Effect of weaning method on body weight,testicular growth,sexual behavior,age of puberty,and testosterone concentration of Beetal bucks

The current study evaluates the effects of early (8th week) and late (16th week of age) weaning of male goat kids on their body growth, testicular growth, sexual behavior, plasma testosterone concentration, and pubertal age. Early (n = 6) and late (n = 7) weaned Beetal bucks were weekly monitored from 18th to 38th week for their body weight, scrotal circumference, testicular volume, testicular echogenicity (via ultrasonography), sexual activities, and plasma testosterone concentration. In comparison to early-weaned, late-weaned bucks showed a marked increase (p < .05) in body weight (11.4 ± 0.8 vs. 13.7 ± 0.6kg), testicular volume (44.1 ± 7.2 vs. 79.8 ± 18.7cm³), scrotal circumference (10.7 ± 0.6 vs. 12.8 ± 0.7cm), and testicular echogenicity (28.3 ± 2.7 vs. 38.3 ± 2.1) from 18th, 28th, 21st, and 24th week onward, respectively. Sexual activities started earlier in late- than early-weaned bucks (22nd vs. 25th week, respectively). Moreover, the sexual behavior index was better (p < .05) after the 34th week in late than early-weaned bucks. The plasma concentration of testosterone (at 39 weeks of age) was relatively more and the onset of puberty was 2–3 weeks earlier (p < .05) in late than early-weaned bucks. In conclusion, age-based early weaning of male kids impairs their testicular growth, sexual behavior, and age at puberty compared to conventional weaning.     

Changes in testicular size,echotexture, and arterial blood flow associated with the attainment of puberty in Dorper rams raised in a subtropical climate

There is a paucity of information on the relationships of testicular morphology, echotextural attributes, and blood flow dynamics with pubertal development of rams raised in a subtropical climate. Forty‐five Dorper rams (24 rams aged 8–11 months and 21 rams aged 12–24 months) were examined using a portable ultrasound scanner connected to a 7.5‐MHz transducer. Computer‐assisted analyses of testicular ultrasonograms utilized commercially available Image ProPlus^® analytical software. Spectral Doppler scans of testicular arteries were performed immediately after scrotal (B‐mode) ultrasonography to determine peak systolic velocity (PSV), end‐diastolic velocity (EDV), resistive index (RI = [PSV?EDV]/PSV), and pulsatility index (PI = [SPV–EDV]/mean velocity) of the blood vessels. The length of the testes (9.7 ± 0.3 compared with 9.0 ± 0.2 cm) and scrotal circumference (33.3 ± 0.5 compared with 31.8 ± 0.4 cm) were greater (p < 0.05) but testicular depth (4.5 ± 0.1  compared with 4.9 ± 0.08 cm) was less (p < 0.05) in sexually mature compared with peripubertal rams. [Corrections added on 9 Jan 2019 after initial online publication: The testicular size values in the sentence were corrected.] There were no differences (p > 0.05) between the two age groups of Dorper rams in blood flow indices of testicular arteries. Mean numerical pixel values (100.5 ± 4.1 compared with 89.2 ± 4.8) and pixel heterogeneity (25.6 ± 0.6 compared with 23.6 ± 0.5) of testicular parenchyma were greater (p < 0.05) in peripubertal than in postpubertal rams. Semen volume was negatively correlated with PI of testicular arteries (r = ?0.57, p = 0.04). In summary, the attainment of sexual maturity in the rams of the present study was associated with significant changes in testicular length and depth, scrotal circumference, and parenchymal echogenicity/hetrogeneity but not in testicular volume and blood perfusion rates. Testicular artery PI can be used to predict the volume of ejaculate in rams.     

Assessment of the accuracy of testicular dysfunction detection in male donkey (Equus asinus) with the aid of colour-spectral Doppler in relation to plasma testosterone and serum nitric oxide levels

This study aimed to determine the usefulness of colour and pulsed Doppler modes for the accurate diagnosis of donkeys suffering from subfertility to determine whether testicular vascularity assessment could be an indicator for sperm functionality. The study sample was composed of 10 male donkeys with normospermia (control group) and 10 donkeys with hypospermia. Animals underwent scrotal circumference measurement, testicular Doppler examination, seminal evaluation, blood sampling and hormonal assay. Semen volume and concentration were significantly (p ≤ .05) lower in the subfertile group (30.25 ± 1.22 ml and 89.44 ± 2.55 × 10⁶/ml) as compared with the control group (82.76 ± 1.65 ml and 452.78 ± 1.25 × 10⁶/ml), and total sperm/ejaculation was significantly (p ≤ .05) higher in the normal donkeys (28.30 ± 2.32 × 10⁹/total ejaculated) as compared with the subfertile group. Intratesticular coloured area showed a marked decline in the hypospermic males. There was no significant difference between the two groups in testosterone level, although the normal group showed an increase in nitric oxide metabolites. Both Doppler indices of the three branches of the testicular artery were elevated significantly (p ≤ .05) in abnormal donkeys, whereas Doppler peak systolic and end-diastolic velocities were increased in the normal group. Male donkeys with subfertility demonstrated lower arterial vascularity parameters in the form of intratesticular coloured area and blood flow rate; therefore, the most optimal parameters for differentiating subfertile hypospermic from normospermic donkeys were found to be the two Doppler indices, velocities parameters, testicular blood flow rate and nitric oxide levels.     

Melatonin improves testicular haemodynamics,echotexture and testosterone production in Ossimi rams during the breeding season

The objective was to determine effects of a single parenteral dose of melatonin on testicular blood flow indices, testicular echogenicity and plasma testosterone concentrations in rams during the physiological breeding season. We hypothesized that melatonin enhances testicular blood flow, echogenicity and plasma testosterone concentrations during the breeding season in rams. During the breeding season, 12 sexually mature Ossimi rams were randomly allocated to either a melatonin group (n = 8) that received 18 mg of melatonin in 1 ml of corn oil (injected SC) or a control group (n = 4) that received 1 ml corn oil only. Blood collection and ultrasonographic assessment of the testes and supratesticular arteries were conducted immediately before treatment (W0) and once weekly for 6 weeks after melatonin injection (W1-W6). Mean plasma testosterone concentrations were greater (p < .05; at least 1 ng/ml) in the melatonin-treated group compared to the control group from W4 to W6 after treatment. A decrease (p < .05) in both resistive index (RI) and pulsatility index (PI) began 1 week after melatonin injection (W1) and persisted until the end of the experiment, with mean RI and PI values in the melatonin group lower (p < .05) than those in the control group on W3 and W4. Furthermore, plasma testosterone concentrations in melatonin-treated rams were inversely correlated to both RI and PI (r = −.7 and −.6, respectively, p < .01). Testicular echogenicity decreased (p < .05) 1 week after melatonin injection (W1) and remained lower (p < .05) in the melatonin-treated group compared to the control group until the end of the study (W6). In conclusion, melatonin administration significantly altered testicular blood flow and echogenicity and increased plasma testosterone concentrations in Ossimi rams during the breeding season.     

Effects of five cryoprotectants on proliferation and differentiation‐related gene expression of frozen‐thawed bovine calf testicular tissue

The cryopreservation of testicular tissue is a potential method for preserving male fertility. However, the effect of cryopreservation on bovine calf testicular tissue is scarce. This study investigated the effect of different cryoprotectants on bovine calf testicular tissue at the molecular level. Testicular tissue from ten immature bovine calves (6 months) was collected after slaughter and cryopreserved in an extender containing different concentrations of the following five cryopreservation solutions (CP): bovine serum albumin (BSA) with 5% dimethyl sulfoxide (DMSO), trehalose with 5% DMSO, DMSO and glycerol and ethylene glycol (EG). After 7‐day cryopreservation, the expression levels of three spermatogonial stem cell (SSC)‐related genes, octamer‐4 (OCT4), KIT ligand (MGF/SCF) and kit oncogene (C‐KIT), were investigated by quantitative PCR (qPCR). The cell viability was highest for the tissues preserved with 30 mg/ml BSA (77.82% ± 1.22) and 40 mg/ml trehalose (74.23% ± 1.16) compared with other groups (p < 0.05), and the level of expression of the three genes was highest with 30 mg/ml BSA (p < 0.05). Compared with other CPs, the 30 mg/ml BSA and 40 mg/ml trehalose have the better cryopreserve protection. The 30 mg/ml BSA is the most viable media for the cryopreservation of testicular tissue from cattle.     

Relationship of age to body weight,scrotal circumference,testicular ultrasonograms,and semen quality in Sahiwal bulls

The objective of the present study was to determine the relationship of age to body weight (BW), scrotal circumference (SC), number of pixels of testicular ultrasonograms (NP), and semen quality in Sahiwal bulls. The study was based on 128 Sahiwal bulls of different age groups (from 0 to >100 months of age). Bulls were evaluated for SC, BW, and NP. Semen was evaluated once a week for five consecutive weeks from regularly collected donor bulls (n = 86) ranging in age from 25–30 to >100 months. Ejaculate volume, sperm motility, sperm concentration, sperm morphology, percent live sperms, sperm plasma membrane integrity, and normal acrosome were compared among different age groups. Mean SC and BW increased (P < 0.05) in a curvilinear manner from birth to >100 months of age. Mean NP of testicular ultrasonograms increased (P < 0.05) from 0 to 24 months and then plateaued until >100 months of age. Body weight, SC, and NP were positively correlated with age from birth until >100 months (r = 0.91, 0.87, and 0.40, respectively). Ejaculate volume (5.7 ± 0.2 vs. 4.6 ± 0.09 ml) and sperm concentration (1,281.6 ± 17.7 vs. 1,115.8 ± 55.9 × 10⁶/ml) increased (P < 0.05) in mature bulls compared to younger ones. However, motility (68.6 ± 0.3%), plasma membrane integrity (50.8 ± 1.0%), and normal acrosome (74.8 ± 0.5%) remained insignificant due to age. In six of eight age groups studied, morphological abnormalities were well within the range (18.1 ± 0.3%). In conclusion, the BW, SC, and NP of testicular ultrasonograms, ejaculate volume, and concentration increased with age. Moreover, semen quality is fairly independent of age except volume and concentration in Sahiwal bulls.     

Effect of caspase inhibitor Z-VAD-FMK on bovine sperm cryotolerance

The aim of this study was to evaluate the treatment of bovine semen with the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-FMK), before or after freezing on semen quality. After the initial assessment, sperm from 4 bulls were pooled (Experiment 1) and cryopreserved in BioXcell containing 0, 20 and 100 μM Z-VAD-FMK. After thawing semen viability, motility, membrane integrity, as well as DNA fragmentation and ΔΨm were evaluated. In Experiment 2, bovine frozen/thawed sperm were incubated for 1 hr with 0, 20 and 100 µM Z-VAD-FMK before assessing the semen quality. The treatment with Z -VAD-FMK before cryopreservation improved post-thawing sperm motility compared to the control group (p < .05), while no differences were recorded in sperm viability and membrane integrity among groups (on average 86.8 ± 1.5 and 69.1 ± 1.4, respectively). Interestingly, at the highest concentration, DNA fragmentation decreased (p < .05), while the percentage of spermatozoa with high ΔΨm increased (p < .05). The results of Experiment 2 showed that 1-hr treatment with Z-VAD-FMK did not affect sperm motility and viability (on average 63.4 ± 5.8 and 83.7.1 ± 1.2, respectively). However, Z-VAD-FMK improved sperm membrane integrity (p < .05) and at the highest concentration tested decreased the proportion of sperm showing DNA fragmentation (p < .05). No differences were recorded in the percentage of spermatozoa with high ΔΨm (on average 57.0 ± 11.4). In conclusion, the treatment with 100 µM of the caspase inhibitor Z-VAD-FMK before freezing increased bovine sperm mass motility and ΔΨm, while decreasing sperm DNA fragmentation. Treatment of semen after thawing with 100 µM Z-VAD-FMK improved sperm membrane integrity and reduced DNA fragmentation.     

The occurrence of reflexes and reactions in cattle following stunning with a captive bolt at the slaughterhouse

The difference in the frequency of the occurrence of reflexes/reactions in 355 cows (average weight 505.5 ± 7.9 kg) and 262 bulls (average weight 735.6 ± 8.4 kg) following stunning with a Matador SS 3,000 B trigger‐activated captive bolt gun (Termet) was determined. The stun shot more than 2 cm from the ideal position was found in 79.6% of animals. Vocalization, corneal reflex, rhythmic breathing, blinking, eyeball rotation and the absence of tongue protrusion occurred more frequently in bulls (p < .05). Spontaneous limb movements and nystagmus occurred more frequently in cows (p < .05). No difference between bulls and cows was determined in the occurrence of a response to painful stimuli or attempts to regain normal posture. No dependency was determined between the frequency of the occurrence of a reflex/reaction and the distance of the stun shot from the ideal point on the skull for any of the monitored signs. This study shows that the occurrence of reflexes/reactions following the stunning of cattle with a captive bolt is not only dependent on the position of the shot if placed within a 9 cm radius from the ideal point on the cattle skull.     

Cryopreservation of Nili‐Ravi buffalo (Bubalus bubalis) semen in AndroMed® extender; in vitro and in vivo evaluation

The study was designed to evaluate AndroMed^® for the freezability and fertility of Nili‐Ravi buffalo semen. Semen was collected from four adult Nili‐Ravi buffalo (Bubalus bubalis) bulls for 3 weeks (replicate). Semen ejaculates from each buffalo bull were divided into three aliquots. One aliquot was used for evaluation of motility, plasma membrane integrity, livability, viability, DNA integrity and normal apical ridge. Remaining two aliquots were diluted (37°C; 50 × 10⁶ spermatozoa/ml) in tris‐citric egg yolk or AndroMed^® extender and cryopreserved in 0.5 ml French straws. After thawing, per cent post‐thaw motility (47.9 ± 0.8, 49.2 ± 1.7), plasma membrane integrity (44.4 ± 1.2, 46.8 ± 1.8) and normal apical ridge (81.4 ± 0.3, 83.2 ± 0.3) were recorded similar (p > .05) in tris‐citric egg yolk and AndroMed^® extender. Higher (p < .05) percentage of sperm livability (70.5 ± 1.4 and 64.4 ± 1.0), viability (67.5 ± 1.5 and 61.5 ± 0.6) and DNA integrity (97.0 ± 0.3 and 93.4 ± 0.21) were recorded in AndroMed^® compared to tris‐citric egg yolk post‐thaw. Values for all the aforementioned spermatozoal quality parameters were observed lower (p < .05) in frozen‐thawed compared to fresh semen irrespective of the experimental extenders. Fertility rates of buffalo semen did not differ (p > .05) either cryopreserved in tris‐citric egg yolk or AndroMed^® extender (45.5% vs. 49%). It is concluded that AndroMed^® is capable in protecting the buffalo bull sperm during freeze‐thawing process and can be adopted safely for routine use replacing the tris‐citric egg yolk extender in artificial insemination programme.     

Assessment of age-related morphological changes in the testes of post-hatch light ecotype Nigerian indigenous chicken

The age-related morphological changes of the testes in light ecotype Nigerian indigenous chicken were evaluated in this study using gross anatomical, histological and histomorphometric techniques. The results showed that the testes of 3- to 9-month-old birds were light pink while testes of sexually mature chicken were creamy white in colour. The left and right testicular weight, length, diameter, circumference and the organosomatic indices increased significantly (p < .05) with increasing age across the groups. Although the mean tubular diameter and epithelial height of the left and right seminiferous tubules increased significantly (p < .05) with age, the tubular diameter, epithelial height and luminal diameter did not vary significantly (p > .05) between the left and right testes of all the groups. The one-cell layer thick germinal epithelium of the left testes at 3 to 6 months old showed islands of cell proliferation that contained spermatogonia and spermatocytes. At 6 to 9 months, the left testes exhibited numerous early spermatids with occasional occurrence of late stage spermatids while the right testes showed scanty early stage spermatids. At 12 to 18 months, the germinal epithelia of both left and right testes were characterized by the presence of Sertoli cells, spermatogonia, primary spermatocytes, numerous early and late stage spermatids as well as spermatozoa. In conclusion, the morphological features highlighted in the present study show that at pre-pubertal periods, the left testes may develop faster than the right testes. However, both left and right testes may participate actively in the production of spermatozoa during the post-pubertal life.     

Extended day length in late winter/early spring,with a return to natural day length of shorter duration,increased plasma testosterone and sexual performance in rams with or without melatonin implants

Sixteen rams were used to quantify the effects of long days, imposed during late winter/early spring, with or without exogenous melatonin, on plasma testosterone concentrations and ram serving capacity. Rams were assigned to two groups: photoperiod‐treated rams (Artificial Photoperiod, AP; n = 8), exposed to 2 months of long days (16 hr of light/day) between 22 December and 22 February, and control rams (Natural Photoperiod, NP; n = 8). At the end of the long‐day period, AP rams were returned to the natural photoperiod, and each ram in the two groups either did (+M) or did not (‐M) receive three subcutaneous melatonin implants. Four groups were created as follows: AP+M (n = 4), AP‐M (n = 4), NP+M (n = 4) and NP‐M (n = 4). Thirty days after of the onset of photoperiodic treatment, AP rams (13.5 ± 2.8 ng/ml) had significantly (p < .05) lower testosterone levels than NP rams (36.7 ± 1.0), and similar differences were not apparent at the end of the photoperiod treatment. A month later, AP rams (24.3 ± 7.9) had higher (p < .10) testosterone levels than NP rams (13.1 ± 5.0), with no effect of melatonin treatment. Fifty days after melatonin implantations, rams were exposed for 20 min to three oestrous ewes. AP rams (2.50 ± 0.42) exhibited significantly (p < .05) more serves than did NP rams (1.11 ± 0.39), and melatonin treatment had no significant effect; however, the interaction between treatments was significant. Time to first serve was significantly (p < .05) shorter in AP (2.30 ± 1.20 min) than it was in NP rams (5.58 ± 0.68 min). In conclusion, exposure to 2 months of long days in late winter/early spring, with a return to natural day length of shorter duration, increased plasma testosterone concentrations and sexual performance in rams with or without exogenous melatonin. This particular management is an option if a non‐hormonal reproductive strategy is scheduled; yet, if the use of exogenous hormones is feasible, melatonin implants increase the mating efficiency of rams.     

m‐carboxycinnamic acid bishydroxamide improves developmental competence,reduces apoptosis and alters epigenetic status and gene expression pattern in cloned buffalo (Bubalus bubalis) embryos

Incomplete or aberrant reprogramming of nuclear genome is one of the major problems in somatic cell nuclear transfer. In this study, we studied the effect of histone deacetylase inhibitor m‐carboxycinnamic acid bishydroxamide (CBHA) on in vitro development of buffalo embryos produced by Hand‐made cloning. Cloned embryos were treated with CBHA (0, 5, 10, 20 or 50 μM) for 10 hr from the start of reconstruction till activation. At 10 μM, but not at other concentrations examined, CBHA increased (p < .05) the blastocyst rate (63.77 ± 3.97% vs 48.63 ± 3.55%) and reduced (p < .05) the apoptotic index of the cloned blastocysts (8.91 ± 1.94 vs 4.36 ± 1.08) compared to untreated controls, to levels similar to those in IVF blastocysts (4.78 ± 0.74). CBHA treatment, at all the concentrations examined, increased (p < .05) the global level of H3K9ac in cloned blastocysts than in untreated controls to that observed in IVF blastocysts. Treatment with CBHA (10 μM) decreased (p < .05) the global level of H3K27me3 in cloned blastocysts than in untreated controls but it was still higher (p < .05) than in IVF blastocysts. CBHA (10 μM) treatment increased (p < .05) the relative expression level of pluripotency‐related genes OCT‐4 and NANOG, and anti‐apoptotic gene BCL‐XL, and decreased (p < .05) that of pro‐apoptotic gene BAX than in untreated controls but did not affect the relative expression level of apoptosis‐related genes p53 and CASPASE3 and epigenetics‐related genes DNMT1, DNMT3a and HDAC1. These results suggest that treatment of cloned embryos with 10 μM CBHA improves the blastocyst rate, reduces the level of apoptosis and alters the epigenetic status and gene expression pattern.     

Associations between endotoxin‐induced metabolic changes and temperament in Brahman bulls,

The influence of temperament on the alteration of metabolic parameters in response to a lipopolysaccharide (LPS) challenge was investigated. Brahman bulls were selected based on temperament score. Bulls (10 months; 211 ± 5 kg BW; n = 6, 8 and 7 for Calm, Intermediate and Temperamental groups, respectively) were fitted with indwelling jugular catheters to evaluate peripheral blood concentrations of glucose, blood urea nitrogen (BUN), non‐esterified fatty acids (NEFA), insulin, epinephrine and cortisol before and after LPS administration (0.5 μg/kg BW LPS). Feed intake was also recorded. Intermediate bulls consumed more feed than the Temperamental bulls during the challenge (p = 0.046). Pre‐LPS glucose (p = 0.401) and BUN (p = 0.222) did not differ among the temperament groups. However, pre‐LPS insulin (p = 0.023) was lower, whereas pre‐LPS NEFA (p < 0.001), cortisol (p < 0.001) and epinephrine (p < 0.001) were greater in Temperamental than in Calm and Intermediate bulls. Post‐LPS glucose was increased in Calm and Intermediate bulls but not in Temperamental bulls (p < 0.001). Insulin concentrations post‐LPS were greater in Calm than in Intermediate and Temperamental bulls (p < 0.001). Concentrations of NEFA post‐LPS were greater in Temperamental than in Calm and Intermediate bulls (p < 0.001). Serum BUN concentration increased post‐LPS, with values being greater in Calm and Intermediate than in Temperamental bulls (p = 0.012). Collectively, these data demonstrate that animal temperament is related to the metabolic responses of Brahman bulls following a provocative endotoxin challenge. Specifically, Temperamental bulls may preferentially utilize an alternate energy source (i.e. NEFA) to a greater degree than do bulls of Calm and Intermediate temperaments. The use of circulating NEFA from lipolysis may reduce the negative metabolic consequences of an immune response by allowing for a prompt answer to increasing energy demands required during immunological challenge, compared with the time required for glycogenolysis and gluconeogenesis.     

Correlation of testicular artery Doppler velocimetry with kinetics and morphologic characteristics of epididymal sperm in dogs

Sperm quality can be affected by a reduction in testicular blood flow, which can be measured by Doppler ultrasonography. The aim of this study was to correlate the Doppler velocimetry of the testicular artery with kinetics of the epididymal spermatozoa in dogs. Twenty-two dogs (44 testicles) were evaluated by Doppler ultrasonography in five regions of the testicular artery before orchiectomy. Spermatozoa were recovered by the epididymal tail compression technique and analysed for kinetics on a computer-assisted semen analysis (CASA system). Morphology (modified Karras) and sperm membrane integrity were analysed by eosin–nigrosine staining. Data were analysed by Pearson's correlation test (p < .01). The mean total motility was 69.0% ± 17.7, progressive motility was 43.7% ± 14.7, average path velocity (VAP) was 127.0 µm/s ± 20.7, curvilinear velocity (VCL) was 221.0 µm/s ± 31.1, and sperm velocity index (SVI) was 389.9 ± 56.1. There were positive correlations between the peak systolic velocity (PSV) in the proximal supratesticular region with the SVI (r = .529), VCL (r = .555) and VAP (r = .473), and a negative correlation with the percentage of slow spermatozoa (r = −.463). The results suggest that the testicular artery blood flow velocity can positively affect the speed of spermatozoa movement. For the first time, we have correlated sperm kinetics with the Doppler evaluation of the testicular artery in dogs.     

Effects of chlorogenic acid-enriched extract from Eucommia ulmoides Oliver leaf on growth performance and quality and oxidative status of meat in finishing pigs fed diets containing fresh or oxidized corn oil

To investigate the effects of chlorogenic acid-enriched extract (CGAE) from Eucommia ulmoides Oliver leaf on growth performance and quality and oxidative status of meat in pigs fed diets containing fresh or oxidized corn oil, a total of 180 barrows (initial body weight: 81.6 ± 2.08 kg) were randomly allocated into 6 diet treatments (5 replicate pens per treatment and 6 barrows per pen) in a 2 × 3 factorial design with corn oil (fresh or oxidized corn oil at 5% inclusion of diet) and CGAE (0, 500 or 1,000 mg/kg of diet containing fresh or oxidized corn oil) as main factors. The experiment lasted for 6 weeks. Dietary oxidized oil reduced average daily gain (ADG, p < .05) and average daily feed intake (ADFI, p < .01) of pigs and pH₂₄ (p < .05), total antioxidant capacity (T-AOC, p < .01), glutathione peroxidase (GPx, p < .05) and sarcoplasmic reticulum Ca²⁺-ATPase (SERCA, p < .05) activities in meat and increased drip loss (p < .01), cooking loss (p < .05), malondialdehyde (p < .01) and carbonyl (p < .01) contents and mRNA expression of superoxide dismutase 1 (SOD1, p < .05) in meat. Dietary CGAE supplementation at 1,000 mg/kg increased (p < .05) ADG and ADFI of pigs and pH₂₄, T-AOC, T-SOD, GPx and SERCA activities and mRNA expression of SOD1 in meat and reduced (p < .05) drip loss, cooking loss, carbonyl and malondialdehyde contents in meat. No interaction effects between oxidized corn oil and CGAE were found in pigs. Overall, dietary CGAE supplementation at 1,000 mg/kg improved growth performance and quality and oxidative status of meat in pigs subjected or not to oxidative stress induced by dietary oxidized oil.     

The effect of astaxanthin supplementation on the post-thaw quality of dog semen

Astaxanthin is a member of the carotenoid family well known for its anti-cancer, anti-diabetic, anti-inflammatory and antioxidant nature. This study was designed to investigate the effects of astaxanthin supplementation of the extender (buffer 2) on post-thaw dog semen quality. Semen from four healthy dogs was collected by digital manipulation twice a week. The ejaculates were pooled, washed, divided into four equal aliquots, diluted with the extender supplemented with different concentrations of astaxanthin (0, 0.5, 1 and 2 µM) and cryopreserved. The results showed that 1 µM astaxanthin was the optimum concentration that led to significantly higher (p < .05) post-thaw motility, kinematic parameters and viability than the other groups. In comparison with the control group, sperm samples supplemented with 1 µM astaxanthin showed significantly higher (p < .05) sperm counts with intact membranes (55.7 ± 0.6% vs. 51.3 ± 0.9%), intact acrosome (58.4 ± 0.7% vs. 53.5 ± 0.6%), active mitochondria (54.9 ± 0.5% vs. 42.6 ± 0.6%) and normal chromatin (67.6 ± 0.9% vs. 61.7 ± 0.6%). Furthermore, astaxanthin-supplemented samples showed significantly lower expression levels (p < .05) of pro-apoptotic (BAX), oxidative induced DNA damage repair (OGG1), oxidative stress-related (ROMO1) genes and higher expression levels of anti-apoptotic (BCL2), and sperm acrosome-associated (SPACA3) genes compared to the control. Thus, supplementation of 1 µM astaxanthin in semen extender results in improved freeze-thaw sperm quality of the dog.     

Relationship between angioarchitecture of the testicular artery and spermiogram parameters in Egyptian buffalo bulls (bubalus bubalis)

The current study aimed to investigate the effect of testicular artery angioarchitecture on the spermiogram parameters in Egyptian Buffalo bulls. Eight adult buffalo bulls aged between 2 and 8 years were used for semen evaluation. For anatomical studies, the masculine gonads were collected after slaughtering 30 adult bulls and prepared for injection by different masses (Urographine^®, Latex and Epoxy) through the testicular artery. The mass activity of the ejaculate was assessed immediately after collection. The sperm motility in fresh bull ejaculate was more than 80%. The overall mean percentage of sperm abnormalities was <18%. The recorded sperm abnormalities were mostly secondary one including distal protoplasmic droplet, fragmented tail, detached head, detached galea capitis and bent tail. The highest percentage of sperm viability was recorded just after sperm collection (alive > 85%). The results revealed that testicular artery can be divided into three parts (abdominal, funicular and marginal parts) along its course. The coils of the funicular part forming a cone-like structure with its base fixed to the head of the testis. Two epididymal branches to the head and tail of epididymis emanate from the funicular part which continues as pars marginalis on the lateral surface of testis before its division into the lateral and medial testicular arteries on approaching the tail extremity of the testis. The increase in the length of the testicular artery with increase in the size of the testes played a great role in the degree of complexity of the architectural vascular patterns. The degree of complexity is affected by the number of coils formed by the vessel. The increase in the convolutions of the vessel will reduce the speed of blood flow to the gonads. Thus in turn will enabling the thermoregulatory mechanism to work more efficiently and will affect the semen value.     

Successful transcervical uterine flushing can be performed without or reduced dose of oestradiol benzoate in cervical relaxation protocol in Dorper ewes

This study assessed the efficiency of cervical relaxation protocol using none, half or full dose (1.0 mg) of oestradiol benzoate in Dorper ewes subjected to non-surgical embryo recovery (NSER). Thirty-six pluriparous ewes received progestogen sponge (60 mg) for 9 days plus eCG administration (300 IU i.m.) 24 hr before sponge removal. Ewes were not mated and were randomly assigned to receive at 16 hr before NSER 37.5 µg d-cloprostenol i.m. and different doses of oestradiol benzoate: 0.0 mg (0EB group; n = 12); 0.5 mg (0.5EB group; n = 12) or 1.0 mg of oestradiol (1.0EB group, n = 12). All ewes received oxytocin (50 IU) i.v. 20 min before NSER, which was performed 8 days after sponge removal. Corpora lutea were counted by transrectal ultrasonography 24 hr before NSER. After procedure, the ewes were kept in natural breeding period to check their post-NSER fertility. NSER was performed in 91.7% (33/36) of the animals with overall fluid recovery efficiency over 97% (p > .05). The cervical transposing with Hegar dilator was longer (p < .05) in 0EB (4.2 ± 0.3 min) compared to 0.5EB (1.7 ± 0.3 min) and 1.0EB group (1.5 ± 0.3 min). The cervical transposing with mandrel/catheter was longer (p < .05) in 0EB (2.4 ± 0.5 min) than 1.0EB group (1.3 ± 0.5 min). Overall duration of uterine flushing was 25.4 min with structure recovery rate of 43.5%, with no difference among groups (p > .05). The post-NSER fertility was higher (p < .05) in 0.0EB (90%) than 0.5EB group (36.4%). In conclusion, NSER can be successfully performed in Dorper ewes by using a cervical relaxation protocol without oestradiol benzoate.     

Morphology and quantification of sheep pineal glands at pre‐pubertal,pubertal and post‐pubertal periods

The pineal gland is a neuroendocrine organ associated with photoperiodic regulation in mammals. The aim of this study was to evaluate the pineal gland at the pre‐pubertal, pubertal and post‐pubertal periods by means of morphology and stereology. The study examined at total of 24 ovine pineal glands collected from healthy female Akkaraman breed. Thick sections (40 μm) were cut and treated with synaptophysin. Following each thick section, six consecutive sections at a thickness of 5 μm were cut. Each thin section was stained with one of the following dyes: Crossman’s modified triple dye, glial fibrillary acidic protein (GFAP), melatonin marker, periodic acid–Schiff, Von Kossa and AgNOR. The pineal gland volume was measured using Cavalieri’s method. The optical fractionator was used to estimate the total number of pinealocytes. The percentage of parenchyma and connective tissue and degree of vascularization were estimated by the area fraction fractionator method. The pineal gland volumes in the pre‐pubertal, pubertal and post‐pubertal groups were 7.53 ± 1.715 mm³, 11.20 ± 1.336 mm³ and 17.75 ± 1.188 mm³, respectively (p < .5). The number of pinealocytes in the pre‐pubertal, pubertal and post‐pubertal groups was 3,244,000 ± 228,076, 4,438,000 ± 243,610, 7,381,766 ± 406,223, respectively (p < .05). The glands of the post‐pubertal group contained the highest amount of connective tissue (11.49 ± 2.103%; p < .5) and the largest GFAP staining area (p < .05). The melatonin staining density was the highest in the pubertal group. The density of lipofuscin staining was higher in the pubertal and post‐pubertal groups.     

Influence of Bacillus subtilis GCB‐13‐001 on growth performance,nutrient digestibility,blood characteristics,faecal microbiota and faecal score in weanling pigs

The present study investigated the influence of Bacillus subtilis GCB‐13‐001 on growth performance, nutrient digestibility, blood characteristics, faecal microbiota and faecal score in weanling pigs. A total of 120 weaning pigs [(Landrace × Yorkshire) × Duroc; 7.73 ± 0.75 kg (28 days of age)] were randomly allotted into three treatments according to their initial body weight (BW) and gender in a 6‐week experiment. There were 8 replication pens in each treatment, with five pigs/pen. Dietary treatment groups were as follows: (a) basal diet (CON), (b) CON + 0.1% Bacillus subtilis GCB‐13‐001 1 × 10⁸ CFU/kg (T1) and (c) CON + 0.1% Bacillus subtilis GCB‐13‐001 1 × 10⁹ CFU/kg (T2). Days 1 to 7, the BW and ADG with T2 treatment were higher (p < .05) than CON treatment, as well as F:G showed trends in linear reduction (p < .1). Days 8 to 21, the BW and ADG were improved (p < .05) in pigs offered T1 and T2 diets compared with CON diet. Days 22 to 42, BW and ADG were higher (p < .05) in pigs fed T2 diet than CON and T1 diets, and the pigs fed T1 diet had higher BW than CON treatment. Overall, the ADG with the T2 treatment was higher (p < .05) than that with the T1 and CON treatments, and pigs offered T1 treatment had higher (p < .05) ADG than CON treatment. Moreover, F:G ratio were significantly decreased (p < .05) by T2 treatment compared with CON treatment. The faecal Lactobacillus counts were improved, and E. coli counts were reduced (p < .05) in pigs fed T2 diet compared with CON at the end of the experiment. In conclusion, supplementation of 0.1% Bacillus subtilis GCB‐13‐001 1 × 10⁹ CFU/kg has shown a beneficial effect in improving BW, increase ADG, decrease F:G ratio.