Characteristics of the menstrual cycle in nonhuman primates. I. Similarities and dissimilarities between Macaca fascicularis and Macaca arctoides.

Comparative studies of reproductive characteristics were carried out on a colony of Macaca fascicularis and M. arctoides. Seasonal differences were not significant between species, and conceptions occurred throughout the year. The occurrence of short cycles ('luteal phase defect') was found in 2.9% of all M. fascicularis cycles and 1.5% of all M. arctoides cycles. Long cycles (40-50 days) were found in 4.0% of all M. fascicularis cycles and 3.5% of all M. arctoides cycles. Gestation lengths averaged 172.4 and 165.3 days, respectively, for M. arctoides and M. fascicularis. Ovulation in M. fascicularis occurred at a day of ovulation/cycle length ratio of 0.48 +/- 0.08 compared with 0.48 +/- 0.09 for M. mulatta data previously published.     

The morphology of follicular development and ovulation in non-human primates.

A colony of Macaca fascicularis have been under continuous observation for the past 4 years to ascertain the follicular morphological changes that occur prior to ovulation and during the development of the corpus luteum. Of 609 experimental cycles, laparoscopy was performed at least once in 44-5% of the cycles. Of the 104 cycles where the ovulatory status was definitely known, 89-4% were deemed ovulatory and 10-6% anovulatory. The presence or absence of ovulation in the previous cycle did not have an effect on the cycle length either for the total cycles or when analysing only cycles over 28 days. Similarly, the occurrence of two consecutive ovulations (in consecutive cycles) on the same vs. opposite ovaries did not have a significant effect on the cycle length. Neither laparoscopic stress or anaesthesia effected the normal cyclicity of the animals. The characteristic changes in follicular morphology are most clearly defined in M. fascicularis. In this species the 24 of 36 hr prior to ovulation are accompanied by discrete changes which occur in a fixed sequence, allowing one to predict the time of ovulation with reasonable accuracy. In S. sciureus ovulation is preceded by extensive bulging at the follicular apex and haemorrhaging at the base of the follicle. Due to this haemorrhaging post-ovulatory follicles in S. sciureus are generally more easily discernible than in fascicularis. The formation of clear areas (stigma) is not as evident in either S. sciureus or G. senegalensis as in the macaque. Actual ovulation has been observed four times in M. fascicularis and twice in S. sciureus.     

Interpretation of serological test results for simian herpes B virus.

In 1992 an annual Simian herpes B virus (BV) screening programme for an experimental group of macaque monkeys (Macaca mulatta and Macaca fascicularis) was initiated with the aim of establishing a specific pathogen free (SPF) colony. In June 1999 one animal was found to be unexpectedly BV positive (non-negative). The investigation of this result highlights some of the issues and difficulties that may be encountered in such a programme.     

Reference blood values of iron metabolism in cynomolgus macaques.

Iron deficiency anemia is a human health problem of global significance, particularly as it affects pregnant women and infants. While the study of nonhuman primates has resulted in valuable knowledge about iron metabolism, hematologic and biochemical reference ranges for the parameters of iron metabolism are difficult to document in healthy monkeys. At our institution, we maintain a large breeding colony of healthy cynomolgus monkeys (Macaca fascicularis). Data compiled after sampling nonpregnant females and male members of this colony are presented as reference ranges for red cell number, hemoglobin, hematocrit, mean cellular volume, mean cellular hemoglobin, mean cellular hemoglobin concentration, serum iron, total iron-binding capacity, serum transferrin, and serum ferritin.     

Evaluation of caged mating behavior in Macaca fascicularis.

Studies were undertaken to characterize the basic behavioral parameters of a breeding colony of Macaca fascicularis under caged conditions. The female Reproduction Performance Index and the Success Ratio were found to correlate with mating behavior relating to cyclicity in females with cycles of less than 30 days. These evaluations proved to be useful in the selection of animals based on sexual vigor.     

Comparative analysis of the fragments of highly repeated DNA of Homo, Pan, Macaca and Cercopithecus

In the present paper the restriction patterns of Homo, Pan troglodytes, Macaca fascicularis and Cercopithecus aethiops, obtained by digestion with Eco RI endonuclease, were compared. All these species showed two common bands at 0.3 and 0.6 Kb. In this region another band at 0.5 Kb was present in Macaca fascicularis and Cercopithecus aethiops. Furthermore Macaca fascicularis, Cercopithecus aethiops and Pan troglodytes showed a series of bands not present in humans. The distance matrix shows a strong similarity between Homo and Pan and a decreasing similarity with Macaca fascicularis and Cercopithecus aethiops. No intraspecific variability was observed.     

Possible influence of measles virus infection of cynomolgus monkeys on the outcome of the neurovirulence test for oral poliovirus vaccine.

Macaque monkeys are susceptible to measles infection which triggers temporary immuno-depression similar to the well known phenomenon in humans. It is known that feral monkeys become infected with measles virus when they are exposed to humans. Since Macaca mulatta and M. fascicularis are species used to assay the neurovirulence of oral poliovirus vaccine, the immunodepression caused by measles infection of the test monkeys could significantly alter the results of the neurovirulence test. The serum titers of measles-neutralizing antibodies were studied in over 1500 monkeys used for neurovirulence tests. A high proportion of the feral monkeys had measles antibodies (51-100%); in contrast, none of 493 M. fascicularis monkeys which had been bred in a primate colony under strict isolation measures was found positive for measles antibodies. An increase in the prevalence of measles in the population of Ontario and Quebec provinces was accompanied with an increase in the proportion of measles-positive monkey and their serum antibody titers were found higher. It was observed that monkeys used in tests that had been performed during high measles prevalence presented with a poliomyelitis of more pronounced severity clinically and histologically. The analysis of 29 tests conducted on type 1 vaccines over several years showed a positive correlation (correlation coefficient = 0.5141, P less than 0.0022) between severity of poliomyelitis and the presence of measles serum antibodies in test monkeys (some animals seroconverted during the test). A similar observation, when type 3 Sabin vaccines were tested in M. fascicularis, was recently reported from another laboratory in Ontario.     

Variable prevalence and functional diversity of the antiretroviral restriction factor TRIMCyp in Macaca fascicularis

The retroviral restriction factor TRIMCyp, derived from the TRIM5 gene, blocks replication at a postentry step. TRIMCyp has so far been found in four species of Asian macaques, Macaca fascicularis, M. mulatta, M. nemestrina, and M. leonina. M. fascicularis is commonly used as a model for AIDS research, but TRIMCyp has not been analyzed in detail in this species. We analyzed the prevalence of TRIMCyp in samples from Indonesia, Indochina, the Philippines, and Mauritius. We found that TRIMCyp is present at a higher frequency in Indonesian than in Indochinese M. fascicularis macaques and is also present in samples from the Philippines. TRIMCyp is absent in Mauritian M. fascicularis macaques. We then analyzed the restriction specificity of TRIMCyp derived from three animals of Indonesian origin. One allele, like the prototypic TRIMCyp alleles described for M. mulatta and M. nemestrina, restricts human immunodeficiency virus type 2 (HIV-2) and feline immunodeficiency virus (FIV) but not HIV-1. The others restrict HIV-1 and FIV but not HIV-2. Mutagenesis studies confirmed that polymorphisms at amino acid residues 369 and 446 in TRIMCyp (or residues 66 and 143 in the cyclophilin A [CypA] domain) confer restriction specificity. Additionally, we identified a polymorphism in the coiled-coil domain that appears to affect TRIMCyp expression or stability. Taken together, these data show that M. fascicularis has the most diverse array of TRIM5 restriction factors described for any primate species to date. These findings are relevant to our understanding of the evolution of retroviral restriction factors and the use of M. fascicularis models in AIDS research.     

Maternal factors affecting reproduction in a breeding colony of cynomolgus macaques (Macaca fascicularis)

A breeding colony of cynomolgus macaques (Macaca fascicularis), composed of imported and colony-born animals and established for 9 years, was evaluated for maternal factors associated with reproductive failure. The factors evaluated included age, gravidity, parity, origin, previous stillbirths, clinical incidents and type of housing. The effects of each factor on pregnancy rate (PR), stillbirth rate (SR), infant mortality rate (IMR) and pregnancy success (PS) were evaluated. The overall colony rates were: PR = 53%, SR = 22%, IMR = 22%, and PS = 60%. Neonatal death rate for the group was 12%. Pregnancy rate was most affected by maternal factors. Clinical incidents occurring during pregnancy were associated with a significant increase in the stillbirth rate, but did not affect infant mortality rate. Maternal age did not affect any of the measures of reproductive output. Pregnancy rate peaked at 6-8 years of age and decreased thereafter, while pregnancy success peaked at 9-11 years of age. Gravidity and parity had a positive linear relationship with pregnancy rate.     

Homologues of the human C and A apolipoproteins in the Macaca fascicularis (cynomolgus) monkey

We used antisera to human A and C apolipoproteins to identify homologues of these proteins among the high-density lipoprotein apoproteins of Macaca fascicularis (cynomolgus) monkeys, and NH2-terminal analysis was used to verify the homology. The NH2-terminal sequence of the M. fascicularis apoA-I is identical with that of another Old World species, Erythrocebus patas, and differs from human apoA-I at only 4 of the first 24 residues. M. fascicularis apoA-II contains a serine for cysteine replacement at position 6 and is therefore monomeric like the apoA-II from all species below apes. Human and monkey apoA-II are not otherwise different through their first 25 residues. About 20% of M. fascicularis apoC-I aligns with human apoC-I through residue 22, and 80% lacks an NH2-terminal dipeptide. Otherwise, the monkey apoC-I differs from the human protein at only 2 of 25 positions. Two forms of M. fascicularis apoC-II were identified. ApoC-II1 is highly homologous with human apoC-II, whereas an NH2-terminal hexapeptide is absent from apoC-II2. ApoC-II2 was the predominant species, and apoC-II1 appears to represent a propeptide from which a hexapeptide prosegment is cleaved at a Gln-Asp bond. Both forms of monkey apoC-II are potent activators of lipoprotein lipase. There are two polymorphic forms of M. fascicularis apoC-III, and their electrophoretic mobilities become identical after treatment with neuraminidase. Except for a glycine for serine substitution at position 10, the first 15 NH2-terminal residues of M. fascicularis and human apoC-III are the same.     

Differences among three macaque species in responsiveness to an observer

Behavioral responses of experimentally naive captive-born juvenile females of three macaque species (Macaca mulatta, M. radiata, M. fascicularis)to a passive human observer were compared across three periods, beginning approximately 6 weeks after animals were transferred from multianimal groups housed outdoors to individual cages in colony rooms and ending approximately 6 months later. Responsiveness was assessed using eight relatively stereotyped behavior patterns characteristic of this genus. Although overall responsiveness declined across periods in all groups, this measure was consistently highest for M. fascicularis.Patterns were also analyzed as indicative of “fear” or “hostility.” Responses of M. fasciculariswere frequent in both categories, although fear scores exceeded those for hostility. M. mulattascored more frequently as hostile than as fearful. Both fear and hostility scores for M. radiatawere extremely low. The only response shown with more than minimal frequency was lip-smacking, a pattern that is usually construed as an appeasing, submissive, or affiliative display. Differences among species in their responses to the observer are generally consistent with other behavioral and psychophysiological data on the same subjects and with patterns of social interaction between conspecifics.     

澄黄滨珊瑚和丛生盔型珊瑚非培养放线菌多样性

【目的】研究澄黄滨珊瑚(Porites lutea)和丛生盔型珊瑚(Galaxea fascicularis)联合放线菌物种多样性。【方法】实验提取两种珊瑚的总DNA,利用放线菌特异性引物对样品总DNA进行扩增,通过构建16S rRNA基因克隆文库和系统发育分析,对三亚鹿回头岸礁区优势物种澄黄滨珊瑚和丛生盔型珊瑚联合放线菌的多样性和群落结构进行研究。【结果】118个从澄黄滨珊瑚克隆文库中随机挑选的阳性克隆子归为58个OTUs,主要分布于酸微菌亚目、棒状杆菌亚目、微球菌亚目、丙酸杆菌亚目和未知类群。丛生盔型珊瑚克隆文库共获得96个序列,归为31个OTUs,主要分布于酸微菌亚目和未知的放线菌类群。多样性指数和稀疏度曲线分析结果显示澄黄滨珊瑚联合放线菌物种多样性比丛生盔型珊瑚更高。【结论】澄黄滨珊瑚和丛生盔型珊瑚拥有较高水平的放线菌物种多样性和复杂的群落结构,并隐藏着大量的高等级放线菌新分类单元。     

Some monoclonal anti-human lymphocyte and class II MHC antigen antibodies do not stain snap frozen Macaca fascicularis lymphocytes or tissue sections.

Experiments were designed to stain snap frozen and fixed Macaca fascicularis peripheral blood mononuclear cell (PBMC) preparations and colon sections assuming that monoclonal mouse anti-human lymphocyte and class II major histocompatibility complex antigen antibodies would cross-react with the monkey counterparts to the human antigens. Most of the monoclonals used in this study did not stain the monkey frozen and fixed tissues in immunoenzymatic protocols despite adequate staining of control human tissues. The fact that snap frozen Macaca fascicularis PBMCs and tissue sections were not always stained is of practical importance for experimental design in Macaca fascicularis using anti-human reagents.     

妊娠晚期食蟹猴与同龄未孕食蟹猴血液生理和生化指标的比较

目的提供妊娠晚期食蟹猴有关的血液学数据。方法采用Sysmex XE-2100血液分析仪、SysmexCA-7000凝血分析仪,及OL YMPUS AU600型全自动生化分析仪,对15只怀孕晚期食蟹猴及10只正常对照非孕食蟹猴进行血常规、凝血三项及17项血生化指标分析。结果妊娠晚期食蟹猴在WBC、NEU%、LYM%、PT、APTT、Fgb、TP、Alb、BUN、CR、CHOL、Glu、Fe、LDK、HDBH上,与同龄未孕食蟹猴相比差异显著（P〈0.05）,其他指标RBC、HGB、HCT、MCV、MCH、MCHC、PLT、ALT、AST、ALP、T.Bili、TG、P相比,于孕晚期有不同程度改变,但无显著性差异（P〉0.05）。结论初步建立了孕晚期食蟹猴的常规生理学数据,为相关的实验研究提供基础资料。     

食蟹猴微卫星DNA标记遗传监测及多态性分析

目的研究国内食蟹猴种群的遗传背景特性,建立食蟹猴种群遗传质量监测方法。方法采用微卫星DNA遗传标记技术对50只食蟹猴种群个体进行遗传质量监测及DNA多态性分析。结果从100个微卫星DNA位点中筛选出20个多态性高的位点,其食蟹猴种群个体的等位基因数目为5-10条,个体间均呈现高度的多态性;其观察等位基因数（Na）为5.0～10.0,有效等位基因数（Ne）为4.6118～8.3404,基因多样性（H）为0.7832～0.8801和香隆信息指数（I）为1.5651～2.1592。结论本实验有效地分析了食蟹猴种群的遗传多态性,为今后筛选特异性微卫星位点来建立食蟹猴种群遗传质量监测方法提供了理论依据。     

Characterization of plasma low density lipoproteins on nonhuman primates fed dietary cholesterol.

LDL from animals of three nonhuman primate species, Macaca mulatta, Macaca fascicularis, and Cercopithecus aethiops, were studied. A standard preparation of 125I-LDL was added to isolated lipoprotein mixtures just prior to separation of plasma lipoproteins by agarose gel chromatography. A relative size index, rI, was determined by dividing the elution volume of the iodinated LDL by the elution volume of the sample LDL, both volumes being determined simultaneously during chromatographic elution. Comparison of rI with molecular weights measured by flotation equilibrium analysis in the analytical ultracentrifuge showed a linear relationship across a molecular weight range of 2.5-8.0 X 10(6), r = 0.985. A regression equation describing this relationship was used to calculate molecular weights of LDL from a group of M. fascicularis that were fed cholesterol-containing diets. In these animals, plasma cholesterol concentration ranged from 100 to over 700 mg/dl and was highly correlated with LDL molecular weight and with the micromolar concentration of the LDL. Using multiple regression analyses, the two variables of plasma LDL could be shown to account for 94% of the variation in plasma cholesterol concentration in the M. fascicularis of this study. Micromolar concentration and molecular weight of LDL were not correlated with each other, suggesting that in M. fascicularis at least two independent types of controls are operative in the response of plasma LDL to dietary cholesterol. The increase in LDL molecular weight was associated with a large increase in cholesteryl ester content and concomitant smaller increases in protein, phospholipid, and free cholesterol. As molecular weight increased, these components appeared to be added to the LDL particles together as discrete increments of fixed composition. The data are consistent with a spherical model of LDL structure with a core of cholesteryl ester and triglyceride and a 21.3 A-thick coat of phospholipid, free cholesterol, and protein.     

Recrudescence of Entopolypoides macaci Mayer, 1933 (Babesiidae) infection secondary to stress in long-tailed macaques (Macaca fascicularis)

Parasites were found in red blood cells of two long-tailed macaques (Macaca fascicularis) imported from Indonesia and housed in the Washington Regional Primate Research Center breeding colony for 7 years or longer. Both macaques developed parasitemias secondary to stress (type D retrovirus in one case and severe trauma in the other). Entopolypoides macaci (Babesiidae) was diagnosed on the basis of morphology from peripheral blood smears stained with Wright's stain. Antibodies against Babesia sp. were detected by immunofluorescence assay (IFA) from one infected macaque, which showed antibody cross-reactions (high titer) to B. bigemina, B. bovis, B. canis, and (low titers) to Plasmodium falciparum. Five feral long-tailed macaques that had been imported recently from the same country had no detectable antibodies. This is the first report of IFA as an aid to diagnose E. macaci in nonhuman primates.     

Outbreak of Mycobacterium bovis in a conditioned colony of rhesus (Macaca mulatta) and cynomolgus (Macaca fascicularis) macaques

We describe a tuberculosis outbreak caused by Mycobacterium bovis in a conditioned colony of rhesus (Macaca mulatta) and cynomolgus (Macaca fascicularis) macaques. Animals in five rooms were exposed, but most (16/27) infections were confined to the room that housed a mixed population of cynomolgus and rhesus macaques. In this room, rhesus (8/8) and cynomolgus (10/11) macaques naturally exposed to M. bovis were infected at nearly identical rates (Fisher exact test, 2-tailed P = 1). The clinical signs of disease and pathologic lesions in infected macaques, however, were moderately different between the two species. Rhesus macaques were more likely (5/8) to exhibit clinical signs of persistent coughing and inappetance, and had more severe pulmonary lesions. By contrast, clinical signs of disease were seen in only 1 of 19 cynomolgus macaques, and overall, the pulmonary lesions were often focal and less severe, although some still had severe involvement of the lungs similar to that seen in rhesus macaques. These differences should be taken into consideration when developing or evaluating a tuberculosis-screening program. On the basis of observations made during this outbreak, we recommend that alternative screening methods such as the PRIMAGAM test and the ESAT-6 ELISA, be incorporated into the screening program to aid in the identification of infected animals.     

Mitochondrial DNA sequence phylogeny of 4 populations of the widely distributed cynomolgus macaque (Macaca fascicularis fascicularis)

We studied the mitochondrial DNA (mtDNA) polymorphism of 304 Macaca fascicularis fascicularis (M. f. fascicularis) individuals, representative of 4 cynomolgus macaque populations (Indochina, Indonesia, Philippines, and Mauritius). By sequencing a 590-bp fragment in the hypervariable II region of the D-loop region, we defined 70 haplotypes. The homologous region was also characterized in 22 Chinese Macaca mulatta and 2 Macaca sylvanus. The phylogenetic analysis confirms the monophyly of M. f. fascicularis and defines 2 haplotype groups inside the M. f. fascicularis clade: one "insular," encompassing 6 Philippines, 2 Mauritius, and 31 Indonesian haplotypes, the other "continental" that contains all Indochinese and 6 Indonesian haplotypes. Continental and insular group divergence time was estimated to be approximately 10(6) years before present (BP). Among Indonesian haplotypes, some have a continental origin. This suggests either direct migration from mainland to Indonesia or that remnant lineages from an ancient population genetically close to the mainland (i.e., in the Sunda Shelf, <550 000 years BP) were subsequently brought southward to Indonesia. The low nucleotide diversity in the Philippines population suggests a bottleneck following colonization by Indonesian individuals, around 110 000 years BP. mtDNA and further observations of nuclear genetic data corroborate the mixed origin (Indonesian/continental) hypothesis of Mauritius individuals and a population bottleneck.     

Construction and validation of a systematic ethogram of Macaca fascicularis in a free enclosure

Behavioral studies in non-human primates have become ideal models for further investigations into advanced cognitive function in humans. To date, there is no systematic ethogram of the cynomolgus monkey (Macaca fascicularis) in a free enclosure. In a field observation of 6012 subjects, 107 distinct behaviors of M. fascicularis were preliminarily described. 83 of these behaviors were then independently validated through a randomized cohort and classified into 12 behavioral categories. 53 of these behaviors were then selected to accurately reflect the daily mundane activity of the species in a free enclosure. These findings systematically document the behavior of M. fascicularis in a free enclosure for use in further investigations.