川金丝猴粪样内3种类固醇激素保存时效分析

川金丝猴(Rhinopithecus roxellanae)是中国特有的灵长类物种,生存于海拔1 400～3 300 m的山地森林地带。采用拾取新鲜粪便的非损伤性途径研究该物种野生种群生理状态,必须首先确定其新鲜粪便在特定保存方法下的保存时效。本项研究探讨在川金丝猴自然分布区夏季可获得的低温(4±1)℃、在95%乙醇中保存条件下,该物种新鲜粪便内3种类固醇激素(睾酮、雌二醇、孕酮)在8个设定天数:0(标准对照)、5、6、7、8、10、20、30 d的保存时效。结果表明,雌性川金丝猴粪样内睾酮和雌二醇在30 d内可稳定保存,孕酮含量在保存10 d时的平均值显著性低于标准参照值(P0.05);雄性川金丝猴粪样中睾酮含量在保存6 d时的平均值显著低于标准参照值(P0.05)。本研究说明,在野生川金丝猴新鲜粪便保存条件一致的情况下,该物种新鲜粪样内睾酮、孕酮、雌二醇3种激素保存时效并不一致,实际运用时需要结合研究目的区别对待。     

大熊猫粪便样本处理与激素萃取方法比较

通过测定圈养动物粪便中的激素水平,监测其发情与怀孕状态和评估其福利状况是目前普遍采用的一种实验手段。然而,现有研究对于圈养野生动物粪便样本的处理及其激素萃取方法存在很大分歧,研究结果也不尽一致。本研究采集了10只(雌∶雄=1∶1)圈养大熊猫(Ailuropoda melanoleuca)的粪便样本,分别采用冷冻干燥-研筛法和冷冻干燥-粉碎法处理粪便样本,然后分别使用80%甲醇、90%乙醇和95%乙醇3种萃取剂进行激素萃取,最后比较2种粪便处理方法和3种溶剂萃取方法所获得的粪便样本中孕酮、雌二醇、睾酮和皮质醇的浓度。结果表明,冷冻干燥-研筛法结合80%甲醇溶液萃取处理的样本中雌二醇和孕酮检测值最高,冷冻干燥-粉碎法结合80%甲醇溶液萃取处理的样本中睾酮检测值最高。冷冻干燥-粉碎法结合90%乙醇溶液萃取处理的样本与冷冻干燥-研筛法结合80%甲醇溶液萃取处理的样本中皮质醇检测值更高,且2种方法的检测结果差异不显著。综合粪便处理难易程度和萃取溶剂性质,建议雌二醇、孕酮和睾酮激素检测根据最高检测浓度的方法进行粪便前处理,皮质醇激素检测使用冷冻干燥-粉碎法结合90%乙醇溶液萃取方法进行粪便前处理。...     

粪便留取量对粪便中细菌结构和功能的影响CSCD

目的探究不同粪便留取量对肠道菌群构成研究结果的影响,为肠道菌群构成研究中粪便样本的采集方式提供参考。方法本研究分别对全便和部分粪便进行预处理,并使用PacBio SMRT测序技术对10名志愿者的20份粪便样本进行了16S rRNA全长测序,通过多元统计学等手段对粪便中细菌的构成、多样性和功能进行比较分析。结果Eubacterium rectale、Eubacterium ventriosum和Allisonella histaminiformans等低丰度物种的相对丰度在不同粪便留取量中有一定差异,其在全便中的相对丰度均显著高于部分粪便(P<0.05)。但Wilcoxon配对检验发现粪便留取量对肠道菌群结构、多样性和功能的影响均无显著性差异(P>0.05)。不同个体间肠道菌群结构存在明显差异,且粪便留取量的影响远小于个体差异。结论全便和部分粪便两种粪便留取量对检测微生物的组成、多样性和功能无显著影响,因此取部分粪便即可满足对肠道菌群研究的要求。     

东方白鹳粪便总DNA五种提取方法的比较

本研究比较东方白鹳(Ciconia boyciana)粪便总DNA的5种提取方法,旨在为后续物种性别鉴定和DNA条形码鉴定提供合适的方法.采用十六烷基三甲基溴化铵法(CTAB法)、十二烷基磺酸钠裂解法(SDS法)、Tiangen试剂盒法、Qiagen试剂盒法和异硫氰酸胍法(GuSCN法)对取自天津动物园的东方白鹳新鲜粪...     

新疆塔里木马鹿粪便激素含量季节性变化的初步研究

目的:检测雄性塔里木马鹿的睾酮,雌鹿的雌二醇、雌三醇、孕酮等粪便激素含量,并比较不同月份之间的变化。方法:利用放射免疫分析法(RIA)在且末和尉犁县马鹿场采集塔里木马鹿粪便,检测了2004年2～12月间塔里木马鹿粪便中的激素水平。结果:粪便性激素水平有明显的季节变化(P<0.05)。雄鹿粪便睾酮水平月间差异显著(P<0.05),浓度在8月有2个高峰。雌鹿粪便雌二醇在3月和6月有2个高峰;粪便孕酮水平在3月有2个高峰,在8月有1个高峰;雌三醇差异不显著,但4月的最高值也显著高于其他各时间点(P<0.05)。结论:利用动物粪便研究激素具有对保护动物完全无伤害、材料收集容易等优点,并且有助于评估动物青春期的健康状况、判断动物排卵方式、早孕检查、产前预测等,因而是目前研究野生珍稀动物性激素的一种较为理想的方法。     

繁殖期雌性蓝狐性激素检测与 性器官组织学观察

为了研究繁殖期雌性蓝狐(Alopexlagopus)类固醇激素含量、发情表现与性器官组织学结构之间的关系,于2019年选取繁殖期不同发情状况的雌性蓝狐,共18只,分为典型发情、非典型发情和不发情3组。通过无损伤取样法采集粪便与尿液并通过放射免疫法(RIA)测定孕酮(P)和雌二醇(E2)的含量;每组选取1只在发情并输精的当天取卵巢和子宫,进行组织学观察。结果显示,所测定典型发情、非典型发情、不发情蓝狐类固醇激素水平,雌二醇在尿液中的含量显著高于粪便,且在不同组间差异明显,尤其在典型发情组更高,达到(11 065.17±546.76)ng/L;孕酮含量尿液与粪样差异不大,孕酮含量在典型发情雌性蓝狐粪便与尿液含量差异不显著,但在不发情雌性蓝狐中差异显著,为(16.61±0.63)μg/L。粪尿测定孕酮和雌二醇激素含量与三组不同情况的发情表现相比相关性显著。粪、尿作为类固醇激素检测样本,虽然含量有差异,但变化趋势一致,都可使用。卵巢与子宫在典型发情雌性蓝狐中体积较大,卵巢可见各级卵泡和多个黄体,子宫黏膜上皮为柱状上皮,排列紧密,固有层内可见大量腺体;非典型发情雌性蓝狐卵巢、子宫发育状况与典型发情雌性蓝狐相类似;不发情雌性蓝狐卵巢、子宫呈静息状态,卵巢中卵泡多处于闭锁状态,无卵母细胞,也无黄体,固有层间质细胞及肌层肌细胞排列更为紧密。说明检测孕酮和雌二醇激素的含量,可以准确判定蓝狐发情的状态。     

圈养林麝粪便类固醇激素保存时效性研究

用非损伤性采样方法对圈养林麝在三个条件:1)未处理,2)90%甲醇,3)0.9%抗生素,15个时间梯度下(20℃)的粪便中类固醇激素及其代谢物的保存时间及方法进行了比较,结果表明:林麝粪便在室温下(20℃),孕酮及其代谢物48 h有明显的上升趋势;雌二醇、睾酮和皮质醇及其代谢物在18 h均出现显著变化;对比抗生素保存,用甲醇保存是一种较好的保存林麝粪便类固醇激素的方法;然而不同激素用甲醇保存的时间不同,孕酮不宜超过1个月,皮质醇不宜超过半个月,睾酮则不宜超过10天,而雌二醇可以保存2个月之久.     

粪便收集方式对异育银鲫表观消化率测定的影响

消化率的高低是评价饲料营养价值的重要指标,但是粪便的不同收集方式会影响消化率的测定结果。研究观察了异育银鲫摄食后的排粪高峰以及排粪持续时间,探讨了粪便的不同收集方式对异育银鲫(Carassius auratus gibelio)(约100 g)表观消化率(Apparent digestibility coefficient,ADC)测定结果的影响。饲料中加入1%三氧化二铬(Cr2O3)作为指示剂,实验采用3种粪便收集方法(积粪器法、虹吸法和后肠取粪法),并且在积粪器法和虹吸法中设置了不同的收粪时间。结果表明,正常投喂的异育银鲫摄食后的排粪高峰出现在摄食结束后的5—10h;饥饿2d的异育银鲫恢复摄食后的排粪高峰出现在摄食结束后的7—9h;饲料的干物质表观消化率(ADC of dry matter,ADCd)和蛋白表观消化率(ADC of protein,ADCp)在排粪初始阶段较低,随着排粪高峰的出现而升高。粪便的不同收集方法和不同收集时间对ADCd和ADCp的测定结果均存在显著影响(P<0.05)。采用虹吸法收集粪便测得的ADCd与在排粪高峰时段内积粪器法的测定值相当;后肠取粪法收集粪便测得的ADCd低于积粪器法排粪高峰期的测定值。采用虹吸法收集粪便测得的ADCp略低于在排粪高峰时段内积粪器法的测定值;后肠取粪法收集粪便测得的ADCp低于积粪器法各时间段内的测定值。因此,在选择挑选新鲜完整粪便的前提下,在排粪高峰期采用虹吸法收集粪便用于ADC的测定是一种方便可靠的方法,反而后肠取粪法由于操作误差容易带来更大偏差。     

环境DNA metabarcoding及其在生态学研究中的应用

环境DNA metabarcoding(eDNA metabarcoding)是指利用环境样本(如土壤、水、粪便等)中分离的DNA进行高通量的多个物种(或高级分类单元)鉴定的方法。近年来,该方法引起了学者的广泛关注,逐渐应用于生物多样性研究、水生生物监测、珍稀濒危物种和外来入侵物种检测等生态学领域。介绍环境DNA metabarcoding的含义和研究方法;重点介绍环境DNA metabarcoding在物种监测、生物多样性研究和食性分析等生态学领域中的应用;总结环境DNA metabarcoding应用于生态学研究领域面临的挑战并对该方法的发展进行展望。     

DNA宏条形码技术在鸟类食性分析中的运用

鸟类在全球广泛分布,不同鸟类物种利用的食物类群存在很大差异,而食性研究是动物营养学和生态学领域的重要研究内容。本文对一些传统鸟类食性鉴别方式及其不足进行回顾,传统鸟类食性鉴别方式包含扎颈法、剖胃法、粪便收集法、相机记录法等。随着测序技术的高速发展,DNA宏条形码技术出现,并广泛应用于动物食性研究。近些年来,该技术也被应用于鸟类食性研究中。本文综述了DNA条形码和DNA宏条形码的操作原理和条件,对鸟类食性研究中的DNA条形码与引物的选择做了详细介绍。对比传统鉴别方法,DNA宏条形码技术降低了物种鉴定难度,减少了人为影响因素,提高了目标样本中物种的鉴定效率,能对粪便、胃容物等混合或不成型样本进行分析。另一方面,在扩增多物种混合的DNA样品中的目标片段时,可能出现偏离,造成结果的不确定性,并且难以根据结果得出较准确各食物组分的比例。未来在使用宏条形码技术对鸟类食性的分析中,可结合其他方法改善对食物的量化以及食物属性的判断。     

白蚁与动物及微生物的共生类型及其机制

综述了白蚁螱客的主要种类、共生关系及相关机制的研究进展。白蚁螱客中,已报道的动物种类达170种。在与动物的共生关系中存在偏利共生（宾主共栖和异种共栖）、互利共生和无关共生三种;在与微生物的共生关系中,存在与内生菌（原生动物、细菌、真菌和放线菌）和外生菌（蚁巢伞菌等）间的互利关系。指出了白蚁与螱客研究中存在的问题,给出了解决方案,并提出了今后可能的研究热点或方向,为白蚁的综合利用（如纤维素酶）及今后研究物种间的协同进化提供了基础资料。     

New amino-dithiaphospholanes and phosphoramidodithioites and their rhodium and iridium complexes

New sulfur derivatives of phosphoramidite ligands were synthesized and the impact of the sulfur unit on the spectroscopic properties of their rhodium and iridium complexes was investigated. The new ligands Bn₂NPSCH₂CH₂S^a(P-S^a) (Bn = benzyl, 4), Bn₂NPSCHCHS^a(CH₂)₃C^aH₂(P-S^a)(C^a-S^a) (6) and Bn₂NP(4-XC₆H₄OMe)₂ (X = S, 7a; X = O, 7b) were converted to the rhodium and iridium complexes trans-[Rh(CO)Cl(L)₂] (L = 4, 6, 7), [RhCl(COD)(L)] (L = 4, 6, 7), [IrCl(COD)(7a)] and [IrCl₂Cp∗(6)]. For comparison, some phosphoramidite complexes of these formulations also were synthesized. The new metal complexes were spectroscopically analyzed. For the carbonyl complexes, the ν_CO IR stretching frequencies were lower than for the corresponding phosphite and phosphoramidite ligands. The ¹J_PRh coupling constants for the rhodium complexes with the new ligands were also smaller than for the respective phosphoramidite and phosphite complexes. Finally, the ¹J_PSe coupling constants of the selenides of the new ligands were lower than those of the phosphoramidite ligands but higher than for PPh₃. The spectroscopic data reveal that the new thio ligands 4, 6 and 7a are more electron donating than phosphites and phosphoramidites but less electron donating than PPh₃.     

Astrocytes and Synaptosomes Transport and Metabolize Lactate and Acetate Differently

Astrocytes transport the monocarboxylate acetate, but synaptosomes do not. The reason for this is unknown, because both preparations express monocarboxylate transporters (MCT). The transport and metabolism of lactate, another monocarboxylate, was examined in these two preparations, and the results were compared to those for acetate. Lactate transport is more rapid in astrocytes than in synaptosomes, but of lower affinity (Kms of 17 and 4 mM, respectively). Lactate (0.2 mM) is metabolized to CO2 more rapidly in synaptosomes than in astrocytes (rates of 0.37 and 0.07 nmol x mg protein(-1) x min(-1), respectively). The reason for this is unclear, but cellular differences in lactate dehydrogenase isotype expression may be involved. Acetate is metabolized to CO2 more rapidly in astrocytes than in synaptosomes (rates of 0.43 and 0.02 nmol x mg protein(-1) x min(-1), respectively). This is likely due to cellular differences in the expression of monocarboxylate transporter subtypes.     

Rapporteur report: Basics and technology and metrology and standards

The first and second sessions of the Workshop focussed on the basics of ultrasound and infrasound, their applications in both industry and medicine, and metrology and protection standards for ultrasound applications.     

Relative and combined effects of ethanol and protein deficiency on strontium and barium bone content and fecal and urinary excretion

To elucidate accumulation of minerals in human iliac arteries with aging, the content of minerals was analyzed by inductively coupled plasma atomic emission spectrometry. Bilateral common, internal, and external iliac arteries of 16 men and 8 women, ranging ages from 65 to 93 yr, were examined. It was found that an extremely high accumulation of calcium and phosphorus occurred in the common iliac artery at old age, being higher than that of the internal and external iliac arteries. It should be noted that the accumulation of calcium and phosphorus is the highest in the common iliac artery among the human arteries examined to date. Regarding sexual differences, the content of calcium and phosphorus in the common and internal iliac arteries was higher in women than in men, whereas their content in the external iliac artery was lower in women than in men.     

Prostaglandin and thromboxane production by human and guinea-pig macrophages and leucocytes

The ability of partially purified human and guinea-pig haematogenous cell populations, when cultured in vitro, to metabolise arachidonic acid (AA) has been studied. Supernatants from 24 hour cell culture have been subjected to analysis for products of AA metabolism by gas chromatography with electron-capture detection.The cell types studied were human peripheral blood monocytes (both glass adherent and non-adherent), neutrophils, eosinophils and leukemic leucocytes; thoracic duct lymphocytes and lung alveolar macrophages. From the guinea-pig, induced and non-induced macrophage or neutrophil enriched peritoneal exudate populations, lymph node cells, peritoneal eosinophils and peripheral blood platelets were examined. Supernatants were assayed for the presence of PGE₂, PGD₂, PGF_2α, TXB₂ and 6-keto-PGF_1α. In all types studied PGE₂ and TXB₂ were the major products formed. The identification of PGE₂ and TXB₂ was confirmed by GC/MS with multiple ion monitoring.The results have been compared with other reports and their possible significance discussed in relation to the proposed role of prostaglandins as mediators and modulators in immunopathology.     

Chemokines and cytokines: axis and allies in asthma and allergy

Allergic asthma can be precipitated by many factors. For the atopic person, fungus, pollen, dust mites, cockroach antigens, and diesel exhaust are all agents that may trigger an allergic attack. Cytokines and chemokines are integral mediators of fungal asthma. From the earliest time points, they recruit and activate the cells required for the clearance of fungus as well as being critical factors involved in the immunopathology of this disease. In the final analysis, it is clear that these mediators can act to the benefit or the detriment of the host.     

Oxidation and nitration of ribonuclease and lysozyme by peroxynitrite and myeloperoxidase

In spite of the many studies on protein modifications by reactive species, knowledge about the products resulting from the oxidation of protein-aromatic residues, including protein-derived radicals and their stable products, remains limited. Here, we compared the oxidative modifications promoted by peroxynitrite and myeloperoxidase/hydrogen peroxide/nitrite in two model proteins, ribonuclease (6Tyr) and lysozyme (3Tyr/6Trp). The formation of protein-derived radicals and products was higher at pH 5.4 and 7.4 for myeloperoxidase and peroxynitrite, respectively. The main product was 3-nitro-Tyr for both proteins and oxidants. Lysozyme rendered similar yields of nitro-Trp, particularly when oxidized by peroxynitrite. Hydroxylated and dimerized products of Trp and Tyr were also produced, but in lower yields. Localization of the main modified residues indicates that peroxynitrite decomposes to radicals within the proteins behaving less specifically than myeloperoxidase. Nitrogen dioxide is emphasized as an important protein modifier.