Egg forensics: An appraisal of DNA sequencing to assist in species identification of illegally smuggled eggs

Abstract

Psittaciformes (parrots and cockatoos) are charismatic birds, their plumage and capacity for learning make them highly sought after pets. The illegal trade in parrots and cockatoos poses a serious threat to the viability of native populations; in addition, species transported to non-endemic areas may potentially vector disease and genetically 'pollute' local native avifauna. To reduce the logistical difficulties associated with trafficking live birds, smugglers often transport eggs. This creates a problem for authorities in elucidating accurate species identification without the laborious task of incubation and hand rearing until a morphological identification can be made. Here, we use 99 avian eggs seized from carriers coming into and within Australia, as a result of suspected illegal trade. We investigate and evaluate the use of mitochondrial DNA (mtDNA) to accurately identify eggs to family, genus or species level. However, Identification of a species based on percentage mtDNA similarities is difficult without good representations of the inter-and intra-levels of species variation.Based on the available reference database, we were able to identify 52% of the eggs to species level.Of those, 10 species from eight genera were detected, all of which belong to the parrot (Psittacidae) and cockatoo (Cacatuidae) families. Of the remaining 48%, a further 36% of eggs were identified to genus level, and 12% identified to family level using our assignment criteria. Clearly the lack of validated DNA reference sequences is hindering our ability to accurately assign a species identity, and accordingly, we advocate that more attention needs to be paid to establishing validated, multi locus mtDNA reference databases for exotic birds that can both assist in genetic identifications and withstand legal scrutiny.

Identificação de Espécies de Animais Silvestres por Meio de Sequências Mitocondriais Utilizados no Combate aos Crimes Contra a Fauna – Mato Grosso – Brasil

“…Na literatura, podem ser encontrados inúmeros exemplos do uso desta região na identificação de vertebrados para fins forenses, incluindo mamíferos 24 , peixes 25 ; répteis 26 , e aves 27,28 , inclusive seus ovos [29][30][31] .…”

Testing of microsatellite multiplexes for individual identification of Cape Parrots (Poicephalus robustus): paternity testing and monitoring trade

“…Molecular forensic methods are widely used to identify suspected illegally obtained wildlife or wildlife products (Comstock, Ostrander & Wasser, 2003; Gupta, Verma & Singh, 2005; Lorenzini, 2005; Dawnay et al, 2009; Lorenzini et al, 2011; Coghlan et al, 2012; White et al, 2012; Mondol et al, 2014; Gonçalves et al, 2015; Presti et al, 2015). One of the most useful molecular forensic tools is genetic fingerprinting using microsatellite markers.…”

Application of STR markers in wildlife forensic casework involving Australian black-cockatoos (Calyptorhynchus spp.)

“…Captive 50 breeding is a major source of individuals to supply aviculture markets, although only a small number of species are sourced from this industry [2]. The total impact of the illegal trade in birds is particularly challenging to quantify in numbers due to its clandestine nature, and the cumulative impact of the mortality rates during capture, transport and pre-and post-sale [2][3][4]. The detrimental impacts of such activities are 55 having a measurable effect on cockatoos and parrots in the wild, both in terms of biodiversity and population number [2,3].…”

“…Species identification with the use of DNA is now widely applied [4,[18][19][20][21][22][23][24], and assigning samples to an individual, population, or geographic origin is gaining momentum [25][26][27][28][29]. 105…”