Movement of cynomolgus and rhesus monkey spermatozoa collected from the lower female reproductive tract

Postcoital (pc) cervical mucus was collected in 73 menstrual cycles of cynomolgus monkeys and in 43 cycles of rhesus monkeys at 2,6,10,30 hr pc. Videomicrography was used to analyze sperm numbers and movement in the mucus. Both cynomolgus and rhesus monkeys had comparable populations of motile sperm in the mucus at 2 hr pc. However, by 6 hr pc, cervical mucus from cynomolgus monkeys contained twice as many total sperm and motile sperm as mucus from rhesus monkeys (P <.05). Mean swimming speeds of the free-swimming cervical sperm were similar for the two species at this time. No motile sperm were recovered in mucus from rhesus monkeys at 30 hr pc. In cynomolgus monkeys, however, 14 of the 26 animals examined at 30 hr pc had motile sperm in their mucus. These sperm exhibited lower percent molility, percent free-swimming sperm, and swimming speed than those sperm observed at 6 hr pc. Uterine sperm were collected by transcervical or transuterine aspiration from cynomolgus monkeys. In the transcervical technique, sperm were successfully obtained in four of nine animals examined at 6 hr and in four of five animals at 30 hr pc. The percentage of motile sperm in the uterine fluid was high, 82% ± 4%, and the swimming speeds (86 ± 2μm/sec) were higher than those observed in cervical mucus. Approximately 5–10% of the uterine sperm exhibited swimming motions similar to the hyperactivated motility seen in most mammals. These findings indicate that the sperm cervical mucus interaction in vivo in cynomolgus monkeys has more similarities to the human situation than does the interaction in rhesus monkeys.     

人工驯养条件下食蟹猴B病毒抗体水平变化规律研究

目的研究人工驯养条件下食蟹猴B病毒抗体水平变化规律,便于有效控制自繁食蟹猴的BV感染率。方法随机选取409只对不同月龄自繁食蟹猴,采用BVELISA法进行BV抗体监测。结果新生仔猴刚出生时均携带不同程度的BV抗体,但随着月龄的增加,BV抗体水平开始下降,至5月龄时BV抗体阳性率降至最低（12．3％）,之后BV抗体水平逐渐升高。结论人工驯养条件下食蟹猴B病毒抗体水平呈由高到低再升高的趋势,5月龄时断奶可最大限度地获得BV抗体阴性猴。     

Study of color variation in the solitary ascidian Halocynthia roretzi, collected in the Inland Sea of Japan

In the vicinity of Yashiro Island in the Inland Sea of Japan, the solitary ascidian (tunicate) Halocynthia roretzi with tunics of various colors were collected. Samples of these animals were sorted into three groups on the basis of visual observation of tunic color. The red group includes animals with dark-red, light-red, or orange tunics. The pink group includes animals with tunic colors ranging between red and white. The white group includes only animals with completely white tunics. Animals in the white group lacked color internally, with the exception of the hepatopancreas and the gonads in breeding season; the epidermis and gill basket were white. In contrast, animals of both the red group and the pink group were colored internally, with red-orange epidermis and yellow gill basket. Alloreactivity was tested by mixed-hemocyte incubation between different animals belonging to the same color group and between animals belonging to different color groups. Alloreactivity between animals of the white group was 56.3%, between animals of the pink group was 60.0%, and between animals of the red group was 69.3%. The relatively high frequency of compatible combinations among the white animals is discussed.     

Normal hematologic values in the cynomolgus monkeys aged from 1 to 18 years

Normal hematological values including erythrocyte count (RBC), hematocrit value (Ht), hemoglobin concentration (Hb), total leucocyte count (WBC), and differential leucocyte count were determined with 206 healthy cynomolgus monkeys (Macaca fascicularis) aged in 1 to 18 years. These animals were born and reared at Tsukuba Primate Center for Medical Science, NIH, Japan. Additionally, 32 cynomolgus monkeys of wild origin estimated to be 5 or more years old were examined for their hematology. The presence or absence of sex- and age-related differences in the values was statistically analysed. As regards infant and juvenile monkeys, there were no significant differences in RBC, Ht and Hb between males and females. However, these values became significantly larger in males than in females after sexual maturation. Lymphocyte count was larger than neutrophil count until 3 to 4 years of age after birth, but this relation was inverted in adult monkeys aged more than 5 years. Segmentation of the nucleus was a prominent finding in neutrophils. Neither sex- nor age-related difference in the number of nuclear segmentation was noted.     

Direct Staining of Reticular Fibers with Gold Chloride

Reticular fibers are selectively stained in paraffin sections of formalin-fixed or Bouin's-fixed tissue as follows: 1% aqueous solution of gold chloride for 20 min, followed by a 10 min immersion in an aqueous solution containing 5% Na₂CO₃ and 0.5% KOH. The sections then are placed in a 5% aqueous solution of KI for 2 min. Counterstaining with a 0.25% aqueous solution of methylene blue chloride is optional. The reticular fibers stain dark pink; the collagen bundles are a light pink to straw color without the counterstain, or a light blue color when the methylene blue is used.     

Respiration rate, heart rate, and body temperature values in cynomolgus monkeys (Macaca fascicularis) during barbiturate anesthesia

Respiration rate, heart rate, and body temperature values were obtained from 14 cynomolgus monkeys (Macaca fascicularis) during neurosurgery under barbiturate anesthesia. Vital sign values markedly declined below baseline during the early stages of surgery, steadily increased as surgery progressed and neared completion, and finally returned to baseline by the end of the postsurgical recovery period. There was considerable variability among the 14 monkeys, but the ranking of each monkey relative to the others remained constant across the period of observation. The findings suggested that the cynomolgus monkey may be more sensitive to barbiturates than the rhesus monkey, and cynomolgus monkeys may exhibit considerable individual differences in their sensitivity to barbiturates.     

Biologic data of Macaca mulatta, Macaca fascicularis, and Saimiri sciureus used for research at the Fiocruz primate center

Physiological parameters of laboratory animals used for biomedical research is crucial for following several experimental procedures. With the intent to establish baseline biologic parameters for non-human primates held in closed colonies, hematological and morphometric data of captive monkeys were determined. Data of clinically healthy rhesus macaques (Macaca mulatta), cynomolgus monkeys (Macaca fascicularis), and squirrel monkeys (Saimiri sciureus) were collected over a period of five years. Animals were separated according to sex and divided into five age groups. Hematological data were compared with those in the literature by Student's t test. Discrepancies with significance levels of 0.1, 1 or 5% were found in the hematological studies. Growth curves showed that the sexual dimorphism of rhesus monkeys appeared at an age of four years. In earlier ages, the differences between sexes could not be distinguished (p < 0.05). Sexual dimorphism in both squirrel monkeys and cynomolgus monkeys occurred at an age of about 32 months. Data presented in this paper could be useful for comparative studies using primates under similar conditions.     

Persistence and disappearance of hyaloid artery in newborn cynomolgus monkeys

Apparently healthy 45 newborn cynomolgus monkeys (Macaca fascicularis) were examined for the persistence and disappearance of the hyaloid artery by using an ophthalmoscope. On the day of birth, the hyaloid artery was observed in all the newborn animals. Between 21 and 45 days of age, the hyaloid artery faded without exception. The persistence of the hyaloid artery in the newborns and its disappearance during an early period of life can be regarded as a normal phenomenon in the developmental process of infant cynomolgus monkeys.     

A comparison of hematological and serum biochemical values between two groups of female cynomolgus monkeys reared under different conditions

The influence of long term rearing conditions on the hematological and serum biochemical values in laboratory-bred female cynomolgus monkeys (Macaca fascicularis) was studied under the following two conditions. Group A: New born infants were nursed by their mothers for at least 12 weeks after birth. After weaning, four monkeys of approximately equal age and body weight were kept in a relatively small cage (90 cm W x 60 cm D x 60 cm H) and reared to five years of age. These rearing conditions are commonly used in the breeding program of our primate center. Group B: One male and five females of wild origin were introduced into a relatively large cage (180 cm W x 180 cm D x 160 cm H). Female monkeys born in these cages were used in this study. All of them were reared by their own mothers and grew well in these cages. Male monkeys born in these cages were removed at about 3.5 years of age. Hematological and serum biochemical values between the two groups in each age class were compared and analyzed by the canonical discriminant analysis (the discriminant analysis with reduction of dimensionality). Age correlated highly with the value of the first canonical variate. The second canonical variate detected a difference in the blood hematology and biochemistry probably induced by the difference in rearing conditions.     

Hour of delivery in cynomolgus monkeys under indoor individually-caged conditions

The hour of delivery was surveyed in 152 cynomolgus monkeys (Macaca fascicularis) which were kept in individual cages placed in completely air-conditioned and artificially lit rooms. The deliveries took place during light hours (05:00–19:00) in 15 animals (10%) and during dark hours (19:00–05:00) in 137 (90%). No significant differences in delivery hour were observed between animals of feral origin and colony-bred F1 animals. In addition, there was no difference according to gravidity.     

Isolation, primary structure, and biological and immunological properties of pink and chum salmon insulins

1. Insulins have been isolated from islet tissue of pink (Oncorhynchus gorbuscha) and chum (Oncorhynchus keta) salmon. The primary structure of chum and pink salmon insulins was found to be identical. Compared to the amino acid sequence of human insulin, the salmon insulins under study differed at 14 positions. 2. Biological activity of pink salmon insulin was 83% of that of standard porcine insulin. 3. The immunological properties of fish insulins were investigated in specific radioimmunoassay (RIA) systems, based on porcine and pink salmon insulins. 4. A significant difference in the antigenic determinants of these fish and mammalian hormones was revealed.     

Noninstrumented enzyme-linked immunosorbant assay for detection of early pregnancy in macaques

A practical, noninstrumented enzyme-linked immunosorbant assay (NELISA) for the measurement of urinary monkey chorionic gonadotropin (mCG) has been developed for the detection of early pregnancy in macaque monkeys for use in both the laboratory and the field. Five rhesus monkeys (Macaca mulatta) and six crab-eating monkeys (Macaca fascicularis) were tested for the presence of mCG in urine on gestational days (GDs) 12 to 35. The mCG NELISA detected pregnancy as early as GD 14, with an average earliest detection at GD 16.5 +/- 1.4 (n = 11). Out of 90 tests, 27 false-negative and zero false-positive tests were obtained, for an accuracy of 70.0%. Without the aid of a spectrophotometer, the presence of mCG in pregnant monkey samples was indicated by a dark green color change. Nonpregnant monkey urine samples, on the other hand, exhibited no color change. These findings suggest that the simple, economical, and reliable urinary mCG NELISA may be useful for diagnosing early pregnancy in these and related species. Because capture and restraint are unnecessary for collecting urine samples, the mCG NELISA has widespread potential for confined and free-ranging animals.     

Delayed response task performance as a function of age in cynomolgus monkeys (Macaca fascicularis)

We compared delayed response task performance in young, middle-aged, and old cynomolgus monkeys using three memory tests that have been used with non-human primates. Eighteen cynomolgus monkeys—6 young (4–9 years), 6 middle-aged (10–19 years), and 6 old (above 20 years)—were tested. In general, the old monkeys scored significantly worse than did the animals in the two other age groups. Longer delays between stimulus presentation and response increased the performance differences between the old and younger monkeys. The old monkeys in particular showed signs of impaired visuo-spatial memory and deteriorated memory consolidation and executive functioning. These results add to the body of evidence supporting the utility of Macaca fascicularis in studies of cognition and as a potential translational model for age-associated memory impairment/dementia-related disorders.     

Macaca fascicularis, a nonpermissive host for the human filarial parasite Loa loa

The ability of the filarial nematode Loa loa to infect 2 species of primates was studied. The primate species selected were closely related to species known to be susceptible. A mandrill (Mandrillus sphinx) and 6 cynomolgus monkeys (Macaca fascularis) were infected by subcutaneous injection of third-stage larvae of human L. loa from Gabon. The mandrill developed microfilaremia with an estimated prepatent period of 147 days, but microfilariae were not detected in any of the cynomolgus monkeys. Thus, mandrills appear permissive to human L. loa, whereas cynomolgus monkeys are not. Serum antibody responses were examined on western blots of adult L. loa antigens. Preinfection sera from all animals gave no reactions, but, after infection, sera from cynomolgus monkeys reacted more intensely and with more antigens than mandrill sera. Antibodies were still detectable in cynomolgus monkeys 15 mo postinfection. These reactions were compared with those found using human infection sera. Reactions with the cynomolgus monkey sera resembled those found with resistant endemic and amicrofilaremic human sera.     

人胎胸腺细胞的异质性荧光及其与T淋巴细胞分化阶段关系的形态学研究

以７６９０－Ｘｕ荧光染色法结合ＷｕＴ３、ＷｕＴ４、ＷｕＴ８致敏红细胞花环实验观察经胸腺细胞分层液分离所得高密度亚群和低密度亚群人胎胸腺细胞的异质性荧光及其膜分化抗原ＣＤ３、ＣＤ４、ＣＤ８的表达。结果表明：不同胎龄胎儿胸腺细胞悬液呈现相似形态和相似分布特征的８种异质性荧光的细胞,其中,墨黑核细胞和桔红核细胞分布于两密度亚群,表型为ＣＤ３－ＣＤ４－ＣＤ８－;少数深蓝核和蓝核细胞分布于低密度亚群．表型为ＣＤ３＋,属成熟的胸腺细胞;大多数深蓝核和蓝核细胞、灰蓝核细胞、灰黄核细胞及部分淡桔黄核细胞分布于高密度亚群,表型为ＣＤ３－、ＣＤ４＋、ＣＤ８＋,为处于中间发育阶段的胸腺细胞。推测这些细胞胞核由深蓝、蓝、灰蓝、灰黄到淡桔黄的荧光光谱的偏移可能与中间发育阶段所发生的生物及理化变化有关。     

Straying of hatchery salmon in Prince William Sound,Alaska

The straying of hatchery salmon may harm wild salmon populations through a variety of ecological and genetic mechanisms. Surveys of pink (Oncorhynchus gorbuscha), chum (O. keta) and sockeye (O. nerka) salmon in wild salmon spawning locations in Prince William Sound (PWS), Alaska since 1997 show a wide range of hatchery straying. The analysis of thermally marked otoliths collected from carcasses indicate that 0–98% of pink salmon, 0–63% of chum salmon and 0–93% of sockeye salmon in spawning areas are hatchery fish, producing an unknown number of hatchery-wild hybrids. Most spawning locations sampled (77%) had hatchery pink salmon from three or more hatcheries, and 51% had annual escapements consisting of more than 10% hatchery pink salmon during at least one of the years surveyed. An exponential decay model of the percentage of hatchery pink salmon strays with distance from hatcheries indicated that streams throughout PWS contain more than 10% hatchery pink salmon. The prevalence of hatchery pink salmon strays in streams increased throughout the spawning season, while the prevalence of hatchery chum salmon decreased. The level of hatchery salmon strays in many areas of PWS are beyond all proposed thresholds (2–10%), which confounds wild salmon escapement goals and may harm the productivity, genetic diversity and fitness of wild salmon in this region     

Clinical pathology data from cynomolgus monkeys from China in which diarrhea was observed during quarantine.

Shin Nippon Biomedical Laboratories, Ltd. (SNBL) imported and quarantined 3,148 cynomolgus monkeys (aged 2.5 to 6.5 years) from China in 2002. The hematology and blood biochemistry data obtained from these monkeys on Day 32 of quarantine were analyzed separately by sex [2,890 animals in which no abnormalities were observed during the 35-day quarantine period (normal group), and 258 animals which exhibited diarrhea 1 to 12 times (diarrhea group)]. The values obtained for all parameters were within the normal range (mean +/- SD), and no significant abnormalities were noted in either sex. The clinical pathology data from 11 animals (6 males and 5 females) exhibiting diarrhea repeatedly (10 to 12 times) were statistically analyzed, and significant differences were noted in PLT and ALP in both sexes. The PLT values of these animals were within the normal group mean +/- 2 SD, and were considered within the normal range. A significant difference was noted in some individual ALP values (males: Nos. 2 and 3, females: Nos. 1, 3, and 4). The clinical pathology data obtained from the normal group in this study basically correspond to the widely reported results already obtained from healthy cynomolgus monkeys, from which it can be concluded that the cynomolgus monkeys from China were generally healthy and presenting no particular abnormality. The clinical pathology data from the normal group will serve as valuable baseline data for experimenters using cynomolgus monkeys.     

Hematologic characterization of naturally occurring malaria (Plasmodium inui) in cynomolgus monkeys (Macaca fascicularis)

Twenty of 47 recently imported cynomolgus monkeys (Macaca fascicularis) were found to have malarial infections. The agent identified was Plasmodium inui. All infections were subclinical in nature. Parasitemias ranged from 10 to 900 parasites/mm3 of whole blood. Pre- and post-treatment hematologic values were evaluated following treatment with chloroquine. Treatment was effective in clearing parasitemias from 13 of 14 infected monkeys. Pretreatment values of hematocrit, hemoglobin, and mean corpuscular volume were significantly different in infected animals compared to noninfected animals. While post-treatment hemoglobin and hematocrit values returned to noninfected control levels, mean corpuscular volume values of infected animals remained significantly lower in the post-treatment period.     

An attempt to predict anergy in tuberculosis suspect cynomolgus monkeys.

Acid-fast microorganisms were identified from the tuberculous lesions of a male cynomolgus monkey (Macaca fascicularis). Twenty-two other cynomolgus monkeys housed in the same room were presumed exposed to tuberculosis (Mycobacterium spp.). In addition to standard intradermal (ID) tuberculin testing, clinicians attempted to evaluate the immune status of these monkeys in order to identify animals exhibiting false negative (anergy) ID tuberculin tests. Twenty-one of the potentially exposed monkeys were immunized with tetanus toxoid (TT). Tetanus antitoxin (TAT) titers were measured before and after immunization. The delayed cutaneous hypersensitivity (DCH) reaction to TT was evaluated using a commercially available human test panel. Some animals did not exhibit a DCH reaction to TT. At necropsy 1 of the 21 animals exhibited tuberculous lesions, and acid-fast microorganisms were identified on direct smears of lymphatic tissue of a second animal. Although reported to be of value in assessing the cellular immune status of rhesus monkeys (Macaca mulatta), the delayed cutaneous hypersensitivity response to tetanus toxoid was not helpful during this outbreak in identifying cynomolgus monkeys infected with M. tuberculosis, or in interpreting suspect ID tuberculin tests.     

Yersiniosis in a breeding unit of Macaca fascicularis (cynomolgus monkeys)

2 outbreaks of acute fatal enteric disease involving 20 animals in a breeding unit of approximately 200 cynomolgus monkeys were diagnosed as yersiniosis; Yersinia pseudotuberculosis was isolated from 50% of the clinically affected animals. Post-mortem findings included enlarged mesenteric lymph nodes with some enterocolitis and necrotic foci in liver and spleen. Approximately 7% of clinically healthy monkeys were found to be excreting Y. pseudotuberculosis and a further 5% Y. enterocolitica. Rectal swabs, though less convenient, were better than faecal samples for the detection of Yersinia spp. in 'healthy' monkeys. Efficiency of the cold saline technique and direct plating for isolating Yersinia spp. were compared. It is thought likely that the infection was introduced into the unit by asymptomatic infected monkeys.