Glucose-6-phosphate dehydrogenase, glutathione peroxidase, total glutatione and reduced nicotinamide adenine dinucleotide phosphate in milk cells of subclinical mastitic cows.

This study aimed to determine the levels of milk cell total protein (TP), reduced nicotinamide adenine dinucleotide phosphate (NADPH), total glutathione (tGSH), activities of glucose-6-phosphate dehydrogenase (G6PD) and glutathione peroxidase (GPx) in subclinical mastitic cows. Milk from each udder was collected and grouped by the California Mastitis Test. Then, a somatic cell count (SCC) was performed, and the groups were re-scored as control (5-87 × 103 cells), 1st group (154-381 × 103 cells), 2nd group (418-851 × 103 cells), 3rd group (914-1958 × 103 cells), and 4th group (2275-8528 × 103 cells). Milk cell TP, NADPH, tGSH levels, G6PD, and GPx activities were assessed. Microbiological diagnosis and aerobic mesophyle general organism (AMG, cfu/g) were also conducted. In mastitic milk, TP, NADPH, and tGSH levels, and G6PD and GPx activities were significantly reduced per cell (in samples of 106 cells). In addition, milk SCC was positively correlated with AMG (r=0.561, p⟨0.001), NADPH (r=0.380, p⟨0.01), TP (r=0.347, p⟨0.01) and G6PD (r=0.540, p⟨0.001). There was also positive correlation between NADPH (r=0.428, p⟨0.01), TP (r=0.638, p⟨0.001) and AMG. NADPH was positively correlated with TP (r=0.239, p⟨0.05), GPx (r=0.265, p⟨0.05) and G6PD (r=0.248, p=0.056). Total protein was positively correlated with tGSH (r=0.354, p⟨0.01) and G6PD (r=0.643, p⟨0.001). There was a negative correlation between tGSH and GPx activity (r=-0.306, p⟨0.05). The microbiological analysis showed the following ratio of pathogens: Coagulase-Negative Staphylococci 66.6%, Streptococcus spp 9.5%, Bacillus spp 9.5%, yeast 4.8%, and mixed infections 9.5%. As a conclusion, when evaluating the enzyme and oxidative stress parameters in milk, it is more suitable to assign values based on cell count rather than ml of milk. The linear correlation between the SCC and AMG, milk cell NADPH, TP and G6PD suggests that these parameters could be used as markers of mastitis.

Expression of genes involved in the embryo-maternal interaction in the early-pregnant canine uterus.

Although there is no acute luteolytic mechanism in the absence of pregnancy in the bitch, a precise and well-timed embryo-maternal interaction seems to be required for the initiation and maintenance of gestation. As only limited information is available about these processes in dogs, in this study, the uterine expression of possible decidualization markers was investigated during the pre-implantation stage (days 10-12) of pregnancy and in the corresponding nonpregnant controls. In addition, the expression of selected genes associated with blastocyst development and/or implantation was investigated in embryos flushed from the uteri of bitches used for this study (unhatched and hatched blastocysts). There was an upregulated expression of prolactin receptor (PRLR) and IGF2 observed pre-implantation. The expression of PRL and of IGF1 was unaffected, and neither was the expression of progesterone- or estrogen receptor ß (ESR2). In contrast, (ESR1) levels were elevated during early pregnancy. Prostaglandin (PG)-system revealed upregulated expression of PGE2-synthase and its receptors, PTGER2 and PTGER4, and of the PG-transporter. Elevated levels of AKR1C3 mRNA, but not the protein itself, were noted. Expression of prostaglandin-endoperoxide synthase 2 (PTGS2) remained unaffected. Most of the transcripts were predominantly localized to the uterine epithelial cells, myometrium and, to a lesser extent, to the uterine stroma. PGES (PTGES) mRNA was abundantly expressed in both groups of embryos and appeared higher in the hatched ones. The expression level of IGF2 mRNA appeared higher than that of IGF1 mRNA in hatched embryos. In unhatched embryos IGF1, IGF2, and PTGS2 mRNA levels were below the detection limit.