太行山猕猴寰椎和枢椎的初步研究

对成年太行山猕猴寰椎和枢椎变量进行了测量;以肱骨最大长为参照,对其变量进行了异速生长分析。结果表明：寰椎全宽及枢椎椎孔横径接近等速生长,其余变量均为负异速生长。     

Efficient Cooperative Restraint Training With Rhesus Macaques

It is sometimes necessary for nonhuman primates to be restrained during biomedical and psychosocial research. Such restraint is often accomplished using a “primate chair.” This article details a method for training adult rhesus macaques to cooperate with a chair restraint procedure using positive and negative reinforcement. Successful training was accomplished rapidly in approximately 14 training days. The success of this training technique suggests that this method represents a refinement to traditional techniques. Further, this method worked effectively for animals previously deemed unfit for traditional pole-and-collar training.     

Evaluating Dominance Styles in Assamese and Rhesus Macaques

Researchers have suggested that several types of agonistic and affiliative behavior covary as a set of species-specific traits, and have used the term dominance style to describe the covariation. We compared measures of dominance style between a group of Assamese macaques (Macaca assamensis) and a group of rhesus macaques (M. mulatta), though kinship information was unknown. Assamese and rhesus female-female dyads each showed a low proportion of counter aggression and a low conciliatory tendency, suggesting that they have despotic social relationships. They also showed a despotic pattern on several other types of agonistic and affiliative behavior, such as approach outcomes and grooming distributions, which is consistent with the covariation of dominance style traits. Assamese male-male dyads showed relatively high levels of reconciliation and counter aggression versus other macaque males portrayed in the literature, suggesting that Assamese males have a tolerant dominance style. Insofar as macaque dominance style depends on the behavior of females, we suggest that Assamese macaques, like rhesus macaques, have despotic social relationships, which contrasts with evidence of a strong correlation between phylogeny and dominance style in macaques. Further, our results indicate that strong male bonding and tolerant dominance relationships among males are independent of female dominance style. Lastly, some measures of agonistic behavior, such as rate of aggression or proportion of bites, are likely altered in competitive environments and thus are not useful indicators of dominance style.     

Identification of Neonatal White Matter on DTI: Influence of More Inclusive Thresholds for Atlas Segmentation

Purpose

Semi-automated diffusion tensor imaging (DTI) analysis of white matter (WM) microstructure offers a clinically feasible technique to assess neonatal brain development and provide early prognosis, but is limited by variable methods and insufficient evidence regarding optimal parameters. The purpose of this research was to investigate the influence of threshold values on semi-automated, atlas-based brain segmentation in very-low-birth-weight (VLBW) preterm infants at near-term age.

Materials and Methods

DTI scans were analyzed from 45 VLBW preterm neonates at near-term-age with no brain abnormalities evident on MRI. Brain regions were selected with a neonatal brain atlas and threshold values: trace <0.006 mm²/s, fractional anisotropy (FA)>0.15, FA>0.20, and FA>0.25. Relative regional volumes, FA, axial diffusivity (AD), and radial diffusivity (RD) were compared for twelve WM regions.

Results

Near-term brain regions demonstrated differential effects from segmentation with the three FA thresholds. Regional DTI values and volumes selected in the PLIC, CereP, and RLC varied the least with the application of different FA thresholds. Overall, application of higher FA thresholds significantly reduced brain region volume selected, increased variability, and resulted in higher FA and lower RD values. The lower threshold FA>0.15 selected 78±21% of original volumes segmented by the atlas, compared to 38±12% using threshold FA>0.25.

Conclusion

Results indicate substantial and differential effects of atlas-based DTI threshold parameters on regional volume and diffusion scalars. A lower, more inclusive FA threshold than typically applied for adults is suggested for consistent analysis of WM regions in neonates.     

A Comparison of Dual-Energy X-Ray Absorptiometry and Somatometrics for Determining Body Fat in Rhesus Macaques

COLMAN, RICKI J., JOHN C. HUDSON, HOWARD S. BARDEN, AND JOSEPH W. KEMNITZ. A comparison of dual-energy X-ray absorptiometry and somatometrics for determining body fat in rhesus macaques. Obes Res. 1999; 7:90–96. Objective : Various approaches have been used to assess fat and fat distribution in nonhuman primates, including measurements of body weight, body dimensions, and estimates derived from these, such as body mass index. Methods such as tritiated water dilution and dual-energy X-ray absorptiometry (DXA) have also been used. The aim of the present study was to evaluate and compare DXA measurements and somatometrics. Research Methods and Procedures : Body composition of 15 adult male rhesus macaques was measured by DXA and somatometrics at four time-points over a 4-year period. Additionally, DXA precision and somatometric variability were analyzed by repeated measurements of the same subjects. Results : DXA estimates of body fat were positively correlated with body weight, body mass index, body circumferences, and abdominal skinfold thicknesses. DXA assessments of soft tissue composition were precise, with coefficients of variation below 3.3% for all compartments analyzed. The majority of the observed variability in somatometrics was explained by subject variance, rather than by inter- or intraobserver variability, or by observer experience level. Discussion : We conclude that noninvasive DXA technology provides precise estimates of nonhuman primate body composition that correlate well with the traditional somatometric measures used in primate studies.     

Extraintestinal Campylobacteriosis in Rhesus Macaques (Macaca mulatta)

Two cases of clinical disease associated with extraintestinal Campylobacter infection were recently encountered in rhesus macaques (Macaca mulatta). The first case was that of a 3-y-old, male, rhesus macaque experimentally infected with SIV, who presented with abdominal pain and a midabdominal mass and was euthanized. Pathology findings included an abscess within the median liver lobe, fibrinopurulent peritonitis, and intestinal serositis with isolation of Campylobacter fetus from the blood, liver, and the hepatic abscess. The second case was that of a 1-mo-old, female, rhesus macaque who died with no apparent history of illness. Gross pathology findings included thin body condition and diarrheic staining of the perineum; histologically, acute multifocal hepatitis with intralesional bacteria was noted. Campylobacter coli was isolated from the liver and colon. Extraintestinal Campylobacter infection is uncommon in humans, usually occurring in immunocompromised subjects and most commonly manifesting as bacteremia. Extraintestinal Campylobacter infections in animals are rare but have been associated with bacteremia and cholecystitis. The macaques presented here were either immunocompromised due to SIV infection (case 1) or more vulnerable due to young age (case 2). These factors likely contributed to the extraintestinal spread of Campylobacter.Campylobacter spp. are curved or spiral, gram-negative, microaerobic, typically motile bacteria with a single flagellum at one or both ends of the cell.²⁵ Campylobacter is one of the most common bacterial causes of gastroenteritis in humans worldwide.³ The Centers for Disease Control and Prevention estimate it to affect more than 1.3 million people in the United States each year.¹¹ Campylobacter spp. colonize the intestinal tract of primates, other mammals, birds, reptiles, and shellfish, but infection is not always associated with clinical signs of disease.^2,23 Contaminated or undercooked poultry represents the largest potential source of human infection.²² Campylobacteriosis is a zoonosis, and other significant sources of human infection include livestock, wildlife, pets, and contaminated water.²² The species most commonly isolated from humans and nonhuman primates are C. jejuni and C. coli; other species like C. fetus are less commonly found.^2,4,9,23 Extraintestinal campylobacteriosis in humans usually occurs in immunocompromised or elderly persons with underlying medical problems and most commonly manifests as bacteremia.²³ There have been a few reports of extraintestinal Campylobacter infections in animals, including bacteremia and cholecystitis in 2 dogs.^17,30     

猕猴结肠镜下活检组织病理学观察

目的通过对猕猴结肠镜检及活检取材,对结肠镜检测方法应用于非人灵长类动物予以评价。方法实验猴在麻醉状态下接受结肠镜检查和活检标本取材,对活检标本进行固定和病理切片观察,并对所检猴进行解剖和组织学观察,比较各取材点的情况。结果结肠镜下及大体标本观察见猕猴肠壁厚度明显小于人体,活检取材部位见黏膜破损、肠出血,个别部位见肠穿孔。病理切片HE染色观察发现,与人体比较,猕猴大肠腺较小而表浅,固有层内淋巴小结数量较少,肌层、黏膜下层明显较薄。结论对猕猴行结肠镜检查及镜下活检取材是可行的,但因猕猴肠壁较人体薄而极易穿孔,故需尽量应用活检杯小的活检钳,并需要充分的肠道准备和有经验的肠镜操作。     

TRIM5 alpha Drives SIVsmm Evolution in Rhesus Macaques

The antagonistic interaction with host restriction proteins is a major driver of evolutionary change for viruses. We previously reported that polymorphisms of the TRIM5α B30.2/SPRY domain impacted the level of SIVsmm viremia in rhesus macaques. Viremia in macaques homozygous for the non-restrictive TRIM5α allele TRIM5^Q was significantly higher than in macaques expressing two restrictive TRIM5alpha alleles TRIM5^TFP/TFP or TRIM5^Cyp/TFP. Using this model, we observed that despite an early impact on viremia, SIVsmm overcame TRIM5α restriction at later stages of infection and that increasing viremia was associated with specific amino acid substitutions in capsid. Two amino acid substitutions (P37S and R98S) in the capsid region were associated with escape from TRIM5^TFP restriction and substitutions in the CypA binding-loop (GPLPA87-91) in capsid were associated with escape from TRIM5^Cyp. Introduction of these mutations into the original SIVsmE543 clone not only resulted in escape from TRIM5α restriction in vitro but the P37S and R98S substitutions improved virus fitness in macaques with homozygous restrictive TRIM^TFP alleles in vivo. Similar substitutions were observed in other SIVsmm strains following transmission and passage in macaques, collectively providing direct evidence that TRIM5α exerts selective pressure on the cross-species transmission of SIV in primates.     

Maternal Anxiety in Rhesus Macaques (Macaca mulatta)

Visual monitoring and scratching were used as behavioral indicators of maternal and social anxiety in small captive groups of rhesus macaques. Young infants were especially at risk from other group members during the first weeks of locomotion away from their mothers. Mothers received aggression from other individuals irrespective of their infants' presence or absence. The rate at which mothers scratched themselves increased significantly when their infants moved away from them and when the infants approached or were approached by individuals who frequently harassed them. The rate of maternal scratching and the rate of glancing at the infant and at other individuals when the infant was away decreased as infants grew older and became less vulnerable to harassment. In contrast, the rate of maternal scratching and visual monitoring of other individuals when the infant was in contact remained stable across the first 12 wk of lactation. The rate of maternal scratching increased when the mother-infant pair was in spatial proximity to the adult male or higher ranking adult females. Although visual monitoring and scratching showed a similar sensitivity to social variables, it is speculated that they might reflect different components of anxiety, namely, anticipation of danger and uncertainty due to motivational conflict. The results of this investigation indicate that a macaque mother's emotional reactivity to a perceived danger for herself and her infant can be measured quite accurately using the rates of visual monitoring and scratching and that the latter represent reliable tools to investigate the emotional correlates of maternal behavior in nonhuman primates.     

Development of Breeding Populations of Rhesus Macaques (Macaca mulatta) That Are Specific Pathogen-free for Rhesus Cytomegalovirus

Development of breeding colonies of rhesus macaques (Macaca mulatta) that are specific pathogen-free (SPF) for rhesus cytomegalovirus (RhCMV) is relatively straightforward and requires few modifications from current SPF programs. Infants separated from the dam at or within a few days of birth and cohoused with similarly treated animals remain RhCMV seronegative indefinitely, provided they are never directly or indirectly exposed to a RhCMV-infected monkey. By systematically cohousing seronegative animals into larger social cohorts, breeding populations of animals SPF for RhCMV can be established. The additional costs involved in expanding the current definition of SPF status to include RhCMV are incremental compared with the money already being spent on existing SPF efforts. Moreover, the large increase in research opportunities available for RhCMV-free animals arguably would far exceed the development costs. Potential new areas of research and further expansion of existing research efforts involving these newly defined SPF animals would have direct implications for improvements in human health.Abbreviations: HCMV, human cytomegalovirus; NHP, nonhuman primate; RhCMV, rhesus cytomegalovirus; SPF, specific pathogen-freeThe impetus for expanding the current SPF definition to include RhCMV is 2-fold. The first is the increasing number of studies involving infection of rhesus macaques with RhCMV as a nonhuman primate (NHP) model of human infection with human cytomegalovirus (HCMV). The second is the recognition that the current SPF protocols result in animals that are also uninfected with RhCMV, such that a relatively minor change in derivation can generate monkeys that meet the current SPF definition and that are uninfected with other endemic viruses, including RhCMV and simian foamy virus.     

Comparison of the Reconciliation Behavior of Stumptail and Rhesus Macaques

Observations were made following spontaneous aggressive incidents in a breeding group of captive stumptail macaques (Macaca arctoides). Participants were observed both during the first 10 min following the aggressive incident, and during matched-control observations. Data on 670 pairs of former opponents were collected, and compared with a sample of 573 such pairs of rhesus macaques (M. mulatta), which had been observed with identical methods in a previous study. Selective attraction between opponents was confirmed for the stumptail monkeys, i.e. both the absolute and the relative rate of nonagonistic body contact between individuals increased after aggression between them. The species was found to be considerably more conciliatory than rhesus monkeys, and to have a remarkably rich repertoire of reassurance gestures. The most characteristic conciliatory behavior is the hold-bottom ritual, in which one individual (usually the subordinate) presents its hindquarters, and the other (usually the dominant) clasps the other's haunches. The high rate of reconciliation among stumptail monkeys extended to all relationship classes, and a correlation with the closeness of social bonds (measured as time spent in association) could not be demonstrated. Because of the generality of the species' high peacemaking tendencies it is assumed that group cohesiveness is of great survival value in the wild.     

模拟HIV性传播特点的SIVmac239恒河猴直肠黏膜感染

目的制备SIVmac239恒河猴(Macaca mulatta)细胞适应株病毒,模拟HIV性传播感染特点进行恒河猴直肠黏膜感染研究,探索引起系统性感染的病毒阈值水平与机体病毒、免疫学之间相关性,为我国艾滋病黏膜疫苗等生物制剂有效性评价提供新的模型构建思路。方法参照HIV性传播自然感染剂量范围,选用SIVmac239连续升高的3种剂量直肠黏膜途径感染两只恒河猴,采取多种方法进行病毒血症和免疫反应特点分析。结果两只恒河猴经2×101TCID50和2×102TCID50病毒滴度2次攻击后45d,经检测均未建立系统性感染,病毒特异性免疫反应均为阴性;第3次2×103TCID50病毒滴度攻击后,M296猴表现出典型的系统性感染特点,并诱导特异性免疫反应。结论确认了HIV性传播过程中的病毒剂量效应关系,为预防性生物制剂的猴体有效性评价提供了新的思路。同时,发现SIVmac239Gag区特异性的T细胞免疫反应在病毒控制过程中发挥了关键作用,对于新一代艾滋病黏膜疫苗的抗原选择具有指导性意义。     

Origins and Long-Term Patterns of Copy-Number Variation in Rhesus Macaques

Mutations play a key role in the development of disease in an individual and the evolution of traits within species. Recent work in humans and other primates has clarified the origins and patterns of single-nucleotide variants, showing that most arise in the father’s germline during spermatogenesis. It remains unknown whether larger mutations, such as deletions and duplications of hundreds or thousands of nucleotides, follow similar patterns. Such mutations lead to copy-number variation (CNV) within and between species, and can have profound effects by deleting or duplicating genes. Here, we analyze patterns of CNV mutations in 32 rhesus macaque individuals from 14 parent–offspring trios. We find the rate of CNV mutations per generation is low (less than one per genome) and we observe no correlation between parental age and the number of CNVs that are passed on to offspring. We also examine segregating CNVs within the rhesus macaque sample and compare them to a similar data set from humans, finding that both species have far more segregating deletions than duplications. We contrast this with long-term patterns of gene copy-number evolution between 17 mammals, where the proportion of deletions that become fixed along the macaque lineage is much smaller than the proportion of segregating deletions. These results suggest purifying selection acting on deletions, such that the majority of them are removed from the population over time. Rhesus macaques are an important biomedical model organism, so these results will aid in our understanding of this species and the disease models it supports.     

促进人工饲养实验猕猴心理康乐

目的促进实验猕猴心理康乐,既能减少动物异常行为的发生、获得可靠的科学数据,同时也是实现这一珍贵非人灵长类实验动物福利的重要内容。实验猕猴社会化集群、环境优化、婴猴饲养方式是影响动物心理康乐几个较大的因素,本文对其研究进展进行了综述,对推进我国灵长类实验动物福利具有积极作用。     

弓形虫（Toxoplasma gondii）的PCR检测方法在笼养猕猴群中的应用

目的建立快速、灵敏、特异及检测结果易判断的PCR方法,并应用于大规模猕猴种群的弓形虫常规检测中。同时比较巢式PCR和单一PCR的一致性。方法根据弓形虫保守基因p30（SAG1）设计了内、外两对进行巢式PCR扩增以及B1基因设计一对引物进行单一PCR扩增,将DNA样本进行10倍倍比稀释,以检测巢式PCR反应的灵敏度;并对医学生物学研究所自繁猕猴共150只进行了弓形虫检测。结果巢式PCR检测法检测限度可达10^-3ng/uL,而且方法特异。两种PCR法检测结果基本一致,其中巢式PCR检测阳性率（10％）稍高于单一PCR检测阳性率（8．67％）。结论巢式PCR和一次PCR方法都可应用于猕猴弓形虫的常规检测中,并提示巢式PCR比单一PCR更敏感、检出率更高。     

Multicolor Flow Cytometry Analyses of Cellular Immune Response in Rhesus Macaques

The rhesus macaque model is currently the best available model for HIV-AIDS with respect to understanding the pathogenesis as well as for the development of vaccines and therapeutics^1,2,3. Here, we describe a method for the detailed phenotypic and functional analyses of cellular immune responses, specifically intracellular cytokine production by CD4+ and CD8+ T cells as well as the individual memory subsets. We obtained precise quantitative and qualitative measures for the production of interferon gamma (INF-) and interleukin (IL) -2 in both CD4+ and CD8+ T cells from the rhesus macaque PBMC stimulated with PMA plus ionomycin (PMA+I). The cytokine profiles were different in the different subsets of memory cells. Furthermore, this protocol provided us the sensitivity to demonstrate even minor fractions of antigen specific CD4+ and CD8+ T cell subsets within the PBMC samples from rhesus macaques immunized with an HIV envelope peptide cocktail vaccine developed in our laboratory. The multicolor flow cytometry technique is a powerful tool to precisely identify different populations of T cells ^4,5 with cytokine-producing capability⁶ following non-specific or antigen-specific stimulation ^5,7.Download video file.^{(83M, mp4)}     

巢式PCR检测食蟹猴和猕猴中SRV和SFV

目的利用巢式PCR方法检测圈养的食蟹猴（Macaca fascicularis）和猕猴（Macaca mulatta）中猴D型逆转录病毒（Simian Type D Retrovirus SRV）和猴泡沫病毒（Simian foamy virus SFV）。方法针对SRV-env和SFV-pol基因的保守区序列设计特异性外引物,然后再设计特异性内引物。将外引物扩增出的片段克隆到PJET1.2blunt载体中作为阳性对照,运用NCBI中BLAST软件比对测序结果。用巢式PCR方法分别检测食蟹猴和猕猴中SRV和SFV。结果发现食蟹猴中SFV感染率为65.2%,SRV感染率为9.5%,猕猴中SFV感染率为60.5%,SRV感染率为12.8%。结论圈养的食蟹猴和猕猴SFV的感染率均较高,SRV感染率很低。     

Immune Activation and Regulation in Simian Immunodeficiency Virus-Plasmodium fragile-Coinfected Rhesus Macaques

Human immunodeficiency virus (HIV) is characterized by immune activation, while chronic malaria is associated with elevated interleukin-10 (IL-10) levels. How these apparently antagonizing forces interact in the coinfected host is poorly understood. Using a rhesus macaque model of simian immunodeficiency virus (SIV)-Plasmodium fragile coinfection, we evaluated how innate immune effector cells affect the balance between immune activation and regulation. In vitro Toll-like receptor (TLR) responses of peripheral blood myeloid dendritic cells (mDC) and monocytes were temporarily associated with acute parasitemic episodes and elevated plasma IL-10 levels. Prolonged infection resulted in a decline of mDC function. Monocytes maintained TLR responsiveness but, in addition to IL-12 and tumor necrosis factor alpha, also produced IL-10. Consistent with the role of spleen in the clearance of parasite-infected red blood cells, coinfected animals also had increased splenic IL-10 mRNA levels. The main cellular source of IL-10 in the spleens of coinfected animals, however, was not splenic macrophages but T cells, suggesting an impairment of adaptive immunity. In contrast to those in spleen, IL-10-positive cells in axillary lymph nodes of coinfected animals were predominantly mDC, reminiscent of the immunosuppressive phenotype of peripheral blood mDC. Concurrent with IL-10 induction, however, SIV infection promoted elevated systemic IL-12 levels. The continuously increasing ratio of plasma IL-12 to IL-10 suggested that the overall host response in SIV-P. fragile-coinfected animals was shifted toward immune activation versus immune regulation. Therefore, SIV-P. fragile coinfection might be characterized by earlier manifestation of immune dysfunction and exhaustion than that of single-pathogen infections. This could translate into increased morbidity in HIV-malaria-coinfected individuals.