A possible predisposition to dilated cardiomyopathy in Huntaway dogs

AIM: To compare the prevalence of dilated cardiomyopathy (DCM) in New Zealand Huntaway dogs with the prevalence of DCM in other breeds of dog.

METHODS: The necropsy database at Massey University was used to identify cases of DCM diagnosed between January 1999 and March 2006. Dogs were considered to have DCM if echocardiographic, gross necropsy, or histological findings were consistent with this diagnosis. The prevalence in Huntaways was then compared with the prevalence observed in all breeds of dog, as well as the prevalence observed in large breeds of dog.

RESULTS: Twelve dogs were identified with DCM. One was diagnosed using echocardiography, while the other 11 were diagnosed by gross necropsy examination. The gross diagnosis of DCM was confirmed histologically in 6/11 dogs. The prevalence of DCM in Huntaways was significantly higher than the prevalence seen in all breeds of dog (p=0.008), and the prevalence in large breeds of dog (p=0.025). All four Huntaways diagnosed with DCM were male, and had an average age of 4 years. Three dogs presented with symptoms attributable to impaired heart function while one presented with symptoms of chronic renal failure. The duration of clinical symptoms prior to presentation ranged between 1 day and 3 weeks.

CONCLUSIONS: The results of this study suggest that Huntaways may be predisposed to the development of DCM. Although the increased prevalence in this breed was significant, only small numbers of affected Huntaways were identified, and additional cases are required to confirm these preliminary findings.

CLINICAL RELEVANCE: Huntaways are the most common working dog in New Zealand. The premature loss of a working dog is expected to have a significant economic impact on farmers. Further investigation of DCM in Huntaways may allow measures to reduce the prevalence in this breed.     

Intrauterine Idiopathic Amputation of the Head of a Porcine Foetus

An anencephalic full‐term porcine foetus accompanied by a mummified head was submitted for examination. The neck almost entirely lacked skin and was covered by granulation tissue as were the exposed parts of the spine and spinal cord. The case represents a rare case of intrauterine amputation. A definitive cause could not be established because the placenta was not available. The most likely cause is strangulation of the neck. Such strangulation could be due to a defect of the allantoamnion with herniation of the foetal head or entanglement by amniotic constriction bands.     

A Case of Successful Pregnancy in a Ewe with Uterus Didelphys

Uterus didelphys is a rare congenital abnormality of the reproductive tract. Although it occurs in various species, there are no published reports describing pregnancy outcome in association with this abnormality. Herein we describe a case of successful unilateral singleton pregnancy in a ewe incidentally found to have uterus didelphys during the course of a biomedical research study. The pregnancy was established using assisted reproductive techniques and interrupted in late gestation, at which point the abnormality was identified. Serial ultrasound assessment of foetal biometry revealed a normal foetal growth trajectory. Despite a 45% reduction in placentome number, total placentome weight was near normal secondary to compensatory placentome growth and development. To our knowledge, this is the first detailed report of normal foetal growth in an animal with uterus didelphys and illustrates the ability of the ovine placenta to adapt to a reduced number of placentomes and maintain foetal nutrient supply.     

Effects of low‐density lipoproteins as additive on quality parameters and oxidative stress following cryopreservation of mithun (Bos frontalis) spermatozoa

Artificial breeding of mithun poses several challenges including lack of standard protocol for cryopreservation of spermatozoa. This is further complicated by harmful effects of hen's egg yolk (EY) as additive in extender. Purified low‐density lipoproteins (LDL) extracted from EY have been shown as beneficial over EY extender for long‐term semen storage in several species. This investigation explored use of LDL versus EY on semen quality and oxidative stress following freezing–thawing of spermatozoa. A total of 25 of 50 ejaculates based on biophysical parameters were selected for the experiment. After diluting with the Tris‐citrate‐glycerol (TCG) extender, each sample was split into three equal aliquots: Group I, control, EY; Group II and Group III contained 8% and 10% purified LDL, respectively. Frozen–thawed samples were evaluated for motility parameters (progressive, and in the bovine cervical mucus penetration test [BCMPT]), viability, sperm and nuclear abnormality, acrosome integrity, and enzymatic (leakage of intracellular contents) and biochemical (oxidative stress) profiles and in vitro fertility (IVF) assay. Study revealed a significant (p < .05) improvement in viability, sperm and nuclear abnormality, acrosome integrity, motility (progressive and in cervical mucus), cholesterol content, and reduction in the leakage of intracellular enzymes in Group II. Moreover, intactness of acrosome and biochemical membranes was protected significantly (p < .05) in addition to significant (p < .05) improvement in binding per cent and binding index in IVF assay in extender containing 8% LDL. These results demonstrate that although cryopreservation of mithun's spermatozoa in EY was comparable with other species, addition of 8% LDL holds a clear advantage over EY or 10% LDL.     

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.     

Effect of Cooling Rate to 5°C,Straw Size and Farm on Fertilizing Ability of Cryopreserved Rabbit Sperm

High fertility and prolificacy in rabbits are currently only achieved using fresh sperm. This study was conducted to determine if the cooling rate to 5°C, the straw size and the farm where artificial inseminations are performed have an impact on the fertilizing ability of rabbit sperm cryopreserved with an extender containing dimethyl sulphoxide (DMSO; 1.75 m ) and sucrose (0.05 m ). Slow cooling to 5°C improved neither fertility rate (58 vs 56% kindling rate for fast and slow cooling, respectively) nor prolificacy (6.5 vs 8.7 total born for slow and fast cooling, respectively; p < 0.05) compared to fast cooling rate to 5°C. The straw size did not have an effect on either fertility or prolificacy (47 vs 57% kindling rate and 6.3 vs 6.8 total born for sperm loaded into 0.25 and 0.5 ml straws, respectively). In addition, similar results were obtained between farms (46–57% kindling rate and 4.9–6.7 total born), although this effect should be studied further. In conclusion, with this extender, slow cooling does not present a beneficial effect on sperm fertilizing ability and either 0.25 or 0.5 ml straws can be used to freeze the sperm, obtaining similar results after artificial insemination. In addition, similar results were obtained between farms when using cryopreserved sperm, and these results were lower than those obtained after artificial insemination with fresh semen. Therefore, new approaches are needed to improve the results obtained when cryopreserved sperms are used before this type of sperm can be used for commercial purposes.     

Echocardiographic evaluation of the effects of medetomidine and xylazine in dogs

The echocardiographic effects of medetomidine and xylazine were evaluated in 6 healthy dogs. Values for echocardiographic variables were significantly different from pre-treatment values after administration of both drugs. The effects of medetomidine were similar to that of xylazine. Because of their cardiac depressant effects, both drugs should be used with care in sick dogs.     

Lythrum hyssopifolia (lesser loosestrife) poisoning of sheep in Victoria

Objective To document an ovine disease attributed to the consumption of Lythrum hyssopifolia (lesser loosestrife). Procedures Historical and histological review of field and experimental cases. Results 1–20% mortality occurred in sheep flocks grazing paddocks where L. hyssopifolia was the predominant green vegetation. Well‐documented disease outbreaks occurred in summer on nine farms across Victoria between 1974 and 2002. Liver damage occurred in all nine outbreaks, with kidney damage in at least eight. Hepatocyte necrosis was usually zonal to midzonal (zone 2) in the liver samples from four farms and periacinar (zone 3) in those from three farms, but some livers showed only single‐cell necrosis. Multinucleate hepatocytes near necrotic areas were a feature in six cases. Proximal tubular epithelium appeared to be the primary renal target and brown granules were often present in renal tubules. Biochemical and histological evidence of liver and kidney damage was obtained from two sheep experimentally pen‐fed harvested L. hyssopifolia. Conclusion Chemicals in L. hyssopifolia are toxic to ovine hepatocytes and renal tubular epithelial cells.     

Effect of Sperm Cryopreservation on the European Eel Sperm Viability and Spermatozoa Morphology

The main objective of the present work was to study the effect of cryopreservation of European eel sperm both on the sperm viability and the spermatozoa head morphology. Spermatozoa morphology was evaluated with computer-assisted morphology analysis after collection in fresh samples, after adding the freezing medium containing dimethyl sulfoxide as cryoprotectant and, finally, after the cryopreservation process and thawing. Cell viability was assessed, in both fresh and thawed samples, by Hoechst 33258 staining. Computer-assisted sperm analysis (CASA) was used to determine the percentage of motile cells and to measure motility parameters in sperm samples. A significant decrease of head perimeter (12.56%) and area (17.90%) was detected from spermatozoa in fresh to thawed samples, indicating that cells do not recover the original size after the cryopreservation process. CASA was used to measure the percentage of motile cells (51.9%) and spermatozoa motility parameters such as curvilinear, straight line and angular path velocities, as well as beating cross frequency. This technique was employed in the fresh sperm samples but proteins present at the freezing medium (L-alpha-phosphatidylcholine) made impossible to use this last technique in thawed samples. When sperm viability was assessed by Hoechst staining, a significant decrease of approximately 15% (73.10 vs 58.26%) of alive spermatozoa was registered from fresh to thawed samples. The percentage of motile cells measured by CASA in fresh samples (51.9%) was lower than the percentage of alive cells determined by Hoechst stainning, suggesting the existence of different batches of spermatozoa in different stages of development, even during the eight to tenth weeks of treatment, when the highest sperm quality was found.