Genetic sex test for the short-beaked echidna (Tachyglossus aculeatus)

Keating, Shannon E.; Fenelon, Jane C; Pyne, Michael; Pinto, Brendan J; Guzmán-Méndez, Irán Andira; Johnston, Stephen D.; Renfree, Marilyn B.; Gamble, Tony

doi:10.1007/s12686-022-01258-3

Cited by 2 publications

(10 citation statements)

References 54 publications

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“…This study verified the sex‐specific RADseq markers identified in Keating et al (2022) to be the most accurate for identifying sex in the short‐beaked echidna with gDNA isolated from blood and hair. In all blood samples, the six male‐specific PCR markers accurately identified gender, as matched to the clinically recorded sex for the corresponding individual, with the exception of one male where no amplification occurred for the CRSPY gene.…”

Section: Discussionsupporting

confidence: 83%

“…Here, we have followed the recommendation of Keating et al (2022), evaluating a combination of markers to accurately determine sex in the short-beaked echidna. In addition, evaluation of markers has been assessed by investigating marker performance with noninvasive samples.…”

Section: Discussionmentioning

confidence: 99%

“…The combination of amplifying multiple sex‐determining markers, targeting numerous sites on the Y‐chromosome, facilitates the confidence and accuracy to verify sex identification in many threatened species (Robertson & Gemmell, 2006). Increasing the number of sex‐specific markers used decreases, but does not entirely eliminate, the chance of a false negative or false positive leading to erroneous sex assignment (Keating et al, 2022). Here, we have followed the recommendation of Keating et al (2022), evaluating a combination of markers to accurately determine sex in the short‐beaked echidna.…”

Section: Discussionmentioning

confidence: 99%

“…Perry et al (2019) identified the AMH gene in both sexes as a potential candidate for a monotreme sex-determining gene. Further, Keating et al (2022) recently identified an additional suite of novel echidna sex-specific genetic markers using restriction-site associated DNA sequencing (RADseq) further increasing the number of sex-specific markers available, and subsequently the accuracy of sex identification in the echidna.…”

Section: Introductionmentioning

confidence: 99%

“…Sex identification determined from noninvasive samples was validated by comparing with gDNA extracted from blood and the clinically recorded gender of each echidna. To improve accuracy for genetic sex identification in the echidna, a suite of male specific sex markers sourced from the studies of Perry et al (2019) and Keating et al (2022) were tested for by PCR; in addition β‐actin ( ACTB ) was used as a positive control to indicate genomic extraction was successful and confirm that echidna DNA was isolated. Each sex marker assay was evaluated for sensitivity and specificity using ROC curves.…”

Section: Introductionmentioning

confidence: 99%

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An evaluation of DNA sample source and molecular markers to determine gender in the short‐beaked echidna (Tachyglossus aculeatus)

Hulse,

Tomsett,

Roser

et al. 2023

Zoo Biology

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The short‐beaked echidna is sexually monomorphic such that gender identification without veterinary intervention is challenging. The aim of this study was to evaluate and compare the most optimal noninvasive genetic source by extracting echidna genomic DNA (gDNA) from fecal scats, plucked hair, and quills to perform genetic sex testing using a range of molecular markers. Sex determination of 14 captive short‐beaked echidnas was determined by amplifying isolated DNA from noninvasive samples, targeting two Y chromosome (male‐specific) genes (mediator complex subunit 26 Y‐gametologue [CRSPY] and anti‐Müllerian hormone Y‐gametologue [AMHY]), in addition to four confirmed sex‐specific RADseq markers. Results of noninvasive samples were compared with blood samples and clinical records. Receiver operating characteristic curves were used to assess accuracy of sex determination of markers for each sample type. The gender of the echidnas was successfully identified on 75% of occasions using fecal samples, 90.6% occasions using hair, and 84.6% occasions with quills. Overall, the male‐specific RADseq markers accurately identified the sex of echidnas with all sample types for 90% of animals; compared with 81.5% using CRSPY, and 82.0% using AMHY to identify sex. Collection of hair, quills, and feces provides a useful alternative to invasively collected samples, however, the accuracy of results depends on sample type and genetic marker selected. We found gender determination in the short‐beaked echidna was most accurate using four male‐specific RADseq markers on gDNA isolated from blood and hair. The noninvasive genetic sexing techniques documented here will inform and facilitate husbandry and genetic management of captive echidna populations.

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Section: Discussionsupporting

confidence: 83%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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An evaluation of DNA sample source and molecular markers to determine gender in the short‐beaked echidna (Tachyglossus aculeatus)

Hulse,

Tomsett,

Roser

et al. 2023

Zoo Biology

Self Cite

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AMHY and sex determination in egg-laying mammals

Shearwin-Whyatt,

Fenelon,

et al. 2024

Preprint

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The sex chromosomes of egg-laying mammals (monotremes), which lack the sex determining gene SRY, evolved independently to those of all therian mammals. Here we characterise the candidate monotreme sex determining gene, the Y-localised anti-Müllerian hormone gene (AMHY) and trace its expression during the period of sexual differentiation. Monotreme AMHX and AMHY gametologues have significant sequence divergence at the promoter, gene and protein level, likely following an original allele inversion in the common monotreme ancestor but retain conserved features of TGF-β molecules. Expression of sexual differentiation genes in the echidna fetal gonad were significantly different from that of therian mammals. AMHY expression was seen exclusively in the male gonad during sexual differentiation, whereas AMHX was expressed in both sexes. Experimental ectopic expression of platypus AMHX or AMHY in the chicken embryo did not masculinise the female urogenital system, a possible result of mammalian specific changes to AMH proteins preventing function in the chicken. Our results provide fundamental insight into the first steps of monotreme sex chromosome evolution and sex determination with developmental expression data strongly supporting AMHY as the primary male sex determination gene.

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Genetic sex test for the short-beaked echidna (Tachyglossus aculeatus)

Cited by 2 publications

References 54 publications

An evaluation of DNA sample source and molecular markers to determine gender in the short‐beaked echidna (Tachyglossus aculeatus)

An evaluation of DNA sample source and molecular markers to determine gender in the short‐beaked echidna (Tachyglossus aculeatus)

AMHY and sex determination in egg-laying mammals

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