Analysis of CHD Gene Polymorphism as a Model Object for Molecular Sexing of Eurasian Eagle-Owl (Bubo bubo)

Kulibaba, R. O.; Liashenko, Yuriy

doi:10.3103/s0095452721040071

Cited by 4 publications

(4 citation statements)

References 30 publications

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“…The CHD-W and CHD-Z genes among avian species are known to have wide diversity (Ciorpac et al, 2016;Kroczak et al, 2021;Kulibaba and Liashenko, 2021). Huang et al (2011) reported differences in the sequences of the CHD-W and CHD-Z genes among 3 Columbidae species (C. livia, C. pulchricollis, and S. tranquebarica).…”

Section: Resultsmentioning

confidence: 99%

“…Although the PCR method provides good results in determining the sex of birds, one of its successes is greatly influenced by the compatibility between the primers used and the DNA template (van der Velde et al, 2017). The CHD-W and CHD-Z genes in avian species are known to have varied nucleotide base sequences (Ciorpac et al, 2016;Kroczak et al, 2021;Kulibaba and Liashenko, 2021). Variations in nucleotide sequence can cause annealing process of primer PCR not run perfectly to the DNA template, which implicate the success of PCR amplification (Morinha et al, 2015;Green and Sambrook, 2019).…”

Section: Introductionmentioning

confidence: 99%

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Appropriate Primer Selection Improves Molecular Bird Sexing Accuracy

Fitriana,

Al Qodry,

De Lucas

et al. 2023

BuletinPeternak

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Birds sexing utilize the Polymerase Chain Reaction (PCR) technique is increasingly being used by researchers and breeders. The PCR technique has high sensitivity, but its success is influenced by the specificity of the DNA template with the oligo primer used. This study aimed to evaluate 5 types of PCR primers P2/P8, 2550F/2718R, CHD1F/CHD1R, 1237L/1272H, and CHD1LF/CHD1LR to determine the sex of Phasianidae, Anatidae, Muscicapidae, and Psittacidae families. This research was conducted by tested primers mentioned above to amplify the target gene chromodomain helicase DNA binding 1 (CHD1) on DNA samples of each pair of males and females from four bird families, respectively. The results indicated that CHD1LF/CHD1LR PCR primer gave the best results and was recommended to determine the sex of four families tested. Some of other primers tested in this study failed to amplify targeted gene correctly, it is important to use appropriate primer to increase bird sexing accuracy.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Appropriate Primer Selection Improves Molecular Bird Sexing Accuracy

Fitriana,

Al Qodry,

De Lucas

et al. 2023

BuletinPeternak

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“…Various laboratory sexing analyses have been performed in studies on owl species, such as snowy owl ( Bubo scandiacus ) 32 , Indian scops owl ( Otus bakkamoena ) 33 , Tengmalm’s owl ( Aegolius funereus ) 34 , Southeast Asian barn owl ( Tyto alba javanica ) 3 , 35 , and Eurasian eagle owl ( Bubo bubo ) 36 . However, before the first molecular sexing of a given species, a primary calibration of the laboratory methods should always be done, and an appropriate primer set should be selected that gives the correct results for the studied species 37 , 38 .…”

Section: Introductionmentioning

confidence: 99%

Comparison of three primer pairs for molecular sex determination in Eurasian pygmy owls (Glaucidium passerinum)

Stehlíková Sovadinová,

Mekadim,

Korpimäki

et al. 2024

Sci Rep

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Bird sex determination is fundamental in various ecological and biological studies, although many avian species cannot be sexed visually due to their monomorphic and/or monochromatic appearance. Thus, reliable laboratory methods for sexing are a prerequisite. Most avian nestlings lack sex-related signs, including the Eurasian pygmy owl (Glaucidium passerinum). We performed laboratory sex determination analysis of this species using blood samples of 242 juveniles and nine adults. It relied on the qPCR of the specific intron from the chromo-helicase DNA-binding protein 1 gene. We tested three primer sets, the P2/P8, 2550F/2718R, and CHD1F/CHD1R, commonly used for bird laboratory sexing. The outcomes were displayed on an agarose gel electrophoresis and a plot from melt curve analysis, which had not been previously conducted in Eurasian pygmy owls. We found that only primer set CHD1F/CHD1R proved reliable, as the only one determined sex with one and two band/s and peak/s on the electrophoresis and the melt curve plot for males and females, respectively. The other two primer pairs failed and depicted one band/peak in all specimens regardless of their sex. Therefore, we recommend performing Eurasian pygmy owls’ laboratory sexing by qPCR with CHD1F/CHD1R primers only.

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“…Variations the sequence of nucleotide bases in the PCR target gene may cause the primers annealing to the template DNA perfectly, which has implicate the success of PCR amplification [17]. CHD-W and CHD-Z genes in avian species are known to have varied nucleotide sequences [5,18,19]. Fitriana et al [20] reported P2/P8, 1237L/1272H, 2550F/ 2718R, and CHD1F/ CHD1R primers gave unsatisfactory PCR results such as the size of PCR band did not match to the reference, extra bands or the absence of PCR products in determining the sex of 5 species of Columbidae family Columba livia, Streptopelia chinensis, Treron griseicauda, Spilopelia chinensis, and Geopelia striata.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of primers targeting chromo helicase DNA-binding gene (CHD) for molecular sexing identification in four bird families

et al. 2023

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Objective: Sex determination of birds is crucial role in breeding and conservation purposes. Genomic approaches by using basis of bird sex polymorphism are accepted to generate an accurate sexing procedure. The PCR method is commonly used and known to provide accurate test results in determining the sex of birds, but its successes is strongly influenced by the compatibility of the primers used with the DNA template. This study aimed to determine the potential use of CHD1LF/CHD1LR targeting CHD gene fragment in 4 species from Phasianidae, Psittacidae, Estrildidae, and Passeridae families. Method: Blood samples (n= 30) from representatives of species in 4 families above were collected for DNA isolation, then PCR test was performed using CHD1LF/CHD1LR primers. The PCR results obtained were compared with positive controls, necropsy examination and information from bird sellers. Result: The results showed that the primer CHD1LF/CHD1LR could be used in Phasianidae, Psittacidae, Estrildidae, and Passeridae families. The PCR product was confirmed to be same as well as the positive control and necropsy results. It is demonstrated that male birds showed a single band (474 bp), while double bands (474 and 319 bp) were observed in female birds. Different accuracies were observed during this study between molecular approach and manual sexing by bird seller i.e: sparrows (46%) and finches (50%), while the accuracy of parakeets was 75%, and quails 80%. Conclusions: Primer CHD1LF/CHD1LR can be used to determine the sex of birds from the families Phasianidae, Psittacidae, Estrildidae, and Passeridae. Our study showed that molecular-based method is valuable and able to reduce error rate to 100% in bird sexing procedures.

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Analysis of CHD Gene Polymorphism as a Model Object for Molecular Sexing of Eurasian Eagle-Owl (Bubo bubo)

Cited by 4 publications

References 30 publications

Appropriate Primer Selection Improves Molecular Bird Sexing Accuracy

Appropriate Primer Selection Improves Molecular Bird Sexing Accuracy

Comparison of three primer pairs for molecular sex determination in Eurasian pygmy owls (Glaucidium passerinum)

Evaluation of primers targeting chromo helicase DNA-binding gene (CHD) for molecular sexing identification in four bird families

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