TPC NEWS Summer 1983 Vol.2 No.2 (Whole
Number 3)
(An
Abridged Translation)
Page-3
Essay: Our research attitudes as sceneshifters of the field of
experimental medicine using nonhuman primates
We are often asked by visitors if we carry out researches here in this
center. Aside from pursuing the
real meaning of "research", I think biomedical researches conducted by
using nob- human primates as a laboratory animal will, so to speak, be compared
to performance of actors who present their creative talents by making the best
use of the speech and settings on the stage, while breeding/rearing of
laboratory primates will be compared to the works of sceneshifters who set the
stage for the actors. It is evident
that we need both for the success of a drama.
The point is whether the work of breeding/rearing involves elements of
research. I believe it does. Just as sceneshifters are required to be very creative, we
are also required to be creative in theory and in practice.
In order to meet and fulfill the requirements and intentions of medical
researches we should be at any time qualified with experience, accurate insight,
thorough knowledge and also enthusiasm.
As far as ideas, systematic methods or scale are concerned, those of our
center are scarcely ever seen before. Therefore,
we often come across with many unexpected and unsolved problems such as,
developing tools and apparatus for the welfare of animals, establishing the
methods of clinical testing and diagnoses, the measures for biohazard control,
and developing and/or improving breeding system from the viewpoints of genetic
control and of getting more prolific monkeys, we carefully study daily-obtained
data and consider these points in all aspects.
In the meantime we concentrate our efforts also to basic researches so as
to analyse biological characteristics of nonhuman primates.
The results obtained with these efforts benefit us also with their
effectiveness at the practical site of breeding, for example, a study on simian
blood groups has already contributed . to the control of genetic variability in
our breeding colony, a method of hormone assay established during the course of
endocrinological studies (particularly of LH) is of practical use 'for
successful breeding. And so the
data of immuna-physiological studies on aging and those of bacteriological and
pathological studies on naturally-occuring diseases have given rise to new ideas
with regard to breeding and rearing.
Furthermore, we have to aim our efforts to proving the usefulness of
simians in various fields of experimental medicines.
In this respect I would sincerely appreciate receiving cooperative
assistances from many other research organs and expert scientists.
Page-4
Breeding topics : Parity and number of offsprings of F1
cynomolgus monkeys
As
you see in the table, we have 79-mature Fl cynos (over 4-year-old),
55 of whom have experienced parturition. Fifty
of 59 over 5-year-old cynos have experienced parturition (84.7 %) (Some of the
59 cynos are not capable of mating). Only
5 of 20 4-year-old cynos who were born after the establishment of this center
had given birth to F2 so far. We
expect the rest 15 will have their offsprings within this year.
Fifty-five of Fl female cynos had 131 F2 in total.
Thirty-two of 50 over 5-year-old cynos (64 %) had twice parturitions and
the best record holder is the one of 13-year-old who had 7 parturitions.
And except the case of 18-year-old cyno who had 5 offsprings out of 5
parturitions, the highest average frequency of parity per Fl. is 4.8 at the age
of 13.
Three
years ago we-have established a new breeding system called "the rotation
line breeding system"(RLBS) which will make it possible for us to get
genetically-controlled progeny whose parental populations were divided into
three subpopulations according to three different countries of origin (Malaysia,
Indonesia or the Philippines). As
of the end of March, the number of F1 bred by the RLBS totalled 1,059
of which 535 were female. I wili
refer to this RLBS some time later.
There
are some cases of F1 cynos who are not able to nurse or hug their
infants. We cope witt) this problem
by doing artificial nursing, foster mother nursing, artificial aids or training
for nursing. Nevertheless this kind of defective motherhood seems to heal
gradually by gaining experiences of parturition. To help promote the normal nursing behavior, it seems that a
certain period for mutual skinships from the time of birth between infants and
playmates as well as between infants and mothers is important, We adopted the
group cage feedinj for cynos under 4 years, accordingly the result is expected
to be quite well in a few years.
Page-5
Natural infections of pathogenic agents in recently imported monkeys :
A survey in cynomolgus monkeys and
squirrel monkeys during the quarantine period in 1982 .
Three
tables are explicitly showing the data we obtained.
(See
the attached page 3).
Page-6
The laundry facility at TPC
Three
laundry staffs wash about 60 sets of work-clothes used in animal quarters a day
as a routine work. Those clothes
are; overalls made of 100 % cotton, hoods, masks and socks.
After steam-sterilized (1.8 kg/cm2, 120% 30 min), soiled
clothes are put into a hot-air-dryer for about 2 hours.
The mend of torn clothes are also done in this room.
Some ideas are made for the work-clothes; the hole for transceiver wires
or the use of heat-stable materials for the cuffs or zippers.
The laundry serves as one of measures to prevent personnals working with
monkeys from possible biohazard in the first place.
Page-7
On the techniques of care for cynomolgus monkeys:
Checkups for general health conditions of mother monkeys and their
infants during nursing period
Routine check-ups for infants are carried out with appetite, activities,
stool conditions, bleeding and body weight.
These check-ups are conducted on the day of birth- and in 1, 2, 3, 6, 9,
12 and 15th or older week of age until the eruption of deciduous teeth.
For mothers, check-ups are carried out from the day of delivery to 3, 6,
9, 12 and 15th week. First we catch
both of them by a catching net. While
fastening mother's hand and holding her legs, we separate the infant from its
mother to put each of them in a separate weighing box. A scale for infants is
graduated in 2 granis and that for
mothers in 10 grams. The phenomena
seen during the separation are interesting.
Almost all infants widely open their arms and legs, scream a little and
excrete about 10 drops of urine. Body
weight changes of an infant which I referred in the article of the last issue is
shown.
Page-8
A case report: Meteorism (Tympanites of the large intestine)
Meteorsm
is sometimes observed in cynomolgus monkeys, especially in large scale indoor
colony of primate breeding such as TPC. This
case of tympanites with twisting right hepatic lobe seems to be very rare .
History
: The case was an adult female cynomolgus monkey who arrived at our quarantine
facility in May 1977 from Malaysia. About
one and a half year after arrival, the monkey showed bloat and diarrhea almost
every day. The animal was dead and autopsied in January 1980.
Autopsy
findings: Macroscopically, large amount of gas accumulated in all part of large
intestine. Congesions were observed
in the lung, right lobe of liver and large intestine.
The torsioned hepatic lobe was markedly swollen.
Microscopically,
the alveolar capillaries of lung were congested and alveoli were filled with
exudated fluid. The sinusaid of
right hepatic lobe was markedly congested.
The liver cord was atrophied and necrosed.
Page-9
On the difference in qualities between indoor colony-bred and
wild-originated cynomolgus monkeys from the viroldgical point of view
It
has been well recognized that wild-originated monkeys generally have serum
antibodies against many viral species. In
. this paper, the author gives a discussion of the presence or absence of
antibodies against measles virus (MV) , herpes simplexvirus (HSV) and simian
virus 5 (SV5) with regard to indoor 'colony-bred and wild-originated cynomolgus
monkeys. These three viruses were selected from the following reason.
Firstly, MV infection is a serious problem when wild-originated macaques
are used for measles vaccine assay, since this virus can easily be transmitted
among non-immune monkeys of a group during transportation from habitat area to a
user' s facility even if the group has only one MV-infected monkey at first.
Secondly, the presence of antibody against HSV may reflect to some extent
the infection with herpes B virus because of the close antigenic relationship
between the two viruses. Thirdly,
svs could be regarded as an indicator for estimating the degree of
contaminations with viruses of simian origin, since this virus has been known to
be one of the most prevalent viruses among macaque monkeys .
The
indoor colony-bred and -reared monkeys
More
than 1, 000 indoor colony-bred and reared monkeys were tested for antibodies
against these viruses every 2 years after 1 year of age.
None of them have these antibodies so far.
Therefore, we think the cynomolgus, monkeys bred and reared at TPC were
virtually free from the infections with -MV, HSV and SV5.
It is very valuable from the viewpdint of laboratory animal science that
our colony-bred mankeyi are free from MV, HSV and SV5 infections. We are now able to use many monkeys havimng no MV, HSV and
svs antibodies for various kinds of experiments.
The
wild originated breeder monkeys
From
1979 through 1982, a total 1,500 wild-originated cynomolgus monkeys of 30
different groups were imported into TPC from the Philippines, Malaysia and
Indonesia. As regards breeders
selected from the 1,500 monkeys, sera-epidemiological follow-up surveys of
infections with MV, HSV and svs were conducted every 2 years since the end of
the quarantine period.
MV:
All of seventy-nine monkeys of 2 groups imported from Indonesia, of 54 monkeys
of 1 group from the Philippines and of 134 monkeys of 3 groups imported from
Malaysia were free from infection with MV.
However, 40 Malaysian monkeys and 60 Indonesian monkeys of the other
imported groups were already positive for MV antibody at the time of arrival.
Remaining 1,100 monkeys of 24 groups became positive without exception by
the end of quarantine period.
As
to 101 monkeys kept for 1 year and 313 monkeys kept for 3 years at TPC, a
significant rise of antibody titer could be noted with 13 monkeys and 6 monkeys
respectively when compared with the antibody titer of previous survey.
On the contrary, a significant decrease of antibody titer was observed
with 33 monkeys and 122 monkeys respectively.
HSV:
About 54 % of 1,500 monkeys had antibody against HSV at the end of the
quarantine period. For 631
Malaysian monkeys, the positive rate was 53 %.
722 monkeys from Indonesia showed a somewhat higher rate, 67 %.
As regards 429 monkeys from the Philippines, the rate was 33 %, that is
the lowest among the monkeys from three different countries.
The positive rate markedly varied with different import-group.
Out
of 89 monkeys which elapsed 1 year after the quarantine, 32 showed a significant
rise of antibody titer. On the
contrary, ,only 1 monkey of the 89 monkeys showed a significant reduction of
antibody titer. Of 295 monkeys
which elapsed 3 years after the quarantine, 39 showed a significant rise of
antibody titer and the other 38 monkeys decreased in antibody titer .
SV5:
For 434 monkeys from the Philippines, the positive rate was only 2 %.
As regards 263 Malaysian
monkeys in 1979, the positive rate was 72 %.
However, 374 Malaysian monkeys imported in 1980 to 1982 showed a positive
rate of only 2 %. Regarding 585
monkeys from Indonesia, showed a higher rate, 77 %.
Out
of 97 monkeys kept for 1 year after the quarantine, 8 showed a significant rise
of
antibody titer and the other 5 monkeys showed a decrease of antibody titer.
Of 305 monkeys kept for 3 years after the quarantine, 9 shdwed a
significant rise of antibody titer and the other 76 monkeys showed a significant
reduction of antibody titer.
Page-11
A contribution by a visiting research fellow: Suitability of cynomolgus monkey
as an animal model of Huntington chorea
I was really lucky being able to do the above experiment because the hope
of using primates for the research of Huntington chorea has been sticking in my
mind. Only
few things are known so fars, and we are urged to find the' proper measures for
this chorea. Using
primates for chorea studies is necessitated because of the structural functions
of brain and limbs . we used Kainate- injection to destroy corpus striatum,
which was very difficult to hit the targeted place because of its smallness of
an infant, We got one successful case which showed typical but transient
symptoms of chorea, However, I found it very hard to expect certain spontaneous
symptoms of Huntington chorea only by injecting Kainate to one side of corpus
striatum.
So I administefed L-Dopa i .v. to another cyno who showed no involuntary
movement but the degeneration of cells. Luckily
involuntary movement was seen t'o this cyno.
The biochemical analyses revealed the decrease of substance P and GABA as
shown in Fig.2. I will carry on my
study further with this point.
Page
12 Overseas topics - Deutsches Primatenzentrum GmbH in Gottingen
German Primate Center is run by a private non-profit company . the
shareholders of which are the state and federal ministries .
Not only breeding primates and keeping them for other scientific
institutes but also such cooperations with other scientific institutes and
scientists as training scientific and technical staffs, organizing field
projects with countries of origin and supplying guest scientists with working
facilities andm exchanging useful informations are the Center's main tasks to be
fulfilled. The establishment of the
Center was projected in the late 1960s and it will be holding 600 rhesus
monkeys, marmosets, and tamarins, 20 baboons and about 60 Saguinus oedipus by
the end of 1983 when the construction of new buildings will be finished.
Staffs will be 75 in number including 25 biomedical researchers.
Page
13 A visit to LENSIP
LENSIP was situated in the most northern part of the Stering Forest,
Orange County. The building of
LENSIP looked like a fascinating maze to me first.
It was founded in 1965, being siffiliated to New York University Medical
Center. The Center, holding 150
chimps and baboons each and 150 Platyrrhini primates, supplies technical
assistances and materials for over one hundred biomedical researchers a year.
Important missions as the WHO Center for Hematology of Primate Animals,
NIR National Chimpanzee Breeding Program and teaching in medical primatology are
to be fulfilled there . This
laboratory, as Dr. M. Jankowski, the Director, commented, is run very actively
with small number of staffs . At
the time we went to LENSIP, Dr. Jankowski kindly gathered his core scientists at
his office who are the professionals of each scientific field.
The highlight of our tour around LEMSIP was to observe a 150 chimpanzees
' colony. Huge hang-type cages in
mono-rail framework and widely spread vinyl sheet on the floor under cages for
disposal of excrements are called "dry-system".
Dr. Mahoney taught us to treat chimpanzees just as we treat our children.
In fact, three 6-month-old chimps were dressed in baby-suits and were
toddling around the play pen being left to the care of a charming
lady-technician.
Anoather intention of our visit to LEMSIP was to have the opportunity to
talk with authorities of simian blood researches.
I was very fortunate to meet and talk to Dr. Socha, and I will never
forget his words, saying, “We will exchange information from now on.”, as an
encouragement in continuing my research.
Page-14
My training course at TPC
I was very interested in the quarantine system.
When 40 Indonesian cynomolgus monkeys kept in wooden transportation cages
arrived at the quarantine facility of TPC, the procedures taken for the
reception of them looked like a ceremony conducted in a fantstic world to me.
Each cynos looked quite fine. Nevertheless
some of the cynos showed diarrheal symptom the next day.
The difference in property of diarrhea is hard to express and the man in
charge taught me how to deal with these cases.
A week’s later they became normal and luckily during 2 months of
quarantine period no death case appeared. I
also learned that transferring them from their individual cages to the other
environment might cause some disorders. Before
I came here I was worried I might get transmissible diseases from monkeys, but I
felt relieved to know that in past 5 years there has been no such cases because
of the biohazard control system here is properly working.
I, as an employee of a pharmaceutical company, do not always want to
adopt quite the same system as this for my primates facility which is small in
its scale, but there are lots of helpful ideas and techniques which I will take
for granted as a "Bible" for my works with monkeys and I am sure that
these ideas will greatly help me in making future plans .
Page
15 Homeland of cynomolgus monkeys - A compound of Mr. Nam Chin Law in Malaysia
His compound is made of 3 buildings , entrances of which . were all
double-doored. In the first
building for monkeys to be exported we saw 3 pig-tailed macaques and 5
squirrel-like animals which are individually caged and at the inner most of the
building 'there were 70 cynos gazing at us anxiously.
We made appraisal by their first appearances; first grade monkeys we
judged were big ,enough and not excreting diarrhea], stools .
Mr. Law checked their conditions and marked the cages by chalk.
He didn' t take any tools to catch monkeys but his hands . we could
choose only 25 monkeys . Thus, Mr.
Law promised us to supply another 15 first- grade monkeys for us later on.
In the second building where they carry younger than 1-year- old monkeys
there were baby pig-tailed macaques whose attitude reminded me of our Center 's
juveniles .
He also feeds birds at his compound.
In the 3rd building many kinds of birds were fed and among them I found
again 50 or 60 squirrel-like animals. I
asked the nami and the answer was "Tree Shrew". which of course, I
know as "Tupaia." It was
not surprising that they were treating them as birds .
This coincidence excited me .
Page
16 Sketches from animal rooms
Page-16
Sketches from animal rooms
1)
As an animal technician of African green monkeys
Their charactears are as
follows;
Body
length: 40 cm
Tail
length: 35 cm
Body
weight: male 3.9 kg, female 3.0 kg
Color
of face: black
Color
of scrotum: bright blue
Color
of penis: crimson
Distance
of between two teats: about 1 to 2 cm and baby sucks the two teats together
Attitude:
green monkeys usually rapidly move their body vertically
Wearing work-clothes in the morning, I start examining general health
conditions of monkeys which has never been a simple work to me.
It requires both experiences and careful attentions.
Their health conditions are not necessarily the same every day.
I come out of animal quarters after cleaning cages and giving them food
(100 g of apples and oranges each). In
the afternoon observational data taken in the morning are transcribed in the
nursery record both in the whole and individual categbry.
Then, we go into the quarters again to help perform medication,
treatments and investigations. After
cleaning the quarter and giving them food (cubed diets of about 100 g), I make
afternoon records. We have got 45
babies already and I am happy to see their faces turn into black gradually.
2)
From kindergarden to high school
After weaned from mothers, a certain period of social adjustment is set
for rearing juvenile monkeys.
Adjustment 1. They have to feed themselves with competition in obtaining
food.
Adjustment 2. In a social group they have to be mentally stable.
Adjustment 3. Physical conditions have to be controlled by themselves.
Each of these adjustment steps might be compared to those of human
beings, from kindergarden to high school. Juveniles
of both species are needed to adapt themselves properly to the society they
belong.