Melatonin reduces apoptotic cells,SOD2 and HSPB1 and improves the in vitro production and quality of bovine blastocysts

Effects of adding different concentrations of melatonin (10^?7, 10^?9 and 10^?11 M) to maturation (Experiment 1; Control, IVM  + 10^?7, IVM  + 10^?9, IVM  + 10^?11) and culture media (Experiment 2; Control, IVC  + 10^?7, IVC  + 10^?9, IVC  + 10^?11) were evaluated on in vitro bovine embryonic development. The optimal concentration of melatonin (10^?9 M) from Experiments 1–2 was tested in both maturation and/or culture media of Experiment 3 (Control, IVM  + 10^?9, IVC  + 10^?9, IVM /IVC  + 10^?9). In Experiment 1, maturated oocytes from Control and IVM  + 10^?9 treatments showed increased glutathione content, mitochondrial membrane potential and percentage of Grade I blastocysts (40.6% and 43%, respectively). In Experiment 2, an increase in the percentage of Grade I blastocysts was detected in IVC  + 10^?7 (43.5%; 56.7%) and IVC  + 10^?9 (47.4%; 57.4%). Moreover, a lower number and percentage of apoptotic cells in blastocysts were observed in the IVC  + 10^?9 group compared to Control (3.8 ± 0.6; 3.6% versus 6.1 ± 0.6; 5.3%). In Experiment 3, the IVC  + 10^?9 treatment increased percentage of Grade I blastocysts with a lower number of apoptotic cells compared to IVM /IVC  + 10^?9 group (52.6%; 3.0 ± 0.5 versus 46.0%; 5.4 ± 1.0). The IVC  + 10^?9 treatment also had a higher mRNA expression of antioxidant gene (SOD 2) compared to the Control, as well as the heat shock protein (HSPB 1) compared to the IVM  + 10^?9. Reactive oxygen species production was greater in the IVM /IVC  + 10^?9 treatment group. In conclusion, the 10^?9 M concentration of melatonin and the in vitro production phase in which it is used directly affected embryonic development and quality.     

Effect of excess degradable intake protein on early embryonic development, ovarian steroids, and blood urea nitrogen on days 2, 3, 4, and 5 of the estrous cycle in mature ewes

Two trials were conducted to determine whether feeding excess degradable intake protein (DIP) during a synchronized estrous cycle and the first 5 d after breeding alters early embryonic development, ovarian steroids, or BUN concentrations in ewes. Ewes were group-fed in Trial 1 (T1) and individually fed in Trial 2 (T2) either 100 (control; T1, n = 15; T2, n = 12) or 200% (high-protein; T1, n = 16; T2, n = 12) of the NRC protein recommendation for maintenance during a synchronized estrous cycle until surgery in the next cycle. Ampullae (AMP), isthmi (IST), and uterine horns (UT) of high-protein and control ewes were removed on d 2 (T1), 3 (T2), 4 (T1), or 5 (T2) after breeding. In T1, jugular blood samples were taken once daily starting on d 2 of the synchronized cycle, and in T2 on d 2, 9, 15, 16, and 17, and in both trials from estrus (d 0) to the day of surgery. Ampullae, IST, and UT flushings were examined microscopically for the presence of embryos, embryo condition, and embryo cell number. There was no trial x treatment interaction (P > or = 0.10), so data for both trials were pooled. Concentrations of BUN were higher (P < 0.05) in high-protein-fed ewes than in control ewes during the synchronized cycle and the first 5 d of the next cycle. Progesterone concentrations of the synchronized cycle did not differ (P > 0.10) between treatments. During the first 5 d of the next cycle, estradiol-17beta concentrations were lower (P = 0.06) in high-protein-fed than in control ewes. Progesterone increased (P < 0.05) to higher concentrations by d 5 in high-protein-fed ewes than in control ewes. More (P < 0.05) embryos were found in AMP of high-protein-fed ewes than in AMP of control ewes on d 4. Fewer (P = 0.05) embryos were found in UT of high-protein-fed ewes than in UT of control ewes on d 4. More embryos were found in UT of high-protein-fed ewes than in UT of control ewes on d 5. Fewer (P = 0.05) embryos were found in IST of high-protein ewes than in the IST of control ewes on d 5. Embryos of high-protein-fed ewes had more (P < 0.05) cells than embryos from control fed ewes on d 5. Feeding ewes excess DIP protein during an estrous cycle and the first 5 d after breeding initially impeded embryo transport; thereafter, embryo transport and development through the oviduct was accelerated.     

Immunogenicity of bovine parainfluenza type 3 virus proteins encapsulated in nanoparticle vaccines,following intranasal administration to mice

Stimulation of long lasting, protective immunity to respiratory viruses is often difficult to achieve with conventional respiratory vaccines. Polymeric nanoparticles, incorporating viral proteins have been shown to offer sustained release of antigen, with consequent prolongued stimulation of the respiratory immune system. In this paper the efficacy of two nanoparticle vaccines (poly-lactide-co-glycolide, PLGA; polymethylmethacrylate, PMMA), incorporating proteins of bovine parainfluenza type 3 virus (BPI-3) was investigated. As a preliminary to experiments in calves, it was considered essential to demonstrate immunogenicity of the experimental vaccine in mice. Mice immunised with PLGA nanoparticles, containing BPI-3 proteins, developed higher levels of virus-specific antibody than mice immunised with the PMMA vaccine or with soluble viral proteins alone. Immunoblotting using serum from the vaccinated mice, demonstrated strong reactions against the major BPI-3 proteins.     

Liver biopsy techniques for adult horses and neonatal foals to assess copper status

Objective To evaluate standing, percutaneous, ultrasound-guided, transthoracic liver biopsy in mares, and transabdominal, laparoscopically-guided, liver biopsy under general anaesthesia in foals, as techniques for obtaining tissue for assessment of copper status. The techniques were evaluated with respect to ease of use and effect on the animal.
Procedure Twenty of 24 Thoroughbred mares and 21 of their foals were biopsied. The animals were part of a larger study of the effect of copper supplementation on copper status and the prevalence of developmental orthopaedic disease. Livers were also collected from unrelated horses and sampled to investigate the variability in the distribution of copper in liver tissue.
Result The biopsy technique caused no lasting effect on the mares, but there was an increased risk of viscus penetration associated with taking multiple biopsy cores. The use of ultrasonography to scan the target area for the liver identified four cases that were not appropriate candidates for liver biopsy, because of large intestine being located in the biopsy area. In the foals there were no serious postoperative adverse effects, nor was there any evidence of problems caused by the procedure when the abdomen was examined post-mortem at 5 months of age. In livers collected to investigate the variability of copper concentration, copper appeared to be relatively evenly distributed through the liver.
Conclusion Standing, percutaneous, ultrasound-guided, transthoracic liver biopsy in mares, and transabdominal, laparoscopically-guided, liver biopsy under general anaesthesia in foals are convenient procedures for obtaining liver tissue for assessing copper status in horses. The use of ultrasound to identify liver tissue is recommended, especially in older mares.     

Evaluation of use of dimethyl sulfoxide for intra-articular lavage in clinically normal horses

The antebrachiocarpal and tarsocrural joints of 10 adult horses were randomly assigned to 1 of 4 groups. Groups were formulated and were treated as follows: group 1, control (arthrocentesis only); group 2, buffered lactated Ringer solution; group 3, 10% dimethyl sulfoxide (DMSO; w/v) in lactated Ringer solution; and group 4, 30% DMSO (w/v) in lactated Ringer solution. Joints were lavaged once with the respective solution. Prior to lavage and on days 1, 4, and 8 after lavage, all horses were evaluated for lameness and joint effusion; synovial fluid total and differential WBC counts, synovial fluid total protein concentration, and mucin clot quality were determined. Horses were euthanatized on day 8, and joints were evaluated grossly, histologically, and histochemically. Significant difference was not observed in effect of lactated Ringer solution, 10% DMSO, and 30% DMSO on any measured variable. At 24 hours after treatment, significant (P less than 0.05) difference in synovial fluid WBC numbers and total protein concentration was detected between control and treated joints. Eighty percent of lavaged joints had effusion 24 hours after treatment, compared with 30% of control joints. Gross, histopathologic, or histochemical differences were not detected between treated and control joints. Results of the study indicate that buffered lactated Ringer, 10% DMSO, and 30% DMSO solutions induce similar inflammatory changes in articular structures and significantly greater inflammatory reaction than does arthrocentesis alone.     

Rhodococcus equi-associated osteomyelitis in foals

Two cases of Rhodococcus equl infection in foals are described, in which osteomyelitis was a feature. Because rhodococcal infection is usually low grade and chronic, and because the signs of early metaphysitis can be subtle, any articular or periarticular swelling in a foal from a farm with a history of rhodococcosis should be strongly suspected to be associated with R equl until proven otherwise.     

Effects of two adenoviruses (type 1 and type 8) on functional properties of bovine alveolar macrophages in vitro.

Cultures of bovine alveolar macrophages were inoculated with type-1 and type-8 adenoviruses, initially isolated from calves with respiratory tract disease, and functional properties of the cells were observed over a period of 10 to 11 days. Both viruses replicated in macrophages; viral titers were low (less than 3.75 log10 TCID50/0.1 ml), and intranuclear inclusions were detected by indirect immunofluorescence in 5 to 10% of the cells from 3 days after inoculation. Highest titers were induced by type-1 adenovirus, which also induced the greatest functional changes. Expression of Fc and complement receptors was reduced by both viruses, although the greatest effects were seen with type 1. Phagocytosis of Candida krusei cells was reduced following type 1 infection, whereas phagocytosis in type-8-infected cells was not different from that of noninfected macrophages. Ability to kill ingested Candida cells also was reduced following type-1 infection, whereas type-8-infected macrophages had lower killing ability only at 2 to 4 days after inoculation. Neither virus had substantial effects on the production of neutrophil chemotactic factors by the macrophages.     

Diagnosis and management of a patent urachus in a white rhinoceros calf (Ceratotherium simum simum).

A 10-day-old female southern white rhinoceros calf (Ceratotherium simum simum) was diagnosed with a patent urachus after urine was observed dribbling from the umbilicus. After being separated from its mother, the animal was sedated with i.m. butorphanol and anesthetized with isoflurane in oxygen for surgical correction of the patent urachus. Mild postoperative complications involved seroma formation and partial skin incision dehiscence, which necessitated three follow-up immobilizations for reevaluation and treatment of the surgical site. Histopathology did not reveal an infectious etiology as the cause for the complications or for the patent urachus. The etiology of the patent urachus in this animal remains undetermined. This report represents the first documented case of a patent urachus in a white rhinoceros.