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Annual report : National Institutes of Health. Division of Research Grants (Volume 1971) online

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several bacteria are responsible for this disease complex. Progress in
reporting has been delayed by the continuing commitment to establish the
mycoplasma-f ree status of the cesarean-der i ved, germfree and barrier rat
stocks, where the infection was discovered for the first time during the
previous year. This problem stimulated the need to devise a means of
detecting active infection in the live rat. To determine this state
previously it was necessary to kill the rat, which is self-defeating in a
production colony. Accordingly, a method of swabbing the oropharynx under
light inhalation anesthesia was tested and found, through followup study,
to be very sensitive. As a result of testing and subsequently destroying
those rats found positive on culture, the mycoplasma infection of germfree
and barrier rats appears to have been eliminated.

73



si gnificance ; Chronic respiratory disease is the gt-eatest cause of
morbidity and eventual mortaiity In laboratory rats. The cesarean-der i ved^
barrier-maintained rat has previously been considered free of all
respiratory infection. We have shown this to be an erroneous assumption.
Accordingly^ it may be necessary to reevaluate all published reports which
could have been influenced by these findings through the use of similar
rats .

Proposed C ourse ; Continuation.

Honors and Awards ; None.

Publ icat ions s None.



7k



Serial No. DRS-LAB-2

1. Laboratory Aids Branch

2. Comparative Pathology Section

3. Bethesda



PHS-NIH

Individual Project Report

July 1, 1970, through June 30, 1971



Project Title: TYZZER'S DISEASE

Previous Serial Number: DRS-LAB-2

Principal Investigator: James R. Ganaway

Other Investigators: Anton M. Allen
Thomas D. Moore

Cooperating Units: None.

Man Years:

Total: 1.0
Professional : 0.5
Other: 0.5

Project Description:

Ob iect ive ; To characterize the etiologic agent. To study the pathogenesis
of the disease through experimental transmission studies. To recommend
measures for the control and/or prevention of the disease in the NIH rabbit
colony.

Methods Employed : Microbiology and pathology

Major Finding s: The disease continued to occur endemically in the NIH
rabbit colony. Study of the etiologic agent, Baci I lus pi 1 i formi s< was
continued. A means was found to obtain large anounts of the vegetating
organism free of spores, but the vegetative form was found to be very
unstable outside the host cell (propagated in embryonated eggs) and it did
not survive the usual methods of preserving microorganisms such as freezing
and storage at -70 degrees centigrade or Lyoph i I izat ion.

Si gnificance : This disease is emerging as a common, often fatal, enteric
infection of a variety of laboratory animals including mice, rats,
hamsters, gerbi Is, rabbits, cats, and monkeys. The etiologic agent is a
most unusual one whose collective characteristics have no equal in the
field of microbiology and, consequently, is unclassified at present. Much
of our present day information on the widespread occurrence of the disease



75



in a variety of animal species was stimulated by our report of the disease
in rabbits, this being the first record of the spontaneous occurrence of
the disease in a species other than the mouse.

Proposed Course ; Continuation.

Honors and Awards ; By invitation. Dr. Ganaway presented a speech,
"Tyzzer's Disease," as one of six contributors to the symposium "Disease of
Laboratory Animals Complicating Biomedical Research," hosted by the
American Society for Experimental Pathology at the Annual FASEB Meeting,
Chicago, Illinois, April 13, 1971.

Publ ications ; Ganaway, J.R., Allen, A.M., and Moore, T.D. Tyzzer's
Disease of rabbits; Isolation and propagation of Bac i 1 lus pi 1 i formi s
(Tyzzer) in embryonated eggs. Infection and Immunity, 3:U29-'t37, 1971.



76



Serial No. DRS-LAB-3

1. Laboratory Aids Branch

2. Comparative Pathology Section

3. Bethesda



PHS-NIH

Individual Project Report

July 1, 1970, through June 50, 1971



Project Title: PASTEURELLA IN GNOTOBIOTIC RATS

Previous Serial Number: DRS-LAB-l*

Principal Investigators: Thomas D. Moore

Anton M. Allen
James R. Ganaway

Cooperating Units: Germfree Unit, RRPS

Man Years:

Total: 1.0
Profess ional : 0.3
Others: 0.7

Project Description:

Obiect i ve : To ascertain the role and incidence of £. pneumotropica in
gnotobiotiC/ barrier -maintai ned rats free of respiratory illness. To
determine the preferred location of this organism in experimentally
inoculated germfree rats.

Methods Employed ; Microbiology and Pathology

Maior Findings : £.. pneumotropica was isolated from the inflamed small
intestine of inbred, gnotobiotic rats maintained in a closed barrier
system. Further investigation and more extensive culturing showed the
presence of this agent in all barrier room rats and mice as well as i n a
number of germfree isolators. An examination of healthy, female
Sprague-Dawley rats from the barrier revealed that the organism had
colonized the intestine with the caecum as the preferred site. The
respiratory tract in these naturally infected rats was free of the organism
while the uterus was culturally positive sporadically. These findings were
generally confirmed in experimentally infected, germfree rats. In this
latter case, occasional isolations were made from the trachea but definite
colonization of the gut occurred with an increase of organisms with time.
Again, isolations from the uterus were sporadic.



77



Significance ; £. pneumotropica has been implicated as an etioiogic agent
in respiratory disease in mice and rats. The organism has been described
as having an affinity for tissues of the respiratory tract although it has
been isolated from brain, uteri, and spleen of mice. More recently, it has
been isolated from eye abscesses in conventional mice here and in two other
colonies. More information on the relationship and role of this organism
in rodent disease is important.

Proposed Course ; Continuation.

Honors and Awards ; None.

Publ i cat ions ; A manuscript is being prepared.



78



Serial No. DRS-LAB-U

1. Laboratory Aids Branch

2. Comparative Pathology Section

3. Bethesda



PHS-NIH

Individual Project Report

July 1/ 1970, through June 50, 1971



Project Title: PREGNANCY TOXEMIA OF GUINEA PIGS

Previous Seri'al Number: None.

Principal Investigator: James R. Ganaway

Other Investigators: Anton M. Allen

Cooperating Units: None.

Man Years:



Total:


0.2


Professional :


0.2


Others:


0.0



Project Description:

Ob iect i ve ; To further characterize the disease; to devise a more simple
means of diagnosis than is presently available; to learn how to control or
prevent the disease.

Methods Employed : Pathology

Maior Findings ; Obesity was found to be the predisposing factor of major
concern. Guinea pigs bred when 900g. body weight suffered heavy losses
(50 percent), then stresses late in pregnancy while those bred when 500g.
body weight were unaffected by the same stress in late pregnancy.
Urinalysis afforded a predictable aid to the early diagnosis of the disease
which was evidenced by a sudden drop in the pH and the appearance of
protein. The disease was further characterized by marked lipemia,
acidosis, ketosis, and death. A similar syndrome was induced, but with
lesser incidence, in obese nonpregnant guinea pigs and was compared to
fast ing ketosi s.

Si gn! f icance ; Pregnancy toxemia is one of the major causes of maternal
fetal wastage in guinea pig production colonies. The above findings
indicate a need for regulating the diet of the guinea pig before and during
pregnancy rather than the present ad 1 i bi turn feeding practice.



79



Proposed Course; Completed

Honors and Awards ; None.

Publ i cat ions ; Ganaway, J.R. and Allen/ A.M. Obesity predisposes to
pregnancy toxemia (ketosis) of guinea pigs. Lab. An. Sci., 21;U0-I»ft, 1971,



80



Serial No. DRS-LAB-5

1. Laboratory Aids Branch

2. Rodent and Rabbit Production Section

3. Bethesda

PHS-NIH

individual Project Report

July 1/ 1970, through June 30, 1971



Project Title: GENETIC ANALYSES AND DEVELOPMENT OF NEW STRAINS FOR
BIOMEDICAL RESEARCH

Previous Serial Number: None.

Principal Investigator: C. T. Hansen

Other Investigators: W. J. McEleney
W. J. Hinkle
P. Phill ips

Cooperating Units: Laboratory of Bacterial Products, DBS

Poultry Research Branch, ARS, USDA, Beltsville, Maryland
Laboratory of Virology and Rickettsi ology, DBS

Man Years:

Total: 1.0

Professional : 0.5

Others: 0.5

Project Description;

Ob iect ive : 1) To maximize the efficiency of the operation, 2) to ensure
that the frequency of desirable characteristics of the Inbred strains
remain stable, and 3) to utilize the existing gene pool to develop superior
models for biomedical research by selective breeding. The techniques used
are necessarily those which are applicable to the particular project.

Methods Employed and Maior Fin dings: Reproductive efficiency has been
improved by determining in a broad way the dietary requirements of various
inbred strains of mice and by changing the mating system used to produce
these animals. The data obtained from these studies have been converted to
automatic data processing procedures and preliminary statistical analyses
have been completed.

Genetic monitoring procedures have been established. Data collected from
these studies show that the age of onset of leukemia and mammary tumors in
mice has remained stable over a long period. Blood pressure monitoring
studies of various strains of rats have revealed considerable variation



81



between strains and have detected additional hypertensive strains. A
genetic analysis of hypertens i veness consisting of various strain
combinations has been undertalcen. The results of this analysis thus far
indicate a complex mode of inheritance. These studies will be continued.

Strain development studies in mice, chickens^ rabbits and dogs are
underway. These studies essentially consist of developing strains
characterized by a high and low frequency of a characteristic by selective
breeding. Selective breeding for sensitivity and resistance to B..
pertussi s has reached the tenth generation. Sensitivity has been increased
from an initial value of 31 percent to 70 percent and resistance to k
percent. This work is being done in cooperation with the Laboratory of
Bacterial Products, Division of Biologies Standards. Work is underway to
develop chicken stocks which are homozygous resistant and susceptible to
leukosis virus. A resistant stock of birds has been developed and these
birds will be used as a tester stock to develop a homozygous susceptible
line. This work is being conducted in cooperation with the Poultry
Research Branch, ARS, USDA, Beltsville, Maryland and the Laboratory of
Virology and Ricketts iology. Division of Biologies Standards. Four lines
of rabbits have been established, each of which is homozygous for a pair of
genes which determine gamma globulin allotypes. Studies are underway to
determine the interaction of these genes in producing the allotypes.

Si gnif icance ! The significance of these projects is to develop, by the use
of genetic techniques, animal models which have an application to
biomedical research.

Proposed Course ; Continuation.

Honors and Awards ; None.

Publ icat ions ; None.



82



Serial No. DRS-LAB-6

1. Laboratory Aids Branch

2. Animal Center Section

3. Poolesvi 1 le



PHS-NIH

Individual Project Report

July 1, 1970, through June 30, 1971



Project Title: HEMATOLOGIC AND BACTER I OLOGIC CHARACTERISTICS OF STORED
CANINE BLOOD

Previous Serial Number: None.

Principal Investigator: Stephen Potkay

Other Investigators: None.

Cooperating Units: None.

Man Years:

Total: 0.2
Profess ional : 0. 1
Other: 0.1

Project Description:



Qb iect i ve : 1) To determine the bacter iologi c status of blood derived fr.
dogs that are normal and healthy as judged by applicable standard clinical
and laboratory criteria, and to evaluate methods of obtaining sterile
blood. 2) To determine the influence of anticoagulants, storage ti
storage temperatures, and storage containers on whole canine blood.



om



I me.



Methods Employed ; Blood (U50-500 ml) was collected into vacuum bottles
containing heparin, ACD, or CPD as anticoagulants. Blood was stored for
four days in heparin and for five weeks in ACD and CPD at three degrees
centigrade. Determinations of the cellular components of the blood were
made weekly and compared to values obtained from blood immediately after
collection. Heparinized blood was examined daily. All blood was tested
for sterility at the end of the periods.

Maior Finding s; Bacterial contamination of the blood did not occur in
association with the collection procedures employed. Changes in cell
populations in refrigerated canine blood appear to be minor. However,
electrolyte shifts become pronounced with increased storage time.
Morphology of the blood cells has not yet been examined.



83



Significance ; The feasibility of suppiying sterile blood from canine
donors for use in research was demonstrated. Storage of heparinized blood
for 2k hours and ACQ blood for up to four weeks does not appear to reduce
its usefulness for some research purposes, based on evaluations of cellular
and electrolytic composition.

Proposed Course s Completedo

Honors and Awards ; None.

Publ ications ; A manuscript is being prepared.



Sk



Serial No. DRS-LAB-7

1. Laboratory Aids Branch

2. Animal Center Section

3. Poolesvi I le

PHS-NIH

Individual Project Report

July 1, 1970, through June 30, 1971

Project Title: A COMPARISON OF PHYSICAL AND HEMATOLOGIC CHARACTERISTICS
OF PUPPIES FED DIETS CONTAINING 9%, 8%, and 6% FAT

Previous Serial Number: None.

Principal Investigator: Stephen Potl<ay

Other Investigators: Joseph J. Knapica
Amos E. Palmer
Terry Pitts

Cooperating Units: Rodent and Rabbit Production Section

Man Years:



Total :


0.


,8


Professional :


0,


.2


Other:


0.


.6



Project Description:

Ob iect ive ; To determine the relative values of feeds containing three
levels of fat as diets for puppies during theweaning and growing periods.

Methods Employed : 1) Matings were scheduled so as to produce litters of
genetic uniformity. 2) Thirty litters of pups (Foxhounds) were equally
divided into three groups which were fed commercial dry diets containing
6 percent/ 8 percent, and 9 percent fat, respectively. Dams were fed the
same diets as their puppies beginning two weeks prior to parturition and
continuing through weaning. 3) Supplemental feeding, which consisted of
reconstituted canned milk, was provided uniformly for puppies to expedite
weaning. The dry feeds were provided ad 1 i bi turn for all bitches and their
litters, k) Evaluations consisted of examining and weighing each bitch
weekly through weaning and each puppy weekly until 20 weeks of age.
Measurement of hematologic values was done by standard methods, samples
were obtained weekly from three randomly selected puppies from each litter
and from their dam.



85



Maior Finding s; The number of litters produced in his study is currently
too small to permit meaningful comparisons. However, data obtained from
one litter receiving the 9 percent fat diet and two litters fed the diet
containing 8 percent fat suggest that the former is markedly superior.

Si gni f i cance ; The majority of conmercially available diets are in
compliance with the minimum nutritional requirements for dogs as stated by
the National Research Council/ Committee on Animal Nutrition. Hov^ever,
these diets differ markedly in providing usable or digestable nutritives to
puppies. The differences are evident from observations of general physical
condition, weight gain, andhematologic values of dogs given these feeds.
Future selection of diets for growing puppies will be based in part on the
information concerning dietary fat content which is derived from his study.

Proposed Course ; Continue until 27 litters have been studied.

Honors and Awards ; None.

Publ i cat ions ; None.



86



Serial No. DRS-LAB-8

1. Laboratory Aids Branch

2. Animal Center Section

3. Poolesvi 1 le



PHS-NIH

Individual Project Report

July 1/ 1970, through June 30, 1971



Project Title: EFFECTS OF CHLORAMPHENICOL ON BARBITURATE AND HALOTHANE
ANESTHESIA IN RHESUS MONKEYS (MACACA MULATTA)

Previous Serial Number: None.

Principal Investigator: Stephen Potkay

Other Investigators: Amos E. Palmer

Man Years:

Total: 0.3
Profess ional : 0.2
Others: 0.1

Project Description:

Object i ve : To determine whether oxybarbi tu rate anesthesia is prolonged
following chloramphenicol administration in rhesus monkeys and to ascertain
the effects of chloramphenicol administration on thiobarbi turate and
halothane anesthesia.

Methods Employed ; Four groups of monkeys, each containing six males and
six females, were used. In Groups 1 and 2 the effects of chloramphenicol
administered intravenously (IV) and intramuscularly (IM) immediately prior
to induction of anesthesia with an oxybarbi turate (pentobarbital sodium)
were studied. Monkeys in Group 3 were anesthetized with a thiobarbi turate
(thiamylal sodium) immediately prior to administration of chloramphenicol
!M. Chloramphyen icol was given immediately prior to thiamylal induced
anesthesia in Group k monkeys. Anesthesia was maintained in his group for
one hour using halothane. Recovery times were calculated as the lapsed
time between administration of the anesthetic and the time at which the
monkey could stand without support. Data were anlayzed by standard
methods.

Maior Finding s: Recovery from pentobarbital anesthesia was prolonged in
monkeys that received chloramphenicol IM or IV. The duration of anesthesia
was not affected by chloramphenicol administration in monkeys anesthetized
with thiamylal or halothane.



87



Significance ; Since it is generally desirable that monkeys become
ambulatory as soon as possible following anesthesia, combined
administration of chloramphenicol and pentobarbital should be avoided.
Rapid recovery of monkeys from thi amyl a1 hal ethane anesthesia^ regardless of
whether chloramphenicol is or is not administered concurrentl y, 1 ndi cates
the suitability of these agents over pentobarbital for anesthesia in
monkeys.

Proposed Course ; Completed,

Honors and Awards ; None.

Publ ications ; A manuscript has been prepared.



88



DIVISION OF RESEARCH SERVICES
Summary of Branch Activities July 1/ 1970, through June 30, 1971

LIBRARY BRANCH Seymour I. Taine, Chief

I . SUMMARY

A new Library Services Adviser program was implemented to broaden services
offered by the Library in an integrated bibliographic and reference effort.

Effective measures to improve conmuni cations were initiated throughout the
Branch. Some significant organizational and personnel changes have been
beneficial for Library operations. Planned physical alterations of the
Lower Level have now been completed. The photocopy service was relocated
in quarters adapted for more effective operations. The Reading Room has
been enlarged, lighted well, and attractively furnished. Relocation of the
Acquisitions Unit has consolidated technical services on the Upper Level.

The Library Advisory Committee continued to assist and support the programs
of the NIH Library,

The case of Williams and Wilkins Company continues against the Government
relating to the photocopy service and copyright law. Mr. Taine, Chief,
Library Branch, appeared as one of the witnesses for the Government at the
trial held September g-l^t.

A decision will be made by the Department of Labor for DHEW regarding the
validity of the composition of the bargaining unit at the time of the
election on October Ik, 1969. The election resulted in Local 2^19 American
Federation of Government Employees AFL-CIO being awarded exclusive
recognition to represent all non-supervisory NIH Library employees. The
NIH and the DHEW have now raised the question of the appropriateness of
including professional personnel in the unit.

The 3M TATTLE-TAPE Security System selected for installation in July has
been in operation during the year to control proper charging and reduce
loss of Library books and journals.

Two Library Consultants to the NIH Library were appointed to advise the
Branch Chief in the areas of Library automation and scope and coverage of
the Library collection. A draft of an overall policy governing scope and
coverage and of the Reference Collection are in preparation.

The FY 1971 Branch Training program emphasized in-house seminars and
lectures for employees of all levels. It was designed to meet the needs of
all Library personnel and was developed on a group basis through group
chairmen and conmittees for close interaction. Career and job counseling
were provided nonprofessional employees. A pilot training course was



89



developed for two selected Library Technicians from the Reference Unit
which led to their passing the difficult Civil Service Commission's subject
matter test in library science and their transfer into the professional
Librarian series.

An IBM study of Library operations has as its purpose the identification of
an existing automated Library system which would meet the needs of the NIH
Library. DCRT's WYLBUR System was implemented in the Acquisitions Unit's
ordering procedures after a detailed experimental study of its feasibility.
The Library's journal control procedures are under review relative to the
feasibility of adaptation of PHI LSOM System developed by Washington
University School of Medicine Library.



90



I I. BRANCH PROGRAMS

A. Objectives

The primary mission of the Library Branch is to operate an efficient/
comprehensive library in support of NIH scientific, medical, and
administrative programs. Activities of the Library include selection,
acquisition, organization, maintenance, and circulation of literature
pertinent to the programs; operation of a photocopy service; provision of
informational, reference, and bibliographical services; provision of
Library services advisory assistance; and provision of a translating
service for foreign scientific and medical literature. To fulfill its
mission, the Library is responsive to changing literature needs of the NIH
investigators, is knowledgeable of current developments in manual and
machine methods of communication and information retrieval, and is alert to
adjustment of procedures for improved Library services.

B. Current P^'ograms

Readers Services

The Circulation staff provides a charging system making available books and
journals which may be borrowed. They issue Library Identification Cards
and Pagemasters and make assignments to locked carrels.

The Interlibrary Loan staff obtains from other libraries, through
Interlibrary loan, literature required by NIH investigators which is not
included in the collection.

The Stacks Maintenance staff maintains the library materials to facilitate
access by the Library clientele.

The Duplication and Audiovisual staff provides a Library Copy Service
allowing greater use of the Library's journals and an audiovisual service
giving access to biomedical literature available as microfilm, microcard,
and microfiche.

Translating Service

The Translation Unit provides oral, recorded and written translations as
requested. Oral translations are emphasized in-house, and written
translations are made for the most part by contractual firms.

Reference and Bibliographic Services

The Library Services Adviser program provides as a new experimental service
an integrated response to the information needs of the NiH scientific
community. This may consist of utilization of substantial external
resources beyond the resources available in the NiH Library, such as
specialized information centers, computerized Information retrieval
systems, and clearinghouses.



91



The Reference staff supplies ready response to questions, verifies
citations, and compiles short reference lists. They man the Reference
Desks in the Reading Rooms on the Upper and Lower Levels-
Senior Reference Librarians answer difficult reference questions and
compile literature searches as requested. They serve as selectors for
Library acquisitions by continually searching journals, publishers'
catalogs and announcements for literature pertinent to the scope of the
Li brary .

Bibliographic Services staff provides bibliographic assistance with
experienced searchers to formulate requests for MEDLARS computer searches


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