Taiwan Monkeys Research Articles

Response of Regenerative Tissues to Plaque: A Histological Study in Monkeys

This study was designed to determine the histologic response of regenerated tissue to plaque, using an adult Taiwan monkey model with chronically-inflamed, surgically-created, periodontal defects. Standardized 2-walled periodontal defects were surgically created at the mesial of the mandibular lateral incisor on one side. Sutures, 3-0 braided silk, were placed for 8 weeks and tissues positioned so that denuded root surfaces were exposed to plaque, with the subsequent development of the chronically-inflamed periodontal tissues. At 8 weeks the defects were subjected to guided tissue regeneration procedures using expanded polytetrafluoroethylene membrane and allowed to heal for 6 months. Contralateral clinically healthy sites, without surgical procedures, were used as controls. In both experimental (membrane) and control sites, silk ligatures were placed to allow plaque accumulation for 2 or 10 weeks. Four monkeys were sacrificed at each time point and specimens processed for histologic and histometric study. The results indicated that plaque-induced inflammation was less at sites treated by guided tissue regenerative procedures when compared to originally clinically-healthy sites exposed to plaque for the same duration. However, histologically the newly formed osseous tissue was compromised under these conditions.

Phylogeny of rheusus monkeys (Macaca mulatta) as revealed by mitochondrial DNA restriction enzyme analysis

Mitochondrial DNA, purified from 36 samples of 23 local populations which are widely distributed in Vietnam, Burma, and 10 provinces of China, has been analyzed to model the phylogeny of rhesus monkeys. The 20 local populations of China may represent nearly all major populations in China. Using 20 restriction endonucleases of 6-bp recognition, we observed a total of 50–61 sites in the various samples. By combining the cleavage patterns for each enzyme, the 36 samples were classified into 23 restriction types, each of which was found exclusively in the respective population from which samples were obtained. By combining the earlier study of Indian rheusus monkeys, phylogenetic trees, which have been constructed on the basis of genetic distance, indicate that rhesus monkeys in China, Vietnam, India, and Burma can be divided into seven groups. Integrating morphological and geographical data, we suggest that rhesus monkeys in China, Vietnam, and Burma may be classified into six subspecies—M. m. mulatta, M. m. brevicaudus, M. m. lasiotis, M. m. littoralis, M. m. vestita, and M. m. tcheliensis-and rhesus monkeys in India may be another valid subspecies.M. m. tcheliensis is the most endangered subspecies in China. Divergence among subspecies may have begun 0.9–1.6 Ma. The radiation of rhesus monkeys in China may have spread from the southwest toward the east. The taxonomic status of the Hainan monkey and the Taiwan monkey require further investigation.

Micro-immunofluorescence antibody responses to trachoma vaccines.

We evaluated antibody responses to trachoma vaccine in Taiwan monkeys using the micro-immunofluorescence assay. In the experiment with B/TW-5 gradient and genetron vaccines, the level of antibody titers could be correlated with the vaccine dose administered. Immunotype-specific protection from disease or modification of disease severity was associated with higher homologous antibody titer. In the experiment with A/G17, A/SA-6, and C/TW-3 genetron vaccines, an extraordinary high antibody response was demonstrated. Again, the high homologous antibody titers were associated with complete or partial protection from eye disease caused by the eye challenge inoculations. Cross reactions between immunotype A and C in the micro-IF test were also reflected in the vaccine protection observed in this experiment. It is concluded that there is an excellent correlation between the micro-IF antibody responses, the mouse toxicity prevention test, and vaccine prevention of monkey trachoma eye disease. Study of micro-IF antibody responses should be a useful tool for development of effective trachoma vaccine.

Effects of intraventricular administration of sympathomimetic agents on metabolic,vasomotor and temperature responses in Taiwan monkeys.

Intraventricular administration of either norepinephrine, tyramine or angiotensin II produced dose-dependent hypothermia, decreased metabolism and lowered cutaneous temperatures in Taiwan monkeys at ambient temperatures of 4 and 22 degrees C. The hypothermia reflected the heat storage consequent to the net effects of decreased heat production and decreased heat loss. In heat (35 degrees C), these agents produced dose-dependent hyperthermia in monkeys. The hyperthermia was brought about mainly by a decrease in dry heat loss.

Experimental transmission of Wuchereria bancrofti to monkeys.

Infective larvae of Wuchereria bancrofti from laboratory-raised Culex pipiens fatigans and Aedes togoi mosquitoes fed on human volunteers in Jakarta, Indonesia (J strain) and Kinmen Island, China (K strain) were introduced into Taiwan monkeys (Macaca cyclopis) by subcutaneous inoculation, by foot puncture, or by permitting infected mosquitoes to feed weekly on the monkeys. Some animals were splenectomized and others were treated with varying regimens of immunosuppressants. Necropsy was done on monkeys that died or were killed and the entire bodies were examined for worms. A total of 78 monkeys (43 males and 35 females) were exposed to infection and parasites were found in 29% of the females and 63% of males. In infections of 38 days or less worms were recovered from the testes of males and the pelt, carcass and lymph nodes of both sexes, but after 42 days of infection most worms were in the testes of males, and a few were recovered from lymph nodes and carcasses of females. Worms recovered at 8-11 days were third-stage, those found between 14 and 38 days fourth-stage, and ones found between 42 and 103 days were young adults. After 148 days most were adults and microfilariae were seen in the uteri of female worms at 160 days and later. The parasites continued to grow in size with time. Microfilariae were detected in the blood of nine monkeys between 8 and 18 months and the patent period varied from 5-21 months. Microfilarial densities were low and erratic, and periodicity could not be determined. The effectiveness of methods of administering infections and the value of various treatment regimens seem uncertain; monkey antilymphocytic sera, however, appeared to have some influence. Parasites were found in 36% of the Taiwan monkeys given the J strain and 54% of those given the K strain. A limited number of M. mulatta (3), M.irus (fascicularis) (3) and Aotus trivirgatus (4) were also given infective larvae and adult W. bancrofti were recovered from the testes of one male M. mulatta and one male M. irus; uterine microfilariae were found in one female worm from the latter monkey. A. trivirgatus were negative. Low numbers of infective larvae recovered from mosquitoes fed on patent monkeys were introduced intermittently into seven clean monkeys and one became microfilaremic between 11 and 17 months postinoculation.

Nasal Adenocarcinoma in a Taiwan Macaque

Although a variety of neoplasms have been reported in nonhuman primates [ 5 ] , tumors of the nasopharyngeal region are rare. Amelobastic odontomas have been found in the Celebes [2] and Rhesus monkeys [6]; oral soft tissue tumors have been found in the spider monkey [4], and bone neoplasms of the cranial-facial region of Rhesus monkeys [ 3 ] . We report a nasal cavity adenocarcinoma with metastasis t o the lung in a Taiwan monkey, Mucaca cyclopis. The macaque was a 10-year-old male that had been housed with 20-25 healthy monkeys in a primate holding room. The other monkeys were used for trachoma virus studies in Taiwan. The macaque had not been used in any study for 18 months prior to discovery of the tumor. A mass about 3 centimeters in diameter was found in the left nasal cavity causing swelling in the left maxilla. Biopsy of the lesion showed it to be a papillary adenocarcinoma. The tumor continued to grow to about 6 centimeters in diameter and eventually ulcerated through the nasal dermis. The mass was surgically removed and the face reconstructed by plastic surgery. Three months later the mass reappeared and was again removed (fig. 1). Because malignant nasal tumors are common in Asians, virologists at the College of Medicine, National Taiwan University, requested that the disease be allowed to progress for several months in order to attempt virus isolation from the tumor. These studies were unfruitful. Thoracic radiographs showed pulmonary metastasis. After erosion of facial bone had become extensive the animal was killed. The tumor had ulcerated through half the facial dermis and was encroaching upon the bony orbit of the left eye. The protruding tumor was gray-tan and friable. In addition, the tumor had replaced the anterior half of the hard palate including the associated dental arcade. Cutting was easy in this area. All lobes of the lung contained foci of various sized metastatic tumors which were harder t o cut than was the primary focus. There were no metastasis in other organs o r structures. The biopsy (fig. 2) taken about a year before the macaque was killed showed papillary adenocarcinoma of interconnecting fronds and islands of neoplastic cells. Mitotic figures were numerous. The neoplastic foci in the lungs tended to be solitary adenocarcinomatous nodules (fig. 3) with numerous mitotic figures. Special stains showed mucin in some areas. Since the history on this monkey included exposure to the trachoma virus only and since it had not been exposed to any other experimentation, speculation on

Reproductive parameters of the Taiwan monkey (Macaca cyclopis)

The following parameters of the Taiwan monkey,Macaca cyclopis, are presented and compared with other species ofMacaca: the menstrual cycle, sexual skin and vaginal desquamation changes during the menstrual cycle, time of ovulation, gestation period, breeding season, body weight of the newborn, age and body weight at menarche, body weight at first conception, spermatozoa count in the ejaculate, and the body weight at sexual maturity of the male. The many similarities in reproductive biology ofMacaca cyclopis, the rhesus monkey (Macaca mulatta), and the cynomolgus monkey (Macaca irus) include sexual skin and vaginal desquamation changes during the menstrual cycle, time of ovulation, gestation period, and placental sign. Body weight ofM. cyclopis is lower thanM. mulatta at birth, at menarche and at the first conception in females and at sexual maturation in males, is heavier thanM. irus at birth and is not different fromM. radiata at birth. No difference in menstrual cycle ofM. cyclopis was observed in animals housed in air-conditioned rooms compared to those housed in nonair-conditioned rooms. Summer amenorrhea was not observed inM. cyclopis but a high incidence of low vaginal desquamation was noted to occur in summer months. The mode of length of the menstrual cycle ofM. cyclopis is shorter thanM. mulatta and other species ofMacaca. The breeding season ofM. cyclopis in the wild extends from the end of September to January. In laboratory conditions their fertile period extends throughout the year.

Antibodies to Epstein-Barr virus in a Burkitt's lymphoma cell line in Taiwan monkeys (Macaca cyclopis).

The prevalence of Taiwan monkeys with antibodies to Epstein-Barr virus, as detected by the indirect immunofluorescence technique, was lower than that of healthy persons in Taiwan.

Antibodies to Epstein-Barr Virus in a Burkitt's Lymphoma Cell Line in Taiwan Monkeys (Macaca cyclopis)

The prevalence of Taiwan monkeys with antibodies to Epstein-Barr virus, as detected by the indirect immunofluorescence technique, was lower than that of healthy persons in Taiwan.

Incidence of neutralizing antibodies to Herpes B virus in the Taiwan monkey (Macaca cyclopis)

A group of 46 recently captured Taiwan monkeys (Macaca cyclopis) was tested for neutralizing antibody for Herpes B virus. Results indicated that 11 (24%) were positive, 25 (54%) were negative, and 10 (22%) were suspect. This is thought to reflect the incidence in nature. Subsequent samples from the same animals, acquired 7 months postarrival, indicated 10 (22%) positive, 27 (59%) negative, and 9 (20%) suspects. A number of animals (18) converted from one category to another, assumedly from loss or acquisition of immunity during the period. A suggestion of geographic difference in incidence of titers was detected on the first sample. Twenty-eight human caretakers and veterinarians were tested with no positives, 25 negatives and 3 suspect.

Experimental trachoma in owl monkeys.

Experimental trachoma in nonhuman primates has been handicapped because trachoma (as opposed to inclusion conjunctivitis) produces very mild clinical disease and minimal microbiologic changes. In a direct comparison between the Old World Taiwan monkey, Macaca cyclopis, and the New World owl monkey, Aotus trivirgatus, the latter was found to be much more susceptible. Owl monkeys manifested severe disease for a period of 2 or more weeks, while none was observed in Taiwan monkeys during the same period of time. Given identical inoculations of 2,000 EID50 in the left eye and 20 EID50 in the right eye, six Taiwan monkeys on the 7th day averaged 21 inclusions in the left eye and one in the right. Comparable figures for six owl monkeys were 641 and 1,396, respectively.

Plasmodium (Vinckeia) watteni sp. n. from the Formosan Giant Flying Squirrel, Petaurista petaurista grandis

A new species of malaria Plasmodium watteni has been found in the Formosan giant flying squirrel, Petaurista petaurista grandis. It is similar to P. booliati from the Malayan flying squirrel except that the trophozoites and schizonts do not exhibit a vacuolated or foamy appearance; the pigment in the trophozoites and gametocytes is more conspicuous and abundant; the mature schizont nearly fills the host cell and has a maximum of 18 merozoites; and the gametocytes fill the host cell. Laboratory rats, mice, hamsters, guinea pigs, the Taiwan monkey (Macaca cyclopis) as well as splenectomized rats and guinea pigs were resistant to experimental transfusion infections. Periodicity studies in the flying squirrel carried out over a 9-day period failed to show synchronism in the asexual cycle of the parasite. During the course of field investigations for parasites of the blood of lower animals in Taiwan, an undescribed species of Plasmodium was found in the Formosan giant flying squirrel, Petaurista petaurista grandis. The senior author first found the parasite in four animals in November 1964 and January 1965 (Sandosham et al., 1965). In further studies since December 1966 additional specimens have been obtained from a number of flying squirrels and the present paper describes the erythrocytic stages of this new species of malaria. MATERIALS AND METHODS Petaurista petaurista grandis (Swinhoe, 1862) is one of the commonest species of the subfamily Petauristainae of the family Sciuridae found on the island of Taiwan. According to Horikawa (1931) this animal inhabits forested hilly areas with elevations of 100 to 2,500 m. It is nocturnal and seen more frequently at night in the tops of trees. In the daytime it retires into large tree holes. Its diet consists of fruits and young leaves. Blood smears were obtained from captured animals or from animals shot from treetops. Thick and thin blood smears were stained with buffered Giemsa at pH 7. Heparinized blood containing the parasite was obtained from one flying squirrel and inoculated into another squirrel in which preinoculation blood smears were found to be negative. Attempts were made to determine the asexual cycle of the parasite by obtaining blood smears at Received for publication 31 May 1968. * The opinions and assertions contained herein are those of the authors and are not to be construed as official or reflecting the views of the Navy Department or the Naval Service at large. This study was supported in part by funding under Public Law 480, Section 104(c), and in part through funds provided by the Bureau of Medicine and Surgery, Navy Department, for Work Unit MF022.03.07-2018. 4-hr intervals for 9 days. The results of this study failed to determine the duration of the asexual cycle of the parasite, and further attempts were unsuccessful because of the high mortality of the flying squirrels in captivity. In an effort to find exoerythrocytic stages of the parasite, serial sections of the livers of five squirrels were examined histologically. No exoerythrocytic stages were found. In other studies parasitized blood was inoculated into 3to 4-week-old laboratory rats, mice, hamsters, guinea pigs and one adult Taiwan monkey (Macaca cyclopis). In addition, experimental infections were attempted in splenectomized rats and guinea pigs. Infection failed to develop in any of these laboratory animals. Plasmodium (Vinckeia) watteni sp. n. (Figs. 1-35) Vertebrate host: Petaurista petaurista grandis (Swinhoe, 1862). Invertebrate host: Not known. Locality: Peiyuan, Tungho Township, Taitung Hsien, Taiwan Province, Republic of China. Date of collection of type host: 1 February 1967. Type material: The morphological description of this species is based on a study of Giemsa-stained blood films obtained at 4-hr intervals for 9 days from a flying squirrel which received a secondary passage of the parasitized blood. The type slides will be deposited in the Department of Medical Ecology, U. S. Naval Medical Research Unit No. 2, Taipei, Taiwan, Republic of China. This malaria parasite is named in honor of Captain R. H. Watten, MC, U. S. Navy, Commanding Officer of the U. S. Navy Medical Research Unit No. 2, Taipei, Taiwan, for his encouragement and interest in parasitology. Description of the erythrocytic stages Host cell (Fig. 1): Uninfected host erythrocytes 4.6 to 7.0 ,L in diameter, average 5.4 Au. Parasitized

COMPARISON OF SIMIAN FOAMY VIRUS STRAINS INCLUDING A NEW SEROLOGICAL TYPE.

CYTOPATHOLOGY noted in spontaneously degenerating cultures of monkey kidney tissue, which was characterized by multinucleated giant cells and foamy appearance1–4, was found to be caused by a transmissible virus that multiplied in such cultures, and antibody to the virus was found in monkeys5–7. Two distinct serotypes of virus with foamy cytopathic effect were isolated from monkey kidney tissue derived from Taiwan monkeys (Macacacyclopsis) and from throat swabbings from such monkeys, and were designated foamy virus Types I and II by Johnston8. Foamy virus isolated from monkey kidney tissue derived from several other monkey species—cynomolgua, vervet (Cercopithecus pygerythrus centralis) and rhesus—was serologically identical to the Johnston Type I and also to the SAI virus of Malherbe and Harwin9, and was unrelated to simian virus Type 5 (SV5) or to the monkey intranuclear inclusion agent10.

A Study Of Filaria In Taiwan Monkeys.

Filarial adult were found in the intermuscular connective tissue of the peritracheal and mandibular region of the Taiwan monkey(Macaca cyclopis). The microfilariae in the peripheral blood demonstrated a marked nocturnal periodicity with a peak between 2100-0100 hours and a low diurnal minimum between 900 and 1300 hours. Staining the microfilaria by various stains demonstrated differences in size. No development of microfilariae was detected in the Culex fatigans, Culex tritaeniohyncus, Aedes albopictus, Anopheles minimus, Armigeres subalbatus or Mansonia uniformis mosquitoes.

NEPHROTOXIC SERUM RENAL DISEASE IN THE TAIWAN MONKEY.

Abstract Experimental renal disease was produced in 5 Taiwan monkeys (Macaca cyclopsis) by the intravenous injection of anti-monkey-kidney serum obtained from rabbits. All developed proteinuria and statistically significant hypoproteinemia. Four monkeys revealed statistically significant reduction of the albumin fraction. Two monkeys had generalised edema, and 3 showed statistically significant hypercholesterolemia. These clinical abnormalities were associated with histopathologic renal lesions which closely simulated those of the nephrotic syndrome as observed in children. In 7 other monkeys receiving rabbit anti-monkey-kidney serum, the disease appeared to be intensified by the administration of ACTH or prednisolone before or during heteronephrotoxic serum injections. Obvious generalized edema associated with ascites and hydrothorax was observed in all animals. In addition to the foregoing changes, statistically significant increase in values of non-protein nitrogen and urea nitrogen in the blood was observed in 6 of the 7 monkeys. Evident gross hematuria was noted in 2 young monkeys. Significantly increased serum α 2 -globulin fraction was observed in 4 monkeys. More prominent hypercholesterolemia was noted in 5. Attempts to produce experimental renal disease by the intravenous injection of anti-monkey-kidney serum obtained from ducks failed in Taiwan monkeys, and ACTH and prednisolone had no significant effect on the clinical and histologic response.

Experimental Whooping cough.

MANY questions about the immunologic mechanism responsible for the permanence of immunity after an attack of whooping cough are still obscure, in spite of a large number of clinical and experimental studies. It was considered worthwhile, therefore, to reinvestigate the problem in animals under more uniform conditions. The following is a report on such studies in Taiwan monkeys in which paroxysmal cough developed after an inoculation of Haemophilus pertussis. Materials and MethodsThe animals used in the experiments were normal young Taiwan monkeys, Macaca cyclopsis, weighing from 550 to 1875 gm. All monkeys were kept under preliminary observation to . . .