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National Institutes of Health(U.S.). Division of R.

Report of program activities : National Institutes of Health. Division of Research Services (Volume 1965) online

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"Safe Handling of Laboratory Animals," produced in cooperation with
Plant Safety Branch. A camera crew from the Communicable Disease
Center spent several days in LAB facilities shooting film for the
movie. As technical advisors, personnel in the Branch assisted with
the script and in scenes. Editing was done by NIH personnel at CDC.
The film has been completed and arrangements for loan may be made
through the PHS Audio-Visual Facility at CDC, Atlanta, Georgia.

D. Problems

A major problem in special services is lack of adequate facilities
to conduct radioactive isotope experiments.

LAB animal surgical facilities are no longer considered adequate to
meet all NIH program needs. The more sophisticated surgical procedures
now used on animals require specialized equipment and facilities.
Either LAB must supply these facilities or services or the Institutes
will turn to establishing and operating their own.

E. Program Plans

The programs will continue as outlined above but will be modified
in response to demands by the Institute for specific services.

Efforts will continue in the design of special purpose buildings
as requested, e.g. a radioactive isotope utilization building and a
surgical suite appendix to Building 28. Planning for the NIH Animal
Center will continue.

PUBLICATIONS AND PATENTS

Publications

Allen, A. M. , Ganaway, J. R. , Moore, T. D. , and Kinard, R.F. :
Tyzzer's disease syndrome in laboratory rabbits. Amer. J. Path .
46: 859-882, May 1965.

Byrne, R. J.: NIH Animal Center. Maryland Veterinarian . 6: 8-9,
May 1964.

Palmer, A. E. : Restraint of monkeys. Maryland Veterinarian .
7: 4-8, April 1965

Presentations

Byrne, R. J.: Clinical, Laboratory, and Epidemiological Aspects
of Arthopod-borne Virus Encephalitis of Horses. New Jersey State
Veterinary Association, March 1965.



4-19



Serial No. 4JL

1. Laboratory Aids Branch

2. Animal Production Section

3. Bethesda



PHS-NIH

Individual Project Report

July 1, 1964 through June 30, 1965



Part A



Project Title: EFFECT OF VARIOUS DAY-LENGTHS ON THE PRODUCTIVITY OF
MICE

Principal Investigator: Cobert D. LeMunyan, Ed.D.

Other Investigators: William R. Clark, Jr., RFPB

Cooperating Unit: Research Facilities and Planning Branch

Man Years (computed for the 12-month period)
Total: 1.1 (APS only)
Professional: .1
Other: 1.0

Project Description:

Objective: Animal breeders have long believed that

reproductive cycles in rodents are influenced
significantly by the length of daylight. The
very small amount of experimental work which has been done in this
field has not concerned itself with laboratory rodents. However, the
empirical evidence is strong enough to justify a controlled experiment
to determine whether the length of day(light) has any influence over
the reproductive performance of breeding mice.

Methods Employed: Six cubicles were constructed within a

room, each capable of housing fifty pairs
of breeding mice. Each cubicle has indivi-
dual temperature and lighting control. For the purposes of this
experiment, the temperature and relative humidity will be held
constant. The length of day is the variable factor; a number of
cubicles will be set at different day lengths and one control cubicle
will follow the natural lighting cycle. For this phase of the project,
light intensity will be held constant by a fluorescent source.

Major Findings: The pilot phase for testing and evaluation

of equipment, methods, and animal heat loads
prior to the actual experimental run was
concluded in August 1964. The equipment was checked out, defective
materials and equipment were replaced, and the caretaker technician



4-20



Part A (Cont'd)

was trained for the final experimental course. The actual experimental
run started in late August 1964 and will be concluded in August 1965.



Significance:



The findings of this research will be
utilized in the planning of the rodent and
rabbit buildings at the NIH Animal Center
in Poolesville, Maryland.



Proposed Course The study of day-length will take one year
of Project: to complete all phases of the lighting cycle.

Following this, it is proposed that light
intensities and sources be studied and the findings be incorporated
in the design of the buildings at the NIH Animal Center.



Part B included



Yes



No X



4-21



Serial No. 4_;_2

1. Laboratory Aids Branch

2. Germfree Animal Production Unit

3. Bethesda



PHS-NIH

Individual Project Report

July 1, 1964 through June 30, 1965



Part A



Project Title: ESTABLISHING AND PRODUCING GERMFREE HAMSTERS

Principal Investigators: Carl E. Miller, D.V.M.

D. B. Fitzgerald, B.S., NIDR

Other Investigators: Howard J. Bohner, B.A.

R. J. Fitzgerald, Ph.D., NIDR

Cooperating Units: Animal Production Section, LAB
Gnotobiotic Section, LM, NIDR

Man Years (computed for the 12-month period)
Total: .2 (GAPU only)
Professional: .1
Other: .1

Project Description:

Objective: To establish hamsters in the germfree

environment and maintain them by production
in the germfree isolators.

Methods Employed: A cesarean delivery of the infants is

performed, followed by an attempt to raise
them either by hand feeding or by foster
nursing on germfree mothers of other species.

Major Findings: The very small size and immaturity of the
infant hamster (16-day gestation period)
makes replacing its mother very difficult.
A method of obtaining hamster milk and sterilizing it without heat
has been developed. A chemical analysis of the milk has been
obtained through a contract with the University of Minnesota. One
newborn hamster was raised outside of an isolator, but this has not
been accomplished inside the isolator as yet.

Significance: The National Institute of Dental Research

considers the germfree hamster the single
most important animal for their program.
The importance of hamsters in other research has increased a great



4-22



Part A (Cont'd)

deal with reports of the hamsters' susceptibility to adenoviruses
and that SV-40 produces tumors in their cheek pouches. The germ-
free hamster will eliminate any complicating infections in this
type of experimental work.

Proposed Course Continuation,
of Project:

Part B included Yes No X



4-23



Serial No. 4^^

1. Laboratory Aids Branch

2. Animal Hospital Section

3. Bethesda



PHS-NIH

Individual Project Report

July I, 1964 through June 30, 1965



Part A



Project Title: BREEDING OF THE STUMP-TAILED MACAQUE MONKEY ( MACACA
SPECIOSA ) IN CAPTIVITY

Principal Investigator: Amos E. Palmer, D.V.M.

Other Investigators: Ruth Kirschstein, M.D. , LP, DBS
William Tullner, Ph.D., EB, NCI

Cooperating Units: Laboratory of Pathology, DBS
Endocrinology Branch, NCI

Man Years (computed for the 12-month period)
Total: .4 (AHS only)
Professional: .2
Other: .2

Project Descriptiont

Objective: To attempt to evaluate the reproductive

potential of the M. speciosa in captivity.
To supply infant M. speciosa to research
laboratories. To introduce a monkey species into NIH laboratories
which is more easily handled than present, commonly held species
(M. mulatta ) .

Methods Employed: Menstrual cycles will be determined and

matings attempted at the expected time of
ovulation. Pregnancies will be ascertained

by either gonadatropin studies or digital palpation. Births will be

recorded and the infants used in research.

Major Findings: Menstruation, as seen in other macaque

monkeys is not detectable in the M. speciosa
presently under study. By mating over pro-
longed periods, without good guidelines to estimate time of ovulation,
we thus far have successfully bred six females. Serum gonadotropins
are detectable with parameters similar to the rhesus monkey. To date
there has been two live births, two pre-term abortions and one still
birth (delivered by cesarean).



4-24



Part A (Cont'd)

Significance: Research laboratories must have primates
at or soon after birth in order to carry
out studies on some evasive agents.
Animals of this nature are not available by commercial suppliers,
thus it is imperative that primates be bred for research needs in
captivity. Because of its nature the M. speciosa is a more desirable
animal for such a program than the M. mulatta presently in use.

Proposed Course Continuation,
of Project:

Part B included Yes No X



4-25



Serial No. 4.4



1. Laboratory Aids Branch

2. Animal Hospital Section

3. Bethesda



PHS-NIH

Individual Project Report

July 1, 1964 through June 30, 1965



Part A



Project Title: EVALUATION OF PROGRAMS FOR IMMUNIZATION OF QUARANTINE DOGSi!

Principal Investigator: Francis J. Judge, D.V.M.

Other Investigators: None

Cooperating Units: None

Man Years (computed for the 12-month period)
Total: .45
Professional: .05
Other: .4

Project Description:

Objective: To determine if the current program of

immunization is of value in a prophylactic
program for the respiratory disease complex
of quarantine dogs.

Methods Employed: Series I : This project compared three groups
of dogs on controlled procedures to a group
receiving the standard sequence of vaccina-
tions. Each group contained 250 dogs. Group I (Standard) received
antiserum at entry and was vaccinated in two weeks. Group II (Serum
Control) received serum only at entry. Group III (Vaccine Control)
received only vaccine two weeks after entry. Group IV received neither
serum nor vaccine.

Series II : Four groups of 250 dogs each were
again used. The first three groups were placed on different immuniza-
tion programs, and the fourth group served as an unvaccinated control.
Group I received vaccine at entry but no serum. Group II received
antiserum at the time of entry and was vaccinated two weeks after entry.
Group III was inoculated with canine globulin concentrate; each dog was
then vaccinated two weeks after entry.

Programs in both series are evaluated by the
morbidity and mortality in each group, as well as the number of returns
to the vendor after one week in each group.



4-26



Part A (Cont'd)

Major Findings: Series I ; The percentage of returns to the
vendor after one week and the morbidity
figures were relatively constant in all
groups. However, in Group IV, which received no immunization, morbi-
dity and returns were highest.

The mortality figures showed the greatest
variation between groups, and ranged from 4.4% for Group II to 13.67o
for Group IV. Group I had a mortality rate of 7.6% and Group III of
10.8%. It must be concluded that the use of serum does decrease the
mortality rate during the 30-day quarantine of dogs.

Series II ; Results are not yet tabulated.

Significance: The above and planned studies are being

performed in an effort to determine a method
of prophylaxis, which will significantly
reduce the mortality and morbidity losses due to the respiratory
disease complex in quarantine dogs. Immune serum may help in reducing
mortality.

At present the morbidity rate in these dogs
ranges from 50 to 70% and no method -of immunization tried has signi-
ficantly lowered this rate.

Proposed Course Further studies to evaluate the different
of Project: products available for distemper immunization

are planned so that the most suitable immuni-
zation program for quarantined dogs may be developed.

Part B included Yes No X



4-27



Serial No. 4.5



1. Laboratory Aids Branch

2. Animal Hospital Section

3. Bethesda



PHS-NIH

Individual Project Report

July 1, 1964 through June 30, 1965



Part A



Project Title: EVALUATION OF SHIGELLA FLEXNERI ATTENUATED VACCINE.

Principal Investigator: Samuel B, Formal, Ph.D., WRAIR

Other Investigators: Amos E. Palmer, D.V.M.

Cooperating Units: Laboratory of Immunology, WRAIR

Man Years (computed for the 12-month period)
Total: ,45 (AHS only)
Professional: .25
Other: .2

Project Description:

Objective: To attempt to develop and evaluate an

attenuated, non-pathogenic form of Shigella
flexneri (of several serotypes) for potential
use as an immunogenic agent to protect rhesus monkeys against these
organisms while under quarantine.

Methods Employed: Hybrid organisms produced by mating Shigella
spp . with E. coli will be given, via the oral
route by stomach tube. Animals will be chal-
lenged with fully virulent organisms at varying intervals to determine
susceptibility to the virulent agents. In later studies, 507o of the
newly arriving rhesus monkeys will be treated, followed by close obser-
vation for differences in incidence of shigellosis between treated
animals and untreated controls.

Major Findings: One dose of the hybrid organisms given orally
affords protection to challenges with virulent
organisms of the same serotype.

Significance: About 15% of the rhesus monkeys imported into

the Primate Quarantine Unit shed Shigella
flexneri organisms of several serotypes. The
clinical syndrome accredited to SMgella spp . is an acute, bloody
diarrhea which costs more in treatment, loss of condition, and deaths
than any other disease encountered in primates. An immunizing agent



4-28



Part A (Cont'd)

of this nature would greatly decrease problems in conditioning primates
and increase the quality of animals issued by the Primate Quarantine
Unit.

Proposed Course Continuation of studies to evaluate the value
of Project: of such vaccines in the quarantine of primates.

A manuscript has been submitted for publication.

Part B included Yes No X



4-29



Serial No. 4. 6

1. Laboratory Aids Branch

2. Animal Hospital Section

3. Bethesda



PHS-NIH

Individual Project Report

July 1, 1964 through June 30, 1965



Part A



Project Title: AN EPIZOOTIC OF MEASLES IN A COLONY OF RHESUS MONKEYS
( MACACA MULATTA )

Principal Investigator: Stephen Potkay, V.M.D.

Other Investigators: James R. Ganaway, D.V.M. , M.P.H., GPS
Roy Kinard, D.V.M. , CPS
Nancy G. Rogers, B.S., LVI, DBS

Cooperating Units: Primate Quarantine Unit, AHS

Comparative Pathology Section, LAB
General Virology Section, LVI, DBS

Man Years (computed for the 12-month period)
Total: Less than .08 (AHS only)

Project Description:

Objective: To determine the etiology of a condition in

the rhesus monkeys of the Primate Quarantine
Unit, which interfered with tuberculosis
test readings.

Methods Employed: Monkeys were examined daily and clinical

findings were recorded. Blood samples were
collected and examined for measles antibody
titer.

Major Findings: Serology showed that the disease was measles

(rubella) and clinical observations were those

of conjunctivitis, maculopapular skin rash,

and varying degrees of facial edema and erythema. The clinical feature

of the natural occurrence of this disease have not been reported previo

Significance: This project has provided a quick method of

determining the presence of measles in the
primate colony.

Proposed Course Completed. A manuscript has been submitted
of Project: for publication.

Part B included Yes No X

4-30



Serial No . 4.7

1. Laboratory Aids Branch

2. Animal Hospital Section

3. Bethesda



PHS-NIH

Individual Project Report

July 1, 1964 through June 30, 1965



Part A



Project Title: CLINICAL ASPECTS OF A FEBRILE HEMORRHAGIC DISEASE OF
MONKEYS IN A QUARANTINE COLONY

Principal Investigator: Amos E. Palmer, D.V.M.

Other Investigators: Anton M. Allen, D.V.M. , Ph.D., CPS
Stephen Potkay, V.M.D.
James Ganaway, D.V.M., M.P.H. , CPS
Ruth Kirschstein, M.D, , LP, DBS
Alexis Shelokcv, M.D. , LVR, DBS

Cooperating Units: Comparative Pathology Section, LAB
Laboratory of Pathology, DBS
Laboratory of Virology and Rickettsiology, DBS

Man Years (computed for the 12-month period)
Total: 1.3 (AHS only)
Professional: .4
Other: .9

Project Description:

Objective: To describe and identify the cause of a

disease which occurred in epizootic propor-
tions among monkeys held in the Primate
Quarantine Unit during November and December 1964.

Methods Employed: Clinical examination, postmortem examination,
clinical medicine, and epidemiological studies
were employed.

Major Findings: The disease was characterized clinically by

fever, anorexia, albuminuria vomition, bloody
diarrhea, cutaneous petechial hemorrhages,

epistaxis, dehydration, and death in most clinically affected animals.

The incubation period in inoculated animals was 48 hours to four days,

deaths occurred in six to 14 days.

Incidence per room ranged from 107o to 607o.
Wide spectrum antibiotics and supportive therapy were ineffective.
Transmission was accomplished by close animal contact and by inoculation



4-31



Part A (Cont'd)

with blood, serum, and tissue suspensions from infected animals.
Several animals survived clinical infections during the epizootic and
after inoculation. The disease was fatal to about 225 monkeys, affect-
ing Macaca mulatta , Macaca irus , and Macaca speciosa .

Significance: The disease is not known to have occurred

previously among primates. Since the etiologic
agent is not yet identified, the importance of
the disease cannot be evaluated at this time. The epizootic stopped
the normal function of the Primate Quarantine Unit for three months.

Proposed Course All work with the disease has been eliminated
of Project: from the facilities of the Primate Quarantine

Unit. Virological studies are continuing in

an effort to identify the agent and to further study the pathogenesis

of the disease.

Part B included Yes No X



4-32



Serial No. 4^8

1. Laboratory Aids Branch

2. Comparative Pathology Section

3. Bethesda



PHS-NIH

Individual Project Report

July 1, 1964 through June 30, 1965



Part A



Project Title: FEBRILE HEMORRHAGIC DISEASE OF MONKEYS

Principal Investigator: Anton M. Allen, D.V.M. , Ph.D.

Other Investigators: James R. Ganaway, D.V.M., M.P.H.
Roy Kinard, D.V.M.
Amos E. Palmer, D.V.M.
Nicola Taurasco, M.D., LVR, DBS

Cooperating Units: Animal Hospital Section, LAB

Laboratory of Virology and Rickettsiology, DBS

Man Years (computed for the 12-month period)
Total: 1.6 (CPS only)
Professional: .6
Other: 1.0

Project Description:

Objective: To study the pathogenesis of the disease and

initiate methods of control or elimination.

Methods Employed: Pathology, clinical pathology, microbiology,
clinical medicine, and epidemiology methods
were employed.

Major Findings: The disease was characterized clinically by
fever, albuminuria, variable leukopenia and
thrombocytopenia, terminal anemia, and death
of most clinically affected animals; pathologically by (1) cutaneous,
pulmonary, renal, hepatic, intestinal, and subperitoneal hemorrhage,
(2) systemic vascular stasis, (3) glomerulo-nephrosis, (4) follicular
necrosis of lymph nodes, thymus, and spleen, (5) splenomegaly due to
engorgement of the cords with plasma, (6) variable focal necrosis of
the heart and adrenal gland, and liver, (7) fatty degeneration and
centralobular cloudy swelling of the liver, (8) bone marrow erythropoetic
hypoplasia. The time from exposure to death was 6 to 14 days. Inci-
dence rates per room were 10 to 607o. Broad spectrum antibiotics includ-
ing chloranphenicol were ineffective. Transmission was accomplished by
close animal contact and by inoculation of infected blood, serum, and



4-33



Part A (Cont'd)

tissue suspensions. Sub-clinical disease with development of
immunity evidently occurred in a few survivors of the epizootic.
The disease was fatal to 225 monkeys, affecting Macaca mulatta, Macacaj
iniS ) and Macaca speciosa .

Significance: The disease is not known to have occurred
previously in the primate colony. The
causative agent has not been identified,
and therefore the health hazard to man is unknown. The epizootic
resulted in the interruption of numerous NIH research programs.

Proposed Course Continuation of studies to identify the
of Project: causative agent and refine the details of

the pathogenesis of the disease.

Part B included Yes No X



4-34



Serial No, 4^9

1. Laboratory Aids Branch

2. Comparative Pathology Section

3. Bethesda



PHS-NIH

Individual Project Report

July 1, 1964 through June 30, 1965



Part A



Project Title: CHRONIC MURINE PNEUMONIA OF THE ALBINO RAT

Principal Investigator: James R. Ganaway, D.V.M. , M.P.H.

Other Investigators: Anton M. Allen, D.V.M,, Ph.D.

Cooperating Units: Animal Production Section, LAB

Man Years (computed for the 12-month period)
Total: .6 (CPS only)
Professional: .2
Other: .4

Project Description:

Objective: To isolate and characterize the etiologic

agents. To perform transmission studies with
each suspected agent. To recommend measures

for prevention and control of this disease complex under colony

conditions.

Methods Employed: Microbiology, immunology, and pathology
methods were employed.

Major Findings: Results of studies to date indicate that the
etiology is of bacterial nature rather than
viral as proposed by other investigators
heretofore. Controlled vaccine studies were carried out in the
General Purpose rat colony using an emulsified univalent bacterial
antigen. The vaccine failed to prevent the disease syndrome.
Transmission studies using three of the more probably suspect agents
in germfree rats housed in plastic isolators are in progress. Other
studies indicate that the etiology of otitis media is separate and
distinct from that responsible for the pathological processes observed
in the lower respiratory tract.



4-35



Part A (Cont'd)

Significance: This disease complex is the greatest cause
of morbidity and eventual mortality in rat
colonies. Affected rats are unfit for

research, particularly when destined for long term or toxicity

studies.

Proposed Course Continuation,
of Project:

Part B included Yes No X



4-36



Serial No. 4.10

1. Laboratory Aids Branch

2. Comparative Pathology Section

3. Bethesda



PHS-NIH

Individual Project Report

July 1, 1964 through June 30, 1965



Part A



Project Title: MUCOID ENTERITIS OF RABBITS

Principal Investigator: James R. Ganaway, D.V.M. , M.P.H.

Other Investigators: Anton M. Allen, D.V.M,, Ph.D.
Thomas D. Moore, Ph.D.

Cooperating Units: Animal Production Section, LAB

Man Years (computed for the 12-month period)
Total: .6
Pro f e s s ional : . 3
Other: .3

Project Description:

Objective: To determine the etiological factor (s), and to

attempt experimental reproduction of the
disease in rabbits. Based on the foregoing,
to recommend measures for control or prevention in the rabbit colonies.

Methods Employed: Routine pathological and bacteriological
methods were used.

Major Findings: This chronic condition in weanling rabbits

smolders endemically in the rabbit colonies.
Although sporadic in occurrence, when it does
appear, usually the entire litter is affected. The agent suspected as
having prime etiological significance, because of its observation in
roughly 95% of the cases, has now been isolated in pure culture.
Characterization of this agent and carrying out of transmission studies
have not been performed due to lack of time and personnel.

A similar but not identical agent of the
mouse has been isolated in pure culture. The agents from both the
rabbit and the mouse occur in the cecum and proximal colon and have
highly characteristic morphology and tinctorial properties. The
agent from the mouse is slightly smaller than the one from the rabbit.
This agent selectively grows in the cytoplasm of hepatic cells of



4-37



Part A (Cont'd)

intravenously inoculated rabbits and results in extensive liver
necrosis and death in 40-45 days.

Significance: Mucoid enteritis is the greatest cause of
morbidity in our rabbit colonies. Though
widely recognized as such throughout the
world, etiology has not been ascertained.

Proposed Course Continuation,
of Project:

Part B included Yes No X



4-38



Serial No. 4.11

1. Laboratory Aids Branch

2. Comparative Pathology Section

3. Bethesda



PHS-NIH

Individual Project Report

July 1, 1964 through June 30, 1965



Part A



Project Title: STUDY OF CAUSES OF DEATH IN STRAIN 13 AND STRAIN 2
INBRED GUINEA PIG COLONIES

Principal Investigator: James R. Ganaway, D.V.M. , M.P.H.

Other Investigators: Anton M. Allen, D.V.M. , Ph.D.
Roy Kinard, D.B.M.

Cooperating Units: Genetics Unit, Animal Production Section, LAB

Man Years (computed for the 12-month period)
Total: .9 (CPS only)
Professional: .3
Other : . 6

Project Description:

Objective: To define the causes of losses in the inbred

colonies of guinea pigs. To isolate the
etiologic agent(s). To recommend measures for
control or prevention of these losses in the colonies.


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