Efficient harvesting methods for early‐stage snake and turtle embryos

Matsubara, Yasutaka; Kuroiwa, Atsushi; Suzuki, Takayuki

doi:10.1111/dgd.12278

Cited by 5 publications

(3 citation statements)

References 18 publications

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“…Their shape became better defined, passing from a snail-like appearance to a spiral shape. This snail-like ultrasonographic aspect recalls the morphology described in relation to the embryos of Elaphe quadrivirgata , an egg-laying species isolated from just-laid eggs [ 33 ]. A similar morphology was also macroscopically described in the case of the common garter snake ( Thamnophis sirtalis sirtalis ), Python sebae, and Japanese mamushin ( Gloydius blomhoffii ) embryos [ 34 , 35 , 36 ].…”

Section: Discussionmentioning

confidence: 67%

Monitoring of the Reproductive Cycle in Captive-Bred Female Boa constrictor: Preliminary Ultrasound Observations

Bertocchi

Bigliardi

Pelizzone

et al. 2021

Animals

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The Boa constrictor is one of the most common reptiles bred in captivity. To achieve a successful breeding season, thorough knowledge of the females’ reproductive activity is necessary. In this regard, information on the Boa constrictor is still rather scarce. The aim of the present study was to monitor the ovarian activity and the embryonic development of boas by ultrasound. We performed brief scans on thirty non-anaesthetized snakes using a portable ultrasound system and a 7.5–10 MHz linear array transducer (Esaote MyLab™ Classic). Ultrasound features, dimensions, and echogenicity of the preovulatory and postovulatory follicles were determined. As gestation progresses, the postovulatory follicle size increases, and the embryonic silhouette becomes increasingly recognizable. During the second month after ovulation, by using color Doppler, early embryos’ heart activity could be evaluated. It is possible to highlight vascular connections between the mother and the membrane covering the embryonic structures. Ultrasound also allows one to identify follicular regression or slugs (nonfertilized eggs) early. The present study suggests that ultrasound could be an excellent noninvasive technique to evaluate the reproductive activity of Boa constrictor, allowing us to precisely identify the correct time for mating, monitor embryo development and viability, and allow the early diagnosis of follicular regression.

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Section: Discussionmentioning

confidence: 67%

Monitoring of the Reproductive Cycle in Captive-Bred Female Boa constrictor: Preliminary Ultrasound Observations

Bertocchi

Bigliardi

Pelizzone

et al. 2021

Animals

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“…Multiple in vivo models exist for studying the vascular responses to biomaterials and drug testing. Examples include zebra fish 11 – 13 , mice, skin flap windows 14 , 15 and snake embryos 16 , 17 . These models are generally complex and are associated with ethical concerns.…”

Section: Introductionmentioning

confidence: 99%

Assessment of flow within developing chicken vasculature and biofabricated vascularized tissues using multimodal imaging techniques

Padmanaban

Chizari

Knop

et al. 2021

Sci Rep

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Fluid flow shear stresses are strong regulators for directing the organization of vascular networks. Knowledge of structural and flow dynamics information within complex vasculature is essential for tuning the vascular organization within engineered tissues, by manipulating flows. However, reported investigations of vascular organization and their associated flow dynamics within complex vasculature over time are limited, due to limitations in the available physiological pre-clinical models, and the optical inaccessibility and aseptic nature of these models. Here, we developed laser speckle contrast imaging (LSCI) and side-stream dark field microscopy (SDF) systems to map the vascular organization, spatio-temporal blood flow fluctuations as well as erythrocytes movements within individual blood vessels of developing chick embryo, cultured within an artificial eggshell system. By combining imaging data and computational simulations, we estimated fluid flow shear stresses within multiscale vasculature of varying complexity. Furthermore, we demonstrated the LSCI compatibility with bioengineered perfusable muscle tissue constructs, fabricated via molding techniques. The presented application of LSCI and SDF on perfusable tissues enables us to study the flow perfusion effects in a non-invasive fashion. The gained knowledge can help to use fluid perfusion in order to tune and control multiscale vascular organization within engineered tissues.

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“…Unlike bird eggs, reptile eggs do not possess a chalaza (strands of albuminous matter that position the yolk and developing bird embryo in the centre of the egg), so the yolk, which has a lower density than the surrounding albumen, floats to the top of the egg and the embryo develops next to the shell membrane. This process occurs soon after oviposition and proceeds with the shell membrane adhering to the shell, creating a white spot on its surface, often referred to as "chalking" (Blanck and Sawyer, 1981;Phillott and Parmenter, 2007;Matsubara et al, 2016;Phillott and Godfrey, 2020). Eggs without a white spot are therefore often interpreted as being unfertilised.…”

mentioning

confidence: 99%

Understanding early reproductive failure in turtles and tortoises

Lavigne,

Bullock,

Shah

et al. 2023

Preprint

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Turtles and tortoises (Order Testudines) are facing an extinction crisis, and ecosystems are at risk of collapsing with the loss of key roles they play. Hatching failure is a crucial barrier to population growth and persistence, but its causes are poorly understood, and it is unknown whether fertilisation rates are declining as many populations become smaller and more female-biased. Here, we first show that fertilisation rates are considered in only a very small proportion of studies of turtle and tortoise hatching success, and those studies that do attempt to measure fertilisation rates use unreliable methods. We also show that studies of hatching success are biased towards marine turtles, as opposed to freshwater and terrestrial species, and wild rather than captive populations. We address the lack of reliable methods for assessing fertilisation rates in turtles and tortoises by developing and testing a microscopy-based method for detecting perivitelline membrane (PVM) bound sperm and embryonic nuclei in the germinal disc of unhatched eggs. We demonstrate that this method can provide unequivocal evidence of egg fertilisation in three different turtle and tortoise species from both captive and wild populations, even after eggs have been left in the nest for the full incubation period. This approach therefore represents a valuable new tool for monitoring egg fertility and embryo survival rates in turtles and tortoises, with the potential to provide important insights into the underlying drivers of reproductive failure in threatened populations.

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Efficient harvesting methods for early‐stage snake and turtle embryos

Cited by 5 publications

References 18 publications

Monitoring of the Reproductive Cycle in Captive-Bred Female Boa constrictor: Preliminary Ultrasound Observations

Monitoring of the Reproductive Cycle in Captive-Bred Female Boa constrictor: Preliminary Ultrasound Observations

Assessment of flow within developing chicken vasculature and biofabricated vascularized tissues using multimodal imaging techniques

Understanding early reproductive failure in turtles and tortoises

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