Development and evaluation of a medium-free incubation method for hatching Chinese soft-shelled turtle (Pelodiscus sinensis) eggs

Zhu, Junxian; Luo, Lei; Chen, Chen; Li, Jiansong; Wang, Yongchang; Wu, Congcong; Wang, Yakun; Hong, Xiaoyou; Liu, Xiaoli; Yu, Lingyun; Chen, Haigang; Wei, Chengqing; Liu, Yihui; Li, Wei; Zhu, Xiaoyan

doi:10.1016/j.aqrep.2023.101643

Cited by 3 publications

(4 citation statements)

References 35 publications

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“…Briefly, the embryos can move within the eggs to select the optimal ambient temperature, thus influencing their own sex fates and maintaining the fitness sex ratios of the population. Most turtles have oval-shaped eggs (Ye et al ., 2019; Zhu et al, 2006), while the eggs of Trionychidae species are small and rounded (Li et al, 2020; Zhu et al, 2023), which may have resulted in limited space for the embryos to move, as well as reduced thermal heterogeneity within the eggs (Cordero et al, 2018; Telemeco et al, 2016), resulting in the loss of the embryo’s ability to self-regulate during the evolutionary process. This also may be one of the driving forces for sex chromosomes formation in Trionychidae species.…”

Section: Discussionmentioning

confidence: 99%

Cross-species genomic survey reveals a single origin of the soft-shelled turtles

Li,

Zhu,

Gao

et al. 2024

Preprint

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Turtles have a have a complicated sex determination system; however, Trionychidae species, as a unique group, have evolved a conserved genetic sex determination, which provides an excellent case to study the evolution of environmental sex determination to genetic sex determination. Analysis of the chromosome-level genome assembly, construction of the first sex chromosomes of Trionychidae, and embryonic transcriptome revealed that P. sinensis has a ZW-type sex determination system, and that the key sex-determining genes, including Dmrt1, Amh, Foxl2, etc., and their homologs, are not located on the sex chromosomes, which rejects the involvement of Dmrt1's SDGs or dosage compensation effect in the sex determination of P. sinensis ZW system and also indicates that the ZW chromosomes in birds, turtles, frogs, and fishes originated from different chromosome pairs in the common ancestors. Subsequently, comparative genomic studies demonstrated that a conserved ZW-type sex determination system exists in Trionychidae species, and inferred that their sex chromosomes originate from the same chromosome pair of the common ancestor. More importantly, we found a single origin of the sex chromosomes in Trionychidae species, approximately 56 million years ago, which is highly coincident with the Paleocene-Eocene thermal maximum, PETM, event. Therefore, we hypothesize that the PETM event and the loss of embryonic self-thermal regulation are two critical dynamics that drive the adaptive evolution of sex chromosomes for the common ancestor of Trionychidae species in response to extreme temperature changes. Finally, we link the ancestral sex chromosome origin time, the species divergence time, the PETM event, and the continental plate movements to paint a picture of a single origin and radiation route for extant Trionychidae species. Collectively, our findings provide novel insights into the mechanisms of sex determination in P. sinensis, and the single origin of sex chromosomes and species in extant soft-shelled turtles.

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Section: Discussionmentioning

confidence: 99%

Cross-species genomic survey reveals a single origin of the soft-shelled turtles

Li,

Zhu,

Gao

et al. 2024

Preprint

Self Cite

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“…The collected eggs were tested for hatchability, and the animal poles of the eggs had obvious white fertilization spots, which meant they had been fertilized and could hatch [76]. Fertilized eggs were hatched in a thermostatic incubator (FHX-400, Shaoguan, China) with the temperature set to 31 • C and the humidity maintained at 75~85% [77]. The gonad-mesonephric complexes of 20 embryos from different developmental stages, including stage 17 (gonadal differentiation normally begins from stage 17) [55,56], stage 20, stage 23, and stage 26, identified according to the established standards [78], were stripped away, respectively.…”

Section: Experimental Turtles and Eggsmentioning

confidence: 99%

Identification of Sex-Specific Markers and Candidate Genes Using WGS Sequencing Reveals a ZW-Type Sex-Determination System in the Chinese Soft-Shell Turtle (Pelodiscus sinensis)

Zhu,

Wang,

Chen

et al. 2024

IJMS

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Male and female Chinese soft-shelled turtles (Pelodiscus sinensis) have sex-dimorphic growth patterns, and males have higher commercial value because of their larger size and thicker calipash. Thus, developing sex-specific markers is beneficial to studies on all-male breeding in P. sinensis. Here, we developed an accurate and efficient workflow for the screening of sex-specific sequences with ZW or XY sex determination systems. Based on this workflow, female and male P. sinensis reference genomes of 2.23 Gb and 2.26 Gb were obtained using de novo assembly. After aligning and filtering, 4.01 Mb female-specific sequences were finally identified. Subsequently, the seven developed sex-specific primer pairs were 100% accurate in preliminary, population, and embryonic validation. The presence and absence of bands for the primers of P44, P45, P66, P67, P68, and P69, as well as two and one bands for the PB1 primer, indicate that the embryos are genetically female and male, respectively. NR and functional annotations identified several sex-determining candidate genes and related pathways, including Ran, Eif4et, and Crkl genes, and the insulin signaling pathway and the cAMP signaling pathway, respectively. Collectively, our results reveal that a ZW-type sex-determination system is present in P. sinensis and provide novel insights for the screening of sex-specific markers, sex-control breeding, and the studies of the sex determination mechanism of P. sinensis.

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“…Artificial hatching can be used to provide kampung chickens because a mother's ability to incubate is limited. It also extends the production period because the time for incubation and childcare is eliminated (Kamal et al, 2024;Yang et al, 2024;Zhu et al, 2023).…”

Section: Introductionmentioning

confidence: 99%

“…The assumptions mentioned above have not been scientifically proven. In addition to gender, chick abnormalities produced from hatching eggs based on egg shape have not been scientifically proven, so there is a need for research related to the influence of egg shape on gender and abnormalities of kampung chicks (Andaruisworo, 2022;Utami et al, 2020;Zhu et al, 2023).…”

Section: Introductionmentioning

confidence: 99%

The Influence of Egg Shape on Gender and Abnormality of Kampung Chicks

Horhoruw

2024

Adv

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This research aimed to determine the influence of egg shape on the gender and abnormalities of hatched kampung chicks, conducted over one month in Wailela Hamlet, Ambon City. Using an experimental method with a quasi-experimental design, the study observed 120 kampung chicken eggs less than five days old. Data were collected by directly observing hatched eggs and chicks using hatching machines and digital scales. The eggs were cleaned, identified, and numbered before placement in a fumigated hatching machine. Ventilation and screening maintained temperature and humidity stability. The chi-square test (X2) analyzed the effect of egg shape on gender and abnormalities. The study's significance lies in its potential to improve breeding practices by understanding the impact of egg shape on hatching outcomes. However, the results indicated that egg shape did not significantly affect the gender or abnormalities of the hatched chicks. Consequently, egg shape cannot be used as a reliable criterion for selecting eggs to determine the gender or abnormalities of kampung chicks. This insight highlights the need for genetic or molecular techniques for gender determination and suggests further research with larger samples to explore other influencing factors.

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Development and evaluation of a medium-free incubation method for hatching Chinese soft-shelled turtle (Pelodiscus sinensis) eggs

Cited by 3 publications

References 35 publications

Cross-species genomic survey reveals a single origin of the soft-shelled turtles

Cross-species genomic survey reveals a single origin of the soft-shelled turtles

Identification of Sex-Specific Markers and Candidate Genes Using WGS Sequencing Reveals a ZW-Type Sex-Determination System in the Chinese Soft-Shell Turtle (Pelodiscus sinensis)

The Influence of Egg Shape on Gender and Abnormality of Kampung Chicks

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