Zoo Biology 00: 1–13 (2012)
Sex Determination in 58 Bird Species
and Evaluation of CHD Gene as a
Universal Molecular Marker in Bird
Milos Vucicevic,1∗ Marija Stevanov-Pavlovic,1 Jevrosima Stevanovic,1 Jasna Bosnjak,1 Bojan Gajic,2 Nevenka Aleksic,1 and Zoran Stanimirovic1 1
Department of Biology, Faculty of Veterinary Medicine, University of Belgrade, Serbia Department of Parasitology, Faculty of Veterinary Medicine, University of Belgrade, Serbia
The aim of this research was to test the CHD gene (Chromo Helicase DNAbinding gene) as a universal molecular marker for sexing birds of relatively distant species. The CHD gene corresponds to the aim because of its high degree of conservation and different lengths in Z and W chromosomes due to different intron sizes. DNA was isolated from feathers and the ampliﬁcation of the CHD gene was performed with the following sets of polymerase chain reaction (PCR) primers: 2550F/2718R and P2/P8. Sex determination was attempted in 284 samples of 58 bird species. It was successful in 50 bird species; in 16 of those (Alopochen aegyptiacus, Ara severus, Aratinga acuticaudata, Bucorvus leadbeateri, Cereopsis novaehollandiae, Columba arquatrix, Corvus corax, C. frugilegus, Cyanoliseus patagonus, Guttera plumifera, Lamprotornis superbus, Milvus milvus, Neophron percnopterus, Ocyphaps lophotes, Podiceps cristatus, and Poicephalus senegalus), it was carried out for the ﬁrst time using molecular markers and PCR. It is reasonable to assume that extensive research is necessary to deﬁne the CHD gene as a universal molecular marker for successful sex determination in all bird species (with exception of ratites). The results of this study may largely contribute to the aim. Zoo Biol. C 2012 Wiley Periodicals, Inc.
Grant sponsor: Ministry of Education and Science of Serbia; Grant number: 46002. ∗ Correspondence to: Milos Vucicevic, Research Assistant, Department of Biology, Faculty of Veterinary Medicine, University of Belgrade, Blvd. Oslobodjenja 18, 11000 Belgrade, Serbia. E-mail: email@example.com
Received 22 May 2011; Revised 5 December 2011; Accepted 13 December 2011 DOI 10.1002/zoo.21010
Published online in Wiley Online Library (wileyonlinelibrary.com).
2012 Wiley Periodicals, Inc.
2 Vucicevic et al.
Keywords: molecular sexing; CHD gene; 2550F/2718R; bird
More than 50% of bird species are monomorphic [Grifﬁts et al., 1998], rendering their sexing based on external morphology impossible. Moreover, even in dimorphic species, sex determination is problematic in chicks [Kahn et al., 1998]. Numerous bird protection programs aimed at the preservation of various species through intensive bird breeding imply that the sex of individuals is accurately identiﬁed [Ito et al., 2003]. In zoological gardens and breeding centers, large numbers of birds are bred and traded, making sex determination extremely important [Vucicevic et al., 2010]. Gender identiﬁcation is relevant to veterinary, medical and ecological sciences, and is helpful in enforcing legislation and resolving paternity disputes [Lee et al., 2010]. As aviculture is constantly advancing, even individual owners wish to determine bird sex. Accordingly, a number of recent studies have focused on the development of efﬁcient molecular methods for sex identiﬁcation, which are gaining undivided attention as an aid in research and conservation of many bird species [Cerit and Avanus, 2007]. Other methods of avian sex determination are based on the observation of sex-speciﬁc behavior and the comparison of different morphological entities [Baker and Piersma, 1999, Jodice et al., 2000, Mendenhall et al., 2010, Tella and Torre, 1993]. Surgical methods (laparoscopy and laparotomy), which enable the direct observation of gonads, although successful in most cases, are aggressive [Grifﬁths and Phil, 2000]. Ultrasonography may also be used in sex identiﬁcation in birds...
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