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The aim of our research was to examine the ability of density gradient preparation BoviPure and swim up method on bull sperm separation and in vitro embryo production (IVP) systems. Frozen/thawed semen from six Simmental bulls was pooled and treated using both methods. The sperm motility, concentration, membrane activity, membrane integrity and acrosomal status were evaluated and compared before and after sperm processing using BoviPure and swim up methods. We also evaluated and compared cleavage rates, embryo yield and quality between the methods. There were significant differences (P < 0.05) between the sperm characteristics before and after BoviPure, but not after swim up method. However, there were significant differences for sperm results among those two mentioned methods. A total of 641 oocytes were matured and fertilized in vitro and cultured in SOFaaBSA. The percentage of cleavage (Day 2) and the percentage of hatched embryos (Day 9) were similar for both methods. However, embryo production rate (Day 7) was significantly higher using BoviPure method (P < 0.05). Also, total cell number and embryo differential staining (inner cell mass and trophectoderm cells) of Day 7 morulas and blastocysts showed that BoviPure treated sperm displayed higher quality embryos compared to swim up method (P < 0.05). Our results indicate that BoviPure method has an enhanced capacity in sperm selection for in vitro embryo production when compared with swim up method. So, we concluded that BoviPure could be considered as a better alternative to swim up method for separating bull spermatozoa from frozen/thawed semen for IVP of bovine embryos.

I n the past two decades many substances of natural or synthetic origin were studied as potential alternatives to antibiotic growth promoters (AGP) and some of them, particularly immunomodulators (IMs) and nutraceuticals (NCs), have shown to be capable of stimulating functions of the immune system and improving general health. At the same time, they were shown to be harmless for animals and the environment. Promising results have been obtained with natural clay minerals, zeolites among which clinoptilolite (CPL) is the best known as zootechnical and biomedical feed ingredient widely reported in scientific literature and used in farm animal nutrition. CPL has a potential to replace AGP due to its unique anti-bacterial properties, safety and efficacy as dietary supplement in food animals unifying potentials of an IM and NC. Currently, there INTRODUCTIONT he phrase "survival of the fittest" as an alternative to "natural selection" has been adopted and published in 1868 by Charles Darwin (1), and nowadays is becoming so true because of the rapid emergence of antibiotic resistant strains of pathogenic microbes of biomedical, particularly veterinary importance. This is a consequence of non-clinical use and misuse of dietary antibiotics for more than sixty years to prevent bacterial infections and to enhance performance in livestock production, mostly in chickens and pigs (2). Namely, the increased sanitary problems in intensive farming of food animals have been overcome thus far by adding sub-therapeutic doses of antibiotic growth promoters (AGP) infeed to enhance production efficiency by increasing growth rate, improv- ing feed utilization and reducing mortality from clinical disease is well documented particularly in swine production systems (3). However, the consumers especially in developed countries are becoming increasingly concerned about drug residues in meat and other animal products (4). In addition, it has been suggested that the continuous use of antibiotics may contribute to a reservoir of drugresistant bacteria (5) which may be capable of transferring their resistance to pathogenic bacteria in both animals and humans (6). As a result, many countries have banned or are banning the inclusion of antibiotics in diets as a routine means of growth promotion. The first experience of an AGP ban in Sweden in 1986 indicated a reduction in growth and an increase in morbidity and mortality rates in weaned pigs, which emphasized the importance of antibiotic use in intensive swine production(7). The total ban of dietary AGP in the EU countries since January 1, 2006 (EU Regulation No. 1831 and the elimination of their use in animal feed and water in the USA effective January 1, 2017 (FDA, CVM 2012, VFD Basics 2017) has had a serious influence on both health and performance of food animals underlining the need to develop alternative dietary and non-dietary strategies to evaluate the potential antimicrobial agents for their replacement (8). According to the World Health Organization, most antibiotics will not ...

The objectives of this study were to examine the influence of body condition of cows on metabolic and antioxidative status, as well as to investigate the relationship between metabolic indicators of lipid mobilization and oxidative stress during transition period. The study was conducted on 24 Holstein-Friesian dairy cows divided into 2 groups according to their body condition score (BCS) as optimal (n = 12; BCS from 3.25 to 3.75) or adipose (n = 12; BCS ≥4). Metabolic status (glucose, triglycerides, total cholesterol, HDL cholesterol, NEFA and BHB), paraoxonase-1 (PON1) and apolipoprotein A-I (ApoA-I) were analysed in sera taken on days -30, -10, -2, 0, 5, 12, 19, 26 and 60 relative to parturition. Adipose cows had significantly higher glucose concentration at parturition being significantly decreased after parturition on days 12 and 19. Total cholesterol and HDL-C concentrations were the lowest at parturition and significantly higher on days 26 and 60 after parturition in both groups of cows. Both investigated groups had significantly higher NEFA concentration from parturition until day 19 after parturition, indicating energy deficit and an increased lipid mobilization after calving. There were no significant differences in BHB concentration during transition period in both groups. No significant differences were found in PON1 activity and ApoA-I concentration during transition period in both groups of cows. However, in adipose cows, although not significantly different, PON1 was decreased from calving until day 19 after parturition indicating a disturbance in antioxidative status in adipose cows. PON1 significantly positively correlated with total cholesterol and HDL-C concentrations and negatively with NEFA indicating a strong relationship of PON1 with lipid metabolism. Significant positive correlation between NEFA and BHB in both groups of cows points out on energy deficit during transition period that cows tend to overcome by lipid mobilization providing alternative source of energy needed for parturition and lactation.

The aim of the research was to determine the influence of preventive application of two different ozone preparations on reproductive efficiency in dairy cows with physiological puerperium. The research was conducted on 404 Holstein cows aged 2-8 years, divided into three groups. The first group consisted of cows treated using 6 intrauterine ozone pearls (RIPROMED OVULI O(3) (®) , Novagen, Italy) at once. The second group consisted of cows treated with ozone foam spray (RIGER SPRAY(®) , Novagen, Italy) application in the body of uterus for 5 s, and the third group was the control group. Cows from the first two groups were observed and treated during early puerperium, 24-48 h after parturition. To assess the reproductive performance of Holstein cows, interval from calving to first insemination (days open to first service, DOFS), interval from calving to pregnancy (days open to pregnancy, DOP), relative pregnancy rate (%), first service conception rate (FSCR, %) and all service conception rate (ASCR, %) were measured. The estimate of hazard ratio (HR) for group B relative to group C for DOFS was 0.62 (p = 0.0002), implying that the relative rate of first service decreased in the control group by 38%. The estimate of HR for group B relative to group C for DOP was 0.65 (p = 0.0006), implying that the relative rate of pregnancy decreased in group C by 35%. Cows treated with Riger spray (group B) became pregnant earlier and had better FSCR and ASCR. It is supposed that the form of ozone preparations and the amount of active substance were decisive. A conclusion may be made that preventive ozone intrauterine application during early puerperal period, especially the foaming form of ozone preparations (spray), improved the reproductive efficacy in dairy cows.

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The metabolism of proteins in the blood serum in Boer and Saanen goats was investigated during puerperium. Twenty Boer goats (10 primiparous and 10 pluriparous) and 10 Saanen goats (five primiparous and five pluriparous) between 2 and 5 years of age were used in this research. Blood for analysis was taken every fourth day from day 3 until day 40 post-partum. Blood samples were collected by jugular puncture. In the obtained blood serum, the concentration of total proteins (PT) and albumin (ALB), and the activity of enzymes aspartate aminotransferase (AST) [the Enzyme Commission number (EC number) 2. 6. 1. 1.], gamma-glutamyltransferase (GGT) (EC 2. 3. 2. 2.), creatine kinase (CK) (EC 2. 7. 3. 2.) and alkaline phosphatase (AP) (EC 3. 1. 3. 1.) were determined by spectrophotometry. These parameters were in physiological ranges in Boer goats and in Saanen goats, without significant differences according to number of kids per doe. According to the research results of the blood serum in goats during puerperium, there were no significant differences in the concentration of ALB. Boer goats had significant higher (p < 0.05) concentration of PT and enzyme activity of AP, CK and GGT. Saanen goats had only enzyme activity of AST significantly higher (p < 0.05). Enzyme activity of alkaline phosphatase was significant higher (p < 0.05) in pluriparous goats in both breeds than in primiparous. The obtained results may represent a contribution to a better understanding of protein metabolism during puerperium in dairy and meat goats and for diagnostic purposes.

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