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Molecular phenotyping of domestic cat (Felis catus) testicular cells across postnatal development – A model for wild felids 家猫(Felis catus)睾丸细胞在出生后发育过程中的分子表型——野生猫科动物的一个模型
Pub Date : 2023-01-01 DOI: 10.1016/j.therwi.2023.100031
M. Bashawat , B.C. Braun , K. Müller , B.P. Hermann

Molecular characterisation of testicular cells is a pivotal step towards a profound understanding of spermatogenesis and developing assisted reproductive techniques (ARTs) based on germline preservation. To enable the identification of testicular somatic and spermatogenic cell types in felids, we investigated the expression of five molecular markers at the protein level in testes from domestic cats (Felis catus) at different developmental phases (prepubertal, pubertal I and II, postpubertal I and II) classified by single-cell ploidy analysis. Our findings indicate a prominent co-labelling for two spermatogonial markers, UCHL1 and FOXO1, throughout postnatal testis development. Smaller subsets of UCHL1 or FOXO1 single-positive spermatogonia were also evident, with the FOXO1 single-positive spermatogonia predominantly observed in prepubertal testes. As expected, DDX4+ germ cells increased in numbers beginning in puberty, reaching a maximum at adulthood (post-pubertal phase), corresponding to the sequential appearance of labelled spermatogonia, spermatocytes and spermatids. Furthermore, we identified SOX9+ Sertoli cells and CYP17A1+ Leydig cells in all of the developmental groups. Importantly, testes of African lion (Panthera leo), Sumatran tiger (Panthera tigris sumatrae), Chinese leopard (Panthera pardus japonesis) and Sudan cheetah (Acinonyx jubatus soemmeringii) exhibited conserved labelling for UCHL1, FOXO1, DDX4, SOX9 and CYP17A1. The present study provides fundamental information about the identity of spermatogenic and somatic testicular cell types across felid development that will be useful for developing ART approaches to support endangered felid conservation.

睾丸细胞的分子特征是深入了解精子发生和开发基于种系保存的辅助生殖技术(ARTs)的关键一步。为了能够鉴定猫睾丸体细胞和生精细胞类型,我们通过单细胞倍性分析研究了家猫(Felis catus)在不同发育阶段(青春期前、青春期I和II、青春期后I和II)睾丸中蛋白质水平的五种分子标记物的表达。我们的研究结果表明,在出生后的睾丸发育过程中,两种精原细胞标志物UCHL1和FOXO1存在显著的共标记。UCHL1或FOXO1单个阳性精原细胞的较小亚群也很明显,FOXO1单阳性精原原细胞主要在青春期前睾丸中观察到。正如预期的那样,DDX4+生殖细胞的数量从青春期开始增加,在成年期(青春期后)达到最大值,对应于标记的精原细胞、精母细胞和精子细胞的顺序出现。此外,我们在所有发育组中鉴定了SOX9+支持细胞和CYP17A1+Leydig细胞。重要的是,非洲狮、苏门答腊虎、中国豹和苏丹猎豹的睾丸对UCHL1、FOXO1、DDX4、SOX9和CYP17A1表现出保守的标记。本研究提供了有关猫发育过程中生精细胞和体细胞睾丸细胞类型的基本信息,这将有助于开发ART方法来支持濒危猫科动物的保护。
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引用次数: 0
Validation of the iSperm for assessing rhinoceros Sperm 用于评估犀牛精子的iSperm验证
Pub Date : 2023-01-01 DOI: 10.1016/j.therwi.2023.100048
Louisa A. Rispoli, Terri L. Roth
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引用次数: 0
Ovarian ultrasound analysis for developing temporal and spatially explicit hormone regimens for induced ovulation and egg deposition in the tiger salamander (Ambystoma tigrinum) 卵巢超声分析用于制定时间和空间明确的激素方案,用于虎螈(Ambystoma tigrinum)的促排卵和卵子沉积
Pub Date : 2023-01-01 DOI: 10.1016/j.therwi.2023.100038
Ruth M. Marcec-Greaves , Carrie K. Kouba , Scott T. Willard , Andy J. Kouba
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引用次数: 0
First detailed description of follicle development in a chimaeroid, the American elephant fish Callorhinchus callorhynchus 首次详细描述嵌合体的卵泡发育,美洲象鱼calorhinchus calorhynchus
Pub Date : 2023-01-01 DOI: 10.1016/j.therwi.2023.100037
S.N. Fuentes , C.A. Awruch , C. Moya , A. Wehitt , M.C.Díaz Andrade

Chimaerods represent an enigmatic lineage of cartilaginous fishes and the least studied group within Chondrichthyans. They have a widespread global distribution with a high degree of endemism. In addition, this group has been considered among the least threatened species of Chondrichthyans, but also the most data-deficient group, with biological information lacking for most species. This study represents the first microanatomical, histological, and ultrastructural description of the different stages of folliculogenesis in Callorhinchus callorhynchus, and the first ultrastructural study about this topic within chimaeroids. Specimens of different maturity stages were collected from landings of the commercial fleet operating in San Matías Gulf, Argentina. Folliculogenesis shows some differences compared to other elasmobranchs. Oogonia were present in immature and mature females. Atretic follicles of different sizes were present in mature female ovaries. There is a significant difference between the macroscopic (∼10 mm) and microscopic (∼ 2 mm) follicular size at the beginning of vitellogenesis. There is no evidence of an epigonal organ. The results are discussed in a physiological and reproductive context. This study provides valuable information that will contribute to the understanding of the reproductive biology of the chimaeroids species and its potential for fishery exploitation recovery.

嵌合体代表了软骨鱼类的一个神秘谱系,也是软骨鱼纲中研究最少的群体。它们在全球广泛分布,具有高度的地方性。此外,这一群体被认为是受威胁最小的软骨鱼类物种之一,但也是数据最缺乏的群体,大多数物种缺乏生物信息。本研究首次在显微解剖、组织学和超微结构上描述了卡罗hinchus calorhinchus calorhynchus卵泡发生的不同阶段,并首次在嵌合体中进行了有关该主题的超微结构研究。在阿根廷圣Matías湾的商业船队上采集了不同成熟阶段的标本。卵泡发生与其他弹性分支有一定的差异。未成熟和成熟雌性均有卵原体。成熟女性卵巢中存在大小不等的闭锁卵泡。卵泡形成初期,肉眼(~ 10 mm)和显微镜下(~ 2 mm)的卵泡大小有显著差异。没有外阴器官的证据。结果在生理和生殖的背景下进行了讨论。本研究为进一步了解拟嵌合体的生殖生物学及其在渔业开发和恢复方面的潜力提供了有价值的信息。
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引用次数: 0
Ovarian cycle, reproductive performance and breeding seasonality of Amazonian yellow-footed tortoises (Chelonoidis denticulatus) in the wild 野生亚马逊黄足象的卵巢周期、繁殖性能和繁殖季节
Pub Date : 2023-01-01 DOI: 10.1016/j.therwi.2023.100022
Pedro Mayor , Sara Hidalgo , Hani R. El Bizri , Thais Q. Morcatty

Studies of the reproductive functional morphology of chelonians are important to understand the reproductive physiology, anatomy, and endocrinology of these species. However, data on the reproduction of this group have been difficult to obtain because of their cryptic habits and as a result the consequent inefficiency of conventional field methods in detecting individuals in the wild. In this study, we obtained reproductive organs from 347 adult female yellow-footed tortoises (Chelonoidis denticulatus), one of the most hunted and traded land vertebrates in South America, donated by local sellers in a wild meat market in the Peruvian Amazon. We described the species’ reproductive cycle and the influence of climatic factors on reproductive performance. Of the sampled females, 116 (33.4%) were gravid and 231 (66.6%) were non-gravid, including 215 vitellogenic (62.0%) and 16 non-vitellogenic (4.6%). The mean number of ovarian follicles per female declined from 28.5 (<10 mm diameter), to 7.1 follicles (20–40 mm) to 3.2 follicles> 40 mm. Gravid females had in average 3.7 shelled eggs in the oviduct, with a mean egg diameter of 53.7 mm. The estimated annual reproductive potential in gravid females was 5.1 eggs per clutch, ranging from one to 22 eggs. There was a strong positive relationship between the diameter of shelled eggs and the straight carapace length of gravid females. The final phase of the folicular growth, the most demanding energetic process in chelonian reproduction, correlated with annual rainfall peak, while oviposition was estimated to occur in the dry season. The yellow-footed tortoise should be considered an opportunistic seasonal breeder, though capable of reproducing throughout the year. Reproductive yields are linked to climatic events that influence food availability in the environment. To improve the sustainable use of this species, adult females should be harvested primarily during reproductive quiescence, from the end of the laying period in the late dry season to the mid-rainy season when large follicles appear, to avoid taking gravid females. Our results are useful not just to better understand the reproductive biology of terrestrial chelonians but can inform the conservation management of harvested species.

研究龟类的生殖功能形态对了解龟类的生殖生理学、解剖学和内分泌学具有重要意义。然而,由于其隐秘的习性以及传统野外方法在野外检测个体的效率低下,很难获得这一群体的繁殖数据。在这项研究中,我们从347只成年雌性黄足龟(Chelonoidis denticulatus)身上获得生殖器官,黄足龟是南美洲最被猎杀和交易的陆地脊椎动物之一,由秘鲁亚马逊地区野生肉类市场的当地卖家捐赠。我们描述了该物种的繁殖周期和气候因素对繁殖性能的影响。其中妊娠116例(33.4%),非妊娠231例(66.6%),其中黄疸215例(62.0%),非黄疸16例(4.6%)。每位女性卵巢卵泡的平均数量从28.5个(直径10毫米)下降到7.1个(20-40毫米),再到3.2个。40毫米。妊娠雌虫平均产蛋3.7枚,卵直径53.7 mm。估计怀孕的雌性每年的繁殖潜力是每窝5.1个卵,从1到22个卵不等。有壳卵直径与妊娠雌鱼的直甲壳长呈极显著正相关。卵泡生长的最后阶段是龟生殖过程中最需要能量的过程,与年降雨量高峰相关,而产卵估计发生在旱季。黄脚龟应该被认为是一个机会主义的季节性繁殖者,尽管全年都有繁殖能力。生殖产量与影响环境中粮食供应的气候事件有关。为了提高该物种的可持续利用,成年雌性应主要在生殖静止期间收获,从旱季后期的产卵期结束到雨季中期出现大卵泡时,以避免捕获怀孕的雌性。我们的研究结果不仅有助于更好地了解陆生龟类的生殖生物学,而且可以为收获物种的保护管理提供信息。
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引用次数: 0
Use of hormones in assessing reproductive physiology of humpback whales (Megaptera novaeangliae) from Juneau, Alaska 激素在评估阿拉斯加朱诺座头鲸繁殖生理中的应用
Pub Date : 2023-01-01 DOI: 10.1016/j.therwi.2023.100050
S. Atkinson , V. Melica , S. Teerlink , K. Mashburn , J. Moran , H. Pearson

Humpback whales (Megaptera novaeangliae) in Southeast Alaska have been studied for over 50 years, and are largely considered a recovery success since the cessation of commercial whaling. Reproductive physiology is an important factor to consider in studying population health and can provide important insights into the drivers contributing to population abundance fluctuations. Validated assays for progesterone and testosterone were used on blubber biopsies from humpback whales (N = 33 whales, 71 samples) near Juneau, Alaska, in 2020 and 2021. Long-term sighting histories were used to confirm detected pregnancies with calf sightings the following year. Blubber samples were divided into two seasonal bins (early and late summer). Pregnant females sampled in both early and late summer of both 2020 and 2021 showed elevated progesterone concentrations compared to other reproductive states (p < 0.05). Progesterone concentrations in adult male whales (0.3 ± 0.2 ng/g) were not significantly different from lactating or resting female whales. Blubber testosterone concentrations in adult male humpback whales ranged from 0.05 to 1.1 ng/g, and mean concentrations were approximately double those of female whales in any reproductive state. Pregnancy was detected in 5 of 11 and 4 of 9 adult females in 2020 and 2021 respectively, yielding summer season pregnancy rates for sexually mature females at 0.45, and 0.44, respectively. Calving rates were 0.36 and 0.22 in 2020 and 2021, respectively, and the annual growth rate for this subpopulation was calculated at 2.6 % per annum. One female had successful pregnancies for four consecutive years. These results demonstrate the synergistic value of combining immunoreactive assays and long-term sighting histories to further knowledge of reproductive physiology in individual humpback whales, which can be expanded to assessing the health of a population or ecosystem.

人们对阿拉斯加东南部的座头鲸(Megaptera novaeangliae)进行了50多年的研究,自商业捕鲸停止以来,座头鲸在很大程度上被认为是恢复的成功。生殖生理学是研究种群健康的一个重要因素,可以为种群丰度波动的驱动因素提供重要的见解。2020年和2021年,在阿拉斯加朱诺附近的座头鲸(N = 33头鲸,71个样本)的鲸脂活检中,使用了有效的孕酮和睾酮检测方法。长期的观察历史被用来确认第二年发现的怀孕和幼崽的观察。鲸脂样本分为两个季节性箱(夏初和夏末)。在2020年和2021年夏初和夏末取样的孕妇与其他生殖状态相比,黄体酮浓度升高(p <0.05)。成年雄鲸的孕酮浓度(0.3±0.2 ng/g)与哺乳期或休息期雌鲸无显著差异。成年雄性座头鲸的鲸脂睾酮浓度在0.05至1.1纳克/克之间,平均浓度大约是处于任何繁殖状态的雌性鲸的两倍。2020年和2021年,11只成虫中有5只怀孕,9只成虫中有4只怀孕,性成熟雌虫夏季怀孕率分别为0.45和0.44。2020年和2021年的产犊率分别为0.36和0.22,该亚群的年增长率计算为每年2.6%。其中一只雌性连续四年成功怀孕。这些结果表明,结合免疫反应分析和长期观察历史,进一步了解座头鲸个体的生殖生理,可以扩展到评估种群或生态系统的健康状况,具有协同价值。
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引用次数: 0
Semen cryopreservation in Bornean Sun Bear (Helarctos malayanus euryspilus) 婆罗洲太阳熊精液的冷冻保存
Pub Date : 2023-01-01 DOI: 10.1016/j.therwi.2023.100057
Boon Nie Yeoh , Zainal Zahari Zainuddin , Mark Wen Han Hiew , Siti Aisyah Sidik , Siew Te Wong , Symphorosa Sipangkui , Abdul Hamid Ahmad

Semen cryopreservation is one common tool for extended genome storage and serves as insurance for declining biodiversity. In the endangered Bornean sun bear, semen was collected with electroejaculation and this is the first attempt at semen cryopreservation for this subspecies. Chilled semen with motility of more than 50 % was extended with Caniplus freezing medium, and cryopreserved with slow freezing protocol. A total of 22 cryopreserved semen straws were evaluated with computer-assisted sperm analysis and conventional semen evaluation tests as well as additional functional tests including evaluation of the acrosome (Rose Bengal Fast Green stain), plasma membrane (hypoosmotic swelling test), and chromatin (toluidine blue stain). Post-thaw semen quality was compromised with poor viability (27.6 ± 11.2 %), motility (8.3 ± 7.1 %), and progressive movement (1.3 ± 2.0 %) but maintained high integrity for the acrosome (71.1 ± 10.8 %), plasma membrane (54.7 ± 9.6 %), and chromatin (89.7 ± 7.6 %). The total post-thaw sperm abnormality was 51.7 ± 10.7 %, predominantly bent tail (28.0 ± 7.6 %) and proximal droplets (20.0 ± 11.8 %). Although this current cryopreservation was not a success, the compromised frozen-thawed semen may be a valuable resource in assisted reproductive technologies such as intracytoplasmic sperm injection. The three additional functional tests demonstrated were simple and affordable and hence are recommended to be part of the routine semen evaluation. Further research is required to develop species-specific cryopreservation protocols and to explore other assisted reproductive technologies in the Bornean sun bear.

精子冷冻保存是延长基因组存储的一种常用工具,是生物多样性下降的保障。对濒危物种婆罗洲马来熊的精液进行了电射精收集,这是该亚种首次尝试精液冷冻保存。用Caniplus冷冻培养基延长活力大于50%的冷冻精液,并采用慢速冷冻方案冷冻保存。使用计算机辅助精子分析和常规精液评估试验,以及附加的功能测试,包括顶体评估(玫瑰孟加拉快速绿色染色)、质膜评估(低渗肿胀试验)和染色质评估(甲苯胺蓝染色),对22根冷冻保存的精液进行评估。解冻后精液质量受到影响,活力(27.6%±11.2%)、活力(8.3±7.1%)和进行性(1.3±2.0%)较差,但顶体(71.1±10.8%)、质膜(54.7±9.6%)和染色质(89.7±7.6%)保持较高的完整性。解冻后精子畸形发生率为51.7±10.7%,以尾弯(28.0±7.6%)和近端滴(20.0±11.8%)为主。虽然目前这种冷冻保存方法并不成功,但冷冻解冻后的精液可能是辅助生殖技术(如胞浆内单精子注射)的宝贵资源。所演示的三个附加功能测试简单且负担得起,因此建议作为常规精液评估的一部分。需要进一步的研究来开发特定物种的低温保存方案,并探索婆罗洲马来熊的其他辅助生殖技术。
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引用次数: 0
Prolactin enzyme-linked immunosorbent assay for rhinoceroses – Another tool for assessing reproductive function and dysfunction in this taxon 犀牛催乳素酶联免疫吸附测定-另一种评估该分类单元生殖功能和功能障碍的工具
Pub Date : 2023-01-01 DOI: 10.1016/j.therwi.2023.100035
Terri L. Roth , Elizabeth M. Donelan , Louisa A. Rispoli , T. Reilly

For decades, progesterone and its metabolites have served as the primary biomarkers for monitoring reproductive activity in rhinoceroses, whereas protein hormones have received little attention despite their potential value in understanding reproductive function and dysfunction in this taxon. The goals of this study were to: 1) identify an enzyme-linked immunosorbent assay (ELISA) effective in measuring rhinoceros serum prolactin, 2) generate representative prolactin data in male and female rhinoceroses in diverse reproductive states, and 3) characterize prolactin throughout pregnancy in a white rhinoceros that exhibited mid-gestation lactation. Our results indicated that an equine prolactin ELISA by CUSABIO® is effective in measuring serum prolactin concentrations in white, black and Sumatran rhinoceroses. Preliminary data suggested that prolactin is lowest in males and acyclic females, but also appeared low during post-partum lactation. In contrast, prolactin concentrations were elevated in pregnant females and moderate in sexually mature females experiencing reproductive cyclicity. Prolactin increased following conception and generally continued to rise throughout pregnancy in the one pregnant white rhinoceros profiled herein. Spikes and dips in prolactin and progesterone, respectively, were documented around the time of mid-gestation mammary development and colostrum production in this individual and may provide some clues into the physiological triggers of this newly described aberrant reproductive condition. In conclusion, we have identified a new tool for studying reproductive activity in rhinoceroses, generated preliminary data, and revealed an intriguing relationship between prolactin fluctuations and premature lactation. This work provides the foundation for larger, focused studies on the role of prolactin in this taxon.

几十年来,黄体酮及其代谢物一直被作为监测犀牛生殖活动的主要生物标志物,而蛋白质激素尽管在了解犀牛生殖功能和功能障碍方面具有潜在价值,但却很少受到关注。本研究的目的是:1)确定一种有效测量犀牛血清催乳素的酶联免疫吸附试验(ELISA); 2)在不同生殖状态下产生具有代表性的雄性和雌性犀牛催乳素数据;3)在妊娠中期哺乳的白犀牛妊娠期间对催乳素进行表征。结果表明,CUSABIO®马催乳素酶联免疫吸附试验可有效测定白犀牛、黑犀牛和苏门答腊犀牛血清催乳素浓度。初步资料显示,男性和无环女性的泌乳素最低,但产后哺乳期也出现低水平。相比之下,催乳素浓度在怀孕的雌性中升高,而在经历生殖周期的性成熟雌性中则升高。催乳素在受孕后增加,并在整个怀孕期间持续上升,在这里描述的一只怀孕的白犀牛。在该个体的妊娠中期乳房发育和初乳产生期间,泌乳素和孕酮分别出现了高峰和低谷,这可能为这种新描述的异常生殖状况的生理触发提供了一些线索。总之,我们已经确定了一个研究犀牛生殖活动的新工具,产生了初步数据,并揭示了催乳素波动和早产之间的有趣关系。这项工作为进一步深入研究催乳素在该分类群中的作用奠定了基础。
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引用次数: 1
Pharmacological semen collection in giant anteaters (Myrmecophaga tridactyla): A feasible option for captive and free-living animals 巨食蚁兽(Myrmecophaga tridactyla)的药理学精液采集:圈养和自由生活动物的可行选择
Pub Date : 2023-01-01 DOI: 10.1016/j.therwi.2023.100030
Gediendson Ribeiro de Araújo , Pedro Nacib Jorge-Neto , Jorge Aparecido Salmão-Júnior , Maitê Cardoso Coelho da Silva , Ricardo Zanella , Antonio Carlos Csermak-Júnior , Fabio de Moraes Francisco , Thyara de Deco-Souza , Cristiane Schilbach Pizzutto
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引用次数: 2
Development of a diluent storage media and effect of cryoprotectants on semen collected from white spotted bamboo sharks (Chiloscyllium plagiosum) and cooled to 5 ºC 稀释储存介质的研制及冷冻保护剂对冷却至5℃的白斑竹鲨精液的影响
Pub Date : 2023-01-01 DOI: 10.1016/j.therwi.2023.100017
Gisele A. Montano , Karen J. Steinman , Amanda McDonnell , Todd R. Robeck
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引用次数: 0
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Theriogenology wild
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