Effect of N‐acetyl‐L‐cysteine Supplementation in Semen Extenders on Semen Quality and Reactive Oxygen Species of Chilled Canine Spermatozoa

The objective of this study was to evaluate the quality of chilled dog semen processed with extenders containing various concentrations of N‐acetyl‐L‐cysteine (NAC). Ejaculates from five dogs were collected, pooled and evaluated for concentration, motility, rapid steady forward movement (RSF‐movement), viability, acrosomal integrity and by the hypo‐osmotic swelling test (HOST). In addition, superoxide anion (O₂^‐?) production, hydroxyl radicals (OH^?) and total reactive oxygen species (tROS) were determined. The pool was divided into five aliquots, which were diluted to a final concentration of 66.66 × 10⁶ spermatozoa/ml with Tris‐glucose‐egg yolk extender containing one of the following concentrations of NAC (0, 0.5, 1, 2.5 or 5 mm ). The semen aliquots were chilled and preserved at 4°C. Semen quality was evaluated after rewarming at 72 h. Sperm motility was significantly higher with the 0.5 mm concentration compared with the control group (p = 0.001). Rapid steady forward movement was higher with the 0.5 and 1 mm concentrations compared with the control and 5 mm group (p < 0.001). Viability and HOST percentages were not significantly altered. Compared with the control, the 5 mm concentration showed significantly reduced percentages of spermatozoa with normal acrosomes (p = 0.049). None of the ROS values at 72 h were significantly affected by the presence of NAC in semen extenders, although all NAC concentrations showed lower O₂^‐? and OH^? values compared with the control. Only the concentrations of 1 and 5 mm inhibited the significant increase of tROS values after 72 h, compared with the fresh semen value. In conclusion, NAC supplementation of semen extenders is beneficial to semen motility of canine spermatozoa during chilling with the 0.5 mm concentration being the most effective, although no significant ROS inhibition was observed at 72 h.     

Effect of mitochondrial uncoupling and glycolysis inhibition on ram sperm functionality

Studies have demonstrated the importance of mitochondria to sperm functionality, as the main source of ATP for cellular homoeostasis and motility. However, the role of mitochondria on sperm metabolism is still controversial. Studies indicate that, for some species, glycolysis may be the main mechanism for sperm energy production. For ram sperm, such pathway is not clear. Thus, we evaluated ram sperm in response to mitochondrial uncoupling and glycolysis inhibition aiming to assess the importance of each pathway for sperm functionality. Statistical analysis was performed by the SAS System for Windows, using the General Linear Model Procedure. Data were tested for residue normality and variance homogeneity. A p < .05 was considered significant. Groups treated with the mitochondrial uncoupler Carbonyl cyanide 3 chlorophenylhydrazone (CCCP) showed a decrease in the percentage of cells with low mitochondrial activity and high mitochondrial membrane potential. We also observed that the highest CCCP concentration promotes a decrease in sperm susceptibility to lipid peroxidation. Regardless the lack of effect of CCCP on total motility, this substance induced significant alterations on sperm kinetics. Besides the interference of CCCP on spermatic movement patterns, it was also possible to observe such an effect in samples treated with the inhibitor of glycolysis (2‐deoxy‐d ‐glucose, DOG). Furthermore, treatment with DOG also led to a dose‐dependent increase in sperm susceptibility to lipid peroxidation. Based on our results, we suggest that the glycolysis appears to be as important as oxidative phosphorylation for ovine sperm kinetics as this mechanism is capable of maintaining full motility when most of the cells have a low mitochondrial membrane potential. Furthermore, we found that changes in the glycolytic pathway trough glycolysis inhibition are likely involved in mitochondrial dysfunction and sperm oxidative unbalance.     

Comparison of three different procedures for anaesthesia of calves with respect to possible pain-associated reactions

The objective of this study was to compare the effects of three anaesthetic protocols with respect to possible pain-associated reactions: injection (INJE) with xylazine (0.2 mg/kg), ketamine (5 mg/kg) and local anaesthesia (procain 2%); a combination of injection and inhalation (KOMB) with xylazine (0.2 mg/kg), ketamine (2 mg/kg), and isoflurane (1.5-3.0 vol%); and inhalation with isoflurane (control group [o]) surgery. During the trial, the anaesthetic level was controlled, and signs of response to surgical stimulation or spontaneous movements, changes in heart rate, mean arterial pressure, plasma L-lactate and cortisol levels were recorded. The dosage of 5 mg ketamine/kg body mass was rarely sufficient to obtain surgical tolerance in group INJE. In order to attain surgical tolerance, it was necessary to administer an average of 8.4 mg ketamine/kg body mass. All animals of the INJEc/o groups started to show signs of spontaneous movements or response to surgical stimulation between five and twelve minutes after the last administration of ketamine. Signs of response to surgical stimulation at the time of skin incision were significantly most frequent (37%) in the INHAc group, while agitation or response to surgical stimulation at least once during the entire time of anaesthesia was significantly (p <0.01) most frequent in the INJEc group (100%). Changes in heart rate and mean blood pressure were not found to be related to surgical stress situations.Ten minutes after the incision, the highest increases in plasma cortisol levels above basal levels of the previous day were determined in the groups INJEc and INJEo (53.5 and 57.7 nmol/l, respectively). However, a significant increase of plasma cortisol levels between the previous day and ten minutes after incision was only found between INHAc and INHAo (30.1 versus 7.5 nmol/l, p < 0.01) but not in the other pairs of groups (INJEc/o and KOMBdc/o). Within the three experimental groups the increase of plasma cortisol levels between the previous day and ten minutes after incision was only significant between INJEc and KOMBc (53.5 versus 28.3 nmol/l, p < 0.01). Among the protocols tested in this study, the combination anaesthesia (KOMB) was associated with the least amount of pain-associated reactions by the calves. Therefore, this protocol should be used preferred to the other two protocols in order to minimise stress and pain for the animals as much as posssible.     

Treatment of virulent footrot with lincomycin and spectinomycin

A mixture of lincomycin and spectinomycin was investigated as a treatment for footrot in sheep. In a controlled clinical trial 92.5% of acute and chronic cases of virulent footrot were cured following a single intramuscular injection of a mixture containing 50 mg lincomycin and 100 mg spectinomycin/ml at a dose rate of 1 ml/10 kg bodyweight. No improvement in clinical response was observed in groups of sheep treated on 3 successive days with this dose rate nor in another group treated once at a dose rate 1 ml/3.3 kg bodyweight. Cure effectiveness of each of the 3 treatment groups relative to untreated controls was 89%, 95% and 95%. Efficacy of lincomycin/spectinomycin was compared with that of penicillin/streptomycin in the treatment of footrot on 2 farms in south western New South Wales. Assessments made 14 to 17 d after treatment showed that on one farm all 122 ewes treated with lincomycin/spectinomycin had recovered while 170 of 175 ewes treated with penicillin/streptomycin recovered in the same period. On the second farm 87 of 90 ewes treated with lincomycin/spectinomycin recovered, compared with 184 of 190 sheep in the same flock treated with penicillin/streptomycin. Supportive footbathing did not seem to improve the clinical response in either treatment group and the paring done was sufficient only to establish diagnosis and to remove grossly overgrown horn.     

Attitudes of veterinarians to undergraduate education in genetics and the use of genetics in veterinary occupations

A questionnaire regarding attitudes to undergraduate education in genetics and the subsequent use of that education was sent to 1000 veterinarians registered in New South Wales. Three hundred replies to the questionnaire were received and analysed. No significant difference in the perceived adequacy of courses at the four universities involved in teaching veterinary science was observed. Opinions on adequacy of education in genetics were not affected by age, sex or years since graduation. The major reason cited for lack of adequacy was that undergraduate genetics courses were not practical. Only 17 respondents stated that they were never approached for information on genetics or animal breeding, while 76 were approached more than 20 times annually for information. Eighty-three respondents claimed to deal with more than 20 cases annually that required some knowledge of genetics. The results of the questionnaire highlight the importance of genetics instruction in undergraduate veterinary education. The questionnaire responses also provide insight into how working veterinarians consider that courses in genetics could be improved.     

Surgical management of an abdominal abscess in a Malayan tapir

A captive Malayan tapir was observed to have inappetence, weight loss, signs of depression, mild dehydration and diar rhoea. Haematological and serum biochemical tests showed anaemia, hypoproteinaemia, hyperfibrinogenaemia and neu-trophilia with a left shift. Ultrasonic examination of the abdomen under anaesthesia revealed a well-encapsulated abscess. The abscess was marsupialised to the ventral body wall. Culture of the pus produced a mixed bacterial growth. Antimicrobial therapy was based on bacterial sensitivity results. Follow-up ultrasonic examinations showed resolution of the abscess. Ninety-one days after surgery the tapir began regurgitating food and water. An abscess originating from the stomach and occluding the lumen of the duodenum was identified at surgery. The abscess ruptured during surgical manipulations and the tapir was euthanased.     

Surgical treatment of osteomyelitis and sequestration of the ilium by a cranial partial hemipelvectomy in an alpaca cria

A 12-week-old female alpaca cria weighing 10 kg presented with a 2-month history of left hindlimb lameness associated with osteomyelitis and sequestration of the shaft and wing of the left ilium. The affected segment of the ilium was surgically removed. Bacteriological culture revealed a small number of mixed aerobic and anaerobic bacteria. The cria recovered well, was discharged from the hospital 8 days after surgery and was paddock sound without complications 8 months later. Osteomyelitis and sequestration of long bones has been reported in alpacas, but further investigation is necessary to define the prevalence and pathogenesis and to develop specific recommendations for prevention and treatment.