Comparison of the efficacy of three concentrations of retinoic acid for transdifferentiation induction in sheep marrow-derived mesenchymal stem cells into male germ cells.

Recent studies have shown the unique role of retinoic acid (RA) in the induction of transdifferentiation in mesenchymal stem cells (MSCs) into germ cells (GCs). This study is the first study that compares the efficacy of three different concentrations of RA for the production of male GCs in vitro. Male sheep marrow-derived MSCs (MMSCs) were treated with the following concentrations of RA: 1 µm (RA1), 5 µm (RA2) and 10 µm (RA3) for a period of 21 days. The production of male GCs was evaluated by the assessment of expressions of GC-specific markers (by RT-PCR, qRT-PCR and immunocytochemistry), morphological characteristics and changes in alkaline phosphatase (ALP) activity. All three concentrations created male GC features. RA treatment upregulated the expressions of VASA and beta1 INTEGRIN and downregulated PIWIL2 and OCT4. DAZL was not expressed by RA treatment. Interestingly, immunocytochemistry detected PGP 9.5 expression in all treatment groups, with the highest expression noted in the RA3 group (P < 0.05). GC-like cells along with increased ALP activity were observed in all treated cultures, too. Finally, results showed that 10 µm RA has the most efficiency for transdifferentiation induction in MMSCs and production of male GCs in vitro.

Molecular typing of Staphylococcus aureus isolated from bovine mastitis based on polymorphism of the coagulase gene in the north west of Iran.

The objective was to investigate the genotypic characteristics and distribution of Staphylococcus aureus in 9 dairy herds of Tabriz and Urmia regions which are located in east and west Azerbaijan provinces, respectively, Iran. In this study 58 S. aureus isolates were recovered from 370 milk samples of cows with clinical and subclinical mastitis. S. aureus isolates were identified on the basis of the cultural and biochemical properties as well as by amplification of the aroA gene specific to S. aureus and then were analyzed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) of the coagulase gene (coa). Amplification of the variable region of the coa gene from these isolates produced five different PCR products ranging in size from approximately 490 bp to 850 bp. To obtain RFLP patterns of the PCR products they were subjected to digestion with restriction endonuclease HaeIII and the fragments separated by gel electrophoresis. Nine coa gene RFLP patterns, numbered I-IX, were observed, with 23 isolates (39.66%) assigned to RFLP pattern I and 14 isolates (24.14%) assigned to RFLP pattern III. Five out of nine patterns were found in both regions and four of nine patterns were only found in one region. The results demonstrated that several variants of the coa gene are present in the studied regions, but only a few of them were predominant, suggesting contagious transmission, a common source, or host adaptation of subset of the population of S. aureus strains. This study also indicated that genetic heterogeneity among S. aureus isolates recovered from bovine mastitis may be exists within and among herds in different regions.

Fibrinogen and ceruloplasmin in plasma and milk from dairy cows with subclinical and clinical mastitis.

The potential using of Acute Phase Proteins (APPs) in the assessment of mammary gland health was studied by examining the levels of Fibrinogen (Fb) and Ceruloplasmin (Cp) in plasma and milk from dairy cows with different grades of mastitis. Plasma samples were taken from jugular vein and milk samples were collected from quarters of cows with subclinical and clinical mastitis, as well as healthy controls. California Mastitis Test (CMT) were performed on each udder quarter of cows for detection of CMT2+ and CMT3+ quarters. CMT (0) and culture negative cases were considered healthy cows. Clinical mastitis, was graded as mild (clots in milk) or moderate (clots in milk and visible signs of inflammation in the mammary gland/s). The concentrations of Fb in the plasma of the cows with subclinical and clinical mastitis were higher than in the plasma of the healthy cows (p<0.01). There was no significant difference in plasma concentration of Cp between healthy and subclinical groups (p>0.05), but differences between clinical and healthy groups were significant (p<0.05). The concentrations of Fb and Cp in the milk of the cows with subclinical and clinical mastitis were higher than in the milk of the healthy cows (p<0.01). The results indicated that measurement of Fb in plasma and milk and Cp only in milk might be suitable for early diagnosis of mastitis in dairy cows.

Preliminary observations on the effect of parity on first day ultrasonic detection of embryo and its organs in bovine.

The aim of this study was to determine the effect of parity on the first day of ultrasonic detection of embryo and its organs using 8 MHz frequency. For this purpose, six nulliparous heifers, five primiparous cows and six multiparous cows of the Holstein breed were selected and mated by a single bull from the herd. From day 16 after mating, cows and heifers were examined by ultrasonography thrice weekly for the day of first detection of the embryo and its organs. The mean of first-day detection in heifers, primiparous and multiparous cows was as follows, respectively: allantois on days 22.3, 23 and 25.8; embryo proper on days 21.8, 23 and 27; heartbeat of embryo on days 26.1, 26.6 and 31.1; amnion on days 31, 31 and 30.8; limb buds on days 29.5, 28.8 and 30.6; stomach (abomasum) on days 32.2, 38.6 and 36.8; spinal column on days 32.6, 33.4 and 37.3; placentomes on days 33.3, 37 and 35.3; eyes on days 35.1, 35.4 and 38.6 and lens on days 40, 41.4 and 44. Although the mean of first day observation of all structures in heifers tends to be earlier than in the other groups, statistically, allantois, the embryo proper and its heartbeat in heifers and primipara were visualized significantly earlier than in multiparous cows. However, there were no differences between heifers and primiparous cows.