Sharks

鲨鱼
  • 文章类型: Journal Article
    珊瑚礁生态系统受到高度威胁,对气候变化的影响极为敏感。多种鲨鱼物种依赖珊瑚礁作为重要的栖息地,因此,在这些生态系统中发挥着许多重要的生态作用。环境压力如何影响常规,与珊瑚礁相关的鲨鱼行为,相对未被探索。这里,我们结合了8年的声学跟踪数据(2013-2020年),从居住的灰礁鲨到印度洋中部查戈斯群岛的偏远珊瑚礁,基于卫星的珊瑚礁环境压力暴露指数。我们表明,平均整个地区,对珊瑚礁的压力增加显着降低了灰礁鲨的居住面积,促进更分散的空间利用,增加远离浅海珊瑚礁的时间。重要的是,这种影响具有长达16个月的滞后效应。这可能会对礁鲨产生重要的生理和保护作用,以及对珊瑚礁生态系统功能的更广泛影响。由于气候变化预计会增加珊瑚礁生态系统的环境压力,了解与地点相关的捕食者如何应对压力对于预测改变捕食者行为的功能意义以及对珊瑚礁鲨鱼和珊瑚礁本身的保护的潜在影响至关重要。
    Coral reef ecosystems are highly threatened and can be extremely sensitive to the effects of climate change. Multiple shark species rely on coral reefs as important habitat and, as such, play a number of significant ecological roles in these ecosystems. How environmental stress impacts routine, site-attached reef shark behavior, remains relatively unexplored. Here, we combine 8 years of acoustic tracking data (2013-2020) from grey reef sharks resident to the remote coral reefs of the Chagos Archipelago in the Central Indian Ocean, with a satellite-based index of coral reef environmental stress exposure. We show that on average across the region, increased stress on the reefs significantly reduces grey reef shark residency, promoting more diffuse space use and increasing time away from shallow forereefs. Importantly, this impact has a lagged effect for up to 16 months. This may have important physiological and conservation consequences for reef sharks, as well as broader implications for reef ecosystem functioning. As climate change is predicted to increase environmental stress on coral reef ecosystems, understanding how site-attached predators respond to stress will be crucial for forecasting the functional significance of altering predator behavior and the potential impacts on conservation for both reef sharks and coral reefs themselves.
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  • 文章类型: Journal Article
    背景:研究单个鱼类在连续时间和空间上的栖息地使用和垂直运动模式是天生的挑战,因此对于各种物种而言,在很大程度上仍然难以捉摸。在鲨鱼中,这尤其适用于体型较小,范围较小的物种,例如spurdog(ScalusacanthiasLinnaeus,1758),which,尽管它对渔业很重要,在生物生物学和生物遥测研究中受到的关注有限,特别是在东北大西洋。
    方法:为了调查女性spurdog的细尺度生态位使用和垂直运动模式的季节性变化,我们使用了来自挪威峡湾的19名孕妇的档案数据,这些孕妇被卫星标记长达365天.我们用内核密度估算了已实现的生态位空间,并进行了连续的小波分析,以确定垂直运动的主导周期。三轴加速度数据用于识别突发事件并推断活动模式。
    结果:怀孕的女性经常在8至14°C的温度下使用300m的浅层深度。振荡垂直矩揭示了持续的diel垂直迁移(DVM)模式,黎明时下降,黄昏时上升。这种严格的正常DVM行为在冬季和春季占主导地位,并且与较高水平的活动爆发有关,而在夏季和秋季,鲨鱼主要选择温跃层上方的温暖水域,只有零星的潜水和爆发事件。
    结论:冬季正常DVM行为的患病率与可能的觅食相关活动爆发增加有关,表明这种运动行为是觅食驱动的。在夏季和秋季,温暖水域的快速启动次数较少,这个季节的栖息地使用可能是由行为体温调节驱动的,然而,其他因素也可能发挥作用。个体和与队列相关的变化表明,运动行为和栖息地的使用与非生物和生物环境之间存在复杂的相互作用。结合正在进行的调查精细水平运动以及性别和年龄差异的工作,这项研究提供了重要信息,以指导新重新开放的渔业的时空分布,并有助于加深对东北大西洋及其他地区spurdog的运动生态学的了解。
    BACKGROUND: Studying habitat use and vertical movement patterns of individual fish over continuous time and space is innately challenging and has therefore largely remained elusive for a wide range of species. Amongst sharks, this applies particularly to smaller-bodied and less wide-ranging species such as the spurdog (Squalus acanthias Linnaeus, 1758), which, despite its importance for fisheries, has received limited attention in biologging and biotelemetry studies, particularly in the North-East Atlantic.
    METHODS: To investigate seasonal variations in fine-scale niche use and vertical movement patterns in female spurdog, we used archival data from 19 pregnant individuals that were satellite-tagged for up to 365 days in Norwegian fjords. We estimated the realised niche space with kernel densities and performed continuous wavelet analyses to identify dominant periods in vertical movement. Triaxial acceleration data were used to identify burst events and infer activity patterns.
    RESULTS: Pregnant females frequently utilised shallow depths down to 300 m at temperatures between 8 and 14 °C. Oscillatory vertical moments revealed persistent diel vertical migration (DVM) patterns, with descents at dawn and ascents at dusk. This strict normal DVM behaviour dominated in winter and spring and was associated with higher levels of activity bursts, while in summer and autumn sharks predominantly selected warm waters above the thermocline with only sporadic dive and bursts events.
    CONCLUSIONS: The prevalence of normal DVM behaviour in winter months linked with elevated likely foraging-related activity bursts suggests this movement behaviour to be foraging-driven. With lower number of fast starts exhibited in warm waters during the summer and autumn months, habitat use in this season might be rather driven by behavioural thermoregulation, yet other factors may also play a role. Individual and cohort-related variations indicate a complex interplay of movement behaviour and habitat use with the abiotic and biotic environment. Together with ongoing work investigating fine-scale horizontal movement as well as sex- and age-specific differences, this study provides vital information to direct the spatio-temporal distribution of a newly reopened fishery and contributes to an elevated understanding of the movement ecology of spurdog in the North-East Atlantic and beyond.
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  • 文章类型: Journal Article
    这里,我们总结了沿海特有的鲨鱼和射线的灭绝风险,架子,西南印度洋和邻近水域的斜坡水域(SWIO+,纳米比亚到肯尼亚,包括SWIO群岛)。该地区是地方性和进化上不同的鲨鱼和射线的热点。近五分之一(70中的n=13,18.6%)的特有鲨鱼和鲨鱼受到威胁,其中一个是极度濒危的,五个是濒危的,七个是脆弱的。另有7人(10.0%)受到威胁,33(47.1%)是最不关心的,17人(24.3%)数据不足。虽然主要威胁是过度捕捞,首先有迹象表明,气候变化通过栖息地减少和近海分布变化,导致灭绝风险增加。通过背投他们的地位,1980年,很少有特有物种受到威胁,但在有针对性的鲨鱼和射线渔业出现后不久,这种情况就发生了变化。南非负有最高的国家保护责任,其次是莫桑比克和马达加斯加。然而,尽管近几十年来南非的渔业管理和执法有所改善,其他地方迫切需要重大改进。为了避免灭绝,并确保该地区特有的鲨鱼和鲨鱼的种群数量旺盛,并保持生态系统的功能,迫切需要严格保护濒危和濒危物种,并对脆弱物种进行可持续管理,几乎受到威胁,和最不关心的物种,以物种级数据收集和减少附带渔获量为基础。
    Here, we summarise the extinction risk of the sharks and rays endemic to coastal, shelf, and slope waters of the southwest Indian Ocean and adjacent waters (SWIO+, Namibia to Kenya, including SWIO islands). This region is a hotspot of endemic and evolutionarily distinct sharks and rays. Nearly one-fifth (n = 13 of 70, 18.6%) of endemic sharks and rays are threatened, of these: one is Critically Endangered, five are Endangered, and seven are Vulnerable. A further seven (10.0%) are Near Threatened, 33 (47.1%) are Least Concern, and 17 (24.3%) are Data Deficient. While the primary threat is overfishing, there are the first signs that climate change is contributing to elevated extinction risk through habitat reduction and inshore distributional shifts. By backcasting their status, few endemic species were threatened in 1980, but this changed soon after the emergence of targeted shark and ray fisheries. South Africa has the highest national conservation responsibility, followed by Mozambique and Madagascar. Yet, while fisheries management and enforcement have improved in South Africa over recent decades, substantial improvements are urgently needed elsewhere. To avoid extinction and ensure robust populations of the region\'s endemic sharks and rays and maintain ecosystem functionality, there is an urgent need for the strict protection of Critically Endangered and Endangered species and sustainable management of Vulnerable, Near Threatened, and Least Concern species, underpinned by species-level data collection and reduction of incidental catch.
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  • 文章类型: Journal Article
    交配系统变异是指遗传一夫一妻制和多妻制之间的光谱,并对性冲突产生重要影响,动物的性选择和个体适应性。从理论上讲,这种变化也可能对人口水平产生重大影响,影响种群生存能力和灭绝风险。这些影响的证据好坏参半,部分原因是,人们认为,要使它们产生明显的人口后果,就需要进行实质性的环境变化。在这项研究中,我们测试了Elasmobranchii(鲨鱼和射线)中一夫多妻制与种群状况之间的关系。Elasmobranchi是一种大型脊椎动物进化枝,在遗传交配系统和种群状态中均表现出实质性的种间变异,以及受到强烈的人为介导的环境变化的影响。我们还通过弹性分支系统发育预测了遗传交配系统过去的宏观进化变化。我们的结果表明,遗传一夫一妻制和一夫多妻制都在Elasmobranchii内独立进化了多次,我们建议这两个极端都代表了在离散的生态和生物条件下更受欢迎的替代适应性策略。然而,没有证据表明弹性枝交配系统变化对种群水平的影响。这些结果很重要,因为它们表明该进化枝的交配系统变化不太可能是灭绝脆弱性的主要决定因素。最终将需要额外的工作,然而,这项研究提高了我们对弹性枝交配系统变化的进化动力学的理解,以及由此产生的人口水平后果的可能性。
    Mating system variation refers to the spectrum between genetic monogamy and polyandry, and has important consequences for sexual conflict, sexual selection and individual fitness in animals. Theoretically this variation could also have substantial population-level effects, influencing population viability and extinction risk. Evidence for these effects is mixed, in part due to the fact that substantial environmental change is thought to be required for them to have visible demographic consequences. In this study we test for the presence of relationships between polyandry and population status in Elasmobranchii (sharks and rays). Elasmobranchii is a large vertebrate clade that exhibits substantial interspecific variation in both genetic mating system and population status, as well as being subject to intense anthropogenically-mediated environmental change. We also predict past macroevolutionary shifts in genetic mating system through elasmobranch phylogeny. Our results show that both genetic monogamy and polyandry have evolved multiple times independently within Elasmobranchii, and we suggest that both of these extremes represent alternative adaptive strategies that are favoured under discrete ecological and biological conditions. Nevertheless, there is no evidence of population-level consequences of mating system variation in elasmobranchs. These results are significant as they suggest that mating system variation in this clade is unlikely to be a major determinant of extinction vulnerability. Ultimately additional work will be required, however this study improves our understanding of the evolutionary dynamics underlying mating system variation in elasmobranchs, and the potential for resultant population-level consequences.
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  • 文章类型: Journal Article
    阐明跨物种的细胞结构和细胞类型进化是理解免疫系统功能和疾病易感性的核心。适应性免疫是软骨和骨鱼的共同祖先的共有特征。然而,这两种下颚脊椎动物淋巴细胞的进化特征仍不清楚。这里,我们介绍了软骨(白斑竹鲨)和骨性(斑马鱼和中国舌底)鱼类的免疫细胞的单细胞RNA测序图谱。跨物种比较表明,不同物种的相同细胞类型表现出相似的转录谱。在竹鲨中,我们鉴定了表达几种模式识别受体的吞噬细胞B细胞群,以及共表达T和B细胞标志物的T细胞亚簇。与骨鱼的功能划分相反,我们显示软骨鱼中淋巴细胞之间的紧密联系和功能专业化差。我们的跨物种单细胞比较提供了一种资源,可以揭示下颌体免疫系统的起源和进化。
    Elucidating cellular architecture and cell-type evolution across species is central to understanding immune system function and susceptibility to disease. Adaptive immunity is a shared trait of the common ancestor of cartilaginous and bony fishes. However, evolutionary features of lymphocytes in these two jawed vertebrates remain unclear. Here, we present a single-cell RNA sequencing atlas of immune cells from cartilaginous (white-spotted bamboo shark) and bony (zebrafish and Chinese tongue sole) fishes. Cross-species comparisons show that the same cell types across different species exhibit similar transcriptional profiles. In the bamboo shark, we identify a phagocytic B cell population expressing several pattern recognition receptors, as well as a T cell sub-cluster co-expressing both T and B cell markers. In contrast to a division by function in the bony fishes, we show close linkage and poor functional specialization among lymphocytes in the cartilaginous fish. Our cross-species single-cell comparison presents a resource for uncovering the origin and evolution of the gnathostome immune system.
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  • 文章类型: Journal Article
    母体投资理论是研究育种雌性如何在后代大小和育龄之间分配资源以实现繁殖成功的研究。在经典权衡模型中,R/K选择和押注对冲选择,母亲对后代投资的主要预测因素是人口密度和资源稳定性。在拥挤的,稳定的环境,K-selected雌性投资于大型后代,其成本相当。在不拥挤的地方,不稳定的环境,r-selectedfemaleinvestedinlargecoubdsatanequivalentcostinafollowingsizeinarelatingsize.在不可预测的资源环境中,押注对冲雌性在育龄期和后代大小方面的投资适度。孕产妇风险管理模型与经典的权衡模型有着深刻的不同。母亲对后代大小的投资,育母大小,和育种数由自主风险因素决定:季节性周期内资源缺口的持续时间,捕食率,和不可预测的灾难性事件。迄今为止,没有一个单一的模式上升到卓越的地位。在这里的鲨鱼,我们发现,物种内和跨物种的孕产妇投资与权衡模型的预测不一致,而是与孕产妇风险管理模型的预测一致.在鲨鱼物种内和整个鲨鱼物种中,后代大小和育龄体是独立的母系投资策略。饥饿的风险有利于对更大后代的投资。捕食的风险有利于对更大的鸡群进行投资。如果实证研究继续证实其预测,通过自然选择,孕产妇风险管理可能会成为多种生殖适应的统一模型。
    Maternal investment theory is the study of how breeding females allocate resources between offspring size and brood size to achieve reproductive success. In classical trade-off models, r/K-selection and bet-hedging selection, the primary predictors of maternal investments in offspring are population density and resource stability. In crowded, stable environments, K-selected females invest in large offspring at an equivalent cost in brood size. In uncrowded, unstable environments, r-selected females invest in large broods at an equivalent cost in offspring size. In unpredictable resource environments, bet-hedging females invest moderately in brood size and offspring size. The maternal risk-management model represents a profound departure from classical trade-off models. Maternal investments in offspring size, brood size, and brood number are shaped independently by autonomous risk factors: the duration of gaps in resources during seasonal cycles, rates of predation, and unpredictable catastrophic events. To date, no single model has risen to a position of preeminence. Here in sharks, we show that maternal investments within and across species do not agree with the predictions of trade-off models and instead agree with the predictions of the maternal risk-management model. Within and across shark species, offspring size and brood size were independent maternal investment strategies. The risk of starvation favored investments in larger offspring. The risk of predation favored investments in larger broods. If empirical studies continue to confirm its predictions, maternal-risk management may yet emerge as a unifying model of diverse reproductive adaptations by means of natural selection.
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  • 文章类型: Journal Article
    以前有报道称,安魂曲鲨鱼(Carcharhinus属)在地中海东部的海洋基础设施周围形成了大型聚集体。虽然这种行为可能会在个人层面提供健身优势,在人类改变的栖息地延长居住对人口持久性的影响尚待评估。在这项工作中,我们调查了以色列燃煤发电和海水淡化站附近鲨鱼的系统地理和人口组成。我们的目的是推断栖息地的使用和聚集行为背后的机制,并强调潜在的保护影响。我们取样,测量,并在2016年至2022年之间释放了70个人,以评估细胞色素C氧化酶I(COI)区域内的遗传变异性,并根据鲨鱼的大小和性别分布分析聚集的结构。此外,我们对在以色列另一个电站收集的死亡参考标本进行了综合计数,以使用上述技术补充物种鉴定。我们的发现表明,成年雌性暗色鲨鱼和雄性沙洲鲨鱼(Carcharhinusobscurus和Carcharhinusplumbeus,分别),每个物种包含两个COI单倍型。在昏暗的鲨鱼中,一种单倍型对应于印度-太平洋血统,另一个与大西洋血统相符。在沙洲鲨鱼中,我们观察到以前在地中海采样的单倍型,红海,西北印度洋,南非,另一种单倍型是我们研究地点独有的,在遗传上比在其他海洋盆地采样的序列更接近前者。这项研究首次表明了系统地理上不同的暗黑和沙洲鲨鱼谱系之间的同胞聚集,这表明地中海东部人类改变的栖息地可能会影响这些物种的分布。根据观察到的偏析模式,我们得出的结论是,该网站不能作为托儿所,分娩,或交配区,并讨论其他值得进一步研究的合理解释。最后,我们强调未来研究的重要方向以及我们的发现对管理和保护的影响。
    Requiem sharks (genus Carcharhinus) have previously been reported to form large aggregations around marine infrastructures in the eastern Mediterranean Sea. While this behaviour may offer fitness advantages at the individual level, the implications of extended residency at human-altered habitats for population persistence have yet to be assessed. In this work, we investigated the phylogeographic and demographic composition of sharks near a coal-fired power and desalination station in Israel. Our aim was to infer habitat use and the mechanisms underlying the aggregation behaviour, and to highlight potential conservation impacts. We sampled, measured, and released 70 individuals between 2016 and 2022 to assess genetic variability within the cytochrome C oxidase I (COI) region and to analyse the aggregation\'s structure based on the sharks\' size and sex distribution. In addition, we performed meristic counts on a reference specimen collected dead at another power station in Israel to supplement species identification using the abovementioned techniques. Our findings indicate size-based sex segregation of adult female dusky and male sandbar sharks (Carcharhinus obscurus and Carcharhinus plumbeus, respectively), with each species comprising two COI haplotypes. In the dusky shark, one haplotype corresponded to an Indo-Pacific lineage, and the other matched an Atlantic lineage. In the sandbar shark, we observed a haplotype previously sampled in the Mediterranean Sea, the Red Sea, the Northwest Indian Ocean, and South Africa, and another haplotype that was unique to our study site and genetically closer to the former than to sequences sampled in other ocean basins. This study provides the first indication of sympatric aggregation amongst phylogeographically distinct dusky and sandbar shark lineages, suggesting that human-altered habitats in the eastern Mediterranean Sea may influence the distribution of these species. Based on the observed segregation pattern, we conclude that the site does not function as a nursery, parturition, or mating area, and discuss other plausible explanations that warrant further research. Finally, we highlight important directions for future research and the implications of our findings for management and conservation.
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  • 文章类型: Journal Article
    完整性在结构支撑中起着至关重要的作用,保护,和鱼类之间的水动力能力。大多数关于鲨鱼皮的研究都集中在外表皮层,而锚定真皮小齿的较大真皮大多被忽视。鲨鱼真皮由两层组成,上层松弛层和下层压实层,保持支撑胶原蛋白和弹性纤维。各种鲨鱼的真皮可能存在形态和组成差异,这可能与其不同的游泳方式和生态有关。这项研究的目的是表征和描述三种鲨鱼的真皮,圆形金鳞茎,Sphyrnamokarran,和Isurusoxyrinchus,每个代表不同的游泳模式。在每条鲨鱼的身体上的16个位置进行了组织学表征;变量,如真皮厚度,丰富的胶原蛋白和弹性纤维,和纤维尺寸进行了量化。结果显示,毛囊的皮肤总体上最厚,胶原蛋白和弹性纤维的最大纤维尺寸,胶原纤维数量增加和弹性纤维数量减少的总体模式。在光谱的另一端,I.oxyrinchus显示沿侧翼区域的真皮最薄,弹性纤维增加,胶原纤维减少。这些发现可能会挑战我们最初的假设,即快速移动的鲨鱼中的刚体和缓慢移动的鲨鱼中的更灵活的身体,并强调鲨鱼外貌的多样性。
    The integument plays essential roles in the structural support, protection, and hydrodynamic capability among fishes. Most research on shark skin has focused on the external epidermal layer, while the larger dermis anchoring the dermal denticles has been mostly ignored. Shark dermis is composed of two layers, the upper stratum laxum and the lower stratum compactum, holding supportive collagen and elastic fibers. There may be morphological and compositional differences in the dermis across various species of sharks that could relate to their different swimming modes and ecologies. The goal of this study was to characterize and describe the dermis among three shark species, Ginglymostoma cirratum, Sphyrna mokarran, and Isurus oxyrinchus, each representing a different swimming mode. Histological characterizations were performed at 16 locations along the body of each shark; variables such as dermal thickness, abundance of collagen and elastic fibers, and fiber size were quantified. Results showed G. cirratum has the thickest skin overall, and the largest fiber size for both collagen and elastic fibers, with overall patterns of increased amounts of collagen fibers and decreased amount of elastic fibers. At the opposite end of the spectrum, I. oxyrinchus showed the thinnest dermis along the flank region, with overall patterns of increased elastic fibers and decreased collagen fibers. These findings may challenge our original assumptions of a rigid body in fast moving sharks and a more flexible body in slower moving sharks and highlight the diversity of the shark integument.
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  • 文章类型: Journal Article
    随着他们的成长,鲨鱼既可以代替丢失的细齿,又可以通过开发新的(从头)细齿而不事先脱落来增殖细齿。细齿的丢失和更换对维持皮肤表面的能量成本具有潜在影响。鲨鱼皮的生物力学功能,以及我们根据沉积物岩心中的化石密度预测鲨鱼丰度的能力。这里,我们寻求更好地理解牙质损失的模式,并展示如何在成熟的鲨鱼中取代牙质。我们使用来自两个物种的身体区域的图像和16种鲨鱼的相似身体区域的图像来说明带有缺失齿的鲨鱼皮肤表面,并量化缺失齿的种内和种间模式。一般来说,鲨鱼在物种和地区之间缺少相似数量的细齿(0%-6%)。然而,也有例外:在光滑的狗鱼中,鼻子区域比大多数后体和鳍区域缺失更多的细齿,普通的脱粒鲨鱼比光滑的狗鱼少了更多的细齿,豹鲨,天使鲨鱼,bonnethead,还有口鲨.树突再生开始于表皮下的冠发育和矿化,接着是王冠的爆发,最后是根的矿化。替代牙齿的牙髓腔最初很大,并在牙本质爆发时被薄薄的釉质壳包围。牙齿喷发后,牙本质到达其最终位置后,牙本质的沉积在内部继续。更换缺失的细齿,在任何时候代表不到6%的皮肤表面,可能不会损害水动力功能,但是通过一生中不断更新皮肤表面,鲨鱼可以通过修复单个小齿的局部损伤来减少表面污染并保持功能复杂的皮肤表面。
    As they grow, sharks both replace lost denticles and proliferate the number of denticles by developing new (de novo) denticles without prior denticle shedding. The loss and replacement of denticles has potential impacts on the energetic cost of maintaining the skin surface, the biomechanical functions of shark skin, as well as our ability to predict shark abundance from fossil denticle occurrence in sediment cores. Here, we seek to better understand patterns of denticle loss and to show how denticles are being replaced in mature sharks. We illustrate shark skin surfaces with missing denticles and quantify both within-species and between-species patterns of missing denticles using images from across regions of the body for two species and images at similar body regions for 16 species of sharks. Generally, sharks are missing similar numbers of denticles (0%-6%) between species and regions. However, there are exceptions: in the smooth dogfish, the nose region is missing significantly more denticles than most posterior-body and fin regions, and the common thresher shark is missing significantly more denticles than the smooth dogfish, leopard shark, angel shark, bonnethead, and gulper shark. Denticle regrowth starts with crown development and mineralization beneath the epidermis, followed by eruption of the crown, and finally the mineralization of the root. The pulp cavity of replacement denticles is initially large and surrounded by a thin shell of enameloid upon eruption of the denticle. After eruption of the denticle, the deposition of dentine continues internally after the denticle reaches its final position. Replacement of missing denticles, representing less than 6% of the skin surface at any one time, may not compromise hydrodynamic function, but by constantly updating the skin surface throughout life, sharks may reduce surface fouling and maintain a functional complex skin surface by repairing local damage to individual denticles.
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  • 文章类型: Journal Article
    背景:由于氧氟沙星(OFL)滥用的严重问题,对OFL的准确、快速检测的需求日益迫切。免疫测定已成为在复杂基质中检测OFL的“黄金方法”,有利于其适用于大规模筛查,快速性,和简单。然而,免疫测定中使用的传统抗体面临挑战,如耗时的制备,不稳定的灵敏度和特异性,和方向性进化的困难。在本文中,为了解决这些问题,我们成功开发了一种基于鲨鱼源单域抗体(ssdAb)的OFL检测方法.
    结果:使用噬菌体展示技术和异源表达系统,OFL特异性克隆1O11、1O13、1O17、1O19、1O21和2O26被成功地分离并以可溶性形式表达。在所有OFL特定的ssdAb中,1O17ssdAb以浓度依赖性方式表现出对OFL的最高结合亲和力。1O17ssdAb的检测限(IC10)计算为0.34ng/mL,检测范围为3.40-1315.00ng/mL,其与其他类似物的交叉反应性经计算小于5.98%,表明高特异性和敏感性。分子对接结果表明,位于1O17ssdAbCDR3区域的100Trp和101Arg对于OFL结合至关重要。在鱼类基质性能测试中,1O17ssdAb在OFL阴性鱼类基质中没有表现出严重的基质干扰,达到令人满意的回收率范围从83.04%到108.82%,重现性高。
    结论:这项研究提供了一种新型高效的OFL检测识别元件,在免疫测定应用中具有显着的潜力,拓宽ssdAbs的应用场景。它为ssdAb和小分子之间的结构-活性关系提供了有价值的见解,为ssdAb在后续应用中的进一步定向修饰和成熟奠定理论基础。
    BACKGROUND: Due to the serious issue of ofloxacin (OFL) abuse, there is an increasingly urgent need for accurate and rapid detection of OFL. Immunoassay has become the \"golden method\" for detecting OFL in complex matrix beneficial to its applicability for a large-scale screening, rapidity, and simplicity. However, traditional antibodies used in immunoassay present challenges such as time-consuming preparation, unstable sensitivity and specificity, and difficulty in directional evolution. In this paper, we successfully developed an OFL detection method based on a shark-derived single-domain antibody (ssdAb) to address these issues.
    RESULTS: Using phage display technology and a heterologous expression system, OFL-specific clones 1O11, 1O13, 1O17, 1O19, 1O21, and 2O26 were successfully isolated and expressed in soluble form. Among all OFL-specific ssdAbs, the 1O17 ssdAb exhibited the highest binding affinity to OFL in a concentration-dependence manner. The limit of detection (IC10) of 1O17 ssdAb was calculated as 0.34 ng/mL with a detection range of 3.40-1315.00 ng/mL, and its cross reactivity with other analogs was calculated to be less than 5.98 %, indicating high specificity and sensitivity. Molecular docking results revealed that 100Trp and 101Arg located in the CDR3 region of 1O17 ssdAb were crucial for OFL binding. In fish matrix performance tests, the 1O17 ssdAb did not demonstrate severe matrix interference in OFL-negative fish matrix, achieving satisfactory recovery rates ranging from 83.04 % to 108.82 % with high reproducibility.
    CONCLUSIONS: This research provides a new and efficient OFL detection recognition element with significant potential in immunoassay applications, broadening the application scenarios of ssdAbs. It offers valuable insights into the structure-activity relationship between ssdAbs and small molecules, laying a theoretical foundation for the further directional modification and maturation of ssdAbs in subsequent applications.
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