Lation [5] and artificial vagina [7]. However, high proportion of ejaculates obtained via rectal massage represented a sperm-rich fraction with minimal seminal plasma contribution and may not represent a `true ejaculate’. Fantastic GLUT2 web motility ejaculates have been characterized by bigger volume and reduce sperm concentration, hence indicating greater high-quality ejaculates with higher percentages of sperm motility were diluted with bigger amounts of seminal plasma. Therefore, despite the fact that the rectal massage approach is practical and represents a somewhat protected choice for semen collection from captive managed bull elephants, an option process that mimics natural ejaculatory responses such as contributions in the accessory sex glands may be warranted to enhance semen collection efficiencies in elephants. As observed herein and inside a previous study [3], a higher proportion of ejaculates obtained through the rectal massage techniqueFigure 2. One-dimensional gel electrophoresis of Asian elephant seminal plasma proteins. Each and every column represents protein profiles from seminal plasma samples obtained from different ejaculates. E1, E2, and E3 denote person Asian elephant bulls. G1, G2, G3, G4, G5, and G6 denote seminal plasma samples obtained from ejaculates exhibiting superior motility ( 65 tMOT). P1, P2, P3, P4, and P5 denote seminal plasma samples obtained from ejaculates exhibiting poor motility (#10 tMOT). MW: molecular weight marker. Band A was detected in 85 of seminal plasma samples from excellent motility ejaculates (a representative gel is shown). Within this figure, protein bands were visualized with Coomassie staining. doi:ten.1371/journal.pone.0071033.gPLOS 1 | plosone.orgLactotransferrin in Elephant Seminal PlasmaFigure 3. Protein separation and peptide mass sequencing of Asian elephant seminal plasma. Seminal plasma proteins have been separated by two-dimensional gel electrophoresis. MAO-B Compound Selected spots had been excised and submitted for mass spectrometry (MALDI-TOF) for protein identification by de novo sequencing of MALDI-PSD tandem mass spectra. Panel A: 2D gel of seminal plasma proteins from an ejaculate exhibiting excellent sperm motility ( 65 tMOT). Exactly the same sample from Panel A can also be shown in Figure two, Lane G5. Cored spots from within the train of this protein (circled; corresponding with band A in Figure two) had been submitted for mass spectrometric evaluation. Panel B: Example of a 2D gel of seminal plasma proteins from an ejaculate exhibiting poor sperm motility (#10 tMOT). Panel C: Mass spectrum benefits from MALDI-TOF. Panel D: Peptide mass sequencing identified the cored proteins as getting homology to lactotransferrin (i = isoleucine or leucine). doi:ten.1371/journal.pone.0071033.gPLOS One | plosone.orgLactotransferrin in Elephant Seminal PlasmaFigure four. Amino acid sequence of predicted African elephant (Loxodonta africana) lactotransferrin determined in the 29 Mammals sequencing project (broadinstitute.org). Alignment of tryptic peptide sequences determined by MALDI-PSD sequencing of SPITC derivitized peptides shown italicized. Residues marked as “i” represent either isoleucine (I) or leucine (L), that are isobaric. Asparagine 546 has probably been deamidated to its corresponding aspartic acid. doi:10.1371/journal.pone.0071033.gconsistently contained spermatozoa with poor motility. Nonetheless, top quality ejaculates have also been obtained utilizing the identical rectal massage strategy [3,4]. This suggests that perhaps other factors, for instance seasonality and/or circulatin.