迁入都江堰野放中心的大熊猫野化放归个体皮质醇水平变化初步研究

动物生存环境的变化引起内分泌变化,而其程度则是动物适应环境的一种能力体现。以迁入成都大熊猫繁育研究基地都江堰野放繁育研究中心6只亚成年大熊猫个体为研究对象,利用酶联免疫吸附法检测其粪便中皮质醇激素,验证饲养环境改变是否引起其激素水平变化。研究发现:一方面,研究个体在迁入后的皮质醇水平较迁入前均呈明显下降趋势;另一方面,不同年龄和性别间,年龄越小则变化越明显,说明环境变化可能是引起其皮质醇变化的原因之一,即年龄越小这种环境引起的应激性越明显。因此,本研究亦可以为今后圈养大熊猫野化放归过程面临的环境适应性提供第一手的资料,进而为合理规划下一阶段大熊猫野化放归提供技术支持。     

粪样在大熊猫研究上的应用

大熊猫(Ailuropoda melanoleuca)数量少,传统取样法难以实施,样品的缺乏给大熊猫研究带来很大困难。而采集大熊猫粪样对大熊猫自身无损伤,粪样易获得和保存。本文总结了应用粪样进行的大熊猫食性与消化吸收、种群与个体识别、类固醇激素、寄生虫与病原微生物等方面的研究进展,对大熊猫粪样的应用前景进行了展望。     

猎豹粪样皮质醇代谢研究

采用放射性免疫法测量猎豹粪样中皮质醇代谢的含量,通过不同时间点的动态采样分析皮质醇含量的变化。在研究的6个雌性猎豹中,粪样皮质醇含量显示它们能够分成3个不同的组,高皮质醇含量的雌性猎豹组(～295.22ng/g粪样,n=2)、中等皮质醇含量雌性猎豹组(～152.29ng/g粪样,n=2)、偏低的皮质醇含量的雌性猎豹组(～94.14ng/g粪样,n=2)。外源压力影响了猎豹的繁殖性能,而皮质醇的代谢水平是反映猎豹来自外源压力的一个重要指标。本研究拟通过猎豹粪样中皮质醇的含量评估动物所处的环境压力水平,为饲养管理水平和饲养环境的改善提供有益的借鉴,进而为解决动物的繁殖问题提供新的思路和方法。     

贮存温度和时间对大熊猫粪便样品皮质醇浓度的影响

通过非损伤性取样比如粪便样品监测野生动物受胁迫的水平已成为一个重要的手段,然而针对从野外采集的样品,其运输和贮存温度条件是否干扰分析结果,尚未有过报道。为评估不同贮存温度和天数对野生动物粪便样品中皮质醇浓度的影响,我们将大熊猫的新鲜粪便置于3 种模拟条件(0℃ 、10℃ 和22℃ ),并进行连续6 d的检测分析。结果表明,贮存天数对粪样中皮质醇浓度有显著影响(F _5,107 = 7.501,P = 0.001),但温度则无显著影响(F _2,107 = 1.094,P = 0.366)。粪样在0℃ 、10℃ 和22℃ 条件下贮存6 d,皮质醇浓度与贮存前的基准值之间均不存在显著差异(0℃ :F _6,41 = 1.274,P = 0.290;10℃ :F _6,41 = 2.027,P = 0.084;22℃ :F _6,41 =1.009, P = 0.434)。结果提示在室温条件下短期运输和贮存(不超过6 d),不会引起大熊猫粪便样品中皮质醇浓度的显著变化。该结果对于野外所采集的粪便样品运输和贮存具有一定的指导意义。     

圈养林麝社会等级与粪样类固醇激素水平的关系

动物社群的社会等级可对个体的资源分配、社群稳定及个体的行为对策产生重要影响,圈养动物的类固醇激素水平与饲养管理有关,可反映动物的社群紧张水平。深入理解圈养动物的社会等级与其类固醇激素水平的关系是进行濒危动物迁地保育和成功驯养的基础。2018年6月15日—8月15日,用焦点取样法对四川马尔康林麝繁育场的25头林麝进行了冲突行为取样及社会等级计算,采用放射免疫分析法检测了同期粪样的类固醇激素水平,分析了林麝社会等级与粪样皮质醇、睾酮及雌二醇水平的关系。结果表明:低等级雌麝的雌二醇水平(289.037±59.710 pg·g^-1,n=11)显著高于高等级雌麝(45.670±27.283pg·g^-1,n=6)(P<0.05),低等级雄麝的睾酮水平(3.863±1.538 ng·g^-1,n=3)和高等级雄麝(8.017±1.295 ng·g^-1,n=5)无显著差异(P>0.05);低等级雄麝(37.891±7.564 ng·g^-1,n=3)和雌麝(37.262±1.544 ng·g^-1,n=11)的皮质醇水平与高等级雄麝(29.947±2.441 ng·g^-1,n=5)及雌麝(37.478±4.628 ng·g^-1,n=6)间的差异不显著(P>0.05)。不区分性别,低等级林麝的皮质醇水平(37.397±1.826 ng·g^-1,n=14)和高等级个体(34.055±2.886ng·g^-1,n=11)间无显著差异(P>0.05)。研究结果表明,圈养林麝雌体的社会等级与其粪样雌二醇水平呈负相关,低等级雌麝的粪样雌二醇水平显著高于高等级雌麝。在麝类驯养实践中,可监测雌麝的社会等级和粪样雌二醇水平变化,预测雌麝的行为健康及繁殖成效。     

唐家河自然保护区大熊猫种群数量及栖息地的移动

本文根据大熊猫季节性活动规律特点,应用海拔高度路线调查法,全面收集唐家河自然保护区大熊猫的比较新鲜粪便,计获１２４个粪样材料,提取ＤＮＡ作ＤＮＡ指纹图,经微机检测,保护区有大熊猫３７只,个体数量与粪样采集量之比为１∶３３５１４,有６个家系,雌兽２１只,雄兽１６只。并搞清了保护区生态环境质量,箭竹遭受病虫害的程度,洪水对栖息地的影响,特别对引起大熊猫数量减少及栖息地移动等原因,作了较为详细的分析。     

大熊猫母兽育幼早期应激水平与母幼行为的关系

大熊猫(Ailuropoda melanoleuca)属于晚熟性动物,育幼早期是幼仔存活和生长发育的关键时期,也是幼仔最易夭折时期。为了解大熊猫产后应激水平及其与母幼行为之间的关系,从行为适应机制的角度提高幼仔存活率,本研究利用酶联免疫吸附法监测7只处于育幼早期(产后0～75日)母兽粪便皮质醇含量,采用焦点动物取样法观测3对母幼个体的行为。结果显示：(1)育幼早期母兽皮质醇含量随幼仔日龄增长总体呈下降趋势(r=-0.950,P<0.001);(2)幼仔断奶前与断奶后相比,母兽皮质醇含量无显著差异(t=-0.935,P=0.361);(3)母兽皮质醇含量与母幼联系(r=0.918,P<0.001)、幼仔尖叫(r=0.777,P=0.001)和休息行为(r=0.731,P=0.002)均呈极显著正相关,与采食行为呈显著负相关(r=-0.608,P=0.016),而与舔幼仔行为不相关(r=0.366,P=0.179)。结果表明,大熊猫母兽育幼早期粪便皮质醇含量变化与幼仔生长发育有关,推测母兽的应激强度可能随幼仔御寒能力及自主活动能力的提升而降低;皮质醇可能参与母性行为调节,有助于...     

雄性小熊猫粪便中睾酮水平的变化与繁殖周期的关系

1999 年12 月至2000 年9 月, 使用放射免疫分析法, 测定了成都动物园和成都大熊猫繁育研究基地的4 只雄性小熊猫粪样中睾酮水平变化情况, 同时对其行为进行了观察。研究结果表明, 雄性粪样中睾酮水平在繁殖期(12 月～3 月) 显著高于非繁殖期(4 月～9 月) (P < 0.01) , 且睾酮水平变化与繁殖行为密切相关。在非繁殖期,雄性小熊猫的睾酮水平处于基础的较低水平时, 其繁殖行为如蹭阴、舔阴、舔标记物和咩叫等几乎没有发生或很少发生; 而在繁殖期, 雄性小熊猫的睾酮水平明显升高, 这些繁殖行为频繁发生。以上结果说明, 睾酮是调节雄性小熊猫繁殖行为季节性变化的生理因子。     

秦岭野生雌性川金丝猴粪便睾酮水平与邀配频次的季节性变化

为研究野生雌性川金丝猴(Rhinopithecus roxellana)不同季节体内睾酮水平与性行为频次的变化及二者的关系,我们以秦岭一野生川金丝猴种群为研究对象,通过长时间连续跟踪观察,完成了该种群中所有87只金丝猴的个体识别。用无损伤取样法采集了三只成年雌性川金丝猴不同季节的粪样,对粪样中的睾酮进行抽提测定,同时对采样个体的性行为进行记录。结果显示:1)成年雌猴在交配盛期与非交配盛期,粪样中睾酮水平均呈明显的周期性波动,交配盛期时睾酮水平的变化周期为24.56 d±4.07 d,非交配盛期时为43.67 d±2.89 d,显著长于交配盛期;2)成年雌猴在交配盛期的粪样睾酮水平为2.85 ng±1.12 ng/g,邀配频次为0.98±1.04次/d,非交配盛期时睾酮水平为1.71 ng±0.77 ng/g,邀配频次为0.28±0.53次/d,与交配盛期相比,非交配盛期的睾酮水平和邀配频次均明显降低;3)交配盛期各成年雌猴的粪样睾酮水平与邀配频次均有较显著的正相关关系(相关系数分别为0.631、0.683和0.659),而在非交配盛期这种相关关系并不存在(相关系数分别为0.091、0.493和0.205)。本研究表明,成年雌性川金丝猴体内睾酮水平、睾酮水平变化周期及邀配行为频次均具有明显的季节性差异,这种季节性差异可能是川金丝猴行季节性繁殖的内在原因和外在表现之一。本研究亦表明交配盛期雌猴的性行为频次在一定程度上可能受体内睾酮水平的控制,而在非交配盛期这种控制作用较弱。     

雄性弯角长角羚和普通大羚羊激素分泌模式与社会等级和季节的关系

为了确定羚羊类固醇激素分泌模式与行为和社群等级的关系,我们检测了成年雄性弯角长角羚(n=15)和普通大羚羊(n=11)的粪样皮质酮和睾酮浓度.采用随意取样和目标取样法记录了共650 h的行为.结果发现,两种羚羊的睾酮水平在雌性的发情时段里都是最高的.两种羚羊皮质激素水平有季节性变化,表现为雨季皮质酮水平高于干旱季节.优势胁迫作用明显存在于普通大羚羊,而在弯角长角羚不明显.没有证据表明从属个体压力的存在,一只弯角长角羚曾经是优势个体,但是后来有两年不是优势个体,与其他雄羚相比,这只雄羚在雌性发情期的睾酮和皮质酮水平有3个交迭出现的峰值.     

Comparison of leucine enkephalin and adrenocorticotrophin effects on adrenal function in fetal and adult sheep

At Day 120-125 of gestation equimolar amounts of ACTH and leu-enkephalin injected in vivo provoked similar rises in plasma cortisol concentrations in chronically catheterized fetuses. There was no concomitant change in plasma DHEA concentrations, or in maternal cortisol concentrations. At term (Days 135-140) 2 out of 5 animals responded similarly to both leu-enkephalin and ACTH injections with a rise in plasma cortisol concentrations, but the other 3 animals, in which basal cortisol concentrations had already risen, showed no response to either agonist. In adult sheep, ACTH provoked a significant increase in the plasma cortisol concentrations, but equimolar amounts of leu-enkephalin were without effect. There was a significant output of cortisol in response to ACTH administration by collagenase-dispersed adrenal cells from term sheep fetuses in vitro. Leu-enkephalin had no effect on cortisol output from dispersed adrenal cells when added by itself, or with ACTH. We conclude that leu-enkephalin is able to function as a stimulator of pituitary-adrenal function during fetal life. The lack of effect of leu-enkephalin on adrenal cells implies that its action is exerted not directly at the adrenal gland, but indirectly at the level of the hypothalamus or pituitary through stimulation of the release of other corticotrophic substances.     

Peripartal endocrine changes associated with retained placenta in dairy cows

Peripheral serum concentrations of prostaglandin F(2)alpha metabolite (PGFM), cortisol, and progesterone (P(4)) were determined in cows that retained placentae (RP; n = 18) and cows that did not retain placentae after parturition (NRP; n = 28). Blood samples collected from Day 15 before until Day 15 after parturition were assayed. Significant increases (P < 0.05) in PGFM concentrations occurred in RP cows between Days 7 and 6 before parturition compared with NRP cows. There were no differences in the interval required to attain peak concentrations of PGFM and the interval required to reach basal concentrations after parturition in both groups. However, the amount of PGFM secreted during the peripartum period was significantly higher in RP cows (P < 0.01). Starting from Day 6 before parturition, serum cortisol concentrations in RP cows increased significantly to peak levels 3 d before parturition (P < 0.05). Cortisol levels decreased to basal concentrations in RP cows on Day 1 postpartum. The peripartal serum P(4) concentrations in RP and NRP cows did not differ. The results indicate that prepartal PGFM and cortisol increases may constitute reliable indicators of retained placenta in dairy cows.     

Rabbit relaxin: the influence of pregnancy and ovariectomy during pregnancy on the plasma profile.

A porcine relaxin radioimmunoassay was developed to evaluate the profile of immunoreactive relaxin in rabbit plasma. Relaxin was nondetectable in pseudopregnant (Days 1, 4, 5-8, 12, and 16), nonpregnant, and male rabbits. However, in pregnant rabbits, relaxin was detected during the peri-implantation period (Days 4-9). Peak concentrations were reached on Day 15 and were maintained until parturition (Day 32). Relaxin concentrations abruptly decreased on Day 1 postpartum to low but detectable concentrations that were unchanged during the first week postpartum. In contrast, progesterone concentrations peaked earlier (Day 13), decreased after Day 25, and were not detectable on Day 1 postpartum. The effect of ovariectomy on the profile of plasma relaxin was evaluated. Four pregnant rabbits were ovariectomized (Day 13) and treated with medroxyprogesterone acetate to maintain pregnancy. As in normal pregnant rabbits, relaxin was observed initially during the peri-implantation period (Days 4-9) and increased to peak concentrations by Day 16. These concentrations were maintained until parturition and abruptly decreased on Day 1 postpartum to low yet detectable concentrations during the first week postpartum. The concentrations of relaxin in the plasma of ovariectomized medroxyprogesterone-treated rabbits were not different from those in three sham controls. These results indicate that the ovary is not a significant source of relaxin in pregnant rabbits. The unique observation of the presence of relaxin during the peri-implantation period suggests that this hormone has a role in preparing the rabbit uterus for implantation. The continued presence of relaxin during the first week postpartum may represent residual hormone, or it may suggest a physiological role during the early postpartum period.     

育幼经历对圈养成年雌性大熊猫尿液皮质醇含量的影响

为研究育幼期不同的育幼方式对圈养成年雌性大熊猫Ailuropoda melanoleuca福利状况的影响情况,了解圈养育幼模式可能引发成年雌性大熊猫的应激问题,完善育幼期大熊猫的饲养管理和兽舍参数设计,提高育幼过程中圈养大熊猫福利状况,选择成都大熊猫繁育研究基地育幼期采取不同育幼方式管理的成年雌性大熊猫为研究对象,利用...     

Localization of placental lactogen-I in trophoblast giant cells of the mouse placenta

The purpose of this investigation was to identify the cellular origin of placental lactogen-I (PL-I) expression in the mouse placenta and to cytologically define the transition from PL-I to PL-II expression during gestation. PL-I mRNA expression was assessed by in situ hybridization, and expression of PL-I and PL-II protein was determined by immunocytochemical analysis. PL-I mRNA and protein were localized to trophoblast giant cells. Trophoblast giant cells ceased producing PL-I at midgestation and began expressing PL-II. PL-I immunoreactivity was present in trophoblast giant cells on Days 9 and 10 of gestation but was not detectable in trophoblast giant cells on Day 11 of gestation. Immunoreactive PL-II-producing giant cells were detected first on Day 10 of gestation, continuing on Day 11 of gestation. Expression of PL-I and PL-II signals a significant functional transition in trophoblast giant cells of the developing mouse placenta.     

Cortisol in fetal fluids and the fetal adrenal at parturition in the tammar wallaby (Macropus eugenii)

Glucocorticoid hormones may play a critical role in initiating parturition in tammar wallabies. In this study, we investigated the concentration of cortisol in fetal fluids and cortisol production by fetal adrenals over the last 3 days of the 26-day pregnancy and within 24 h postpartum. The fetal adrenals almost doubled in size between Days 24 and 26 of pregnancy, and their cortisol content increased over 10-fold during this period, from 10 pg to over 100 pg per adrenal pair. After birth, neonatal adrenals continued to grow, but cortisol content fell dramatically to 20 pg. The prepartum increase in adrenal cortisol was reflected by a substantial rise in cortisol concentrations in yolk sac fluid, allantoic fluid, and fetal blood, which were below 10 ng/ml on Day 24 and rose to over 40 ng/ml by Day 26. Cortisol concentrations in neonatal blood decreased postpartum, mirroring decreased cortisol content in neonatal adrenals. Cortisol production by the fetal adrenal was stimulated in vitro by ACTH and prostaglandin E2, suggesting that the in vivo increase may be stimulated by release of ACTH from the fetal hypothalamic-pituitary axis and prostaglandin E2 from the placenta. These results indicate that increasing cortisol production by the fetal adrenal is a characteristic of late pregnancy in the tammar wallaby and support the suggestion that fetal cortisol may trigger the initiation of parturition in this marsupial species.     

Effect of restraint stress on plasma concentrations of cortisol, progesterone and pregnancy associated-glycoprotein-1 in pregnant heifers during late embryonic development

The aim of the present study was to evaluate the effect of restraint stress, which is commonly practised in the field, on plasma concentrations of cortisol, progesterone (P4) and bovine pregnancy-associated glycoprotein-1 (boPAG-1) in pregnant heifers between Days 30 to 40 of gestation. Twelve Holstein-Friesian heifers between Days 30 (Day 0 of experiment) and 40 (Day 10 of experiment) of pregnancy in a Hungarian dairy farm were used in the present study. The heifers were exposed to an acute stressor consisting of immobilisation (restraint stress) in a crush for 2 h (Group 1, n = 6) on Day 2 (Hour 48) and for 2 × 2 h (Group 2, n = 6) on Days 2 and 3 (Hour 72) of the experiment.Transrectal ultrasonography (7.5 MHz linear-array rectal transducer) was performed daily from Day 0 to Day 10 of the experiment to detect embryonic heartbeat or the fate of the conceptus. Blood samples were withdrawn before each ultrasonographic examination. Additional blood samples were withdrawn by 1 and 2 h (at Hours 49 and 50 in Groups 1 and 2 and Hours 73 and 74 in Group 2) of the onset of applying the stressor. Plasma cortisol, P4 and boPAG-1 concentrations were measured by radioimmunoassay. Acute restraint stress significantly (P < 0.001) increased the plasma cortisol level in pregnant heifers at 1 h of the exposure to the stressor at Days 2 (48 h) and 3 (72 h) of the experiment. On the other hand, the restraint stress did not affect the concentration of P4 and boPAG-1 concentrations in both groups. In conclusion, restraint stress for 2 h during early pregnancy in heifers increased blood cortisol, but it did not affect the concentrations of P4 and boPAG-1 between Days 30 to 40 of gestation.     

Serum luteinizing hormone, 13,14-dihydro-15-keto-prostaglandin F2alpha and cortisol profiles during postpartum anestrus in Brahman and Angus cows

Pluriparous suckled Brahman and Angus cows were utilized to evaluate the effect of breed, day after calving and endogenous opioid peptides (EOP) on hormonal profiles during postpartum anestrus. On Days 17 and 34 after calving, blood samples with and without heparin were collected at 15- and 30-min intervals, respectively, for a 7-h period via jugular cannula. Two hours after the start of blood sampling, cows of each breed were administered either 1 mg/kg iv naloxone or saline. Three hours later, all animals received 10 ng/kg iv GnRH. On Day 34 after calving cows received 0.2 IU/kg iv ACTH. Mean LH, basal LH and area under the LH curve increased (P < 0.01) from Day 17 to Day 34 after calving. Height of LH pulses increased (P < 0.05) by Day 34 after calving. Brahman cows had higher (P < 0.05) mean LH, basal LH, LH pulse frequency and area under the LH curve than Angus cows. Naloxone increased postchallenge area under the LH curve in treated cows above that of control cows (P < 0.06). Naloxone also increased the postchallenge area under the LH curve above that of the prechallenge level (P < 0.01). No breed differences in the response to the naloxone challenge were observed. The LH response to naloxone challenge occurred earlier on Day 34 than on Day 17 after calving but the amount of LH released was similar between days. The GnRH-induced LH release was greater in Brahman than in Angus cows (P < 0.04). Mean cortisol concentrations and area under the cortisol curve decreased (P < 0.05) between Day 17 and Day 34 after calving. Mean cortisol concentrations and area under the cortisol curve were lower (P < 0.01) in Brahman than in Angus cows. Cortisol secretion after ACTH treatment was similar between Brahman and Angus cows. The cortisol response after ACTH challenge was positively correlated (r=0.68; P < 0.001) to the prechallenge area under the cortisol curve. Under optimal environmental conditions Brahman cows have a greater LH release and their anterior hypophysis is more sensitive to GnRH challenge than the Angus cows.     

Contrasting seasonal modulation of the stress response in male and female rainbow trout

Between June and September the magnitude of the plasma cortisol response of maturing male and female rainbow trout to confinement was indistinguishable. A progressive increase in confinement-induced cortisol levels in both sexes occurred during May to September, associated with the seasonal rise in water temperature. Between September and January a reduction of >50% in the magnitude of the cortisol response to confinement in male fish (but no decline in females) coincided with declining water temperature and significant increases in plasma 11-ketotestosterone and elevated plasma testosterone levels. Plasma oestradiol-17β levels were significantly greater in females than males throughout the study period and this difference was maximal between September and January. However, plasma testosterone was also elevated in females during this period and levels overall were higher than those in male fish. Previous studies have shown oestradiol-17β and testosterone to have diametrically opposed effects on stress responsiveness in trout, with the former enhancing, and the latter suppressing, the cortisol response to a stressor. The relative roles of androgens, estrogen and water temperature in modulating the stress responsiveness of rainbow trout are discussed.     

Ingestion and ejection of hooks: effects on long-term health and mortality of angler-caught yellowfin bream Acanthopagrus australis

Ninety juvenile yellowfin bream Acanthopagrus australis were angled from holding tanks, allowed to ingest nickel-plated, carbon-steel J-hooks and released (with their lines cut) into individual experimental tanks during 2 experiments in order to assess their (1) long-term (up to 105 d) health, mortality and rate of hook ejection and (2) short- and medium-term (< 42 d) temporal changes in health during hook ingestion. Equal numbers of control fish were scooped from holding tanks and similarly monitored in experimental tanks. Of 20 hook-ingested fish released during Expt 1, 3 died within 8 d, providing a non-significant mortality of 15%. Between Day 6 and Day 56 post-release, 13 of the surviving individuals ejected their hooks, which were typically oxidized to about 94% of their original weight and often broken into 2 pieces. At Day 105, there were no significant differences between the 20 control and 17 hook-ingested/-ejected fish in terms of their ability to digest and assimilate food (measured as changes in apparent digestibility coefficients), stress (measured as concentrations of plasma cortisol and glucose) or of morphological parameters that included weight (Wt) and maximum height (MH), maximum width (MW) and maximum girth (MG). During Expt 2, 3 individuals that still contained ingested hooks and 3 controls were sampled on each of 9 occasions between Day 3 and Day 42 post-release. All fish were sampled for blood cortisol and glucose and were then euthanized before being weighed and measured for total length (TL), MH, MW and MG. Hook-ingested individuals were also X-rayed to determine the position and orientation of hooks. There were no significant differences in plasma glucose between hook-ingested and control fish. Irrespective of the treatment of fish, concentrations of cortisol were elevated on some sampling occasions, indicating variable, acute stress. The MH and MG of fish were not significantly different between groups. Significant differences were detected for MG and Wt, with hook-ingested fish having weights similar to those of the control fish but a relatively greater MW (owing to stomach distension from ingested hooks) until 2 wk post-release, after which both morphological parameters generally declined. There was no significant temporal progression of hooks in the stomach of treatment fish; however, some hooks reorientated to positions that may have precluded passage along the digestive tract. We conclude that, for the J-hooks examined, cutting the line is an appropriate strategy that results in the greater majority of released hook-ingested yellowfin bream surviving with minimal negative long-term effects.