Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

Expression of the Vascular Endothelial Growth Factor (VEGF‐A) and its Receptors in the Umbilical Cord in the Course of Pregnancy in the Pig

The umbilical cord (UC) and the placenta are important organs through which respiratory gases, nutrients, wastes and biologically active substances are exchanged between the maternal and the foetal system. A rapid placental vascularization observed in the second half of pig pregnancy is positively correlated with the mRNA expression of the vascular endothelial growth factor (VEGF). Based on these findings, we hypothesized that VEGF may have a stimulatory effect in the dynamically growing UC. To further understand the role of the VEGF–VEGFR system during UC development, mRNA and protein expression as well as the cellular localization of VEGF‐A, VEGFR‐1 and VEGFR‐2 in UC were examined on days 40, 60, 75 and 90 of pregnancy and after physiological delivery in the pig (day 114 of pregnancy). Real Time RT‐PCR analysis showed an increase in the mRNA levels of VEGF120 and VEGF164 from day 90 of pregnancy. VEGFR‐1 mRNA expression was significantly increased on day 75 of pregnancy. No significant changes in VEGFR‐2 mRNA expression were detected. In turn, western blot analysis revealed an increase in VEGF‐A protein expression on day 40, compared to the later days of pregnancy. A rapid increase in the VEGFR‐1 protein level was noted on day 75 and 90 of gestation. No significant changes in VEGFR‐2 protein expression were detected on any of the analysed days of pregnancy. Immunohistochemical staining enabled detection of VEGF–VEGFR system, in endothelial and tunica media cells of the umbilical vessels and in allantoic duct and amniotic epithelium on all analysed days of pregnancy. Positive reactions for VEGF‐A and VEGFR‐1, but not VEGFR‐2, were also observed in myofibroblasts. In conclusion, this data shows that members of the VEGF–VEGFR system are temporally and spatially well localized for playing key roles during umbilical cord formation and its intensive growth observed after day 75 of pregnancy.     

Ontogenetic movements of juvenile blacktip reef sharks: evidence of dispersal and connectivity between coastal habitats and coral reefs

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Leukotrienes Affect Secretory Function of Ovarian Cells In Vitro*

The aim of this study was to determine which cells are the source of production and target for leukotriene (LTs) action within the bovine ovary. Luteal (CL, days 14–16 of the oestrous cycle), steroidogenic cells (LSC) and endothelial cells (LEC) of the bovine corpus luteum (CL), and granulosa cells (GC) were isolated enzymatically, cultured in a monolayer and incubated with LTC₄, LTB₄, Azelastine (an antagonist of LTC₄) or Dapsone (an antagonist of LTB₄). Then cells were collected for determination of mRNA expression for LT receptors (LTRs) and 5‐lipoxygenase (5‐LO) by real time RT‐PCR, and media were collected for determination of prostaglandin (PG)E₂, F_2α, progesterone (P4; LSC only), endothelin‐1 (ET‐1; LEC only) and 17‐β oestradiol (E2; GC only). The greatest mRNA expression for LTR‐II and 5‐LO were found in LEC, whereas LTR‐I mRNA expression did not differ among cell types. The level of PGE₂ increased after LTs treatment in each type of ovarian cell, excluding LTC₄ treatment in LEC. The secretion of PGF_2α was also increased by LTs, but decreased after LTB₄ treatment of LSC. In GC cultures, both LTs stimulated E2 secretion; in LEC cultures, LTB₄ stimulated whereas LTC₄ inhibited P4 secretion; in LEC cultures, LTC₄ stimulated but LTB₄ inhibited ET‐1 secretion. The results show that LTs are produced locally and are involved in PGs production/secretion in all examined cells (LSC, LEC and GC) of bovine ovary. Leukotriene treatment modulate secretion of E2, by GC, P4 by LSC and ET‐1 by LEC, which indicates that LTs are involved in regulation of ovarian secretory functions.     

Caecal disease in equids

Objective To review the breed, age, gender, clinical and laboratory findings, treatment and outcome of horses with caecal disease presented to a referral centre. Design Retrospective study of 96 cases. Procedure The breed, age, and gender of the study population were compared with the corresponding hospital population for the same period. The means (± SD) for clinical and laboratory findings were recorded for each caecal disorder. Treatment was categorised as medical or surgical, and outcome was recorded. Results Caecal diseases included impaction (40% of total cases), rupture associated with concurrent unrelated disease (13%), rupture with parturition (9%), rupture with no associated disease (5%), infarction (11%), torsion (9%), abscess or adhesion (7%), tumour (3%), and miscellaneous conditions (3%). The breed or gender of affected horses did not differ from the hospital population, although horses > 15 years were more frequently represented (P < 0.05). This age group was specifically more predisposed to caecal impaction (P < 0.05), as were Arabian, Morgan, and Appaloosa breeds (P < 0.05). In horses with caecal impaction transrectal examination was the most useful diagnostic procedure; 90% of affected horses treated medically were discharged while horses treated by typhlotomy alone, or typhlotomy and blind end ileocolostomy, had survival rates to discharge of 71% and 86%, respectively. Horses with caecal rupture associated with concurrent un-related disease showed no signs of impending rupture; all were receiving phenylbutazone, all were euthanased, and 50% had caecal ulceration at necropsy. Of horses with caecal rupture with parturition 56% had prior dystocia; in two-thirds the site of rupture was the ileocaecal junction and all were euthanased. Horses with caecal rupture with no associated disease died or were euthanased; rupture was idiopathic. Horses with caecal infarction usually had signs of abdominal pain and abdominal fluid changes consistent with peritonitis; transrectal examination was nonspecific, and typhlectomy was successful in seven of eight horses. Horses with caecal torsion had signs of severe, acute abdominal pain and typhlectomy was successful in three of five horses. Diagnosis of caecal adhesion or abscess was assisted by transrectal palpation in two of seven horses and surgical treatment was successful in two of five horses. A caecal tumour was diagnosed in three horses aged 20 years or older that presented with chronic weight loss. Other caecal diseases were uncommon. Conclusion Caecal disease is uncommon in equids but some specific features of the history and physical findings can alert the veterinarian to the possibility of caecal involvement in horses with gastrointestinal dysfunction. Medical or surgical therapy can be effective in horses where caecal rupture has not occurred.     

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.     

The occurrence of equine arteritis virus in Australia

This paper reports the first isolation of equine arteritis virus (EAV) in Australia and serological evidence of exposure to EAV in Australian horses. Twelve Standardbred stallions imported from North America were found to shed EAV in semen. One hundred and seven stallions were tested for serum antibodies to EAV and 73% of Standardbred stallions tested were seropositive as compared to 8% of Thoroughbred stallions. Serum antibody was detected in 71% of Standardbred mares, 6% of Standardbred racehorses and 1% of Thoroughbred mares and racehorses. Examination of stored serums demonstrated that EAV had been present in Australia since at least 1975.     

Surgical resection of a dysgerminoma in a mare

A mare was referred for further evaluation of a mass found in the left caudal abdomen during a routine postpartum reproductive palpation. The mare was clinically normal with no history of health problems. Ultrasonographic examination of the mass confirmed its presence, but the origin of the mass could not be accurately determined. Routine haematology and serum biochemistry results were within normal limits. The mare was initially treated conservatively with antibiotics, but the mass continued to increase in size, so it was surgically excised. The mass involved the left ovary. The mare showed transient abdominal pain after surgery, but developed no other complications and was in foal 7 months later. On histology, the mass was diagnosed as a dysgerminoma, a rare ovarian tumour of germ cell origin.