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.     

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.     

Surveillance for highly pathogenic avian influenza in migratory shorebirds at the terminus of the East Asian-Australasian Flyway

AIM: To determine if migratory birds arriving in New Zealand in the Southern Hemisphere spring of 2004 were infected with the highly pathogenic avian influenza (AI) virus, H5N1.

METHODS: Cloacal and faecal samples were collected from migratory red knots following their arrival in New Zealand in October 2004. Two species of resident sympatric birds, wrybill and mallard duck, were sampled prior to, and following, the arrival of migratory birds.

RESULTS: No AI viruses were isolated from migratory or resident shorebirds. Non-pathogenic AI viruses were isolated from six resident mallard ducks, comprising the endemic subtypes H4 (n=2), H7 (non-pathogenic), H10, and H11 (n=2).

CONCLUSIONS: Highly pathogenic AI H5N1 virus was not detected in migratory shorebirds or sympatric water birds in the Firth of Thames, New Zealand, in 2004-2005, despite the possible proximity of migratory birds to outbreaks of the disease in East Asia in 2004.     

Antioxidative Effects of Astaxanthin against Nitric Oxide‐Induced Oxidative Stress on Cell Viability and Gene Expression in Bovine Oviduct Epithelial Cell and the Developmental Competence of Bovine IVM/IVF Embryos

The aim of the present study was to elucidate the fundamental mechanism of bovine oviduct epithelial cell (BOEC) co‐culture on developmental capacity of bovine in vitro oocyte maturation/in vitro fertilization (IVM/IVF) embryos. We examined the effects of astaxanthin against nitric oxide‐induced oxidative stress on cell viability by MTT assay, lipid peroxidation (LPO) by using thiobarbituric acid (TBA) reaction for malondialdehyde (MDA) and the expression of antioxidant genes (CuZnSOD, MnSOD and Catalase) or apoptosis genes (Bcl‐2, Caspase‐3 and Bax) by RT‐PCR in BOEC. We also evaluated the developmental rates of bovine IVM/IVF embryos co‐cultured with BOEC pre‐treated with astaxanthin (500 μm ) in the presence or absence of sodium nitroprusside (SNP, 1000 μm ) for 24 h. Cell viability in BOEC treated with SNP (50–2000 μm ) lowered, while astaxanthin addition (50–500 μm ) increased it in a dose‐dependent manner. Cell viability in astaxanthin plus SNP (1000 μm ) gradually recovered according to the increase in astaxanthin additions (100–500 mm ). The LPO in astaxanthin group (50–500 μM) gradually decreased in a dose dependent manner and among SNP or astaxanthin plus SNP group, SNP alone and astaxanthin (50 μM) plus SNP shown a significant increase than other groups (p < 0.05). Expression of apoptosis or antioxidant genes was detected by RT‐PCR. Bcl‐2 and antioxidant genes were detected in astaxanthin or astaxanthin plus SNP group, and Caspase‐3 and Bax genes were only found in SNP group. When bovine IVM/IVF embryos were cultured for 6–7 days under co‐culture system such as BOEC treated with astaxanthin in the presence or absence of SNP, the developmental ability to blastocysts in 500 μm astaxanthin group was the highest of all groups. These results suggest that astaxanthin has a antioxidative effect on cell viability and LPO of BOEC, and development of bovine IVM/IVF embryos due to the induction of antioxidant genes and suppression of apoptosis genes.     

Feline nasal and paranasal sinus tumours: clinicopathological study, histomorphological description and diagnostic immunohistochemistry of 123 cases

Histological examination was performed in 123 cats with primary nasal and paranasal sinus tumours; 117 had undergone surgical biopsy and six necropsy. Special stains and immunohistochemistry were performed on poorly differentiated cases. Ninety-two percent (113/123) of the tumours were malignant. There was an increased risk for old cats (mean age of 10.9 years), and a male predilection (59% males). Clinical signs and breeds varied with the histological type of tumour. Thirty-nine percent (48/123) of the cases presented with nasal discharge, 21% (26/123) with dyspnea, 20% (24/123) with facial swelling, and 15% (19/123) with epistaxis. Forty-three percent (53/123) of the tumours were of epithelial origin. Adenocarcinomas (18/53) and squamous cell carcinomas (17/53) were the most common epithelial tumours. Fifty percent (26/53) of the epithelial tumours originated from the pseudo-stratified respiratory epithelium of the nasal cavity, 28% (15/53) from the stratified squamous epithelium of the vestibule, 9% (5/53) from olfactory epithelium, 9% (5/53) from submucosal glands and 4% (2/53) from minor salivary glands. Malignant lymphoma (35/123) was the most common tumour. Seventy-one percent (25/35) of the malignant lymphomas were B-cell tumours and 29% (10/35) were T-cell tumours. Six cases of malignant lymphomas were proved to be epitheliotropic T-cell lymphomas. This is the first report of a primary nasal epitheliotropic T-cell lymphoma in cats.     

Cloacal papillomatosis in the absence of herpesvirus and papillomavirus in a sulphur-crested cockatoo (Cacatua galerita)

CASE HISTORY A 21-year-old male sulphur-crested cockatoo (Cacatua galerita) was presented following the sudden appearance of blood associated with the passage of faeces and urates.

CLINICAL AND PATHOLOGICAL FINDINGS: There was fresh blood-staining of the feathers around the vent. The dorsal mucosal wall of the proctodeum was erythematous and roughened in appearance. An endoscopic biopsy was performed, and histological examination revealed multiple fronds of epithelium; the mucosa varied from simple to pseudostratified columnar epithelium, with diffuse hyperplasia of goblet cells. The underlying connective tissue stroma was well vascularised and was infiltrated with mixed inflammatory cells, comprising granulocytic cells and macrophages. PCR testing for both herpesvirus and papillomavirus, using consensus primers, was negative.

DIAGNOSIS: Cloacal papillomatosis.

CLINICAL RELEVANCE: This case manifested typical clinical signs and histological lesions of cloacal papillomatosis in the absence of demonstrable herpesvirus or papillomavirus. Veterinarians need to consider this disease in the differential diagnosis of blood in the droppings of parrots and cockatoos even in countries where psittacine herpesviruses are exotic diseases.     

Bilateral ovarian adenocarcinoma in a mare causing haemoperitoneum and colic

Abstract

CASE HISTORY: A 13-year-old Thoroughbred mare was presented with a history of mild colic over 3 days. This colic had acutely exacerbated and was unresponsive to analgesic treatment, and was referred to Massey University Veterinary Teaching Hospital.

CLINICAL FINDINGS: On examination the heart rate was 100 beats per minute, and mucous membranes were pale and tacky. A large mass was detected on transrectal palpation in the caudal abdomen to the left of midline. Explorative laparotomy revealed severe haemoperitoneum and several masses that were associated with the reproductive tract. The mare was then subject to euthanasia. On post-mortem examination, adjacent and attached to each ovary were soft, lobulated dark red masses up to 200 mm in diameter. Similar masses were present in the omentum and on the peritoneal surface of the diaphragm and the serosa of the spleen and liver. Histopathology revealed that the neoplastic component of the masses comprised proliferating cuboidal to columnar cells forming disorganised acini and cords separated by dense collagenous stroma. Immunohistochemistry showed the neoplastic cells were positive for cytokeratin AE1/AE3 and vimentin, but negative for cytokeratin 7 and inhibin α.

DIAGNOSIS: Bilateral ovarian adenocarcinoma with transcoelomic metastasis and terminal decompensation due to rupture of a neoplastic mass and consequent haemoperitoneum.

CLINICAL RELEVANCE: To the authors' knowledge, bilateral ovarian adenocarcinoma has not been previously reported in a horse. Ovarian adenocarcinoma should be considered when horses present with haemoperitoneum and colic. Further research is required on the immunohistochemical differentiation of adenocarcinoma of ovarian and intestinal origin in the horse.