction:

Host

Bison - American Buffalo, Bison bison

Organism

Brucella abortus

first isolated in 1930's from the testicle of bull on National Bison Range, Moiese, MT

Serological positive animals found in 1917 in Yellowstone National Park

Disease

Reproductive disease that causes abortions (late term) that have been documented in the wild.

Problem

Bison which inhabit the Greater Yellowstone area

approximately 4000 animals with up to 50% seropositive for brucellosis

Cattle grazing in areas adjacent to the park may be susceptible to infection - Wyoming, Montana, and Idaho.

These states could lose their brucellosis-free status.

Vaccine strain

Brucella abortus RB51 - rough derivative of virulent Brucella abortus strain 2308. Multiple passages on Rifampin led to the loss of the O-polysaccharide side chain of the LPS. Therefore vaccination with this strain does not lead to the production of antibodies which will interfere with sero-diagnostic test for brucellosis. RB51 provides protection against virulent challenge with strain 2308 in a variety of species including cattle, goats, swine, elk and mice. In cattle, RB51 produces similar protection to that achieved with vaccination with S19 without vaccinal titers. In Pregnant animals, RB51 has also been found to be less pathogenic than S19 in that it induces fewer abortions.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

The purpose of this experiment was to evaluate the safety and pathogenesis of Brucella abortus strain RB51 in adult and young bison from a previously exposed herd.

Materials and Methods:

Animals

North American buffalo (Bison bison) were obtained from a reactor herd in Kansas. The herd contained 3 reactor animal as measured by conventional brucellosis serology. The herd was composed of 10 adult males, 7 calves and 14 adult females. The animals were shipped to and housed at Texas A&M University, College Station, TX throughout the experiment.

Vaccine

Brucella abortus Strain RB51 was obtained from Colorado Serum Co. and rehydrated according to the manufacturer's instructions.

Dose

Adult males and calves received 1-3×1010 colony forming units subcutaneously (standard calfhood dose in cattle). Adult females received 1×109 colony forming units subcutaneously (standard adult dose in cattle).

Experimental Design

The adult males and calves plus 5 non-pregnant cows were divided into 2 groups; group 1 was slaughtered at 13 weeks post vaccination, and group 2 was slaughtered at 16 weeks post vaccination.

Pregnant females were monitored until parturition, and delivery status was recorded. Live calves remained with the cows, and dead or weak calves were cultured for Brucella.

Tissue collection

The following tissues were collected aseptically at slaughter: liver, spleen, various lymph nodes, and reproductive tracts.

Bacterial culture

All of the tissues were homogenized in sterile distilled water, and the homogenates were plated on Brucella selective media.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×
Serology

Standard western blot analysis using RB51 and smooth field strain cell lysates were performed on all pre- and post-vaccination serum samples.

Results

Culture data

Table 1. Culture data from adult males, non-pregnant females and calves at 13 and 16 weeks post vaccination with RB51.

Weeks post-vaccination

Brucella abortus strain recovered

 

Field strain*

RB51

13

1 adult male+

none

16

none

none

* rifampin sensitive, smooth organism

+ animal number 3

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

Results

Serology Data

Table 2. Western blot analysis of serum samples taken before vaccination and at slaughter using cell lysates from Brucella abortus smooth strain 2308 and rough strain RB51.

Animal number

Pre-vaccination

Post-vaccination

 

2308

RB51

2308

RB51

1

-

-

+++

3*

+++

+

+++

++

5

+++

+

+++

++

8

-

+/-

+

++

9

-

+/-

+

++

11

-

-

-

++

13

++++

++

++++

+++

16

+/-

-

+/-

+++

19

-

+

-

+++

20

-

+/-

-

+++

21

-

-

-

+++

22

-

+/-

-

+++

24

-

+/-

-

++

25

-

-

-

++++

26

-

-

-

++

28

-

-

-

+++

29

-

+

-

+++

30

-

++

-

+

31

-

-

-

+/-

* culture positive animal (field strain)

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

Results

Fetal Pathogenesis

Table 3. Delivery status of female bison vaccinated with 1 × 109 colony forming units of RB51.

No. animals

Abortion

Live birth

Dystocia

Pending

9

0

5

1*

3

* culture negative for RB51 and field strain

Note: another pregnant animal was necropsied at 16 weeks post vaccination, and both the cow and the calf (150 days) were culture negative for RB51 and field strain.

Future Studies:

Determination of the vaccine efficacy of strain RB51 in female bison.

Group 1. Controls - saline subcutaneously

Group 2. RB51 vaccinates (this study) + another vaccination this year

Group 3. RB51 vaccinates (Idaho) + another vaccination this year

Group 4. RB51 vaccinates (this year)

All of the animals will be vaccinated in September.

The animals will be bred between October and November.

Animals will be challenged in the conjunctival sac with Brucella abortus strain 2308 (1 × 107 colony forming units). Delivery and culture status will be monitored.

Conclusions:

  1. Vaccination with RB51 in adult or young bison does not result in sero-conversion on standard brucellosis diagnostic tests.

  2. Vaccination with RB51 does not result in any gross pathological lesions in calves or adult males.

  3. RB51 does not appear to be pathogenic in adult males, non-pregnant females, or calves as measured by increased or prolonged colonization.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×
  1. RB51 does not appear to be pathogenic to adult pregnant females when administered to animals from a reactor herd.

  2. Further studies are necessary to determine the vaccine efficacy in bison.

Acknowledgments:

APHIS Brucellosis Research Committee

Dr. Jack Rhyan - NVSL

Dr. Mike Gilsdorf - USDA

Dr. Steve Olsen - NADC

Dr. Joe Templeton - Texas A&M University

Dr. Fred Enright, Sue Hagius, Joel Walker and William Flahive - Louisiana State University

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

EVALUATION OF THE VACCINE EFFICACY OF RB51 ADMINISTERED ORALLY IN ELK

Philip H. Elzer1, Gerhardt G. Schurig2, Fred M. Enright1, and Donald S. Davis3.

1  

Department of Veterinary Science, Louisiana State University Agricultural Center, Baton Rouge, LA 70803;

2  

College of Veterinary Medicine, Virginia Tech, Blacksburg, VA 24061; and

3  

Department of Veterinary Pathobiology, Texas A&M University, College Station, TX 77843.

Wild ungulates are susceptible to the infection and disease known as brucellosis. Brucella abortus can infect elk (Cervus elaphus canadensis ); and under experimental procedures, elk have transmitted the disease to cattle. There is circumstantial evidence that elk may have transmitted brucellosis to cattle under natural conditions. Large numbers of brucella-infected elk are found in the winter feedground areas of western Wyoming. The largest concentration of these elk frequent the winter feedgrounds of the National Elk Refuge, Jackson, Wyoming, which is administered by the U.S. Fish and Wildlife Service, Department of the Interior. To a lesser extent, brucella-infected elk also exist in Yellowstone National Park. Wild, free-ranging bison (Bison bison ) are also known to harbor B. abortus. These animals continue to hamper the efforts of brucellosis eradication. Therefore the purpose of this study was to orally vaccinate elk with B. abortus strain RB51 to mimic oral vaccination of large numbers of animals on the winter feedgrounds.

Brucella abortus RB51 is a stable, rough variant of strain 2308 which is rifampin resistant and has been demonstrated to induce protection against virulent brucella challenge in swine, goats, mice, and cattle. RB51 does not produce any O-side chain antigens in its lipopolysaccharide; therefore animals vaccinated with RB51 do not illicit antibodies against the O-side chain. This is of great benefit in that all of the standard tests used to diagnosis brucellosis measure antibodies specific for the O-side chain. Animals vaccinated with RB51 will not make antibodies which react in the standard diagnostic tests thus ''vaccine-induced titers" and "false positives" would not cloud eradication efforts.

A preliminary study using RB51 as an oral vaccine was first performed in cattle. Briefly, twenty brucella-naive crossbred heifers were divided into 2 groups. Group 1 (saline controls) received their normal ration of hay/pellets with karo syrup and saline poured over it; and group 2 (vaccinates) received their normal ration laced with > 1010 colony forming

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

units (cfu) of RB51 resuspended in karo syrup and saline. All of the animals were individually housed during the vaccination and monitored until they ingested all of their rations. The animals were pasture bred; and at approximately 180 days gestation, they were challenged with 1 × 107 cfu of virulent B. abortus strain 2308. Strain 2308 has an established virulence in cattle, bison, and elk and typically induces abortions in infected animals. The delivery status of the animals was noted; and all calves or fetuses were necropsied immediately after birth or abortion with selected tissues bacteriologically examined. Uterine swabs and milk samples were taken from the cows and also cultured for brucella. Four weeks after parturition, the cattle were necropsied and tissues taken for culture. The results are as follows: the saline controls had 7/10 abortions whereas the oral vaccinates had 3/10 abortions; the challenge strain was recovered from 80% of the controls as compared to only 20% of the vaccinates. This study indicated that RB51 could be used as an oral vaccine and when administered by this route stimulated protective immunity against virulent challenge.

The oral exposure of elk with RB51 to protect against infection and abortion was investigated. Brucella-negative female elk were obtained from a site in South Dakota and transported to North Dakota for the oral vaccination study.

Female elk were pasture bred and orally exposed to RB51 (or saline as a placebo) in December to mimic the feed ground situation as that would be the practical time and site of vaccination. The females were divided into two groups; group 1 received saline and group 2 received at least 1010 cfu of RB51 placed into their mouths following scarification with a float. At one month post-vaccination, 2 out of 26 vaccinates were blood culture positive for RB51. The pregnant elk were transported to Texas A&M University 8 weeks post-vaccination. At midgestation all of the elk were challenged conjunctivally with 1 × 107 cfu of virulent B. abortus strain 2308. The animals were monitored for abortions, weak, and live births. All of the calves were necropsied soon after birth, and tissues were collected for culture. The adult females were necropsied, and tissues were cultured for brucella.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

Table 1. Vaccine efficacy of RB51 administered orally to female elk.

Delivery Status

Nonvaccinated saline controls

RB51 vaccinates

9 abortions

5 abortions

2 stillborn

0 stillborn

4 weak (died)

0 weak (died)

0 live

4 live

Table 2. Culture status of elk females and calves following challenge with virulent Brucella abortus.

Culture Results

Nonvaccinate saline controls

RB51 vaccinates

11/15 for strain 2308 (80%)

3/9 for strain 2308 (30%)

Conclusions

RB51 administered orally to female elk provided partial protection against virulent B. abortus challenge compared to non-vaccinated controls as demonstrated by abortion and colonization. Protection was measured by the number of aborted fetuses or calves which died after birth and the colonization of the fetuses and females with the challenge strain. In the non-vaccinated control group, 100% of the calves died and 80% of the animals were culture positive for strain 2308. However, in the RB51 vaccinated group, 55% of the calves died and 45% of them were born healthy; and only 30% of the animals were culture positive for strain 2308. Based on these findings, RB51 should be further investigated as a possible oral vaccine in elk which frequent feedgrounds.

Acknowledgments

We wish to acknowledge the following for assistance in this project:

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

Louisiana State University (Sue Hagius and Joel Walker), ND Governor's Office; Windcave National Park, Ross Rice; ND Elk Breeders Association; North American Elk Breeders Association; ND State Veterinarians, Robert Velure and William Rotenberger; Mitchell Charles; John Murphy; SD State Veterinarian, Daryl Thorpe; SD AVIC, Lynn Tesar; WY Game and Fish (Drs. Tom Thorne, Terry Kreeger, Beth Williams and Walt Cook);

USDA/APHIS/VS (Drs. John Kopec and Mike Gilsdorf) and USDA/APHIS/VS Cooperative Agreements.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

ISSUES IN VACCINATION FOR BRUCELLOSIS

Fred M. Enright, Department of Veterinary Science, Louisiana State University Agricultural Center, Baton Rouge, LA 70803

WHAT MIGHT THE USE OF AN EFFICACIOUS VACCINE ACCOMPLISH?

WHAT WILL THE USE OF SUCH A VACCINE FAIL TO ACCOMPLISH?

Factors which modify the effectiveness of any brucellosis vaccine.

Host susceptibility

  • in herd differences based on genetics, sex, age

  • inverse relationship between degree of susceptibility and level of protection afforded by the vaccine

Duration of immunity

  • necessity for repeated vaccination

Level of exposure

  • most clearly demonstrated in the level of protection demonstrated by experimental challenge v. field challenge

What differences may be expected in field exposure under the following circumstances?

  • a dry lot packed with pregnant dairy cows

  • a 100-acre pasture containing 50 pregnant beef cows

A brief history of brucellosis vaccines and the development of Strain 19 vaccine

  • initially sought a vaccine to prevent late-term abortions in cattle

  • use of virulent B. abortus isolates to infect (vaccinate) heifers prior to breeding

    1. partial protection against abortions

    2. cattle infected with these vaccines shed B. abortus in milk and could infect other cattle

  • 1923-1924: Buck discovers Strain 19

  • by the early 1930s had demonstrated the effectiveness of S-19 in adults and calves

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×
  • 1940s: S-19 became the official vaccine in the National Brucellosis Program

TRADE-OFFS WITH S-19

  • Initially used to vaccinate all females regardless of age

    Problems: vaccine infections; vaccine titers

  • 1950s and 1960s limited use to heifer calves

  • by 1970s Dr. Nicoletti rediscovered adult vaccination

  • 1990s replacement of S-19 with RB51

The development of S-19 forced scientists to develop and/or modify how S-19 was used.

  • lower doses for adults

  • lower vaccination age of calves

  • different routes of vaccination

What did we learn?

  • very young cattle not protected as well as adults

  • still had vaccine titer problems

  • oral vaccination/conjunctival vaccination yield better protection

Why live vaccines?

Vaccination of elk and bison with S19

  • Elk reproduction

    • females become sexually mature by 2 years of age

    • males not actively involved in breeding until 3 years of age

    • breeding until 3 years of age

    • breeding season mid-September to mid-October

    • gestation 8.5 to 9 months

    • calving mid-May to mid-June

Brucellosis in elk

  • brucellosis in elk causes abortions/premature deliveries

  • 50-70% of female elk that become infected with B. abortus lose their first calf following infection

  • like cattle, most abortions occur during the last 1/3 of gestation

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×
  • retention of placentas and other forms of infertility associated with brucellosis in cattle do not occur in infected elk

  • transmission of brucellosis from infected elk to susceptible cattle occurs

  • experimental animals were closely confined and transmission was associated with delivery of infected elk calves

  • transmission occurs during late winter and early spring

"Brucellosis transmission from elk to cattle is extremely unlikely to occur at any other time or circumstance, including normal calving in traditional elk calving ranges (Thorne et al., 1991)."

  • elk normally seek remote secluded areas to calve

  • elk have been incriminated epidemiologically in spreading brucellosis to at least 4 cattle herds adjacent to the GYA (circumstantial)

Response of elk to S-19

  • 27% of S-19 vaccinated elk aborted S-19 infected fetuses

  • 38% of 66 vaccinated female elk vs. 69% of 35 nonvaccinated lost their calves following challenge with virulent B. abortus

S-19 Vaccination of Elk

Recovery of 2308 at necropsy* of elk challenged 6 to 1 weeks post hand vaccination

 

Number of animals

Number from which 2308 recovered

Vaccinates

11

6 (55%)

Controls

8

7 (88%)

Vaccinates

6

0 (0%)

Controls

2

2 (100%)

Vaccinates

12

7 (58%)

Controls

6

4 (66%)

* Necropsies performed 24 to 55 weeks post challenge.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

Cumulative results of hand vaccinated elk challenged 6 to 10 weeks post vaccination

 

Number of animals

Number from which 2308 recovered

Vaccinates

29

13 (45%)

Controls

16

13 (81%)

Recovery of 2308 at necropsy from hand vaccinated elk challenged 1 to 2 years post vaccination

 

Number of animals

Number from which 2308 recovered

Vaccinates

16

7 (44%)

Controls

14

5 (36%)

Vaccinates

8

3 (37%)

Controls

4

2 (50%)

* Necropsies performed 27 to 37 weeks post challenge.

Cumulative results of hand vaccinated elk challenged 1 to 2 years post vaccination

 

Number of animals

Number from which 2308 recovered

Vaccinates

24

10 (42%)

Controls

18

7 (38%)

These studies suggest that elk are more susceptible to B. abortus than cattle.

S-19 vaccination of adult elk is more effective than 2-19 vaccination of elk calves.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

Experimental Infection of Bison in Their Second Trimester of Pregnancy

  • Infected bison (10/12) aborted 47.5 days post challenge

  • Infected cattle (11/12) aborted 69.2 days post challenge

This difference is most likely due to the levels of bacteremia. More B. abortus in the bloodstream at an earlier interval following challenge in bison than in cattle.

S-19 vaccination of adult bison (90-120 days of gestation)

  • hand injection with 5.3 × 108 cfu S-19 aborted 29/48 (60%)

  • ballistic injection with 1.7 × 109 cfu S-19 aborted 34/44 (77%); nonvaccinated bison aborted 6/46 (15%)

  • 63% (30-48) of the hand injected bison demonstrated seroreactivity 12 months post vaccination

  • 80% (36/45) of the ballistically injected bison demonstrated seroreactivity 12 months post vaccination.

  • 2% (14/837) nonvaccinated contact bison sharing winter pastures with the vaccinates seroconverted

  • 1 vaccinated cow was chronically infected with S-19 and aborted a second fetus 13 months post vaccination

These vaccinated adult bison were challenged with 1.0 × 107 cfu of B. abortus strain 2308 approximately 13 months post vaccination.

Protection against abortion

Hand vaccinated

57% (16/28)

Ballistically vaccinated

79% (19/24)

Controls

4% (1/27)

Protection against infection

Hand vaccinated

30% (9/30)

Ballistically vaccinated

44% (12/27)

Controls

0% (0/30)

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

Vaccination of Bison Calves with S-19

  • 8-10 months old bison calves were vaccinated

  • these calves vaccinated by hand or ballistically or given saline only; were challenged with 1.0 × 107 cfu of B. abortus strain 2308 as pregnant adults about 2 years post vaccination

Results

  • 5% (5/96) S-19 vaccinated calves remained seropositive for 24 months

  • S-19 vaccinated bison calves were not protected against either abortion or infection

Conclusions

  1. Bison are very susceptible to B. abortus infection

  • an attenuated vaccine strain resulted in prolonged infection, persistent seroreactivity, abortions, and exposure of contact controls

  1. S-19 vaccinated adult bison demonstrated significant resistance to infection and protection from abortions

  2. S-19 vaccination of 8-10 month old bison calves failed to increase their resistance to infection or protect against abortion

While not satisfactory, the elk and bison S-19 studies suggest that:

  1. Adults are most effectively protected against infection.

  2. This partial protection against infection translates into increased protection from abortions–and may result in decreased exposure to other susceptible animals.

  3. Calfhood vaccination of bison is not effective.

  4. Calfhood vaccination of elk may not be effective–was the diminished protection due to the extended intervals (1-2 years) between vaccination and challenge or due to the inability to immunize elk calves?

With reference to any vaccine for brucellosis in wildlife–S-19, RB41, and those not discovered:

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

Do the constraints related to vaccine safety imposed by its use in commercial livestock also apply to its use in wildlife?

Specifically, are vaccine-induced abortions ever acceptable in wildlife populations?

Perhaps the number of abortions induced in pregnant elk and bison by S-19 are excessive.

We have, however, already been shown that bison abortions due to S-19 were adequate to expose and perhaps even immunize 2% of a nonvaccinated population of contact bison.

I now feel that an acceptable vaccine which occasionally causes abortions in vaccinated animals can serve to enhance immunity in nonvaccinated members of the herd.

Finally, I would like to answer the two questions stated in the beginning of my presentation:

  1. What will an effective vaccine accomplish?

  • It will limit the spread of virulent B. abortus in the population (herd)

  • It will enhance resistance to infection within the population (herd)

  • Thus, with time, numbers of new infections are reduced

  1. What will such a vaccine fail to accomplish?

  • It will not eliminate the disease within the herd

I will end by quoting Dr. Paul Nicoletti:

''The control of brucellosis depends largely on two of the main principles of disease management: prevention of exposure to susceptible animals, and increasing resistance of the population through vaccination. The best results are achieved through a combination of these, but vaccination, especially in large cattle populations, is far more effective."

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

Oral RB51 Vaccination of Elk: Tissue Colonization and Immune Response

A Study Conducted by the Wyoming Game and Fish Department in Collaboration with Louisiana State University and Virginia Polytechnic Institute

Methods

  • 40 elk captured from the National Elk Refuge; card tested negative

  • Transported to Sybille Wildlife Research Unit; re-tested several times

  • 34 elk vaccinated

  • 15 adults (10.5)

  • 15 yearlings (7.8)

  • 4 calves (2.2)

  • 6 controls (6.0)

  • Elk vaccinated orally with 6 × 109 cfu Brucella abortus, strain RB51 every other day for three vaccinations

  • Mucosa excoriated prior to vaccination

  • Two elk necropsied every other week post vaccination for blood and tissue culture and histopathology

  • Remaining elk bled every other week for serology and hemoculture

Results (preliminary)

  • All elk remained serologically negative for field strain Brucella abortus (i.e., no vaccine crossreaction with standard tests)

  • Last positive hemoculture, 54 days post vaccination

  • Last positive tissue culture, 68 days post vaccination (3/43 tissues positive)

  • Last detectable RB51 titer (ELISA 1:50), 10 weeks post vaccination

Conclusions (preliminary)

  • Oral vaccination of elk resulted in tissue colonization

  • Oral vaccination resulted in an immune response

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

RB51 Vaccination of Elk: Safety and Efficacy

A Study Conducted by the Wyoming Game and Fish Department in Collaboration with University of Wyoming, Louisiana State University and Virginia Polytechnic Institute

Methods

  • 45 female elk calves captured from the National Elk Refuge in 1995; card tested negative

  • Transported to Sybille Wildlife Research Unit; re-tested several times

  • Elk vaccinated in May 1995

  • hand: 1 × 109 cfu (n=16)

  • biobullet: 1 × 108 (n-16)

  • 13 controls

  • Elk were bred in fall 1996

  • All elk challenged in March 1997 with 1 × 107 cfu Brucella abortus strain 2308 intraconjuctivally

  • Elk observed daily thereafter for abortion

  • Elk necropsied after delivery/abortion

Results (preliminary)

  • Hand: 14/16 aborted (88%)

  • Biobullet: 12/16 aborted (75%)

  • Controls 13/13 aborted (100%)

Conclusions (preliminary)

  • There was no difference in abortion rate between vaccinates and controls (P ≥ 0.10)

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

BRUCELLOSIS IN THE GREATER YELLOWSTONE AREA: WHAT IS THE PROBLEM?

Paul Nicoletti, D.V.M., M.S., Professor, College of Veterinary Medicine, University of Florida, Gainesville, FL 32611-0880

One would have to be the equivalent of Rip Van Winkle to not know of some of the controversies surrounding this subject. Passions and opinions are many and intense. Numerous articles have been published and the broadcast media has been busy and seen by millions.

The conflict between the natural wildlife and those who wish to protect them and those of private interests, especially of ranchers, is a classic example of a problem in the United States. Whether it is overpopulation of raccoons in Pinellas County in Florida, or too many deer on Long Island, or brucellosis in Yellowstone Park bison, resolution of these conflicts is difficult. There is a direct confrontation between the concepts of doing whatever is necessary to eradicate a disease and to leave natural forces to function.

My credibility to address this group is based upon a near lifetime career of specialization in brucellosis, an episode near Gainesville which also involved bison brucellosis, serving as an expert witness in trials and hearings and distance. Everyone knows that the further one is form the problem, the more expertise can be claimed.

Brucellosis is characterized in natural animal hosts by abortion, retained placenta, and pathologic lesions in males. The susceptibility to infection and severity among wildlife hosts have been studied and some results are conflicting. It is quite clear that under natural conditions, brucellosis in bison is of little consequence in fecundity. Bison are not shaggy cows and their behavior, physiology, and responses to infectious agents may be unique.

Control and hopeful eradication of brucellosis in domesticated livestock are based upon quarantine, vaccination, and slaughter of seropositive animals. Clearly, these methods are far more difficult to apply in wildlife hosts.

The USDA and states depend upon selected surveillance systems to identify possible infected herds of cattle as part of the national brucellosis eradication program. In beef cattle, this surveillance largely relies upon blood samples which are collected at slaughter. This system provides data upon which classification of states depends. Classification of states as

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

"free" of cattle brucellosis allows more freedom in cattle commerce. The most affected states by the wildlife brucellosis issue of Wyoming, Idaho, and Montana are classified free. The threat of reducing this state status if one or more cattle herds become infected causes terror among ranchers. Some states have threatened boycotts of cattle movements. These scenarios are harsh and without scientific merit. Surely, disease control officials can have more wisdom in handling disease. The surveillance system which is used to detect a problem should also be evidence of the lack of a problem and there is still no evidence that bison of the YNP have been responsible for any transmission of brucellosis to area cattle. There is anecdotal evidence of transmission from wildlife to cattle in the National Elk Refuge feeding grounds area to a few herds. The elk are known to have a high prevalence of clinical infection and seropositivity.

It is important to understand the differences between seropositivity, infection, and disease. Many surveys among free-living bison have found a rather high seroprevalence of brucellosis. When specimens are examined bateriologically, only about 20 percent of those with antibodies are culture positive. Further, it is rare to isolate the bacteria from female reproductive organs. It is interesting that the highest percentage of culture positive bison is among young animals and bulls.

The migration and subsequent slaughter of over 1000 bison during the severe winter of 1996-1997 caused enormous outcries among many persons and groups. A further 600 or so starved within the park boundaries. The migration is apparently assisted by snowmobile paths and it has often been suggested that snowmobiles be banned from the park or use be restricted.

It seems epidemiologically correct to suggest that the elk feeding grounds in Wyoming present a far greater risk in disease prevalence and management than the bison of YNP. A project to vaccinate some of the elk with biobullets of stain 19 has been in progress for several years with reduction in seroprevalence among the elk.

Some Observations

  1. It is hyperbole to suggest that if brucellosis cannot be eradicated from the GYA, that efforts to eradicate the disease from domestic animals have been wasted. Many believe that measures which would be necessary to eliminate brucellosis from the wild animals would eliminate the hosts.

  2. The work eradication and the state classification system must be modified to conform with reality.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×
  1. The excessive attention to the bison and much less attention to the elk are driven by attitudes of ranchers towards the two species. Clearly, there must be some attitudinal changes or ranchers face possible eventual loss of privileges of using public lands for cattle grazing.

  2. There is much agreement that the bison population within YNP needs more management. It remains very questionable if this should include possible vaccination to prevent brucellosis. There is no satisfactory vaccine, delivery system or evidence of a disease problem.

During testimony in the rather famous Parker lawsuit, Judge Bremmer asked "Dr. Nicoletti, what would you do with the problem of brucellosis in the Greater Yellowstone Area?" My reply was "Your honor, I don't know." While I have several observations and opinions, I feel that my answer puts me among a rather large company of others.

I appreciate the invitation to attend this conference and to present this paper. I trust that meetings such as this will educate and perhaps eventually, lead to some compromises and solutions to some very complex issues.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

Safety and Efficacy of Existing Vaccines to Prevent Brucellosis in Bison

Steven Olsen, DVM, Ph.D., United States Department of Agriculture, Agricultural Research Service, National Animal Disease Center, Ames, IA 50010

Protection and lasting immunity against brucellosis is achieved with vaccines containing live bacteria which stimulate a strong cell-mediated immune response. Factors enhancing cell-mediated immunity following administration of live vaccines may include prolonged antigenic stimulation due to proliferation of the vaccine strain within the host and internal antigen processing with more efficient presentation with major histocompatibility antigens (Class I) associated with cellular immune responses. Mouse models of brucellosis indicate that antibodies may have a minor role in short-term protection.1,2 However, studies in cattle have demonstrated a poor correlation between the vigor of the humoral response and protection.3 This is supported by data from cattle experiments in which vaccinated animals which were seronegative prior to midgestational challenge with a virulent Brucella abortus strain were protected against infection and abortion at a time of maximum susceptibility. Additionally, it is customary for animals which abort to have very high titers against brucellosis despite having failed to mount an effective immune response which prevented localization in placental and fetal tissues.

An ideal vaccine against brucellosis would persist long enough to induce good immunity without persisting into adulthood, would not cause clinical illness, and would not induce serologic responses which interfere with detection of animals infected with virulent field strains of B. abortus. Typically, vaccines against brucellosis are more efficacious in preventing abortions than preventing infection. Vaccination of wildlife with live vaccines would also have to consider potential detrimental effects on nontarget species, such as predators, which may inadvertently be infected with the vaccine strain.

Studies evaluating the safety and efficacy of brucellosis vaccines in bison are limited. When B. abortus strain 19 was administered by hand (1.7 × 109 colony-forming units (CFU)) or ballistic methods (7.7 × 109 CFU) to bison heifer calves, 5% of vaccinated calves had titers on brucellosis

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
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serologic tests at 2 years of age.4 Following challenge with 1 × 107 CFU of virulent B. abortus strain 2308 during pregnancy, calves vaccinated with strain 19 averaged 25% abortions as compared to 30% abortions in heifers vaccinated with saline (controls). Only 9% of heifers calfhood vaccinated with strain 19 were protected against infection as compared to 17% of nonvaccinated controls. No statistical differences in abortion or infection rates were detected between bison calfhood vaccinated with strain 19 and nonvaccinated controls.

A second study evaluating strain 19 as a vaccine for adult bison indicated that a high percentage (58%) of pregnant animals aborted following vaccination.5 When challenged with B. abortus strain 2308 during following pregnancy (13 months after vaccination), the percentage of abortions was less in strain 19-vaccinated bison as compared to nonvaccinated bison (33% versus 96%, respectively). In a similar manner, protection against infection was greater in strain 19 vaccinated bison as compared to nonvaccinates (39% versus 0%, respectively). In addition to its abortogenic effects, the strain 19 vaccine also induced persistent serologic titers on brucellosis surveillance tests and chronic infections in bison vaccinated as adults.

Research at the National Animal Disease Center has identified a new vaccine for cattle, B. abortus strain RB51, that is efficacious in preventing abortion and infection.6,7 This vaccine does not induce antibody responses which cause positive responses on brucellosis surveillance tests8,9 and therefore does not impair the identification of Brucella-infected cattle under field conditions. Research projects to evaluate strain RB51 as a vaccine for bison have been initiated at our facility.

A preliminary study to evaluate strain RB51 vaccination (1010 CFU) of bison indicated that the vaccine is clinically safe in bison calves and does not induce positive responses on brucellosis surveillance tests.10 Antibody responses against the vaccine strain were detected using a dot-blot test which has been demonstrated to have a high sensitivity and specificity in cattle.11 Adverse clinical signs were not detected following vaccination of bison with strain RB51. The vaccine strain was still present at 16 weeks after vaccination in bison whereas cattle typically clear strain RB51 from the draining lymph by 12 to 14 weeks. These bison were raised to maturity and pasture bred. Data obtained following challenge at midgestation with 1 × 107 CFU of B. abortus strain 2308 suggested that strain RB51 induces some protection in bison. However, as nonvaccinated bison were not included in the challenge portion of the study, conclusions cannot be made on the efficacy of strain RB51 in bison without additional studies.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

Additional studies have been completed evaluating calfhood vaccination of bison with 1010 CFU of strain RB51. These studies have provided further evidence that strain RB51 persists longer in bison when compared to cattle but does not appear to cause adverse clinical signs. Data from these studies suggests that strain RB51 localizes in lymphatic tissues and induces cell-mediated immune responses. Data from biosafety experiments have indicated that the strain RB51 vaccine is not shed from bison following vaccination.

The strain RB51 vaccine may have similar problems in adult bison as the strain 19 vaccine. When administered to pregnant bison at a 109 CFU dosage, strain RB51 appears to induced abortion in some animals.12 This dosage is safe in pregnant cattle.13 Ongoing studies will determine if adverse clinical or biosafety effects may limit the use of strain RB51 in adult bison bulls.

At the present time, the strain RB51 vaccine is the most likely candidate for use to prevent brucellosis in bison. Continued research efforts will be required to verify the efficacy of strain RB51 to prevent brucellosis in bison. Addition research will also be required to develop delivery methods and guidelines for the use of strain RB51 in management programs to reduce or eliminate Brucella infections in bison.

References

1. Winter, A.J., Duncan, J.R., Santisteban, C.G., Douglas, J.T., Adams, L.G. Capacity of passively administered antibody to prevent establishment of Brucella abortus infection in mice. Infect. Immun. 57: 3438-3444, 1989.

2. Cloeckaert, A., Jacques, I., Bosseray, N., Limet, J.N., Bowden, R., Dubray, G., Plommet, M. Protection conferred on mice by monoclonal antibodies directed against outer-membrane-protein antigens of Brucella. J. Med. Microbiol. 34: 175-180, 1991.

3. Nicoletti, P. Vaccination. In: Animal Brucellosis. K. Nielsen and J.R. Duncan eds. 1990. pp 283-300.

4. Davis, D.S., Templeton, J.W., Ficht, T.A., Huber, J.D., Angus, R.D., Adams, L.G. Brucella abortus in bison. II. Evaluation of strain 19 vaccination of pregnant cows. J Wildlife Dis 1991; 27:258-264.

5. Davis, D.S. Summary of Bison/Brucellosis Research conducted at Texas A&M University 1985-1993. Proceedings of North American Public Bison Herds Symposium. July 27-29, 1993, Lacrosse, WI, pp 347-161.

6. Cheville, N.F., Stevens, M.G., Jensen, A.E., Tatum, F.M., Halling, S.M. Immune responses and protection against infection and abortion in cattle experimentally vaccinated with mutant strains of Brucella abortus.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
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Am. J. Vet. Res. 54: 1591-1597, 1993.

7. Cheville, N.F., Olsen, S.C., Jensen, A.E., Stevens, M.G., Palmer, M.V. Effects of age at vaccination on efficacy of Brucella abortus strain RB51 to protect cattle against brucellosis. Am. J. Vet Res 57: 1153-1156, 1996.

8. Stevens, M.G., Hennager, S.G., Olsen, S.C., Cheville, N.F.. Serologic responses in diagnostic tests for brucellosis in cattle vaccinated with Brucella abortus strain 19 or RB51. J. Clin. Microbiol. 32: 1065-1066, 1994.

9. Olsen, S.C., Evans, D., Hennager, S.G., Cheville, N.F., Stevens, M.G. Serologic Responses of Calfhood-Vaccinated Cattle to Brucella abortus strain RB51. J. Vet. Diagn. Invest. 8: 451-454, 1996.

10. Olsen, S.C., Cheville, N.F., Kunkle, R.A., Palmer, M.V., Jensen, A.E. Bacterial survival, lymph node changes, and immunologic responses of bison (Bison bison) vaccinated with Brucella abortus strain RB51. J. Wildlife Dis. 33: 146-151, 1997.

11. Olsen, S. C., Stevens, M.G., Cheville, N.F., Schurig, G. Experimental use of a dot-blot assay to measure serologic responses of cattle vaccinated with Brucella abortus strain RB51. J. Vet. Diagn. Invest. (In Press)

12. Palmer, M.V., Olsen, S.C., Jensen, A.E., Gilsdorf, M.J., Philo, L.M., Clarke, P.R., Cheville, N.F. Abortion and placentitis in pregnant bison (Bison bison) induced by the vaccine candidate Brucella abortus strain RB51. Am. J. Vet. Res. 57: 1604-1607, 1996.

13. Palmer, M.V., Olsen, S.C., Cheville, N.F. Safety and immunogenicity of Brucella abortus strain RB51 vaccine in pregnant cattle. Am. J. Vet. Res. 58: 472-477, 1997.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

Lesions and Sites of Tissue Localization of Brucella abortus in Female Bison from Yellowstone National Park: Preliminary Results

Jack C. Rhyan,1 Keith Aune,2 Thomas J. Roffe,3 Thomas Gidlewski,1 Darla R. Ewalt,1 and Michael Philo4

1  

U.S. Department of Agriculture, Animal and Plant Health Inspection Service, Veterinary Services, National Veterinary Services Laboratories, P.O. Box 844, Ames, IA 50010;

2  

Montana Department of Fish Wildlife and Parks, Research and Technical Services Bureau, Montana State University Campus, Bozeman, MT 59717;

3  

U.S. Department of the Interior, U.S. Geological Survey, Biological Resources Division, National Wildlife Health Center, Bozeman Station, Montana State University Campus, Bozeman, MT 59717;

4  

U.S Department of Agriculture, Animal and Plant Health Inspection Service, Veterinary Services, Western Region, 9439 Owl Way, Bozeman, MT 59715

Introduction

Brucella abortus produces abortions in cattle, bison (Davis et al., 1990; Rhyan et al., 1994; Williams et al., 1993) and elk (Thorne et al., 1978). Metritis and retained placentas have also been associated with the infection in cattle and bison (Corner and Connell, 1958; Williams et al., 1993). Seminal vesiculitis, orchitis, and epididymitis have been observed with B. abortus infection in male cattle and bison (Corner and Connell, 1958; Creech, 1930; Tunnicliff and Marsh, 1935; Williams et al., 1993; Rhyan et al., 1997). In a recent study, B. abortus was isolated from two or more tissues from six of seven young bison bulls that had recently seroconverted (Rhyan et al., 1997). The purpose of this study was to determine the most frequent sites of tissue localization of B. abortus in female bison from Yellowstone National Park (YNP).

Materials and Methods

Between February 1995 and January 1997, specimens were collected from 26 seropositive adult female bison. Twenty-five of the animals were killed after leaving YNP, and one animal was killed by YNP personnel because it had a retained placenta and was in close proximity to the northern border of YNP. The cow had recently aborted as evidenced by

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
×

the early date (March of 1995) and the lack of mammary gland development. No fetus or calf was found. Additionally, specimens were collected from a term fetus and placenta that were found near Gardiner, Montana, in April of 1996. Tissue specimens were collected from all animals for culture in accordance with the recommendations published by the Greater Yellowstone Interagency Brucellosis Committee (GYIBC, 1996). Additionally, portions of the uterus and placenta from the cow killed in YNP and portions of lung and placenta from the fetus found near Gardiner, Montana, were fixed in 10 percent neutral buffered formalin and routinely processed for histopathologic examination. Selected tissues were also stained using a previously described immunohistochemical technique (Rhyan et al., 1997) that employs a polyclonal antibody developed against B. abortus (Palmer et al., 1996).

Tissues were cultured using a previously described technique (Rhyan et al., 1997) in which each piece of tissue was individually minced, macerated with an equal volume of PBS in a stomacher, and further processed in a glass tissue grinder. The resulting slurry was then poured in aliquots onto the following media: tryptose agar with five percent bovine serum and antibodies (TSA), TSA with ethyl violet, Ewalts medium, and Farrel's medium. Plates were incubated with added CO2 at 37 C for 2 weeks. Cultures were identified and biotyped using the techniques of Alton et al. 1988).

Sero status of the animals was initially determined using the card test and was confirmed with the following tests: standard plate, standard tube, rivanol, complement fixation (CF), buffered acidified plate antigen (BAPA), and particle concentrate fluorescence immunoassay (PCFIA). All animals chosen for this study were positive on multiple serologic tests.

Results

At present, cultures have been completed on 16 of the adult bison and on the fetus. Brucella abortus was isolated from tissues of 7 of the 16 animals. The most common culture positive tissues were the supramammary lymph nodes (7/7), retropharyngeal lymph nodes (5/7), and iliac lymph nodes (5/7). Brucella abortus was isolated from 15 specimens including the placenta and feces from the bison with the retained placenta. The organism was also isolated from 15 sites cultured from the term fetus and placenta found near Gardiner. Histologically, lesions from both placentas and the fetus consisted of necropurulent placentitis and mild pleocellular bronchointerstitial pneumonia. Immunohistochemical staining

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
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revealed large numbers of brucellae in placental trophoblasts and in phagocytes present in placental and uterine exudate. Fetal lung also contained brucellar antigen in exudate in airways.

Discussion:

The preliminary results of this study suggest that the supramammary, iliac, and retropharyngeal lymph nodes are the most frequent sites of tissue localization of B. abortus in female bison from YNP. Additionally, the results from the cow that had recently aborted suggest widespread infection in that animal at the time of abortion. The presence of B. abortus in the feces probably resulted from ingestion of portions of the infected placenta and/or licking off the infected fetus. Similar findings in cattle have been reported. The placentitis and fetal pneumonia with large numbers of organisms in placental trophoblasts are consistent with lesions produced by B. abortus in cattle (Payne, 1959), goats (Meador et al., 1986), and captive bison (Davis et al, 1990).

REFERENCES

Alton GG, Jones LM, Angus RD, Verger JM: 1988, Techniques for the brucellosis laboratory. Institut National de la Recherche Agronomique, Paris, France, 190 pp.


Corner AH, Connell R: 1958, Brucellosis in bison, elk, and moose in Elk Island National Park, Alberta, Canada. Can J Comp Med 22:9-20.

Creech GT: 1930, Brucella abortus infection in a male bison. North Am Vet 11:35-36.


Davis DS, Templeton JW, Ficht TA, et al.: 1990, Brucella abortus in captive bison I. Serology, bacteriology, pathogenesis, and transmission to cattle. J Wildlife Dis 26:360-371.


Meador VP, Tabatabai LB, Hagemoser WA, Deyoe BL: 1986, Identification of Brucella abortus in formalin-fixed, paraffin-embedded tissues of cows, goats, and mice with an avidin-biotin-peroxidase complex immunoenzymatic staining technique. Am J Vet Res 47:2147-2150.


Palmer MV, Cheville NF, Tatum FM: 1996, Morphometric and histopathologic analysis of lymphoid depletion in murine spleens following infection with Brucella abortus strains 2308, RB51, or an htrA deletion mutant. Vet Pathol 33:282-289.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
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Payne JM: 1959, Pathogenesis of experimental brucellosis in the pregnant cow. J. Pathol Bacteriol 78:447-459.


Rhyan JC, Quinn WJ, Stackhouse LL, et al.: 1994, Abortion caused by Brucella abortus biovar 1 in a free-ranging bison (Bison bison) from Yellowstone National Park. J Wildlife Dis 30:445-446.

Rhyan, JC, Holland SD, Gidlewski T, et al.: 1997, Seminal vesiculitis and orchitis caused by Brucella abortus biovar 1 in young bison bulls from South Dakota. J Vet Diagn Invest 9: IN PRESS.


Thorne ET, Morton JK, Blunt FM, Dawson HA: 1978, Brucellosis in elk. II. Clinical effects and means of transmission as determined through artificial infections. J Wildl Dis 14:280-291.

Tunnicliff EA, Marsh H: 1935, Bang's disease in bison and elk in Yellowstone National Park and on the National Bison Range. J Am Vet Med Assoc 86:745-752.


Williams ES, Thorne ET, Anderson SL, Herriges JD Jr: 1993, Brucellosis in free-ranging bison (Bison bison) from Teton County, Wyoming. J Wildlife Dis 29:118-122.

Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
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Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
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Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
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Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
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Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
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Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
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Suggested Citation:"Appendix B: Meeting Agendas and Presentations." National Research Council. 1998. Brucellosis in the Greater Yellowstone Area. Washington, DC: The National Academies Press. doi: 10.17226/5957.
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Next: Appendix C: Other Diseases in GYA Wildlife »
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Brucellosis, a bacterial disease, was first noted in the Greater Yellowstone Area in 1917 and has been a chronic presence there since then. This book reviews existing scientific knowledge regarding brucellosis transmission among wildlife, particularly bison, elk, and cattle, in the Greater Yellowstone Area. It examines the mechanisms of transmission, risk of infection, and vaccination strategies. The book also assesses the actual infection rate among bison and elk and describes what is known about the prevalence of Brucella abortus among other wildlife.

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