Background: Porcine embryonic development is widely utilized in the medical industry. However, the blastocyst development rate in vitro is lower compared to in vivo . To address this issue, various supplements are employed. Extracellular vesicles (EVs) play the role of communicators that carry many bioactive cargoes. Additionally, the contents of EVs can vary on the estrous cycle. Methods: We compared the effects of adding EVs derived from porcine uterine fluid (UF), categorized as non-EV (G1), EVs in estrus (G2) and EVs in diestrus (G3). After in vitro culture (IVC) was performed in three different groups, cleavage rate and blastocyst development rate were examined. In addition, glutathione (GSH) and reactive oxygen species (ROS) levels were measured 2 days after activation to assess oxidative stress. Results: Using NTA and cryo-TEM, we confirmed the presence of EVs with sizes ranging from 30 nm to 200 nm, that the particles were suitable for analysis for analysis. In IVC data, the highest cleavage rate was observed in G2, which was significantly different from G1 but not significantly different from the next highest, G3. Similarly, the highest blastocyst development rate was observed in G2, which was significantly different from G1 but not significantly different from the next highest, G3. Conclusions: These results indicate that estrus derived EVs contain biofactors beneficial for early blastocyst development, including GSH which protects the blastocyst from oxidative stress. Additionally, although diestrus-derived EVs are expected to have some effect on blastocyst development, it appeared to be less effective than estrus-derived EVs.

A 10-year-old intact female Cocker Spaniel weighing 15.3 kg presented with dysbasia. The dog had hyperglycemia and glucosuria at presentation. Neurological examination and magnetic resonance imaging on the following day indicated the onset of cervical disk disease. Four days later, the patient returned due to sudden vomiting and anorexia with severe dehydration. Abnormal laboratory findings included severe hyperglycemia with ketonuria. Plasma progesterone concentration was very high at presentation. She was hospitalized for critical care, and insulin administration with fluid therapy was initiated. Mean blood glucose concentration showed a gradual decline, and hyperglycemia with glucosuria disappeared four days after initiation of insulin therapy. This case demonstrated that high plasma progesterone concentration during diestrus can contribute to development of diabetes mellitus in intact female dogs.

Serial ultrasonography was conducted on Miniature Schnauzer bitches, on purpose to observe the ultrasonographic appearance of normal ovaries and ovarian structures during the estrous cycle. The size of ovaries was increased from on Day-12 (Day-0=ovulation day) to on Day-8 and there was no significantly different between both ovaries. The ovaries were recognized by its proximity to the caudal renal pole and appeared moderately echogenic oval shape with a smooth contour. The size of follicles was increased from on Day-12 to on Day-0 and there was no significantly different between both ovaries. The number of follicles was increased from on Day-12 to on Day-0 and there was no significantly different between both ovaries. The follicles were small anechoic fluid-filled structures in early of proestrus, more increased, and indistinguished from each follicles in late of proestrus. The size of corpora lutea was increased from on Day-0 to on Day-8 and there was no significantly different between both ovaries. The number of corpora lutea was increased from on Day-0 to on Day-38 and there was no significantly different between both ovaries. The corpora lutea were small anechoic cavity and thin hyperechoic wall in early of diestrus, became more hyperechoic, and increased homogenous structures. The results of this study would be useful for differential diagnosis between normal and abnormal structures of ovaries.