人类甲型肝炎动物模型（狨猴）的研究

本文报道以不同剂量的不同HAV毒株,经静脉和胃肠道分别接种普通狨猴和白须狨猴后,其血清ALT、AsT和LDH活性均见升高。对酶活性比接种前增加,大于十3SD值的6只狨猴作肝活组织检查,见有与急性病毒肝炎相一致的组织病理变化。此外,用免疫电镜法检出狨猴粪便中HAV颗粒,狨猴接种HAV后36天,血清IgM类和总抗HAV抗体均阳转。上述结果提示,普通狨猴有可能作为研究甲型肝炎的理想动物模型。     

引进的棉顶狨猴群简介

棉顶狨猴(Cotton-top tamarin;Saguinus oedipus oedipus:以下简称棉狨)产于南美哥伦比亚,是珍稀濒危(犭胥)狨猴的一种。狨猴体型小,易于实验室笼养,是肿瘤、心血管病、肝炎、计划生育等研究中较为理想的模型动物。近年来,建立实验室狨猴繁殖群已在国内外引起重视。1988年7月,本所从英国引进6只(3雌3雄)育龄棉狨,计划由此建立实验室繁殖群落。经半年余适应性饲养管理,目前个个健康活泼,3只雌猴均有妊娠迹象。现将基本情况报告于后。     

棕肩狨猴人工饲养与繁殖的初步研究

本文对棕肩狨猴在实验室笼养条件下,进行了饲养繁殖与育仔的初步研究,发现控制好一定的温度、相对湿度、光照、营养和良好的管理,对于该狨猴的繁殖与育仔的成功具有十分重要的作用。该品种狨猴在国内繁殖成功尚属首次。     

野生白须狨猴的体内寄生虫

本文介绍了对我所引进的野生白须狨猴体内寄生虫检查初步结果,在14只白须狨猴静脉血液涂片检查中,均发现有微丝蚴;经对死亡6只白须披猴的尸检,发现感染最严重的是线虫,其次是蛇舌形虫、棘头虫。同时对寄生虫直接导致狨猴死亡的可能性和防治方法作了初步探讨。     

普通棉耳狨猴仙台病毒感染的报道

仙台病毒在分类上属副粘病毒科、副粘病毒属、副流感病毒 1型。仙台病毒过去的名称包括新生儿肺炎病毒 ,日本血凝病毒和Ｄ型流感病毒等。在国内普通实验小鼠群中仙台病毒感染非常普遍 ,而未见仙台病毒感染普通狨猴的报道。 1999年夏季 ,天津某高校一个狨猴实验室中暴发仙台病毒感染 ,动物发病率达 80 % ,病死率约 10 %。流行大约持续了一个月。狨猴发病初始均表现为精神不振 ,嗜睡 ,惧冷 ,食欲下降。继续发展表现为鼻腔出现分泌物 ,呼吸加快 ,发热 ,气喘 ,呼吸困难 ,多数病狨从发病至死亡在 3- 15ｄ。解剖发现病死狨猴肺叶呈杨梅色 ,切开时…     

精制甲型肝炎灭活疫苗的狨猴保护实验

目的　检测精制甲型肝炎 (甲肝 )灭活疫苗的保护效果。方法　甲型肝炎病毒 (YN5株 )经Vero细胞培养制成精制甲肝灭活疫苗 ,采用普通狨猴进行疫苗的免疫原性和保护性研究。结果　 7只狨猴均有特异性抗体产生 ,无血清酶活性升高和肝组织学病理改变 ,接种疫苗的狨猴能抵抗甲肝病毒强毒 (Han ZD)株的攻击。但是 ,对照组狨猴均出现血清酶异常升高和肝组织学病理改变。结论　甲肝灭活疫苗 (YN5株 )具有良好的免疫原性和保护效果。     

狨猴生殖工程与基因修饰研究进展

人类疾病相关基因修饰动物模型已成为研究人类疾病分子机理和治疗靶点的重要途径和材料.狨猴是最小型实验灵长类动物,其体型小、繁殖快、遗传进化及生理生化方面与人类接近,是人类疾病动物模型研究的理想实验动物之一.因此,利用高效的生殖工程技术和基因编辑技术创制基因修饰狨猴具有巨大的生物医药价值.本文就狨猴生殖工程技术和基因修饰进行综述,旨在开拓人类疾病动物模型,推动疾病机理和新药创制的突破性发展.     

介绍一种灵长目实验动物——狨猴

狨猴属灵长目(Primates),狨科(Callithriadac),约有30个种及许多具有生育力的杂交种。最早在西方国家作为玩赏兽饲养,至60年代初才开始小群繁殖,以估计其用于生物学、医学研究的可能性。60年代后期便被广泛用于科学研究。目前以普通狨猴(Callithrix jaccus)繁殖情况最佳,用量也最多(见图1)。 成年狨猴体重300—500克,性情温顺,容易饲养,繁殖力高,一年可生二胎,一胎可产仔2—3只,怀孕期平均142天,流产率和死产率各仅1％左右,14—16周断乳,生育力无季节性差     

白须狨猴血液学初步测定

目前,白须狨猴(Saguinus myatax)在生物医学研究中已成为一种重要的实验动物,尤其在人类甲型肝炎的研究中,已被公认为最理想的实验动物模型。然而,对于这种小型灵长类动物的血液学数值所知甚少,国内至今未见报道。为了给人类甲型肝炎动物实验作好准备,我们对引进的野生白须狨猴进行血液学部分项目测定。     

基于高通量测序技术测定普通棉耳狨猴粪便微生物多样性

目的应用高通量测序技术分析普通棉耳狨猴粪便菌群的结构与组成,为进一步开发利用新型实验动物奠定基础。方法采集4只成年雄性普通棉耳狨猴粪便,用细菌16SrRNA通用引物扩增V3~V4区,采用IlluminaMiSeq测序平台对普通棉耳狨猴粪便微生物进行研究。结果共测序获得315511条有效序列与596个OTU。普通棉耳狨猴粪便中的细菌共鉴定出9个门、14个纲、26个目、50个科、82个属和64个种和226个OTU。其中,1)优势门是拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes),平均含量分别为54.52%和25.39%2)丰度最高的纲为拟杆菌纲(Bacteroidia)和厌氧菌纲(Negativicutes),平均含量为54.5%和17%。3)乳杆菌目(Lactobacillales)和拟杆菌目(Bacteroidales)的丰度最高,为50.01%和20.52%。4)普雷沃菌科(Prevotellaceae)和双歧杆菌科(Bifidobacteriaceae)所占丰度最高,平均含量分别为43.14%和11.33%。5)双歧杆菌属(Bifidobacterium)和拟杆菌属(Bacteroides)的丰度较高,平均含量分别为11.33%11.12%。6)有益菌群双歧杆菌属丰度较高,但在所检测样本中都含有。7)丰度最高的前10个种,归类为7个科、5个纲。这10个种占到33.16%,其他53种总和仅占到0.74%,其他未鉴定出菌种,且相对丰度还较高,需要进一步研究。8)PICRUSt功能预测分析:氨基酸转运等代谢功能和蛋白质翻译、折叠遗传信息处理功能丰度较高。结论应用高通量测序技术,较全面的检测了普通棉耳狨猴粪便菌群,普通棉耳狨猴粪便细菌组成具有丰富的多样性,其中还有许多未被分类鉴定且相对丰度较高的细菌,需要进一步研究。     

Spontaneous pathology of the common marmoset (Callithrix jacchus) and tamarins (Saguinus oedipus, Saguinus mystax)

Background  Marmosets and tamarins are increasingly used in research, but their pathology remains poorly defined compared with Old World primates.

Methods  Necropsy records of 129 marmosets and 52 tamarins were reviewed; none was used experimentally.

Results  The most common marmoset lesions were dehydration, emaciation, nephritis, colitis, and inanition. The most common tamarin lesions were dehydration, ascites, emaciation, and congestive heart failure. Colitis and heart disease were the most common cause of death in marmosets and tamarins, respectively. Immature marmoset and tamarin deaths often occurred within the first month of life. Immature marmosets usually died from inanition, stillbirth, and colitis; immature tamarins from atelectasis, stillbirth, heart failure, and colitis. Lymphoma was the most common neoplasm for both marmosets and tamarins.

Conclusion  The findings were similar to prior reports with differences in frequency and severity. We report the first case of endometriosis in a marmoset.     

Monitoring ovarian function in marmosets and tamarins by the measurement of urinary estrogen metabolites

Practical aspects of urinary estrogen analysis were considered with regard to establishing simple and reliable methods for monitoring ovarian function in marmosets and tamarins. Changes in the hormone:creatinine ratio in small volumes of urine from the common marmoset were significantly correlated with changes in 24-h excretion. Comparison of the metabolism and excretion of estrogens during the ovarian cycle in the common marmoset and cottontop tamarin revealed interesting species differences. High concentrations of conjugated estrone were measured in marmoset plasma, but estradiol 17β was the predominant estrogen in urine. In contrast, estrone was the most abundant estrogen measured in tamarin urine. Both species excreted very little estriol. Sulfates and glucuronides were present in urine in similar proportions before ovulation in the marmoset, although after ovulation sulfates were the more abundant. Conversely, most of the estrogens in tamarin urine appeared to be conjugated as glucuronides. Direct assay for estrone sulfate was applied to the measurement of urinary estrogen excretion during the ovarian cycle in a marmoset. The results compared well with those for total estradiol 17β after hydrolysis and ether extraction. The use of direct assays for conjugated estrogens in small volumes of urine is suggested as a practical method for monitoring ovarian function in marmosets and tamarins.     

An extensive monoclonal antibody panel for the phenotyping of leukocyte subsets in the common marmoset and the cotton-top tamarin.

New World monkeys are valuable animal models to study human diseases. To determine the phenotype of cells involved in immune responses, we used flow cytometry to screen a large panel of anti-human monoclonal antibodies (mAb) for cross-reactivity with cells of the common marmoset and the cotton-top tamarin. Certain antigens (e.g., CD2, CD8, CD20) are well conserved. However, CD10, CD23, and CD33 showed a clear discrepancy in their reaction patterns in both species, indicating that significant differences on the epitope level occurred during evolution. Epstein-Barr virus-transformed B-cell lines were shown to be a valuable tool for screening B-cell-specific reagents. In some cases, fluorescein isothiocyanate (FITC) and phycoerythrin (PE) modification of mAbs had a negative effect on the binding capacity, which stressed the importance of choosing the right label. Despite the fact that some CD antigens were not detected, adequate numbers of cross-reactive mAbs were identified to perform extensive studies on immunological functions in both the common marmoset and the cotton-top tamarin.     

Yersinia enterocolitica infections in non-human primates.

Yersinia enterocolitica was isolated from ten non-human primates in the Netherlands. The following species were represented: Potto, Senegalgalago, wooly monkey, black spider monkey, common marmoset, cottonhead tamarin, pigtailed macaque and lesser whitenosed guenon.     

Prevalence of ocular disease in a colony of tamarins and marmosets.

In a large tamarin and marmoset experimental colony, 526 animals were examined by biomicroscopy and ophthalmoscopy for the presence of ocular disease. In 109 animals, there were 147 abnormalities involving the eyelids, cornea, iris, lens, retina or optic nerve. Most abnormalities were unimportant in terms of ocular function, but a few did cause loss of vision and included diffuse, progressive retinal atrophy and a severely traumatized globe. The survey indicated that while usually minor, ocular disease in the tamarin and marmoset was widespread.     

Pregnancy weight gain: marmoset and tamarin dads show it too

Paternal behaviour is critical for the survival of offspring in many monogamous species. Common marmoset (Callithrix jacchus) and cotton-top tamarin (Saguinus oedipus) fathers spend as much or more time caring for infants than mothers. Expectant males of both species showed significant increases in weight across the pregnancy whereas control males did not (five consecutive months for marmoset males and six months for cotton-top tamarin males). Expectant fathers might be preparing for the energetic cost of fatherhood by gaining weight during their mate's pregnancy.     

A Monoclonal Antibody Selection for Immunohistochemical Examination of Lymphoid Tissues From Non-human Primates

Non-human primates (NHPs) offer valuable animal models for basic research into human diseases and for the preclinical validation of new therapeutics. Detailed in situ examination of the involved cell types using immunohistochemistry is often hampered by the lack of cross-reactive antibodies (Abs). In the current study, we have tested a large panel of monoclonal antibodies raised against human leukocyte differentiation and activation markers for cross-reactivity on cryosections of lymphoid tissue from six NHP species. In total, we have tested 130 Abs against 69 antigens expressed in tissues from one great ape species (chimpanzee/Pan troglodytes), two Old World species (rhesus macaque/Macaca mulatta and cynomolgus macaque/Macaca fascicularis), and three New World species (common marmoset/Callithrix jacchus, cotton-top tamarin/Saguinus oedipus, and owl monkey/Aotus triviogatus). We have found a large panel of cross-reactive Abs: 93 of 102 (91%) in chimpanzee, 97 of 125 (78%) in rhesus macaque, 70 of 109 (64%) in cynomolgus macaque, 69 of 116 (60%) in common marmoset, 40 of 81 (49%) in cotton-top tamarin, and 35 of 80 (44%) in owl monkey. The availability of a reliable panel of cross-reactive markers is important to gaining further insight into immunological processes in disease-affected tissues from NHP species. (J Histochem Cytochem 57:1159–1167, 2009)     

Islet hyperplasia in callitrichids

Five callitrichids (three common marmosets -Callithrix jacchus -, a black tufted-eared marmoset-C. penicillata-, and a saddle-back tamarin -Saguinus fuscicollis) were diagnosed with islet hyperplasia by histopathology and immunohistochemistry. All were privately-owned, unrelated callitrichids ranging from 2- to 4-year-old. Relevant findings were anorexia (3/5), vomiting (2/5), ptyalism (1/5), polyuria/polydipsia (1/5), respiratory distress (1/5), hyperglycemia (2/3) and glycosuria (1/1); hyperglycemia and glycosuria were associated with pregnancy in a common marmoset and resolved after reducing simple carbohydrates in diet. All five animals died, three of them after few premonitory signs; in two cases, other concurrent diseases unrelated to islet hyperplasia were considered the cause of death. Additional animals from two facilities had high weight (4), physical obesity (3), polyuria/polydipsia/polyphagia/uriposia (1), hyperglycemia (1), and/or glycosuria (2). Pathologic findings in the deceased callitrichids were: islet hyperplasia (5/5); hemosiderosis (5/5); lipomatosis (4/5) of several tissues (atria, 3/5; pancreas, gall bladder, intestine, esophagus, and thyroid, 2/5; liver, 1/5); pancreatic necrosis or steatonecrosis, and/or acute pancreatitis (3/5); and vacuolation of hepatocytes and renal tubular cells most likely consistent with hepatorenal lipidosis (2/5). The islets of Langerhans were more numerous and larger than in a control, and morphologically normal in all cases, except in a common marmoset that had a few cells with a foamy cytoplasm and shrunken hyperchromatic or picknotic nucleus. Insulin (5/5), glucagon (3/5), and somatostatin (3/5) immunohistochemistry revealed that most cells stained positively for insulin diffusely in their cytoplasm (5/5) (staining restricted to the vascular pole of β-cells in the control). These findings suggest that obesity, insulin resistance and/or type II diabetes may be implicated and thus a prospective study on these diseases in callitrichids is necessary to determine their etiopathogenesis.     

Ultrasonographic and hormonal monitoring of pregnancy in the saddle back tamarin,Saguinus fuscicollis

Ultrasonography was used in six saddle back tamarin females (Saguinus fuscicollis) to diagnose pregnancy, monitor the patterns of uterine growth and embryonic/foetal development and examine the incidence loss of single embryos/foetuses. Pregnancy was reliably diagnosed 17 days after conception, 10 days earlier than by plasma progesterone measurement. The patterns of uterine and embryonic/foetal growth paralleled those reported for the common marmoset, including a delay in embryonic development. The results support the hypothesis of retardation of organogenesis in most callitrichid species. Individual embryos could be reliably identified from day 50 of pregnancy; a loss of single embryos/foetuses after this stage did not occur. All pregnancies were carried to term, resulting in five times twins and one singleton. The smaller litter size compared to the common marmoset may be due to loss of single embryos at earlier stages of pregnancy or to a lower ovulation rate.     

Comparative analysis of masseter fiber architecture in tree-gouging (Callithrix jacchus) and nongouging (Saguinus oedipus) callitrichids

Common marmosets (Callithrix jacchus) and cotton-top tamarins (Saguinus oedipus) (Callitrichidae, Primates) share a broadly similar diet of fruits, insects, and tree exudates. Common marmosets, however, differ from tamarins by actively gouging trees with their anterior teeth to elicit tree exudate flow. During tree gouging, marmosets produce relatively large jaw gapes, but do not necessarily produce relatively large bite forces at the anterior teeth. We compared the fiber architecture of the masseter muscle in tree-gouging Callithrix jacchus (n = 10) to nongouging Saguinus oedipus (n = 8) to determine whether the marmoset masseter facilitates producing these large gapes during tree gouging. We predict that the marmoset masseter has relatively longer fibers and, hence, greater potential muscle excursion (i.e., a greater range of motion through increased muscle stretch). Conversely, because of the expected trade-off between excursion and force production in muscle architecture, we predict that the cotton-top tamarin masseter has more pinnate fibers and increased physiological cross-sectional area (PCSA) as compared to common marmosets. Likewise, the S. oedipus masseter is predicted to have a greater proportion of tendon relative to muscle fiber as compared to the common marmoset masseter. Common marmosets have absolutely and relatively longer masseter fibers than cotton-top tamarins. Given that fiber length is directly proportional to muscle excursion and by extension contraction velocity, this result suggests that marmosets have masseters designed for relatively greater stretching and, hence, larger gapes. Conversely, the cotton-top tamarin masseter has a greater angle of pinnation (but not significantly so), larger PCSA, and higher proportion of tendon. The significantly larger PCSA in the tamarin masseter suggests that their masseter has relatively greater force production capabilities as compared to marmosets. Collectively, these results suggest that the fiber architecture of the common marmoset masseter is part of a suite of features of the masticatory apparatus that facilitates the production of relatively large gapes during tree gouging.