Impact of Ultra‐Rapid Freezing on the Motility,Morphology, Viability and Acrosome Integrity of Epididymal Cat Sperm Diluted in Sucrose‐Based Extenders

The objective of the present study was to investigate the influence of different sucrose‐based extenders on the motility, morphology, viability and acrosomal integrity of epididymal cat spermatozoa cryopreserved by ultra‐rapid freezing method. Nine cats were castrated, and collected semen was diluted 1 : 1 with Dulbecco`s phosphate‐buffered saline‐BSA1%‐based extender supplemented with different sucrose concentrations (0, 0.25, 0.4 and 0.6 m ). After ultra‐rapid freezing, samples were thawed and sperm motility, morphology, viability and acrosome status were assessed. At thawing, the number of progressively motile (p < 0.01) and morphologically normal (p < 0.01) sperm was higher in the sucrose‐supplemented groups than in the sucrose‐free group. Viability of spermatozoa cryopreserved without sucrose was significantly reduced. In extender supplemented with 0.4 m sucrose, spermatozoa viability showed higher values (57.0 ± 4.7; p < 0.01). No significant differences were detected among groups for sperm acrosome integrity. Results support that cat sperm survive after ultra‐rapid freezing using sucrose as a cryoprotectant, and the best results were achieved when 0.4 m of sucrose was used. This is the first report on sperm ultra‐rapid freezing of cat sperm and further studies on extenders, sperm management or cryovials should be carried out to improve sperm cryosurvival.     

Morphometry and abnormalities of the feet of Kaimanawa feral horses in New Zealand

Objective The present study investigated the foot health of the Kaimanawa feral horse population and tested the hypotheses that horses would have a large range of foot morphology and that the incidence of foot abnormality would be significantly high. Procedures Abnormality was defined as a variation from what the two veterinarian assessors considered as optimal morphology and which was considered to impact negatively on the structure and/or function of the foot. Fifteen morphometric variables were measured on four calibrated photographic views of all four feet of 20 adult Kaimanawa feral horses. Four morphometric variables were measured from the lateromedial radiographs of the left forefoot of each horse. In addition, the study identified the incidence of gross abnormality observed on the photographs and radiographs of all 80 feet. Results There was a large variation between horses in the morphometric dimensions, indicating an inconsistent foot type. Mean hoof variables were outside the normal range recommended by veterinarians and hoof care providers; 35% of all feet had a long toe conformation and 15% had a mediolateral imbalance. Abnormalities included lateral (85% of horses) and dorsal (90% of horses) wall flares, presence of laminar rings (80% of horses) and bull-nose tip of the distal phalanx (75% of horses). Both hypotheses were therefore accepted. Conclusions The Kaimanawa feral horse population demonstrated a broad range of foot abnormalities and we propose that one reason for the questionable foot health and conformation is lack of abrasive wearing by the environment. In comparison with other feral horse populations in Australia and America there may be less pressure on the natural selection of the foot of the Kaimanawa horses by the forgiving environment of the Kaimanawa Ranges. Contrary to popular belief, the feral horse foot type should not be used as an ideal model for the domestic horse foot.     

A prospective study on fitness,workload and reasons for premature training ends and temporary training breaks in two groups of riding horses

Little is known about wastage in riding horses and the factors like fitness and workload that may reduce injuries and maximise welfare. To evaluate fitness, workload and reasons for premature training ends (PTEs) and temporary training breaks (TTBs) during a nine week training period, two groups of riding horses were used: Group A consisting of 58 horses used for student equitation courses (32 with training prior to admission and 26 without) and Group B consisting of 26 horses owned by two riding schools (school-I and school-II). To assess fitness, all horses performed a standardised exercise test (SET) at the start (SET-I) and end of the training period (SET-II) measuring heart rate (HR bpm) and speed (m/s). In addition, all horses were monitored daily during the training period for their health and workload.In Group A, trained horses had significantly lower HRs in SET-I (P = 0.05) compared to untrained horses and in SET-II, trained horses tended to have lower HRs than untrained horses, though this was not statistically significant (P = 0.057). During the training period all horses received an identical workload. A total of 19.0% of Group A horses ended the training period prematurely for veterinary reasons (PTEV); of those untrained horses had earlier a PTEV in the training period (after 2.8 ± 1.3 weeks) than trained horses (after 4.1 ± 1.5 weeks, P = 0.030).In Group B, school-I and school-II horses did not differ significantly in fitness level nor in workload. More school-II horses ended the training period prematurely for veterinary reasons (n = 7; 70%) compared to school-I horses (n = 4; 25%, P = 0.032), although seven (63.6%) of these horses were still continuously used in riding lessons. In both groups (A and B), small injuries (without a temporary training break) were significantly associated with premature training ends for veterinary reasons later on: in Group A small injuries preceded 27.3% of the PTEVs (P = 0.005) and in Group B small injuries preceded 54.5% of the PTEVs (P = 0.030).In conclusion, as all horses in each subgroup had the same workload, the occurrence of PTEV seemed not associated with the workload. In Group A horses, level of fitness seems to be an important factor for the point in time injuries will occur during the training period. In all horses, injuries were more likely when a temporary training break was not taken following seemingly minor injuries. Since a lot of injured Group B horses were used in riding lessons against veterinary advice, this may indicate that riding school owners have different perception on welfare and if true this may cause serious welfare problems.     

Social and Breed Effects on the Expression of a PGF2α Induced Oestrus in Beef Cows

Social organization and breed effects following PGF2αwere studied in mature Angus, Brahman and Senepol cows allocated into two groups (each A = 5, B = 5 and S = 5). Variables including interval to oestrus onset (IEO), oestrous duration (DE), total mounts received (TMR), and oestrous intensity (IE) were derived via HeatWatch®. Breed‐type influenced IEO (B = 42.6 ± 6.7 h; S = 54.6 ± 6.0 h; and A = 27.8 ± 5.8 h; p < 0.003). Within breeds, dominant B (69.4 ± 13.3 h) and S (65.5 ± 7.4 h) cows were slower (p < 0.05) to be detected in oestrus than subordinate (38.1 ± 4.4 h) and intermediate (40.6 ± 6.0 h). However, within A, dominant cows (16.4 ± 12.5 h) were detected in oestrus earlier (p < 0.05) than intermediate (44.3 ± 9.2 h) and subordinates (32.7 ± 5.1 h). Angus (21.5 ± 2.4 h) and B (22.1 ± 3.0 h) cows had longer (p < 0.01) DE than S (9.1 ± 2.8 h). Dominants (20.4 ± 3.0) and intermediates (20.2 ± 2.3 h) cows had longer DE (p < 0.04) than subordinates (12.1 ± 2.1 h) although the interaction breed × social order showed that dominant S had shorter DE than dominant A and B (10.1 ± 3.3; 34.8 ± 6.0 h; and 20.0 ± 6.4 h, respectively; p < 0.001). Angus cows had less TMR than B (p < 0.02) and tended to be less than S cows (p < 0.06). Overall, greatest (p < 0.008) IE occurred in the first 9 h after onset of oestrus with no breed effect (p > 0.05). Dominant cows tended (p < 0.10) to have less TMR (3.2 ± 0.7 mounts) than subordinate (4.1 ± 0.4 mounts) and intermediate (4.7 ± 0.6 mounts) throughout, especially 3–6 h after oestrus onset (p < 0.07). Breed and social order both influence PGF2α‐induced oestrus behaviour.     

Fibrin–alginate hydrogel supports steroidogenesis,in vitro maturation of oocytes and parthenotes production from caprine preantral follicles cultured in group

This study aimed to establish a culture system that improves the in vitro development of caprine preantral follicles. In a first experiment, follicles were encapsulated as a single unit per bead and cultured singly or in groups or with five follicles in the same alginate (ALG) bead for 18 days. In a subsequent experiment, the “five follicles per bead” design was chosen to culture in ALG, fibrin–alginate (FA) or hyaluronate (HA) for 18 days. In a third experiment, we chose the five follicles per bead in FA to culture for 30 days. The culture set‐up of five follicles per ALG bead increased antrum formation and follicle diameter compared to the other culture designs (p < .05). Moreover, under this condition, 44.44% of the oocytes from in vitro cultured preantral follicles reached meiotic resumption. A significant increase of follicle diameter occurred in attachment system and FA (p < .05), but the ALG condition reached the highest among all groups on day 18 (p < .05). Follicles encapsulated in matrix produced more estradiol and progesterone than attachment system (p < .05). The expression of MMP‐9 mRNA was higher in FA than in other groups (p < .05) and similar to antral follicles from in vivo control (p > .05). Only FA group resulted in oocytes matured. After 30 days, oocytes from preantral follicles in vitro grown in FA developed to eight‐cell parthenotes. In conclusion, a culture system using FA supported the development of caprine preantral follicles cultured in group and included in the same bead of hydrogel, improving the oocyte maturation and producing parthenotes.     

The effects of allopurinol on the ultrastructure of ischaemic and reperfused large intestine of sheep

Objective To test the possible inhibitory effect of allopurinol on reperfusion injury, caused by oxygen-derived free radicals, of sheep large intestine.
Design An ultrastructural study on caecal tissues from control and treated groups.  

Animals

Fifty sheep in four ischaemic and reperfused (treatment) groups and one control group. Three of the treatment groups were subdivided for half to be injected with allopurinol and the other half with its solvent, potassium hydroxide (KOH).
Procedure Ischaemia of the caecum was induced in the four treatment groups for 60 minutes by clamping the apex. Allopurinol and its KOH solvent were injected intravenously in three treatment groups prior to ischaemia. Samples were collected before and 1 hour after induction of ischaemia and 1 min, 1 h and 8 h after reperfusion. Tissues were processed and examined with an electron microscope.
Results Untreated and solvent injected sheep showed minor ultrastructural changes following ischaemia. With reperfusion, there was severe mitochondrial, goblet cell and basement membrane damage. Tissues from allopurinoltreated sheep were preserved and appeared similar to tissues from the control group.  

Conclusion:

Pre-treatment with allopurinol prevented damage to tissues whereas untreated or allopurinol solventtreated showed severe damage following reperfusion. It is believed that allopurinol, an analogue of hypoxanthine and xanthine, prevents reperfusion injury by competitively binding with xanthine oxidase. This reduces or inhibits the xanthine oxidase mediated conversion of hypoxanthine to xanthine thereby preventing the formation of oxygen-derived free radicals.     

Effects of supplemental dietary vitamin C on quality of semen from Nile tilapia (Oreochromis niloticus) breeders

The objective of this study was to evaluate the effects of vitamin C on growth and quality of semen from Oreochromis niloticus breeders. One hundred and sixty males were fed with different levels of vitamin C (0, 261, 599 and 942 mg/kg diet). The higher weight values were recorded for 599 (166 g) and 942 (175 g) mg of vitamin C/kg diet. Sperm motility, vigour and concentration were higher with 599 and 942 mg of vitamin C/kg diet. The semen volume, gonadosomatic index and plasma protein data from the last week showed a direct relationship with increasing levels of vitamin C. No changes were observed in the hepatosomatic index and blood glucose. The haematocrit and erythrocyte showed higher values estimated by equations derived at 850 and 638 mg vitamin C/kg diet, respectively. The leucocytes were inversely proportional to the increasing levels of vitamin C. After 100 days of feeding, animals fed the diet containing 942 mg vitamin C/kg diet had higher sperm motility, linearity, curvilinear velocity, straight line velocity and average path velocity (p < .05). Higher values of beat cross‐frequency were observed in broodfish fed diets containing 942 and 599 mg vitamin C/kg. The different vitamin C levels did not cause differences in straightness, lateral head displacement and sperm morphology. For Nile tilapia males on intensive rearing and handling conditions, vitamin C levels between 599 and 942 mg/kg may be used for a better performance and quality of semen.     

Chemical Activation with a Combination of Ionomycin and Dehydroleucodine for Production of Parthenogenetic,ICSI and Cloned Bovine Embryos

The aim of this study was to evaluate the potential of dehydroleucodine (DhL), a new drug isolated from a medicinal herb used in Argentina, for activation of bovine oocyte. Several DhL concentrations and exposure times after ionomycin (Io) treatment were tested. The optimal DhL treatment, found for parthenogenetic development, was employed to produce bovine embryos by intracytoplasmic sperm injection (ICSI) and somatic cell nuclear transfer (SCNT). The best parthenogenic embryo developments were observed with 5 μm Io for 4 min followed by 5 μm DhL concentration and after 3‐h exposure time (52.3% cleavage; 17.4% morulae; 7.3% blastocyst; n = 109). This treatment generated no significant differences with standard Io plus 6‐dimethylaminopurine (DMAP) treatment in preimplantation embryo development. In our conditions, the embryo development reached after ICSI and SCNT assisted by the DhL treatment did not differ in terms of cleavage and blastocyst development from activation with standard Io plus DMAP treatment (p > 0.05). In conclusion, DhL utilization to activate oocytes and induce development of parthenogenotes, ICSI‐embryos or SCNT‐embryos is reported here for first time.