Optimizing injection time of GFP plasmid into sheep zygote

Microinjection of exogenous DNA into the cytoplasm of matured oocytes or zygotes is a promising technique to generate transgenic animals. However, the data about the microinjection time and procedure in sheep are limited and have not treated in detail. To obtain more in-depth information, the Sarda sheep oocytes from abattoir-derived ovaries were subjected to IVM and IVF. Then, the GFP plasmid as a reporter gene was injected into the cytoplasm of MII oocytes (n: 95) and zygotes at different post-insemination intervals (6–8 hpi, n: 120; 8–10 hpi, n: 122; 10–12 hpi, n: 110 and 12–14 hpi, n: 96). There were no significant differences in the cleavage rates between the groups. However, blastocyst rate of injected zygotes at all-time intervals was significantly lower than injected MII oocytes and control group (p < 0.05). Interestingly, the proportion of GFP-positive embryos was higher at 8–10 hpi compared with other injected groups (4 % versus 0 %, p  < 0.01). Among these, the proportion of mosaic embryos was high and two of those embryos developed to the blastocyst stage. In conclusion, we settled on the cytoplasmic microinjection of GFP plasmid at 8–10 hpi as an optimized time point for the production of transgenic sheep and subsequent experiments.     

Clinical presentation and management of suspected ribavirin toxicosis in a dog

A 5-month-old pit bull terrier was presented for evaluation of progressive lethargy, vomiting, diarrhea, and anorexia 45 hours after ingestion of 625 mg/kg body weight (BW) (9000 mg) of the antiviral medication, ribavirin. Abnormalities that were detected included dehydration, tachycardia, elevated liver enzymes, and prolonged prothrombin time. The dog was discharged after 5 days of aggressive supportive care consisting of intravenous fluids, antiemetics, gastroprotectants, hepatoprotectants, dextrose supplementation, and vitamin B/K1 supplementation.     

Influences of Sex and Age on the Hematological Profile of the Jundiá (Silver Catfish) Rhamdia quelen

In this study, sex and age influenced the hematological profiles of Jundiá (Silver Catfish) Rhamdia quelen. Females showed lower levels of hemoglobin, young fish increased lymphocyte counts, and older fish increased hematocrit values. These results indicate that, depending on age and sex, the fish have disparate hematological profiles. For this reason, it is important to consider the sex and age of an R. quelen when examining the impact of environmental and management factors on this species in terms of their hematological profiles.

Received May 24, 2015; accepted March 24, 2016 Published online August 2, 2016     

Neutrophil myeloperoxidase measurements in plasma, laminar tissue, and skin of horses given black walnut extract

OBJECTIVE: To compare measurements of myeloperoxidase (MPO) in plasma, laminar tissues, and skin obtained from control horses and horses given black walnut heartwood extract (BWHE). ANIMALS: 22 healthy 5- to 15-year-old horses. PROCEDURES: Horses were randomly assigned to 4 groups as follows: a control group given water (n = 5) and 3 experimental groups given BWHE (17) via nasogastric intubation. Experimental groups consisted of 5, 6, and 6 horses that received BWHE and were euthanatized at 1.5, 3, and 12 hours after intubation, respectively. Control horses were euthanatized at 12 hours after intubation. Plasma samples were obtained hourly for all horses. Laminar tissue and skin from the middle region of the neck were harvested at the time of euthanasia. Plasma and tissue MPO concentrations were determined via an ELISA; tissue MPO activity was measured by use of specific immunologic extraction followed by enzymatic detection. RESULTS: Tissues and plasma of horses receiving BWHE contained significantly higher concentrations of MPO beginning at hour 3. Laminar tissue and skin from horses in experimental groups contained significantly higher MPO activity than tissues from control horses. Concentrations and activities of MPO in skin and laminar tissues were similar over time. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, BWHE administration causes increases in MPO concentration and activity in laminar tissue and skin and the time of increased MPO concentration correlates with emigration of WBCs from the vasculature. These findings support the hypothesis that activation of peripheral WBCs is an early step in the pathogenesis of acute laminitis.