A comparison of plucked feathers versus blood samples as DNA sources for molecular sexing

Abstract

ABSTRACT. Feathers are increasingly collected as a nondestructive source of DNA for avian genetic research. Although feather samples are not optimal in some important ways than more robust blood or tissue samples, feather sampling requires less training for field workers, results in shorter handling times for the organism, generates no hazardous wastes, and requires simpler storage procedures. Along with these largely positive attributes comes a set of challenges, particularly the relatively low copy number of DNA present in feather samples. We compared the utility and reliability of feathers to the more traditional blood samples as sources of DNA for polymerase chain reaction (PCR)-based molecular sexing of Black-capped Chickadees (Poecile atricapilla). DNA from 102 individuals was extracted separately from both single rectrices and from blood samples, and the sex of each bird was then determined using standard PCR-based methods. We found complete agreement between sex determinations based on feather versus blood DNA extractions. Slight variations in lab protocols were necessary to obtain consistent results from these two DNA sources; and we briefly discuss other sources of error that could occur in feather-based molecular sexing studies. This controlled comparison of feather versus blood samples demonstrates that plucked rectrices provide a highly reliable source of DNA for molecular sexing of wild birds. SINOPSIS. Una comparación entre el uso de plumas versus muestras de sangre como fuentes de ADN para estudios moleculares de determinación de sexoSe está incrementando la modalidad de usar plumas como una fuente no destructiva de ADN para llevar a cabo investigación genética en aves. Aunque las muestras tomadas de plumas, en algunas instancias no sonóptimas que muestras más robustas como sangre y tejido, el uso de estas no requiere adiestramiento especial, toman menos tiempo en la manipulación del ave, no generan desperdicios peligrosos y requieren un almacenaje sencillo. Aunque hay atributos positivos en el uso de plumas, también existe el problema de obtener una muestra baja de ADN. Examinamos la utilidad y confiabilidad de muestras de plumas en comparación con técnicas más tradicionales como obtención de muestras de sangre como una fuente de ADN para el sexado de Poecile atricapilla utilizando PCR. A tales efectos se extrajo ADN, de rectrices, de 102 individuos y un número similar de muestras de sangre para determinar el sexo usando PCR. Encontramos resultados similares con ambos métodos. Sin embargo, fueron necesarios pequeñas variaciones en el protocolo de laboratorio para obtener resultados consistentes, de ambas fuentes de ADN. Discutimos algunas fuentes de error que pueden ocurrir cuando se utilizan plumas para la determinación del sexo utilizando técnicas moleculares. Este estudio demuestra que las rectrices son una fuente confiable para obtener buenas muestras de ADN y poder determinar el sexo en aves silvestres.

Application of Two Molecular Sexing Methods for Indonesian Bird Species: Implication for Captive Breeding Programs in Indonesia

“…the primers may match one CHD gene slightly less well that the other. Harvey et al (2006) supported our result give evidence that feathers can provide sufficient DNA for molecular sexing reactions. This low cost, speed, and ease of collection, storage, and transport of feather samples are the major advantages (Duan & Fuerst 2001).…”

Exonic versus intronic SNPs: contrasting roles in revealing the population genetic differentiation of a widespread bird species

“…In our study, about 79% of DNA extracts were from feather samples (n ¼ 136) and we found that, as expected, shorter fragments yielded significantly higher PCR efficiencies (Table 2). This is not unexpected because feathers usually contain less DNA (see, for example, Bush et al, 2005) that is sometimes more degraded than in blood and tissue samples (Harvey et al, 2006). Another factor affecting the PCR efficiency is storage condition.…”

Rapid sex determination of a wild passerine species using loop‐mediated isothermal amplification (LAMP)

“…We chose blood over feathers, for example, because bird blood is a better tissue for obtaining high amounts of DNA; nucleated erythrocytes provide much more DNA compared to relatively low amounts of DNA recovered from feather samples (Harvey, Bonter, Stenzler, & Lovette, 2006). We chose blood over feathers, for example, because bird blood is a better tissue for obtaining high amounts of DNA; nucleated erythrocytes provide much more DNA compared to relatively low amounts of DNA recovered from feather samples (Harvey, Bonter, Stenzler, & Lovette, 2006).…”