The evaluation of xenotransplantation of feline ovarian tissue vitrified by needle immersed vitrification technique into male immunodeficient mice.

In this study, the efficiency of the "Needle Immersed Vitrification" technique was tested on cryopreserved feline ovarian tissue. For vitrification, ovarian fragments (0.5-1.5 mm2) from each ovary were collected; the grafts were exposed to 7.5-15% ethylene glycol and 7.5-15% dimethyl sulfoxide at room temperature and stored in liquid nitrogen at least 1 week. Morphologic examinations, expression of genes such as B cell lymphoma 2, B-cell lymphoma-2-associated X protein, Bone morphogenetic protein 15, zone of polarizing activity, zona pellucida C protein and DNA (cytosine-5)-methyltransferase 1, ultrastructural analysis and viability tests were carried out from collected grafts. Light microscopy examinations revealed the percentage of morphologically normal primordial follicles in a fresh group which was significantly higher than the treatment groups (p < 0.001). Terminal deoxynucleotidyl transferase dUTP nick end labeling and anti-caspase-3 staining observed in oocytes, follicle cells, interstitial tissue showed higher rates of apoptosis for post-vitrification and -transplantation groups than freshly grafted ovarian tissues. Furthermore, we observed significant downregulation of zone of polarizing activity and zona pellucida C protein gene expression in vitrified ovarian tissue grafts than in the fresh grafts (p < 0.05). In conclusion, we suggest that the needle immersed vitrification method is a convenient, cheap, and feasible vitrification method for cat ovarian tissues. However, further studies need to be performed to determine more optimal vitrification solutions and equilibration times for the needle immersed vitrification method in order to adapt it for cat ovaries.

The genetic diversity of bovine papillomaviruses (BPV) from different papillomatosis cases in dairy cows in Turkey.

Papillomaviruses (PVs) are epitheliotropic viruses that cause benign proliferative lesions in the skin (warts or papillomas) and mucous membranes of their natural hosts. In bovines specifically, 13 types of Bovine papillomaviruses (BPVs) are currently described in the literature, although the actual number may be greater than 20. BPV types are classified into four genera based on homology within the genomic regions of the L1 ORF, the most conserved sequence. This study conducted molecular typing of BPV in dairy cows with different papillomatosis cases and investigated the presence of co-infections across distinct BPV types in the same sample. After carrying out PCR using degenerate primers and type specific primers, 35 BPV suspected samples were detected as positive for BPV and these samples were used for typing using sequence analysis/PCR with type-specific primers. This analysis identified BPV-1, -2, -3, -4, -6, -7, -9 and -10, new putative types (BPV/BR/UEL6-like viruses) and the previously described putative type viruses (BAPV-6) in the 35 BPV-positive samples. In addition, co-infections across different BPV types were widely detected in the BPV-positive samples.  This study shows that PCR assays using degenerate primers to amplify partial fragments of the L1 gene followed by sequencing is useful for genotyping BPV. However, results need confirmation using type-specific primers in order to consider co-infections. In addition, this study identified a new putative type (in the same cluster as BPV/BR/UEL6-like viruses) and the previously described putative type viruses (BAPV-6) in teat papillomatosis of Turkish dairy cows. The study shows that it is essential to identify BPV types and their prevalence/distribution, and also to determine the clinical consequences of infection for the development of prophylactic and/or therapeutic procedures.

Detection and characterisation of sheep-associated malignant catarrhal fever infection from ruminants by using tegument and gB gene sequences of OvHV-2.

In this study, positive blood and organ samples were obtained from different mixed herds of sheep and cattle against ovine herpesvirus 2 (OvHV-2) infection. Target-positive DNA was sequenced and compared with worldwide distributed OvHV-2 sequences. Tegument gene (422 base pairs) and glycoprotein B (gB) gene (2800 base pairs) amplicons of OvHV-2 genome were used for understanding of epidemiology of malignant catarrhal fever (MCF) infection in Turkey. The results of nucleotide sequencing of polymerase chain reaction (PCR) products indicated presence of sheep-associated form for MCF infection in Turkey. Although the obtained sequences were genetically different from each other, it was found that genetic variations were limited.

Effects of blood beta-hydroxybutyric acid levels on the response to Ovsynch in primiparous cows

The aim of this study was to investigate the effects of low and moderate blood β-hydroxybutyric acid (BHBA) levels in primiparous cows during the early postpartum period on follicle development and pregnancy rate after the Ovsynch protocol. Holstein primiparous cows with low (n = 85) and moderate (n = 71) BHBA concentrations were used as experimental animals. At 7- 8 weeks postpartum, blood samples were collected and the Ovsynch protocol was performed on the same day. Ultrasonographic examination was performed to determine the dominant follicle (DF) diameter and presence of corpus luteum on the day of the first and second gonadotropin releasing hormone (GnRH1 and GnRH2) injections and prostaglandin F2 alpha (PGF2α) injection as well as 24 and 48 h after the GnRH1 and PGF2α injections. The ovulatory response to GnRH1 and pregnancy rate at day 30 and day 60 post insemination were also evaluated. No significant differences were observed between groups in the DF diameter on the day of GnRH1 and PGF2α injections, and at 24 and 48 h after these injections, as well as in the ovulation and atresia response to GnRH1 (P > 0.05). Significant differences were observed between groups in DF diameter (low-BHBA group, 13.8 ± 0.2 mm; moderate-BHBA group, 13.2 ± 0.2 mm; P = 0.039) at the time of insemination, and in pregnancy rates at 30 days (low-BHBA group, 38.82%; moderate-BHBA group, 21.13%, P = 0.017) and 60 days after insemination (37.6 and 18.3%, respectively; P = 0.008). Our findings suggest that moderate blood levels of BHBA (0.80-1.19 mmol/L) in primiparous cows during the early postpartum period affect the response to the Ovsynch protocol.(AU)

Effects of blood beta-hydroxybutyric acid levels on the response to Ovsynch in primiparous cows

The aim of this study was to investigate the effects of low and moderate blood β-hydroxybutyric acid (BHBA) levels in primiparous cows during the early postpartum period on follicle development and pregnancy rate after the Ovsynch protocol. Holstein primiparous cows with low (n = 85) and moderate (n = 71) BHBA concentrations were used as experimental animals. At 7- 8 weeks postpartum, blood samples were collected and the Ovsynch protocol was performed on the same day. Ultrasonographic examination was performed to determine the dominant follicle (DF) diameter and presence of corpus luteum on the day of the first and second gonadotropin releasing hormone (GnRH1 and GnRH2) injections and prostaglandin F2 alpha (PGF2α) injection as well as 24 and 48 h after the GnRH1 and PGF2α injections. The ovulatory response to GnRH1 and pregnancy rate at day 30 and day 60 post insemination were also evaluated. No significant differences were observed between groups in the DF diameter on the day of GnRH1 and PGF2α injections, and at 24 and 48 h after these injections, as well as in the ovulation and atresia response to GnRH1 (P > 0.05). Significant differences were observed between groups in DF diameter (low-BHBA group, 13.8 ± 0.2 mm; moderate-BHBA group, 13.2 ± 0.2 mm; P = 0.039) at the time of insemination, and in pregnancy rates at 30 days (low-BHBA group, 38.82%; moderate-BHBA group, 21.13%, P = 0.017) and 60 days after insemination (37.6 and 18.3%, respectively; P = 0.008). Our findings suggest that moderate blood levels of BHBA (0.80-1.19 mmol/L) in primiparous cows during the early postpartum period affect the response to the Ovsynch protocol.