The only known egg of the Night Parrot? A molecular and morphometric assessment of an alleged egg from the Tanami Desert

Olsen, Penny; Austin, Jeremy J.; Murphy, Stephen; Dally, Gavin

doi:10.20938/afo33211214

Cited by 3 publications

(2 citation statements)

References 9 publications

(11 reference statements)

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“…The fact that the thicker wedge-tailed eagle eggshell (10.953 g) remained intact may alternatively indicate that the digest buffer was incubated for too long inside the eggshell, and that eggshell integrity may be maintained if shorter incubation times are used (e.g. 10 min, as suggested by Olsen et al, 2016), or if a more dilute buffer is used.…”

Section: Sampling Strategy: Successes and Failuresmentioning

confidence: 99%

“…Eggshell also provides a unique opportunity to gain historical insight into avian evolution in warm climate regions where other sources of DNA (such as skeletal remains) are typically poorly preserved (Grealy et al, 2017). While DNA can be retrieved from the membranes of fresh eggshell (Rikimaru & Takahashi, 2009;Trimbos, Broekman, Kentie, Musters, & de Snoo, 2009), and small amplicons have been successfully sequenced from the membranes of museum eggshell (Lee & Prys-Jones, 2008;Olsen, Austin, Murphy, & Dally, 2016); thus far, intracrystalline DNA is the only DNA recovered from eggshell that has resulted in the retrieval of genomic-scale information.…”

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Avian mitochondrial genomes retrieved from museum eggshell

Grealy

Bunce

Holleley

2019

Molecular Ecology Resources

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Avian eggshell is a bio‐ceramic material with exceptional properties for preserving DNA within its crystalline structure, presenting an opportunity to retrieve genomic information from extinct or historical populations of birds. However, intracrystalline DNA has only been recovered from the large, thick eggshell of palaeognaths; members of their more‐diverse sister group (neognaths) lay smaller, thinner eggs that may not exhibit the same propensity for DNA preservation. Here, we use three 40–60‐year‐old museum eggshell specimens of Australian neognath birds to determine the minimum mass of eggshell from which intracrystalline DNA can be retrieved, and to characterize the yield and quality of such DNA. In doing so, we describe the first protocol for successful extraction of intracrystalline DNA from neognath eggshells, with the view to unlocking the potential of vast museum egg collections for genetic research. We were able to retrieve DNA fragments over 200 bp in length from 10 mg of eggshell powder from all three specimens, and demonstrate that expanding the existing blow‐hole can allow sufficient material to be collected for DNA extraction while minimizing damage to the appearance and structural integrity of the egg. Furthermore, we were able to reconstruct near‐complete mitochondrial genomes at a coverage of 40‐83X through shotgun sequencing of these extracts on three NextSeq lanes. Given the current extinction and extirpation rates of many avian species world‐wide, genetic data from eggshell could provide a rapid and cost‐effective approach to examining temporal changes in avian diversity, which is not only becoming crucial for conservation management, but also serve to deepen our understanding of genome‐wide evolutionary processes.

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Section: Sampling Strategy: Successes and Failuresmentioning

confidence: 99%

mentioning

confidence: 99%

Avian mitochondrial genomes retrieved from museum eggshell

Grealy

Bunce

Holleley

2019

Molecular Ecology Resources

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Genetic barcoding of museum eggshell improves data integrity of avian biological collections

Grealy

Langmore

Joseph

et al. 2021

Sci Rep

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Natural history collections are often plagued by missing or inaccurate metadata for collection items, particularly for specimens that are difficult to verify or rare. Avian eggshell in particular can be challenging to identify due to extensive morphological ambiguity among taxa. Species identifications can be improved using DNA extracted from museum eggshell; however, the suitability of current methods for use on small museum eggshell specimens has not been rigorously tested, hindering uptake. In this study, we compare three sampling methodologies to genetically identify 45 data-poor eggshell specimens, including a putatively extinct bird’s egg. Using an optimised drilling technique to retrieve eggshell powder, we demonstrate that sufficient DNA for molecular identification can be obtained from even the tiniest eggshells without significant alteration to the specimen’s appearance or integrity. This method proved superior to swabbing the external surface or sampling the interior; however, we also show that these methods can be viable alternatives. We then applied our drilling method to confirm that a purported clutch of Paradise Parrot eggs collected 40 years after the species’ accepted extinction date were falsely identified, laying to rest a 53-year-old ornithological controversy. Thus, even the smallest museum eggshells can offer new insights into old questions.

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