Effects of Cryopreservation and Constituents of Semen Extenders on Mitochondrial Function of Bull Spermatozoa

This study investigated the effects of semen extender constituents and cryopreservation on bovine spermatozoal mitochondrial function. Three yearling Holstein bulls were used. Two ejaculates per bull were collected and pooled on a weekly basis for five weeks and extended in four treatments: 1) sodium citrate egg yolk extender with antibiotics (lincomycin, spectinomycin, gentamicin and tylosin); 2) ¡°1¡± with glycerol; 3) ¡°2¡± without antibiotics; and 4) ¡°1¡± without antibiotics. Each was divided into portions for analyses before freezing and after cryopreservation. The pre-freeze and thawed semen were transferred to a 37¡ãC water bath, the same assays were performed. In experiment 1, resazurin reduction (RD) was measured spectrophotometrically at sequential 25 minute intervals for 125 minutes. In experiment 2, specific activities of cytochrome c oxidase (CytoCox) and citrate synthase (CS) were measured spectrophotometrically immediately post-thaw and after 125 minutes of incubation. In experiment 3, ATP was measured using luciferin-luciferase assay simultaneously with RD. Total and progressive motilities (TM and PM), progressive (PV), curvilinear (VCL) and pathway (VAP) velocities were measured simultaneously with RD, ATP content and CytoCox and CS using computer assisted semen analysis system (CASA). In experiment 4, the NADH dehydrogenase (ND1) gene of mtDNA was sequenced before and after cryopreservation using PCR. Data were analyzed by least square methods; mean differences were delineated by Tukey¡¯s test. In experiment 1, RD differed among treatments (P\u3c0.05). Cryopreservation decreased (P\u3c0.05) RD, TM, PM, PV, VAP and VCL. Resazurin reduction correlated with PM (r=0.45, P\u3c0.05) and TM (r=0.2, P\u3c0.05). In experiment 2, incubation time and incubation with Triton X100 were sources of variation in CytoCox and CS specific activities (P\u3e0.05). Only CS from spermatozoa incubated with Triton X100 correlated with RD (r=0.22, P\u3c0.05). CytoCox and CS did not correlate with motility parameters. In experiment 3, spermatozoal ATP was not different (P\u3e0.05) among treatments. However, cryopreservation decreased (P\u3c0.05) ATP. Spermatozoal ATP correlated with motility parameters (r¡Ý0.65) and RD (r=-0.30) (P\u3c0.05). In experiment 4, the frequency of amino acid change was higher (P\u3c0.05) post-thaw in the treatment containing only antibiotics. Cryopreservation, more than extender constituents impacted mitochondrial function of bovine spermatozoa

Hematology and serum biochemistry variables in apparently normal Arabian Oryx (Oryx leucoryx)

Background and Aim: Hematology and serum biochemical analyses are integral parts of the clinical evaluation of sick animals. This is especially true regarding the clinical care of wildlife species, where clinical signs and historical data relating to the particular illness are often not available. Therefore, this study was designed to report various hematology and serum biochemistry parameters in Arabian oryx (Oryx leucoryx). Materials and Methods: Various hematology and serum biochemistry parameters were determined in 49 Arabian oryx of various ages and sexes. Hematology parameters included total red blood cells (RBC), packed cell volume (PCV), hemoglobin concentration, mean corpuscular volume, mean corpuscular hemoglobin (MCH), MCH concentration, and total leukocyte count (white blood cell) using an automated hematology analyzer. Serum biochemistry variables included glucose (Trinder method), total protein (biuret method), albumin using the Bromcresol Green (BCG) method, and blood urea nitrogen (colorimetric method). In addition, serum electrolyte concentrations of calcium, magnesium, and phosphorus were determined using colorimetric methods. Results: There was a significant difference in RBC count, PCV, and serum glucose concentration between adult and young Arabian oryx. The RBC count was significantly higher in males than in females, whereas the serum glucose concentration was significantly higher in females. Conclusion: Results of this study showed significant differences in RBC, PCV, and serum glucose concentration between apparently normal young and Adult Arabian oryx. Similar differences were also detected between normal males and females. Knowledge of these data could prove vital in the clinical evaluation of the health status of this wildlife species

Female reproductive tract anatomy of the endangered Arabian oryx (Oryx leucoryx) in Jordan

Female reproductive anatomy of the Arabian oryx is unknown. In this study, reproductive tracts of seven female Arabian oryx (aged 2 to 7 years) were examined to characterize their reproductive anatomy. Observations and measurements were obtained in situ from dead animals during necropsy. Animals were allocated into two groups: cycling (n=3; follicles or corpora lutea present) and not-cycling (n=4; follicles or corpora lutea absent). Different reproductive tract segments for each animal in both groups were measured using a digital caliper. The mean, SD and range for each reproductive tract segment were generated and compared between groups. Female oryx reproductive anatomy share some anatomical characteristics with that of domestic ruminants except that the oryx uterus has no distinct uterine body and the cervix has two internal openings for each respective uterine horn. In addition, there were more than 8 rows of caruncles within each uterine horn. There were significant differences in the length and width (P<0.05), but not in height, of both the right and left ovaries between cycling and not-cycling animals (P>0.05). Posterior and anterior vaginal lengths varied between cycling and not-cycling groups (P<0.05). Length of right and left oviducts, left and right uterine horns, cervix and vulva did not vary between cycling and not-cycling groups (P>0.05). Defining this unique morphology of female Arabian oryx reproductive anatomy will help in the development of appropriate reproductive techniques in order to propagate this endangered species and control its reproduction

Investigation of scrotal circumference, testicular dimensions and semen characteristics of the vulnerable Arabian oryx (Oryx leucoryx): an approach supports the future use of artificial insemination

Data regarding semen collection and characteristics in Arabian oryx are not available so far. Therefore, the current study aimed to evaluate scrotal circumference, testicular dimension and semen characteristics in four age groups of this species. Eleven male Arabian oryx bulls were divided into four groups according to age: group 1: 3 years old (n = 2); group 2: 4 years old (n = 2); group 3: 5 years old (n = 2) and group 4: 10 years old (n = 3). After immobilisation, scrotal circumference, testicular dimensions, volume and weight and gonadosomatic index (GSI) were estimated for each animal. Semen was collected from each male on a weekly basis for 3 weeks. Immediately after collection, semen was evaluated for volume, pH, osmolality total motility (TM), forward progressive motilities (PM) using a phase microscope. Furthermore, TM, PM, progressive velocity (PV), curvilinear velocity (VCL) and pathway velocity (VAP) have been evaluated using computer-assisted sperm analyser (CASA). Sperm concentration was evaluated using a haemocytometer, whereas sperm morphology was detected using phase contrast microscopy and staining methods by eosin-nigrosin and Spermac stains. Sperm viability was assessed using eosin-nigrosin stain under light microscopy and Sybr14/PI stain under fluorescence microscopy. The current findings indicated a positive correlation between body weight versus scrotal circumference, testicular weight, testicular volume and GSI. However, age did not have a positive association with percentage of normal sperm in Arabian oryx. Conclusively, the current findings provide a valuable data for future use of artificial insemination, which will be crucial for propagation and conservation of Arabian oryx