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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 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.

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The aim of this study was to evaluate whether the addition of cysteamine to the in vitro culture media enhances the yield, hatching rate, total cell number and inner cell mass/total cell number ratio of bovine embryos. A total of 933 bovine oocytes collected from ovaries of 60 slaughtered donors were subjected to in vitro maturation and in vitro fertilization. Following fertilization, embryos were cultured in synthetic oviductal fluid without glucose. After 24 h embryos were transferred into synthetic oviductal fluid with 1.5 mM glucose and 0 (control), 50, 100 and 200 µM of cysteamine. After 48 h, the embryos were transferred into synthetic oviductal fluid with glucose but without cysteamine and cultured until Day 9. The number of cleaved embryos on Day 2, the total number of blastocysts on Day 7 and the number of hatched blastocysts on Day 9 were calculated. Differential staining of inner cell mass and trophectoderm cells of blastocysts were performed on Day 7 and Day 9 of in vitro culture. Supplementation of in vitro culture media with 100 µM cysteamine increased the blastocyst yield (P < 0.05) without affecting the hatching rate. Furthermore, the embryos cultured in the presence of 100 µM cysteamine had significantly higher number of inner cell mass cells (P < 0.05) and the proportion of inner cell mass cells (P < 0.05) compared with the controls. The results of the present study demonstrated that the addition of 100 µM cysteamine to the in vitro culture media improved blastocyst production rate and enhance embryo quality, which could lead to the improvement of the in vitro culture system for bovine embryos.