In vitro Maturation of Oocytes from Santa Ines Ewes Subjected to Consecutive Sessions of Follicular Aspiration by Laparoscopy

The success of embryo production in vitro depends upon the use of an efficient oocyte retrieval technique, and the best results have been obtained by laparoscopic aspiration. The aim of this study was to evaluate the effect of consecutive sessions of follicular aspiration on the quantity, quality and in vitro maturation competence of oocytes obtained from ewes subjected to hormonal stimulation. Six Santa Ines ewes underwent nine sessions of follicular aspiration by laparoscopy with a 7‐day interval between sessions, totalling 56 aspirations. After 24 h of culture, oocytes were stained and classified according to the stage of nuclear and cytoplasmic maturation. Oocyte retrieval rate was 61.4 ± 2%, resulting in a total of 249 oocytes. No significant variation was observed between sessions (p > 0.05). The average number of oocytes retrieved from each ewe was 6.4 ± 2 per session and 42 ± 4 in total. No significant difference was observed between the frequencies of the different stages of nuclear maturation: 32.72% mature, 40.74% immature and 26.54% degenerated/indeterminate oocytes; however, a significant difference was observed between the frequencies of the different stages of cytoplasmic maturation: 10.7% mature, 73.25% immature and 16.05% degenerated/indeterminate oocytes. No significant difference was observed in nuclear or cytoplasmic maturation between the weeks of procedure. We conclude that after nine consecutive sessions of follicular aspiration, the quantity and quality of retrieved oocytes remained unchanged as well as the levels of nuclear and cytoplasmic maturation obtained, demonstrating the viability of this technique for repetitive follicular aspirations on the same donor.     

Analgesia for pelvic limb surgery. A review of peripheral nerve blocks and the extradural technique

ObjectivesTo describe the anatomy and approaches reported for peripheral nerve blockade (PNB) of the pelvic limb in dogs and cats and to consider the role of PNB in relation to the extradural technique.Databases usedThis review was conducted using the terms ‘nerve block’, ‘extradural’ ‘dog’ and ‘cat’ entered into Pubmed and Google. Results were filtered manually to narrow the field to pelvic limb nerve blocks. The reference lists of retrieved papers were scrutinized to identify further studies for inclusion.ConclusionsSuccessful PNB techniques require thorough anatomical knowledge for the establishment of reliable landmarks, puncture sites, the direction and depth of needle insertion, and relevant structures to be avoided. To date, clinical evaluations have been made in subjects undergoing stifle surgery where the sciatic nerve has been blocked in combination with various approaches to the femoral nerve. Currently the bulk of literature examines new approaches to these nerves and each of these is described. To date there are no veterinary studies directly comparing one approach versus another, and therefore one is unable to draw conclusions of superiority. The role of PNB’s versus the extradural technique is discussed.     

Motility Characteristics and Fertilizing Capacity of Boar Spermatozoa Stained with Hoechst 33342

Flow cytometry sorting of spermatozoa using fluorescence dye Hoechst 33342 is the only effective sex selection methodology validated in numerous laboratories. This study was carried out to determine the effect of Hoechst 33342 on the motility and fertility of stained boar spermatozoa. Experiment 1 evaluated motility parameters (percentage of motile spermatozoa, velocity, angularity and oscillation) of boar spermatozoa stained with Hoechst 33342 by a computer‐aided sperm analysis (CASA) instrument. Spermatozoa (30 million/ml) were divided into five treatment groups and stained during 1 h at 35°C with 9, 18, 27, 60 and 90 μM of H33342. There were no differences in sperm motility patterns nor percentages of motile spermatozoa incubated in the presence of 9, 18 or 27 μM. Percentage of motile spermatozoa and motility parameters decreased significantly (p < 0.05) at 60 μM of Hoechst 33342. Spermatozoa were immotile at concentration of 90 μM. In experiment 2, pregnancy rates, farrowing rates and litter size from sows (n = 275) artificially inseminated (AI) with either Hoechst 33342 stained (27 μM) or unstained (control) spermatozoa were determined. Sows inseminated with stained spermatozoa had no significant lower pregnancy rate (88.33%) as compared with controls (90.32%). Staining neither affected farrowing rates (85.0 vs 87.7%) nor total number of piglets born (10.56 ± 0.32 vs 10.47 ± 0.24, stained and controls, respectively). No phenotypical abnormalities were registered among the newborn piglets. The data suggest that incubating spermatozoa with Hoechst 33342 at levels required for X‐ and Y‐bearing chromosome sperm sorting, does not impair sperm viability or their fertility after AI.     

The Effect of Two Antibiotic Mixtures on Haemophilus somnus, Campylobacter fetus ssp. venerealis, Mycoplasma bovis, and Ureaplasma diversum in Frozen Bovine Semen

A two-step semen-extending protocol was compared to a one-step protocol in its efficiency in inhibiting growth of Haemophilus somnus, Campylobacter fetus ssp. venerealis, Mycoplasma bovis , and Ureaplasma diversum in experimentally infected semen. In both protocols, the effect of an antibiotic mixture of 500 μg gentamycin, 100 μg tylosin, 300 μg lincomycin, and 600 μg spectinomycin (GTLS) was compared to a mixture of 500 IU penicillin, 500 IU streptomycin, 150 μg lincomycin, and 300 μg spectinomycin (PSLS). The one-step extending method was as effective as the two-step extending method. Both antibiotic mixtures were equally effective in controlling C. fetus . For H. somnus and U. diversum , the PSLS mixture was more effective than the GTLS mixture. It was striking that both antibiotic mixtures had no effect in decreasing the numbers of M. bovis .     

Muscle strain injuries of the hindlimb in eight horses: diagnostic imaging,management and outcomes

Objective To describe the clinical presentation, ultrasound findings, management and outcome in horses with muscle tear injuries of the hindlimbs. Design Retrospective case series Procedure Medical records of eight horses were reviewed and information on signalment, history, presenting complaint, physical examination findings and further diagnostic tests were recorded. Diagnosis of muscle injury was determined by the presence of abnormal ultrasound findings, compared with the contralateral limb, and, when required, nuclear scintigraphy. Follow-up information was obtained via telephone interviews with owners, trainers and referring veterinarians. Results Muscle tears causing lameness were identified in the middle gluteal (3), semitendinosus (1), semimembranosus (2) and gracilis (2) muscles. Tears were classified by ultrasound imaging as partial (6) or complete (2). The degree of lameness did not appear to be indicative of the extent of injury or of completion of healing. Long-term follow-up was available for seven horses and the outcome was favourable in six cases. Conclusions and clinical relevance Ultrasonography is useful in the diagnosis and assessment of moderate to severe muscle strain injuries. The prognosis appears to be favourable in most cases, although recurrence of injury and lameness can delay the return to athletic activity and an inferior outcome with persistent gait abnormality may occur.     

Post anaesthetic myopathy/neuropathy in horses undergoing magnetic resonance imaging compared to horses undergoing surgery

REASONS FOR PERFORMING STUDY: Patient positioning and long anaesthetic duration required for magnetic resonance imaging (MRI) may result in a higher frequency of post anaesthetic myopathy/neuropathy syndrome (PAMNS) as compared to horses undergoing anaesthesia for surgery. HYPOTHESIS: Equine anaesthesia for MRI is associated with a higher frequency of PAMNS than anaesthesia for nonemergency, nonabdominal surgery. METHODS: Anaesthetic and medical records of horses (n = 633) undergoing MRI or surgery between January 2001 and January 2005 (inclusive), were reviewed. Information obtained included patient details (breed, sex, age, bodyweight), area of body scanned or involved in surgery, body position, anaesthetic and inotropic agents administered, anaesthetic duration, adverse events during anaesthesia and outcome at 7 days. Data were examined by cross tabulation and Chi-square or Fisher's exact test of association. The influence of individual variables was examined by univariant and multivariant analysis models. RESULTS: There were no statistically significant differences between the 2 groups in parameters examined, except that horses in the MRI group were heavier (P<0.0001) and anaesthetic duration longer in the surgery group (P<0.004). Eight horses (2.3%, 95% Confidence interval [CI]: 1.1-4.2) in the MRI group had clinical signs of PAMNS in the post anaesthetic period, whereas only 2 horses (0.98%, 95% CI: 0.2-2.8) in the surgery group were affected. This was not statistically significantly different (odds ratio = 2.7, 95% CI: 0.8-13, P = 0.3). Two horses undergoing MRI were subjected to euthanasia due to the severity of PAMNS. CONCLUSIONS: There was no difference in the occurrence of PAMNS between the 2 groups. POTENTIAL RELEVANCE: The risk of performing general anaesthesia for diagnostic procedures such as MRI may not be greater than that for a surgical procedure. However, the benefits should be carefully weighed against the risks involved.     

Effect of Oxytocin Treatment on Artificial Insemination with Frozen–Thawed Semen in Murciano–Granadina Goats

The site where the semen is deposited appears to be one of the most important factors affecting pregnancy of inseminated goats. In Murciano–Granadina (MG) goats, post-cervical insemination is achieved in a limited number of females. An effective way to increase fertility rate could be by increasing post-cervical inseminations. Effect of exogenous oxytocin application to facilitate the cervical penetration and its effect on kidding rate and prolificacy in MG goats were investigated. Oestrus was synchronized using progesterone-impregnated sponges for 11 days. Females were randomly divided into three groups (n = 190) and received either an i.v. injection of 100 or 200 IU of oxytocin or saline solution 15 min before being inseminated. Data on semen deposition depth were recorded for each animal using a catheter scaled in centimetres (up to 4 cm). Depth of semen deposition was affected by the oxytocin treatment (p < 0.05). Oxytocin enhanced cervical passage only with the dose of 200 IU compared with the control group, increasing the deposition depth (2.9 cm vs 1.9 cm). No significant effect of oxytocin treatment on kidding rate and prolificacy was detected. Depth of semen deposition affected kidding rate (p < 0.01). In conclusion, oxytocin treatment improved the depth of semen deposition in AI of MG goats, but kidding rate and prolificacy was not affected. More studies must be conducted to assess the minimal effective dose required for sufficient cervical dilation, and to determine the effects of such doses of oxytocin on uterine motility, sperm transport and fertility in goats.