This study aimed to determine the effect of microfluidic sperm sorting chip on embryo development and quality in the sperm treatment step in in vitro embryo production in cattle. Only A‐quality oocytes obtained from the ovaries of Holstein cattle were included in the study. These oocytes were first placed in in vitro maturation medium, and matured oocytes were randomly divided into two groups at the 24th hour of maturation. Oocytes in the first group with the Microfluidic Sperm Sorting Chip (MFSC, n = 154) were put into a fertilization medium with spermatozoa prepared with microfluidic sperm sorting device. Oocytes in the second group (Con, n = 169) were fertilized with spermatozoa prepared by the routine sperm treatment step of the commercial company. The rate of cleavage (85.71% vs. 76.33%, respectively) and of reaching the blastocyst (44.15% vs. 32.54%, respectively) in the MFSC group were higher than the control group. In addition, it was determined that the numbers of ICM (45.8 ± 2.04 vs. 39.2 ± 1.85, respectively), TE (122.13 ± 2.19 vs. 115.0 ± 2.61, respectively), TC (167.93 ± 2.89 vs. 154.2 ± 2.62, respectively) increased in the MFSC group compared to the control group. A statistically significant difference was found in the number of cells with apoptosis per embryo (5.14 ± 0.77 vs. 11.91 ± 0.79) and apoptotic index rates (3.06 ± 0.47 vs. 7.72 ± 0.55%) of the MFSC and Con groups. As a result, we concluded that using microfluidic sperm sorting chips during sperm treatment in bovine IVEP increases the rate of reaching the blastocyst, embryo development, and quality and reduces the possibility of apoptosis in developing blastocysts. For this reason, it is also thought that the use of microfluidic sperm sorting devices during sperm treatment in bovine IVEP may be a new alternative in this field.

This study aimed to investigate the effect of putrescine supplementation to maturation medium during in vitro embryo production in cattle on maturation and embryo development/quality. Oocytes obtained from the ovaries of Holstein cattle were used in the study. Obtained cumulus-oocyte complexes were evaluated according to morphological structure, cytoplasmic features, and cumulus cell number, and only Category-I ones were used in the study. Before the in vitro maturation step, oocytes were randomly divided into two groups. In the first group (Putrescine group, n = 159), 0.5 mM putrescine was added to the maturation medium before in vitro maturation. No addition was applied to the maturation medium of the second group (Control group, n = 149). Cumulus expansion degrees of oocytes following maturation (Grade I: poor, Grade II: partial, and Grade III: complete) were determined. In addition, the meiosis of oocytes after maturation was evaluated by differential staining. Then the oocytes were left for fertilization with sperm and finally, possible zygotes were transferred to the culture medium. After determining the developmental stages and quality of the embryos after in vitro culture, only the embryos at the blastocyst stage were stained with the differential staining method to determine the cell numbers. When the cumulus expansion degrees of the groups were evaluated, the Grade III cumulus expansion rate in the putrescine group was higher than the control group (74.21% and 60.4%; respectively) and the Grade I expansion rate (11.95% and 26.17%; respectively) was found lower (p < .05). When the resumption of meiosis was evaluated according to the cumulus expansion degrees, it was determined that the rate of resumption of meiosis increased as the cumulus expansion increased. In addition, the cleavage rates of oocytes and reaching the blastocyst in the putrescine group were found to be higher than in the control group (p < .05). Moreover, inner cell mass, trophectoderm cells, and total cell counts were found to be higher in blastocysts obtained after the putrescine supplementation to the maturation medium compared to the control group (p < .05). As a result, it was determined that the putrescine supplementation to the maturation medium during in vitro embryo production in cattle increased the degree of cumulus expansion and the rate of resumption of meiosis. In addition, putrescine supplementation was thought to increase the rate of reaching the blastocyst of oocytes due to better cell development in embryos.

Background: Pyometra is a bacterial and hormone-induced reproductive disease that occurs in the post-estrus luteal phase in intact queens. Pyometra is more common in the diestrus period due to the high progesterone concentration (in queens that mated, spontaneously ovulated, or were induced to ovulate). However, it can also be seen due to the use of exogenous hormones such as progesterone for the suppression of estrus. More research is needed in cases of pyometra in queens, as well as in bitches. Because, considering that the pathogenesis and characteristics of feline pyometra is similar to bitches, studies on pyometra-affected bitches are taken as reference in studies and applications on queens. From this point of view, the aims of this study were to reveal the changes in complete blood count, blood gas, and serum biochemistry parameters in feline pyometra cases and to determine the correlation between the mentioned parameters. Materials, Methods & Results: In the study, a total of 25 female cats of different breeds were used, between the ages of 6 months and 7 years, 15 were diagnosed with pyometra, and 10 healthy. Anamnesis, clinical findings, and ultrasonographic examinations were used in the diagnosis of pyometra. Abdominal ultrasonography was performed on queens brought to the clinic with complaints such as anorexia, polydipsia, polyuria, abdominal tension, and fever. The control group (n =10) consisted of queens that were introduced to the clinic and were reproductively healthy. Before any treatment in queens with pyometra and the control group, 1 mL blood samples were taken from v. cephalica to evaluate complete blood count, blood gases and serum biochemistry parameters. In complete WBC, Lym, Mon, Gra, RBC, Hb, HCT, MCV, MCH and PLT parameters and, blood gas parameters such as pH, pCO2, pO2, sO2, Na, K, Cl, lactate, glucose, HCO3, and BE were also evaluated in taken blood samples. Biochemical parameters BUN, creatinine, ALT, AST, ALP, amylase, T.BIL, D.BIL, P, CHOL, TG, LDH, TP, CPK, ALP, Ca, GGT were measured in serum samples. After examination and laboratory analysis, ovariohysterectomy was performed on queens as a treatment. Granulocyte, WBC, HCT and MCH levels of the pyometra group were higher (P < 0.05) and Lym levels were lower (P < 0.05) compared to the control group. According to these results, pH, HCO3, and BE were lower (P < 0.05) in queens with pyometra than those in the control group, while Na and lactate parameters were higher (P < 0.05). According to the results of biochemical analysis, it was determined that BUN, creatinine levels, GGT, and LDH enzyme activities were found to be higher in the pyometra group compared to the control group, while the Ca level was found to be low (P < 0.05). A positive correlation was observed between BUN and creatinine and LDH, WBC, granulocyte, HCT, and lactate, and a negative correlation between lymphocytes, pH, and BE in the correlation analysis performed on queens with pyometra and control group. However, a positive correlation was observed between creatinine and LDH and HCT, and a negative correlation between lymphocyte, pH and BE. Discussion: There is not enough information about pyometra in queens. As a result, it was determined that there were significant changes in complete blood count, blood gases and serum biochemical parameters in queens with pyometra in this study. These changes were generally thought to be related to dehydration and sepsis or endotoxemia. In addition, it was evaluated that prerenal azotemia occurring in pyometra affected queens may cause renal dysfunction. For this reason, it is thought that the results obtained in the presented study may contribute to the diagnosis, treatment, and prognosis of pyometra cases in queens. Keywords: female cats, blood gas analysis, complete blood count, correlation, serum biochemistry.