TPC NEWS Vol.4, No.1, 1985 (Whole Number 7)

( English Summary )

 

Page-3 Essay: What I think after my contribution to Adv. Sci. & Com. Med. Vol., 28

     Recently, Advances in Veterinary Science and Comparative Medicine Vol. 28 (Academic Press) has been published.  Its main theme is 'Research on Nonhuman Primates".  Eleven papers appeared in the volume edited by Dr. A. G. Hendrickx, Associate-Director of California Regional Primate Research Center.  Nine papers of them were written by American Scientists, one by Indian Scientist and one by us, Japanese (Honjo, Cl-m and Terao).  As Dr. Hendrickx and I are acquainted with each other, he asked me to contribute to the book, in July 1982.  I sent my manuscript to him at the end of May 1983, and the time when I read the first proof was the middle of June 1984.  Then, I received the book issued at the end of November.  Time had flown like an arrow.

     At first I was requested by Dr., Hendrickx to write a review article concerning artificial breeding of nonhuman primates.  I, however, tried to give an explanation of our general concept, as precisely as possible, regarding the process of establishing nonhuman primates as laboratory animals, while I made it an important point to discuss the problems of breeding.  Although this is, more or less, diagrammatic expression, we conceive that the process of "laboratory conditioning" of nonhuman primates is the process which be developed gradually from the stage of Microbiologic Quality Control to the stages of Physiologic Quality Control, Reproductive Quality Control and Genetic Quality Control.  Furthermore, we think that the final stage of this process is to search and fix a certain genetic trait that will be useful to specified experimental purposes in genetically well-controlled breeding colony.  However, our primate center has not yet come to this final stage.

     By the way, in the section concerning "Reproductive Quality Control", we showed a lot of data demonstrating that the breeding system established for wild-originated cynomolgus breeders can satisfactorily be applied to breeders of successive generations (F1, F2...).  I am sure that these data are considerably essential.  In the section of "Genetic Quality Control” the data shown were not so abundant since our experience and knowledge about this stage were not yet sufficient.  However, it was proved by using several genes as markers that the Rotation Line Breeding System by different countries of origin (RLBS) which we designed five years ago was well functioning.  Any way I believe we were able to give an outline of the principal concept of our works.  Probably our paper will be the first one which represents the concept or way of genetic quality control in large-scale breeding colony of nonhuman primates.

     Now, I am afraid that there may be another problem, that is, how relevantly I could have expressed the way of thinking of us, Japanese Scientists, to foreign readers.  As to a certain term, I had to make several exchanges of letters, telegrams and of direct talk with Dr. Hendrickx.  When we make a presentation of experimental data to foreigners in a special paradigm of science or technology, I think it is comparatively easy to get common understanding.  But I really feel that it is not so easy to understand the deep meaning of assertions or dissertations written by foreigners, whose scientific background, whose way of thinking or grasping things and even whose way of feeling may be fairly different from that of us, Japanese.  Indeed therefore, I think we need international cooperation of study.  At all events, since I received the book, I have felt keenly that if we would understand and cooperate each other, going over the difference in the way or conception of learning, the firm base for the stable progress of science and technology in human society would be established.

 

Page-4  Breeding Topics:  Birth of a cynomolgus monkey of the 5th-Generation and a Green Monkey of the 3rd-Generation

     (1) On October 15, 1984, we had the first birth case of F4 cynomolgus monkey.   The baby was a male, weighing 298g.  Both father and mother monkey are of the fourth-generation (F3).  Mother monkey well treated her placenta, cutting the umbilical cord after she gave birth to her baby.  She cleaned baby's skin and had sufficient milk secretion.  Everything associated with her parturition was normal.

     This case is a typical example of successful mothering of colony-bred and -reared monkeys. The rearing record of this mother monkey will be a good reference: She weighed 310g at birth and was weaned 16 weeks after birth.  Following seven months of paired feeding with a male peer of about the same age, she was kept in a group feeding cage with other four monkeys for two and a half years.  After she was confirmed to draw regular cycles of menstruation, she was mated with a male breeder for 3 days at her optimal mating time and became pregnant by the 2nd timed- mating.  At 18 weeks of gestation age she was grouped with other parous female breeders in order to make her observe and learn the other monkey’s behaviors of parturition and infant-nursing.  Thus she had opportunities to see deliveries of her seniors six times before her own delivery.

     (2) On January 2, 1985, a colony bred and -reared green monkey (Fl) was delivered of a male baby (F2) at last.   This mother green monkey was weaned at the age of three-months and began to live with a male monkey of about the same age.   While being reared, she also had several chances to observe mating behaviors, deliveries and nursing behaviors of wild-originated breeders from her own cage.  Now she is nursing well her baby.

 

Page-5 A Survey of Campylobacter Infection in the Cynomolgus Monkeys

     Campylobacter is known as a causative agent of diarrhea in man.  Recently Campylobacter infection has become a serious problem in nonhuman primate colonies.  Thus, we have tried to isolate Campylobacter from stools of cynomolgus monkeys in our colony so that we know epidemiology of Campylobacter infection. The results are shown in Tables 1 and 2.

    As for imported monkeys, the infection rate decreased with the increase of feeding period at TPC.  These wild originated monkeys were usually kept in individual cages except at the mating time.  Of colony-bred and -reared monkeys, 0 year old infants showed high positive rate (51.4%).  With respect to stool condition, there was no definite relationship between the infection rate and the property of stools as far as newly imported monkeys were concerned.  Regarding the monkeys fed for a long time at TPC and the colony-bred and -reared monkeys, however, the positive rate of Campylobacter infection in diarrheal stool was higher than that in normal stools.

     Table 2 shows the infection rate of Campylobacter in monkeys who were suffering from diarrhea for more than 6 weeks.  Also here, infant monkeys had a higher positive rate.

     Species and types of Campylobacter we isolated in this survey were as follows; Campylobacter jejuni C.fetus subsp. fetus, C. coli and C. laridis.

     We have to make further studies on the relationship of Campylobacter infection and diarrhea.  Moreover, we have to consider and establish some adequate measures to eradicate these pathogenic agents from our monkey colony.

   Table l.  Infection rate with Campylobacter in cynomolgus monkeys.

   Table 2.  Infection rate based on the ages with Campylobacter in monkeys suffering from diarrhea for more than 6 weeks.

 

Page-6  Five Big News at TPC in 1984

1.   A male cynomolgus monkey of the 5th-generation was born in October.

    Birth weight: 289g.

    Gestation length: 169 days.

    Maternal behaviors are normal.

2   A F1-female green monkey became pregnant at the age of 2 years and 11 months as a result of mating for three days with a male of wild origin.

3.  The first foster nursing in the green monkey colony at TPC was successful, (See, TPC News Vo;l.3 No.2 "Breeding Topics)

4.  The parturient period of the squirrel monkeys has expanded.

    Only July         in 1981

    July to October    in 1982

    June to October   in 1983

    May to September in 1984

5   The incestuous relation was evidenced in a gang cage of cynomolgus monkeys.   The gang cage was holding one adult male and eleven females of which six were daughters of the adult male.  The five females were mothers of the six daughters.  One of six daughters became pregnant and gave birth to a male baby at the age of three years and nine months.  Thus, this baby is both a son and a grandson of the adult male.

Page-6  Quarantine Report of Monkeys Recently Imported

     Three tables are showing the data we obtained.   ( See attached page )

 

Page-7 Introduction of the Equipment: Domestic-generator

     TPC is provided with the electric power of 1000 KVA by Tokyo Electric Co..  Occasionally, the electric failure happens because of some reasons such as constructions carried out by the power company, typhoon and thunderbolt etc..  It will do great damage and confusion not only to the lighting, air conditioning of animal facilities without windows but also to important laboratory equipment such as ultracentrifuges, fraction collectors and freezers.  Domestic generator was equipped to prevent such disorder.

     When an electric failure happens, several bulbs of 60 watts in each animal room are turned on instantly by the electricity sent from a storage battery.  At the same time the engine of dynamo starts to function by compressed air.  After forty seconds from the occurrence of electric failure, the generator can provide about one-third of the electric power needed to drive the most important equipment set at TPC.

     The electric generator has a capacity of 750 KVA, yielding power by a diesel engine of 900 H.S..  The engine can be worked continuously for 36 hours with light oil as fuel which is pumped up from the underground tank.  As the engine makes big noise and heat, soundproof walls and cooling system are fixed.

     Since TPC was opened, this domestic generator has proved its usefulness twice in the cases of electric failure for a long time by typhoon.  "Readiness is all.'  We made test-operations of this generator once a month following a careful examination for its mechanical structure.

 

Page-8  Case Report:  Malignant lymphoma in a green monkey Adult T-cell leukemia (ATL) in man results from ATL virus (HTLV-1) infection.

     In Southern Kyushu of Japan, ATL patients are frequently found.  The shape of their lymphocytes is distinctively peculiar.

     As for nonhuman primates, Miyoshi et al, first discovered that wild Japanese monkeys were infected with ATLV-like virus (STLV) at high ratio.  But pathogenicity of this virus to man as well as oncogenicity to monkeys were not known at all.

     Recently, we have found the first case of ATL-like disease in the African green monkey and made clear that STLV was oncogenic to nonhuman primates, in collaboration with Dr. Hayami and his colleagues of The Institute for Medical Science, the University of Tokyo.

Case: A female African green monkey, weighing 2.3 kg.  Her estimated age was more  than 4 years old.  She was parous twice at TPC.  She lost her appetite and activities 7 days before death.  By the abdominal palpation, swelled spleen was recognized.

Hematological Findings: Total leucocyte count was 14,100/m3 of which 52.5% were abnormal lymphocytes, Neutrophils 24.5%, lymphocytes 69.5%, monocytes 5.5%, basophils 0.5%.  Total erythrocyte count was 1 , 770 ,0u/mm , hematocrit value 13.1%, hemoglobin content 4.8g/100m1.

Virological and Immunocytological Findings: Anti-STLV titer was 1:160.  Surface markers of the peripheral lymphocytes;  Leu 2a+ (53%), Leu 3a- (40%), Tac+ (50%), IgG+ (29%) on surface and in cytoplasma of lymphocytes.

Autopsy and Histopathological Findings: Enlargement of the lymph node, splenomegaly and a little enlargement of the liver were noticed.  Grayish white nodules of needle head size were scattered or forming cluster in the heart muscle and the kidney.

     Neoplastic cells infiltrating diffusively into lymph nodes were pleomorphic, for example, some of them were medium-sized cells with globular nuclei, some were binucleate cells and others were multi neucleate giant cells. The neuclei of these neoplastic cells showed various shapes such as round, convolution, lobulation and distortion.  Some neoplastic cells had big nucleoli.  Cell divisions were rarely observed.  Proliferation of fibrous tissue was marked and necrosis was also observed. Compartmentalization of neoplastic cells surrounded by fibrous tissue was a characteristic finding.  Infiltration of the neoplastic cells was observed in the liver (Glisson's sheath, sinusoid ), kidney, heart, lung, uterus and bone marrow.

 

Page-9  A Comment on My Research: Bacterial Diarrhea of Nonhuman Primates

     This author who is in charge of bacteriological examinations on nonhuman primates at TPC describes bacterial diarrhea of nonhuman primates from three view points: cause, prevention and treatment.

     He points out Shigella, Salmonella, Yersinia, Campylobacter, Escherichia coli, Proteus and Pseudomonas aeruginosa as main causes of naturally occurring diarrhea of monkeys, explaining their specific traits epidemiologically.  Then some effective ways he has tried to prevent and treat the infections of these causative agents are described, He state also that at present, however, to eliminate completely the carriers of Campylobacter from laboratory primates colonies is difficult.  Further studies of epidemiology and discussions on bacterial diarrhea of nonhuman primates are needed.

 

Page-12  A Special Article:  Needs of genetic studies with nonhuman primates

     Recent progress of biomedical sciences has strengthened the view that fundamental biological functions of living organisms including man could be determined by means of the functions of the genes.  Thus it now becomes exceedingly valuable to obtain information on the genes relevant or harmful environmental agents a$ well as inherited human diseases, For medical and clinical studies, experimental systems examining the gene functions associated with diseases are urgently needed. For this purpose the animal model is indispensable.  Nonhuman primate models are extremely valuable as a human analogue especially for dealing with neurobiological problems owing to the close phylogenetic relationship of nonhuman primates to human.

     Despite of their importance, nonhuman primates have not been utilized so widely as a laboratory animal as compared with mice, since the animal supplies are not easy and their genetic backgrounds are heterogeneous.  Furthermore, due to the limitation of formal genetic analysis by the breeding technique in these animals little genetic information concerning mutant genes, which is necessary for advanced research, is available.

     Advances in genetic research in nonhuman primates, however, will find a way out of such difficulties as mentioned above.  For example, further progress in the recombinant DNA technique could shed the light on the problem of isolation of a mutant line, since the technique detects mutant genes at DNA level without formal genetic analysis.  Therefore, future genetic research in nonhuman primates will be very promising.  Accordingly, to develop a nonhuman primate model for biomedical research, progress in the genetic studies is obviously needed.

     For further genetic studies in nonhuman primates, several fundamental problems must be solved.  Clarifying genetic status of nonhuman primates in relation to man is important for utilization of the animal model as a human analogue.  Another important subject is the establishment of breeding colonies of nonhuman primates, which is prerequisite to genetic studies.  Appropriate consideration and judgment about the maintenance of breeding populations, as well as preservation and distribution of genetic information are strongly desired.  In addition, to establish the technological back-up systems which have such works actually performed.

     I think that national and international collaboration is indispensable to realize these researches and works.  I am greatly expecting TPC as a center of international cooperation and its endeavors to make a great advance in genetic studies with nonhuman primates.

 

Page-14  A Clinical Commentary b a Cub Veterinarian : A monkey whose tongue was injured

     One day I received a wireless call informing that a male monkey aged 1 year was wounded and lost activities in a group cage, moreover, there occurred prolapse of the rectum with him.

     Immediately I anesthetized the monkey for treatment.  His wound in the back was not serious.  As for the rectal prolapse of about 3cm length, I could push it back slowly into the anus, since it had not degenerated.  Next, when I opened his mouth to examine the oral mucosa, I nearly shouted to see something dark-red that was squirming in his mouth.

     It was his own tongue.  An upper half of the tongue was sliced off., In addition to that, its inner part was lacerated.  I had never read such a case in any textbook of veterinary surgery or primate medicine.  How should I suture these wounds?

    I cut off the end of a cylindrical plastic case for an injection needle and put it between the monkey's upper and under canine teeth to keep the mouth opening.  I pulled the tongue out with an intestine forceps.  Then I began to suture the lacerated wound using a short needle and a fine silk thread.  There was no difficulty to suture it.  Problem was to suture the part sliced off.  After serious considerations, I decided to fold back the tongue double and to suture.  In this way the tongue might be rather shortened, but I thought it would be better than the muscle of the tongue would remain exposed.  As a matter of fact, it was so difficult to fold the tongue being originally straight.  With very hard effort I finished suturing.  The tongue became ill-shaped.

     In the afternoon, I saw the monkey recovered from anesthesia ate fruits as usual without showing any painful look, though he left some hard diets.  I gave him fruits, nutrients medicine and baby biscuits for about a week.  After ten days, he got well.

     By the way, who was an assailant?  I found a female monkey whose mouth was smudged with blood, in the same group.

     Now, this monkey with the short tongue is being kept in an individual cage.   He is very well but he would not be able to pronounce "R" sound well any more .

     I would like to ask you, readers of TPC News, to tell me the better treatment and your experience in the case similar to this.

 

     Page-15  The Third TPC Symposium Held under the Joint-Auspices of the Japanese Committee of U.S.-Japan Science Cooperative Program on Laboratory Primates

     The third TPC symposium titled "Establishment of Genetic Control in Laboratory Primates Colonies' was held at Tsukuba Center for Institutes on December 7 in 1984.  The aims of this symposium were to put in order genetic knowledge of nonhuman primates obtained so far and to discuss how to practically use these knowledge for genetic control of laboratory primates colonies.  Furthermore, we discuss the strategy of genetic studies using nonhuman primates in the future. Seven fruitful lectures including that of Dr. Smith from the University of California and lively discussions were made.

     According to the opinions of attendants, this symposium was successful in providing some practical strategies applicable to genetic control of nonhuman primates breeding colony.  However, it is needless to say that, to obtain more fruitful results, further studies, for example, studies on the establishment of inbreeding- strain and on the problems of inbreeding depression are needed.  Cooperation of researchers of both basic and applied sciences are strongly desired.  The record of this symposium will be published in Japanese J. Med. Sci. Biolo., Vol., 38 (1985) .

 

Page-16  A Memory of a Trainee: From a diary I picked up

     One day a visiting researcher from Kyoto University picked up a diary on the floor of a cynomolgus monkey room.

     “'..... Ten years have passed since I was brought here from Indonesia.  This center, being compared to "another planet" by the Director of the Regional Primate Research Center at the University of Washington, is very nice.  Here, we, monkeys, are all equal.  There in no need to contend with each other for the post of boss.  We little suffer from hunger or thirst, far from that, we are given even apples and oranges every day.  It is quite comfortable, the room temperature is not so hot or cold as in Indonesia.  Cages are clean and water is so, too.   We need not feel anxious about dysentery or viral diseases.  When we suffer from diarrhea, we are given bananas containing a dose of medicine immediately.  This is quite a paradise on earth.

     But, I wonder if I keep being here, I will end my life as a good-for-nothing.  I want to be a monkey with distinct individuality, to become bigger than any other monkeys and to be alive longest.  Here, however, all the monkeys look alike in their body size and character.   I think I have capacities to be a boss.   But I am not given any chance to appeal my ability to others.  Will I have to end my life as an obscure monkey?

     Although I want to have a time to think deeply alone, I am always observed by many eye.  0h, I shall be mad!  I heard, Dr. Honjo, Director of this center, once wrote an essay titled "Do monkeys have a mind or a heart?'  in TPC News. What a thing he said.  I desire a pretty lady monkey for my mate.  Why am I always mated with the same one?  I do have a right to select my mate.  I wonder there are any means to tell her my feeling.  She is very charming I think....  I can't sleep tonight.....  Light went out.  Night has come.  I feel sleepy.  Tomorrow, I will think of it again”.

 

Page-17 Sketches fr6m Animal Rooms: Three essays

     An animal technician and two fresh animal caretakers describe what they think and feel through their daily works.

(1) It is not too much to say that the animal technicians of TPC have mastered the techniques to handle well whatever monkeys we are keeping in this center.

     We, animal technicians, ate taking care of the monkeys by rotation in different four buildings, Quarantine, Breeding, Rearing building and Housing of candidate- and retired-breeders. When my charge was changed from quarantine to breeding eight months ago, I had to tackled my new job as if I were a freshman of this center.

     Though the way to be expert animal technician is difficult, I work with monkeys, feeling a sense of fulfillment and freshness every day.

(2)   Two months have passed since I began to work for this center.  My present tasks are feeding of animals and cleaning of animal rooms.  I heard this period was important to cultivate a good eye for animal health-conditions.  So I am doing my best in daily works, enjoying to take care of monkeys.

(3)  I am a fresh animal caretaker.  One day I saw a monkey injured seriously.  He died in spite of all the effort and medication we had taken for him.  His death gave me a chance to think about the value of life.  Science that time I have tried to observe monkeys more carefully, because a very little wound may result in serious illness.