Effect of putrescine supplementation to in vitro maturation medium on embryo development and quality in cattle.

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.

Immunohistochemical and qPCR determination of the expression and serum level of anti-Müllerian hormone in pre-pubertal, intact and ovarian remnant syndrome detected bitches.

The aim of this study was to determine the serum concentrations, ovarian presence and expression of anti-Müllerian hormone (AMH) in pre-pubertal, bitches with signs of ovarian remnant syndrome (ORS) and intact bitches. In addition, we aimed to verify the suitability of serum AMH concentrations for diagnostic purposes in sterilized bitches and/or in suspected cases of ORS in the field of veterinary medicine. For this purpose, 36 healthy female dogs divided into six groups: proestrus, oestrus, dioestrus, anoestrus, pre-pubertal and ORS. Serum AMH concentrations were determined by electrochemiluminescence immunoassay, and ovarian presence and distribution of AMH was confirmed by immunohistochemical and qPCR techniques. According to the results of qPCR, while the expression values of AMH were at the highest concentrations in the proestrus and oestrus, there was a statistically significant decrease in these values at the later stages of the cycle (p < 0.05). According to hormone analysis, the serum AMH values of the ORS group had decreased significantly compared with the proestrus and oestrus (p < 0.05). Although serum AMH levels of ORS group were increased compared with anestrus and pre-pubertal groups, this increase was statistically non-significant (p > 0.05). Immunohistochemically, AMH expression was first observed in the granulosa cells of primordial follicles in folliculogenesis. Expression values were the highest in the proestrous and oestrus groups, but values from bitches in later stages of the cycle were statistically significant decrease in comparison with these groups (p < 0.05). As a result, AMH concentration and expression were found to be higher in proestrus and oestrus than in other periods (p < 0.05). In addition, the measurable level of AMH concentration in bitches with ORS is an indication that it can be used in the diagnosis of ORS.

Transcriptomic evaluation of metals detected in placenta.

This research aims to assess the concentration of metals in human and canine placentas from the same geographic area and to investigate how these metal levels influence gene expression within the placenta. Placentas of 25 dogs and 60 women who had recently given birth residing in Ankara, Turkey were collected and subjected to metal analysis using ICP-OES. Placentas with detectable metal levels underwent further examination including Next Generation Sequencing, transcriptional analysis, single nucleotide polymorphism investigation, and extensive scrutiny across various groups. For women, placentas with concurrent detection of aluminum (Al), lead (Pb), and cadmium (Cd) underwent transcriptomic analysis based on metal analysis results. However, the metal load in dog placentas was insufficient for comparison. Paired-end sequencing with 100-base pair read lengths was conducted using the DNBseq platform. Sequencing quality control was evaluated using FastQC, fastq screen, and MultiQC. RNA-sequencing data is publicly available via PRJNA936158. Comparative analyses were performed between samples with detected metals and "golden" samples devoid of these metals, revealing significant gene lists and read counts. Normalization of read counts was based on estimated size factors. Principal Component Analysis (PCA) was applied to all genes using rlog-transformed count data. Results indicate that metal exposure significantly influences placental gene expression, impacting various biological processes and pathways, notably those related to protein synthesis, immune responses, and cellular structure. Upregulation of immune-related pathways and alterations in protein synthesis machinery suggest potential defense mechanisms against metal toxicity. Nonetheless, these changes may adversely affect placental function and fetal health, emphasizing the importance of monitoring and mitigating environmental exposure to metals during pregnancy.

Live in same region, respond differently: Canine and human response to pollutants in placental accumulation.

Polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), polybrominated diphenyl ethers (PBDEs), and polycyclic aromatic hydrocarbons (PAHs) are endocrine-disrupting chemicals (EDCs). Their presence in the environment is particularly concerning in cases of fetal exposure, which is the most vulnerable period of life for both human and animals who share the same environment. Placenta, as a sample collected using noninvasive methods to screen EDCs, is a good indicator for potential fetal exposure. Although recent studies indicate that companion animal may correspond to human exposure, species-specific anatomo-morphological and metabolic differences are controversial. In this study, placenta samples of 60 women and 25 dogs living and giving birth within the same region were evaluated for the presence of PCB, OCP, PBDE, and PAH residues; where, socio-demographic factors were also assessed to identify the possible sources. Gas chromatography-mass spectrometry method was validated for the matrix, and among 45 screened and targeted pollutants, only 18 were found in human placentas. While the most frequently detected pollutants were DDTs, followed by PAHs and PCBs in decreasing order, the pollutants with the highest concentrations were PAHs, followed by PCBs and DDTs. Only five of the target contaminants were detected in the dog placentas. These results indicate that; as dogs have different bioaccumulation capacities and higher excretion rates than humans, the life-long effects of exposure to endocrine compound and possible consequences related to adverse health outcomes are expected to vary and concentrations cannot be directly correlated.

The effect of anti-müllerian hormone and progesterone concentrations on superovulation response and embryo yield in goats.

We aimed to evaluate the relationship of anti-Müllerian hormone (AMH) and progesterone concentrations with superovulation response in goats and to determine donors exhibiting better superovulation response by measuring AMH concentrations. For this, blood samples were collected from multiparous Angora goats (n = 24) for measuring the progesterone and AMH concentrations on the day the synchronization protocol was initiated (Day 0), on the day of the first FSH administration (Day 9), on the day the progesterone source was removed (Day 11), and on the day of uterine flushing. Descriptive statistics (mean, standard deviation, median, minimum value, maximum value, and percentile) were given for superovulation response and embryo yield. To compare the differences between the two groups, the Student's t-test was used. The relationship between two continuous variables was assessed by the Pearson Correlation Coefficient. The AMH cutoff values in superovulation responses were evaluated by ROC analysis on the day the synchronization protocol was initiated. A strong positive correlation was found between the AMH concentrations measured on the day the synchronization protocol was initiated (Day 0), on the day of the first FSH administration (Day 9), and on the day of removal of the progesterone source (Day 11) and the count of total corpus luteum (CL), total oocyte/embryo, transferable embryo, and Code I quality embryo (P < 0.05). Furthermore, AMH concentration increased on the day the synchronization protocol was initiated, the donor's superovulation response increased as well. The cutoff value was 4.74 ng/ml, as assessed by the ROC curve analysis conducted for selecting donors exhibiting better superovulation responses. The sensitivity and specificity of the selected cutoff value were found to be quite high (P < 0.01). However, a positive correlation was noted between the progesterone concentrations measured on the day of uterine flushing and total CL count, total oocyte/embryo count, transferable embryo count, and Code I quality embryo count (P < 0.01). In conclusion, it was determined that an increase in AMH concentrations in goats led to an increase in the total CL count, embryo count, and embryo quality and that AMH measurement could be used to identify donors that responded better to superovulation. Additionally, a positive correlation was found between the progesterone concentration measured on the day of uterine flushing and the total CL count, transferable embryo count, and embryo quality.

Aglepristone and cloprostenol combination in the termination of late-term pregnancy in queens.

OBJECTIVES: Termination rates for the highly recommended aglepristone (AGL) treatment are low in late-term pregnancy in queens. We studied the effects of an AGL and cloprostenol (CLO) combination in the termination of late-term pregnancy. METHODS: Pregnant queens were assigned to two groups. Queens in the AGL group (n = 10) received AGL 10 mg/kg, twice, 24 h apart. Queens in the AGL-CLO group (n = 9) were additionally injected with a single dose of CLO (5 µg/kg) 24 h after the second dose of AGL. Progesterone, 17beta(ß)-oestradiol, cortisol, oxytocin and prostaglandin F2alpha (PGF2α) metabolite were measured in sera obtained at days 0, 1 and 2, and on the day of abortion. RESULTS: Average gestational age in both groups was similar (AGL 38.61 ± 0.91 days vs AGL-CLO 39.39 ± 1.35 days; P >0.05). Termination rates were 80% and 100% in the AGL and AGL-CLO groups, respectively (P <0.05). Fetal expulsion time was significantly longer (P <0.001) in the AGL group (96.9 ± 6 h) compared with the AGL-CLO group (69.8 ± 3.3 h). Duration of abortion was 19.8 ± 2.6 h and 12.6 ± 1.4 h in the AGL and AGL-CLO groups, respectively (P <0.05). Both treatments were well tolerated. Significantly (P <0.05) lower serum progesterone concentrations were observed in both groups at the day of abortion and concentrations in the AGL-CLO group (4.19 ± 0.80 ng/ml) were lower than in the AGL group (9.89 ± 2.21 ng/ml; P <0.05). CONCLUSIONS AND RELEVANCE: AGL and CLO combination increases pregnancy termination rate in late-term pregnant queens. In addition, CLO contributes to a decrease in luteal function in AGL-treated late-term pregnant queens.