Semen Collection and Polymerase Chain Reaction‐Based Sex Determination of Black‐Headed and Straw‐Necked Ibis

Abstract

This study aimed to develop a polymerase chain reaction (PCR)-based sexing and effective semen collection methods for black-headed and straw-necked ibis species. However, most birds are not sexually dimorphic, that is, the sexes appear similar. Therefore, the gender should be determined before semen collection. DNA was extracted from the blood samples of 11 black-headed and 4 straw-necked ibis. The sex was determined after PCR amplification of the EE0.6 region of W-chromosome. The PCR products were separated using gel electrophoresis. A single band indicated the presence of the EE0.6 region and that the individual was a female, while no band indicated that the individual was a male. Further, the single bands from seven specimens were amplified. Semen collection was performed by massage or a combination of massage with electro-ejaculation and was attempted during all four seasons. The semen was successfully collected in March from male straw-necked ibis using the massage method. Limited motility, viability and concentration of straw-necked ibis sperm were observed. The sperm length was 180 μm and that of the nucleus was 30 μm with acrosome located at the tip of the nucleus. Thus, the PCR-based sexing proved to be an accurate molecular sexing method for black-headed and straw-necked ibis. Furthermore, we successfully collected semen and observed the stained sperm nucleus and acrosome of the straw-necked ibis sperm. We propose that the use of this PCR methodology can be applied as a routine method for sex determination and semen collection in ibis species for future ecological research. However, considering our limited success, further studies on semen collection method are required.

Comparative Examination of Capercaillie (Tetrao urogallus L.) Behaviour Responses and Semen Quality to Two Methods of Semen Collection

“…For semen collection from Muscovy drake [ 30 ], [ 31 ], as well as from nondomestic bird [ 32 ], the electro-ejaculation method has been applied. However, this procedure causes considerable stress to male, since he has to be anesthetized or physically restrained by human, therefore this method of semen collection is practiced rather sporadically.…”

Transgenerational Effects on Calf Spermatogenesis and Metabolome Associated with Paternal Exposure to the Fukushima Nuclear Power Plant Accident

“…The acrosomal integrity of the delivered calf sperm was assessed by staining with fluorescein isothiocyanate-conjugated peanut agglutinin (FITC-PNA; Wako, Japan) according to the procedure described previously [13].…”

A proteomic analysis of Nipponia nippon (ID#162)

“…; Kaneko et al . ), it is necessary to identify the global protein expression within each tissue in a Crested Ibis’ body and to provide a biochemical, molecular biological and physiological baseline for the Crested Ibis. The genome analysis for Crested Ibis is reported to be complete (http://news.xinhuanet.com/english2010/sci/2011-04/15/c_13830955.htm); however, it cannot be used for proteomic analysis, as it has not been placed in the public domain.…”