Use of Powdered Egg Yolk vs Fresh Egg Yolk for the Cryopreservation of Ovine Semen

Egg yolk is a common additive to sperm cryopreservation diluents. Because of its animal origin, however, it also represents a potential risk of microbiological contamination in the diluent. This potential contamination can be avoided by using powdered egg yolk, instead of fresh egg yolk, as it is pasteurized. This study was conducted to determine ram sperm cryosurvival was affected by the type of egg yolk used (powdered egg yolk or fresh egg yolk) and by yolk concentration (10, 15 or 20%) in the diluent. Microbiological analyses were also performed to quantify the microbiological contamination in the diluents containing the two types of egg yolk. Sperm cryosurvival was determined by motility and morphology analyses after thawing. Motility parameters were assessed using a computer-assisted sperm analysis (CASA) system, and the percentage of sperm with a normal apical ridge was evaluated using a differential interference contrast microscope. No significant differences were observed between diluents in the percentage of sperm with normal apical ridge. However, higher percentages of total motile cells were observed for samples containing powdered egg yolk (69%) compared to samples containing fresh egg yolk (60%). However, sperm in diluents containing fresh egg yolk, exhibited higher values for average-path velocity, straight-line velocity and beat cross frequency and lower values for amplitude of lateral head displacement (p <0.05), compared to cells in diluents containing powdered egg yolk. Microbiological contamination was similar (<200 CFU/ml) in both diluents, and no bacterial growth was observed in either, when antibiotics were added. Therefore, powdered egg yolk can be effective used in diluents for the freezing of ram semen. However, the in vivo fertility of sperm frozen in diluents containing powdered egg yolk should be tested, as some motility parameters were different for sperm treated with powdered egg yolk compared to fresh egg yolk.     

Death of a neonatal lamb due to Clostridium perfringens type B in New Zealand

ABSTRACT

Case history and clinical findings: A flock of 20 sheep was kept within three paddocks on a single property. None of the animals in the flock had been vaccinated against any disease for at least three years. Abdominal bloating and haemorrhagic diarrhoea were observed in Lamb 1 at 24 hours-of-age. The lamb subsequently died within an hour of the onset of clinical signs. Lamb 2 was 3-days-old when observed to be recumbent with opisthotonus. The lamb was treated with dextrose, vitamins B1 and B12, and penicillin G, but died 4 hours later.

Pathological findings: Examination of Lamb 1 revealed markedly increased gas within the peritoneum and within dilated loops of intestine. The intestines were dark red and contained large quantities of haemorrhagic fluid. Histology of the intestines revealed peracute mucosal necrosis with minimal accompanying inflammation. The intestinal lumen contained cell debris, haemorrhage, and myriad large Gram-positive bacilli. The intestines of Lamb 2 did not appear bloated or reddened. However, multiple fibrin clots were visible within the pericardial sac. Histopathological examination revealed small foci of necrosis within the mucosa of the distal intestine. The necrotic foci were often associated with large numbers of large Gram-positive bacilli.

Immunohistochemsitry and molecular biology: Intestinal samples from Lamb 1 were processed for Clostridium perfringens immunohistochemistry, which revealed large numbers of intralesional, positively immunostained rods. Fragments corresponding to the expected sizes for genes encoding alpha, beta, and epsilon C. perfringens typing toxins were amplified by PCR from DNA extracted from formalin-fixed sections of intestine.

Diagnosis: Lamb dysentery due to C. perfringens type B.

Clinical relevance: C. perfringens bacteria have a worldwide distribution, but disease due to C. perfringens type B has only been diagnosed in a small number of countries and has never been reported in New Zealand or Australia. C. perfringens type B produce both beta toxin and epsilon toxins, therefore both haemorrhagic enteritis and systemic vascular damage can develop. As many animals are exposed to C. perfringens without developing disease, there must be additional unknown factors that resulted in disease in these particular sheep. Vaccines that specifically protect against C. perfringens type B are available and may be recommended for use in smaller non-commercial flocks, as in the present case.     

Retrospective assessment of tolerability and efficacy of zoledronate in the palliative treatment of cancer-bearing dogs

Zoledronate is a bisphosphonate frequently used for the treatment of hypercalcaemia of malignancy and tumour-associated bone pain in dogs, however, there is a paucity of information regarding its use in veterinary medicine. The aim of this retrospective study was to report the tolerability of zoledronate in the palliative treatment of cancer-bearing dogs and secondarily to to assess the efficacy of zoledronate for the treatment of hypercalcaemia of malignancy. Thirty-seven dogs (22 with tumour-associated bone pain and 15 with hypercalcaemia of malignancy) that received 114 zoledronate infusions were included. Tolerability was assessed by the absence of post-zoledronate hypocalcaemia or other adverse events as defined by Veterinary Cooperative Oncology Group-Common Terminology Criteria for Adverse Events criteria. Efficacy was assessed by comparison of available ionized calcium levels before and after zoledronate administration in hypercalcaemic dogs. In 79% of zoledronate infusions, no adverse events were reported. The majority of adverse events which occurred in the other 21% of infusions could be attributed to concurrent chemotherapy or the underlying neoplastic disease. There was a small but significant increase in creatinine following treatment with zoledronate, however, none of the dogs developed clinically significant renal disease. In eight hypercalcaemic dogs with available ionized calcium following zoledronate administration, ionized calcium decreased rapidly within 7 days following treatment with zoledronate. Zoledronate is well-tolerated with few recorded adverse events, however, monitoring of serum creatinine is advised. Zoledronate seems to be effective in the treatment of hypercalcaemia of malignancy.     

Compensatory growth and body composition of juvenile black rockfish <Emphasis Type="Italic">Sebastes schlegeli</Emphasis> following feed deprivation

ABSTRACT:   Compensatory growth, feeding rate, feed efficiency and chemical composition of juvenile black rockfish (mean weight 1.43 g) were investigated for 35 days after a 14-day feed deprivation treatment under four feeding conditions: one group continuously fed (control) and the other three groups fasted for 5 days (F5), 10 days (F10) and 14 days (F14). All fasted fish were re-fed from day 15. Only F5 achieved the same body weight as the control, indicating that complete compensation occurred in F5. The specific growth rate (SGR) of F5 was the highest at day 21 and then decreased thereafter, showing higher values than the control at days 21, 28 and 42. In contrast, although SGRs of F10 and F14 were higher than that of the control during the whole refeeding period except day 21, they did not catch up the control in body mass, indicating that only partial compensation occurred in F10 and F14. The feeding rate (FR) of all groups except F14 changed in a pattern similar to SGR (Spearman's rank correlation, r _s > 0.9), suggesting that SGR varied depending on FR. Similar feeding efficiencies (FEs) were found in the four groups and they did not vary significantly during the whole refeeding period, suggesting that FE was not the factor affecting SGR. At day 14, the ratios of lipid to lean body mass in F10 and F14 were lower than those in the control and F5, and there was no difference between the control and F5. At day 49, however, only F14 showed a lower value than the other three groups, and there was no difference among the three groups. These results indicate that juvenile black rockfish fasted for 5–14 days can exhibit compensatory growth after refeeding, but timing and degree vary depending on the duration of feed deprivation.     

Bluetongue virus serotypes 1 and 3 infection in Poll Dorset sheep

Objective To study the clinical signs following bluetongue virus serotypes 1 and 3 infection in Poll Dorset sheep.
Design A clinical and pathological study.
Procedure Twenty Poll Dorset sheep were inoculated with bluetongue virus serotypes 1 or 3, each inoculum having a different passage history. The sheep were examined daily and their clinical appearance and rectal temperatures recorded. Heparinised and non-heparinised blood samples were taken at intervals for virological and serological study. Gross pathological findings were recorded for several sheep at necropsy and tissue samples were collected from three sheep for virological studies.
Results All inoculated sheep developed clinical disease. The clinical signs and gross pathological changes varied considerably but were consistent with damage to the vascular endothelial system. There was a decline in the titres of infectious bluetongue virus and of antigen in tissues collected between 7 and 12 days after infection.
Conclusions The severity of disease was related to the speed of onset and duration of pyrexia and not the development or titre of viraemia. Generally, those animals with sensitive mouths, depression, coronitis, recumbency and reluctance to move were the most debilitated. Whole blood was the most reliable source of infectious virus from acutely and chronically infected and convalescent animals. However, tissue samples particularly spleen, collected from dead or killed animals suffering from either peracute or acute forms of disease were most appropriate for the rapid confirmation of a clinical diagnosis.     

Unguided bronchoalveolar lavage techniques and residual effects in dogs

Objectives To investigate the use of unguided bronchoalveolar lavage techniques in dogs without fibreoptic bronchoscopy, using an adapted single vascular catheter and a double-lumen catheter made from two single vascular catheters.
Animals Sixty-nine dogs were examined with the single-catheter technique and 110 dogs with the double-catheter technique.
Design A prospective study.
Procedure Sixty-nine and 220 samples, collected with the single catheter and the double catheter respectively, were examined cytologically Lungs of 69 dogs were examined grossly and histologically. Radiographic examination was performed on 11 dogs.
Results The double-catheter technique produced samples with significantly higher cellularity (P < 0.01) and fewer red blood cells (P < 0.01) than the single-catheter technique. Repeat samples collected with a double catheter were not significantly different (P > 0.01) in any value. A reference range for nucleated cell counts of 62 to 1210 – 10⁶/L was calculated from 57 clinically and histologically normal dogs. The major residual effects of the technique were localised pulmonary oedema, and alveolar distension with collapse and congestion of distant parenchyma. Thoracic radiographs revealed increased lung opacity for at least up to 7 h after the procedure.
Conclusions The cellularity of the bronchoalveolar lavage fluid obtained was adequate and sufficient fluid was retrieved when the single catheter was located in a proper position. However, the double catheter obtained better samples more quickly and easily, with less damage to the respiratory tract.     

Subject-based problem-based learning in the veterinary science course at the University of Queensland

Objective
To assess students' perceptions of subject-based problem-based learning in the fourth year of the veterinary science course at the University of Queensland.
Design
A questionnaire-based study.
Procedure
Subject-based problem-based learning was introduced into parts of two fourth year subjects in a 5-year veterinary science course. The problem-based learning exercise used modified clinical cases and was computer-assisted. Students worked in groups of two to four, and small group discussion sessions were tutorless. Lectures were replaced by large group discussion and feedback sessions, led by the teacher, with approximately 85 students.
Results
There was a significant increase in the percentage of students who strongly agreed that they had better understanding of the subject, and had learned to apply principles from this class in new situations. The only consistent criticism by students was directed at the extra time required compared to traditional lecture-based subjects.
Conclusion
Students' perceptions of the learning outcome were very favourable for problem-based learning when compared to the lecture-based subject.     

Flow cytometric evaluation of disseminated intravascular coagulation in a canine endotoxemia model

Sepsis is associated with substantial morbidity and mortality in dogs. Alterations in hemostasis by systemic inflammation play an important role in the pathophysiology of sepsis. To evaluate the functional hemostatic changes in sepsis, we evaluated coagulation profiles and flow cytometric measurement of P-selectin (CD62P) expression on platelets, as well as platelet-leukocyte aggregation from a lipopolysaccharide (LPS)-induced endotoxemia model in dogs (n = 7). A sublethal dose of LPS [1 mg/kg body weight (BW)] induced thrombocytopenia and increased activated partial thromboplastin time (aPTT), prothrombin time (PT), and D-dimer concentrations. Flow cytometry analysis showed a significant increase in P-selectin expression on platelets between 1 and 24 h of a total 48 h of the experiment. In addition, platelet-leukocyte aggregation was significantly increased in the early stage of endotoxemia (at 1 and < 6 h for platelet-monocyte aggregation and at 3 h for platelet-neutrophil aggregation). Our results suggest that CD62P expression on platelets and platelet-leukocyte aggregation, as measured by flow cytometry, can be useful biomarkers of disseminated intravascular coagulation (DIC) in canine sepsis. These functional changes contribute to our understanding of the pathophysiology of hemostasis in endotoxemia.