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Luther, I., 2016. Semen characteristics of free-ranging African elephants (Loxodonta africana) and Southern white rhinoceros (Ceratotherium simum simum) using Computer-aided sperm analysis, Electron microscopy and Genomics as diagnostic tools. Thesis presented to the University of the Western Cape

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Location: Africa - Southern Africa - South Africa
Subject: Reproduction
Species: White Rhino


Original text on this topic:
The survival of free-ranging (in situ) African elephant and Southern white rhinoceros populations are currently being challenged on a daily basis in Africa. Reproductive health is considered a vital component of species conservation. Conservation of the last mega land mammals may ultimately require intervention by breeding management or combined with assisted reproductive technologies (ART). There is a strong case for gathering baseline information, both physiological and biological, of any species, as opportunities arise. During this study a total number of 21 ejaculates collected over two seasons from 12 free-ranging African elephant bulls were characterised, as well as 10 ejaculates collected from 10 free-ranging Southern white rhinoceros bulls from two populations. Ejaculates were collected from adult bulls by means of electroejaculation under anaesthesia. Routine semen analysis was combined with Computer-aided sperm analysis (CASA), Computer-aided sperm morphology analysis (CASMA), Transmission electron microscopy (TEM) and Genomics as diagnostic tools. Additionally, sperm functionality within different media was investigated and sperm subpopulation classification according to the motion pattern displayed. The results presented is based on the evaluation and classification of ≈ 45 000 individual African elephant spermatozoa and ≈ 18 000 individual Southern white rhinoceros spermatozoa. The average elephant ejaculate contained a total number of 47 x 10⁹ spermatozoa (volume of 56 38mL x concentration of 818 750 x 10⁶/mL) that recorded a total motility of 81 29% of which 62 26% were progressively motile. CASA recorded velocities for curvilinear velocity (VCL 241 58μm/s), straight-line velocity (VSL 173 181μm/s) and average path velocity (VAP 201 54μm/s), and kinematics at straightness of track (STR 86 85%), linearity of track (LIN 67 16%), amplitude of lateral head displacement (ALH 4 0.75μm) and beat cross frequency (BCF 21 3Hz). Structural analysis revealed 68 11% of the spermatozoa were viable (intact plasma membrane) and 77 11% maintained acrosome integrity. Ejaculates contained 55 14% morphologically normal spermatozoa, CASMA measured sperm head lengths at 6.83 0.26μm and width 3.32 0.18μm (total head area of 20.17 1.96μm) of which 38.95 0.92% is covered by an acrosomal cap. The average rhinoceros ejaculate contained a total number of 1.1 x 10⁹ spermatozoa (volume of 24 24mL x concentration of 83 96 x 10⁶/mL) that recorded a total motility at 82 8% of which 28 23% were progressively motile. CASA recorded velocities for VCL (85 29μm/s), VSL (44 25μm/s) and VAP (69 30μm/s, and kinematics at STR (63 14%), LIN (51 16%), ALH (2 0.16μm) and BCF (16 6Hz). Structural analysis revealed 73 10% of the spermatozoa were viable (intact plasma membrane) and 76 4% maintained acrosome integrity. Ejaculates contained 62 14% morphologically normal spermatozoa, CASMA measured sperm head lengths at 5.5 0.17μm and width 2.9 0.19μm (total head area of 14.8 1.43μm) of which 36.3 0.59% is covered by an acrosomal cap. Based on a Boolean argument and CASA data exploration it was possible to derive elephant and rhinoceros CASA cut-off criteria to sort between activated and hyperactivated motile spermatozoa. For the genomic component of this study, the CatSper1 (Loxodonta africana) gene was identified,sequenced and verified in a free-ranging (natural) African elephant population. Multivariate analysis(MVA) was applied to examine the associations between the semen and sperm parameters and the traits they accounted for in this study. Our understanding of wildlife reproductive sciences can substantially progress as the analytical techniques applied and the combination thereof is expanded. This investigation presents a new set of comprehensive semen and sperm threshold values for future investigations.

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