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Summary An animal experiment can be defined as any procedure in which a live animal is used with the aim of testing a scientific assumption, gathering information, obtaining or testing substances or observing the effect of a particular measure on the animal, as well as using an animal in behavioral experiments. The Ethics Committee (EC) determines how animal testing shall be conducted and, in accordance with the Law on Animal Welfare, provides expert supervision of animal testing, organizes training courses for persons conducting testing, provides expert opinions to the minister regarding ethical and scientific justification of animal testing, and submits annual reports to the minister. The data used in this paper were derived in the course of work of the EC in the period between 2016 and 2018. The meetings of the EC were held on a regular basis and in accordance with the Rules of Procedure. Positive Opinions were issued for most of the submitted Applications (87.5%), while a smaller number was returned (6.25%) or directed to the Council for further consideration (6.25%). In several cases, the EC also issued Opinions that certain experiments are beyond its scope of authority and thus beyond its formal legal capacity. Based on the Application of a candidate, the EC also approved changes in previously issued Opinion to ensure consistency in the documentation. The Committee regularly submitted reports and there were no objections to its work. In accordance with the abovementioned, the performance of the EC can be considered efficient and successful, but it can be further significantly improved by more active effort of the individual members of the EC within their home institutions to educate candidates and raise awareness of the importance of the work performed by the EC.

Summary Chorioallantoic membrane (CAM) is an extraembryonic membrane very frequently used for in vivo studies in various researches. Since researches require a fast method for quantifying the CAM angiogenic response, there is a need to develop a new precise and unbiased method of quantification of angiogenesis in CAM, which would be easy to perform and suitable for analysis of a large number of samples. The objective of this paper is to apply a new method of quantification of angiogenesis in investigation of the development of blood vessels in the CAM, in particular days of embryonic life considered essential for CAM development. The present research included 75 fertilized eggs of heavy hybrid Ross 308. CAM sampling for stereological analyses was in key phases of embryonic development, namely on the 12th, 15th and 19th day. The results of the present investigation show that the increase in embryonic age results in increase in circulation index, which is also an indicator of angiogenic processes developing in CAM. The lowest value of circulation index (0.1952) was recorded on the first sampling day (E12), while the highest value (0.2666) was recorded on the last sampling day (E19). This method may be applied in researching different factors which affect angiogenesis in CAM.

Introduction: Hormonal and metabolic changes, as well as energy imbalance, can affect health, production and reproductive performance of dairy cows. In the present study, we evaluated phagocytosis and respiratory burst neutrophil activity during the transition period and early lactation and compared it with biochemical and hematological parameters in dairy cows. Methodology: Simmental cows (n = 21) were enrolled in the study. Whole blood samples were collected weekly from 3 weeks pre- calving until 6 weeks post calving. Basic metabolic and blood parameters were assessed by routine laboratory analyses, while neutrophil functions were analyzed by commercial test kits. Results: Optimal neutrophil response was observed pre and post calving. The highest value was recorded in the 6th week after calving (89.54 ± 7.61%) and being significantly higher (p < 0.01) as compared to values recorded at two and one week before and one week after calving. The percentage of activated neutrophils was high during the entire study period: from 70.80 ± 5.22% at the beginning of the study to 89.54 ± 7.61% at the end of the study. During the study period, production of Reactive Oxidative Species by neutrophils was positively correlated with β-hydroxybutyrat and non-esterified fatty acids values (0.454** and 0.423**, respectively) and calcium levels (0.164* and 0.212**, respectively). Conclusions: The most prominent changes in all parameters had no influence on phagocytic and respiratory burst activity of neutrophils. Neutrophil function is preserved at the optimal level during the transition period and early lactation in Simmental cows.

Morphological characteristics of skeletal muscles of slow- and medium-growing strains of chickens are very important for meat quality and comparison with fast-growing strains. The aim of this paper was to evaluate morphological parameters of breast and thigh muscles of slow- and mediumgrowing strains in a free-range system. The slow-growing strains used in the experiment were autochthonous breeds Sombor crested and Banat naked neck, and the medium-growing strain was Red-bro. The tissue samples were taken from the thigh muscle and muscles of the breast of 10 chickens of each breed. Samples were stained with hematoxylin - eosin and enzyme succinate - dehydrogenase (SDH). The following morphological parameters were observed: diameter of muscle cells, nucleo-cytoplasmic ratio of muscle cells, volume density of connective tissue within the muscle and the presence of red, white and intermediate muscle cell types. Between strains, the type of muscle or genotype didn’t have significant effects on diameters of muscle cells and nucleo-cytoplasmic ratio in the muscle cells. Results indicated that genotype had significant effect on volume density of the connective tissue in breast muscles. Red muscle cells were, in all strains, significantly more represented in m. biceps femoris than m. pectoralis superficialis. Genotype had significant effect on ratio between connective tissue and muscle cells and no significant effects on other morphological parameters. [Projekat Ministarstva nauke Republike Srbije, br. TR 31033] Biotechnology in Animal Husbandry (2016), 32(1): 27-35

The aim of this paper was to examine the histological characteristics of leg muscles of pheasants hatched from eggs of a different colour. From muscle samples (M. biceps femoris) of 56-day old pheasants hatched from eggs of different colour (dark brown, light brown, brown/green, blue/green) histological preparations were made. Following parameters were examined: diameter of muscle cells, volume density of connective tissue in muscles, nucleo-cytoplasmatic ratio of muscle cells. Results showed that diameter of muscle cells was smaller in pheasants hatched from blue/green eggs compared to all other examinated groups. There was no differences in volume density of connective tissue in muscles between groups. Nucleo-cytoplasmatic ratio of muscle cells was higher in pheasants hatched from blue/green eggs compared to all other examinated groups. From obtained results it can be concluded that pheasants hatched from blue/green eggs had weaker muscle development than pheasants hatched from eggs of other eggshell colour. Cause of this could be related to structural differences of eggshells of various colour. This leads to weaker development of embryos and chicks hatched from blue/green eggs which reflects on differences in development of leg muscles.

Cardioprotective effects of fullerenol C-60(OH)(24) nanoparticles (FNP) were investigated in pigs after a single treatment with doxorubicin (DOX). Semithin and ultrathin sections of myocardial tissue routinely prepared for transmission electron microscopy were analyzed. Extensive intracellular damage was confirmed in cardiomyocytes of DOX-treated animals. By means of ultrastructural analysis, a certain degree of parenchymal degeneration was confirmed even in animals treated with FNP alone, including both the oral and the intraperitoneal application of the substance. The cardioprotective effects of FNP in animals previously treated with DOX were recognized to a certain extent, but were not fully confirmed at the ultrastructural level. Nevertheless, the myocardial morphology of DOX-treated animals improved after the admission of FNP. Irregular orientation of myofibrils, myofibrillar disruption, intracellular edema, and vacuolization were reduced, but not completely eliminated. Reduction of these cellular alterations was achieved if FNP was applied orally 6 h prior to DOX treatment in a dose of 18 mg/kg. However, numerous defects, including the inner mitochondrial membrane and the plasma membrane disruption of certain cells persisted. In FNP/DOX-treated animals, the presence of multinuclear cells with mitosis-like figures resembling metaphase or anaphase were observed, indicating that DOX and FNP could have a complex influence on the cell cycle of cardiomyocytes. Based on this experiment, further careful increase in dosage may be advised to enhance FNP-induced cardioprotection. These investigations should, however, always be combined with ultrastructural analysis. The FNP/DOX interaction is an excellent model for the investigation of cardiomyocyte cell death and cell cycle mechanisms.

Four groups of broiler chicken hybrid ( Ross 308) eggs were incubated under different temperature and light conditions to assess the effects of both variables on musculature development. During the incubation, the first group was the control and the eggs were incubated at a temperature of 37.8 °C without light, the second group was exposed to increased temperature, while the third group was exposed to monochromatic green light. The fourth group of eggs was incubated under simultaneously changed temperature (according to the regime of the second group) and light (according to the regime of the third group) conditions. Tissue samples from the breast muscle ( M. pectoralis superficialis) were taken on embryonic days 9, 12, 15, 19 and 20, as well as on days 1, 3, 7, 14, 21 and 42 after hatching. Histological preparations were used to assess muscle cell diameter, connective tissue volume density, and the nucleocytoplasmic ratio of muscle cells. Our findings indicate that thermal and light manipulations during incubation increase diameter and decrease nucleocytoplasmic ratio in postnatal skeletal muscle cells. No differences were found among groups with regard to connective tissue volume density. These results can be explained by the influence of thermal and light manipulations during incubation on prolonged myoblast proliferation, as well as on the greater postnatal development of skeletal muscle cells.

Fullerenol (C-60(OH)(24)) is present in aqueous solutions in the form of polyanion nanoparticles with particles size distribution within the range from 15 to 42 nm. In this research it is assumed that these features could enable fullerenol nanoparticles (FNPs) to bind positively charged molecules like doxorubicin (DOX) and serve as drug carriers. Considering this, fullerenol/doxorubicin nanocomposite (FNP/DOX) is formed and characterized by ultra-performance liquid chromatography tandem mass spectrometry, dynamic light scattering, atomic force microscopy and transmission electron microscopy. Measurements have shown that DOX did not significantly affect particle size (23 nm). It is also assumed that FNP/DOX could reduce the acute cardiotoxic effects of DOX in vivo (Wistar rats treated i.p.). In this study, quantitative real time polymerase chain reaction results have shown that treatment with DOX alone caused significant increase in mRNA levels of catalase (p LT 0.05) enzyme indicating the presence of oxidative stress. This effect is significantly reduced by the treatment with FNP/DOX (p LT 0.05). Furthermore, mRNA levels of antiapoptotic enzyme (Bcl-2) are significantly increased (p LT 0.05) in all treated groups, particularly where FNP/DOX was applied, suggesting cell resistance to apoptosis. Moreover, ultrastructural analysis has shown the absence of myelin figures within the mitochondria in the heart tissue with FNP/DOX treatment, indicating reduction of oxidative stress. Hence, our results have implied that FNP/DOX is generally less harmful to the heart compared to DOX.

The aim of this paper is to examine the effect of different sound stress duration on adrenal gland of broiler chickens of different age. The experiments were conducted on 90 HYBRO broiler chickens, divided into nine groups (each group consisting of 10 birds) by different age and lenght of sound stress duration. The chickens were exposed to sound stress in sound attenuated building using a fire alarm bell (95 dB) for stress sessions lasted 120 min every day. The histological structure of adrenal gland was analyzed by light microscopy. The results indicate that adrenal gland is sensitive on this kind of stress and sound as stressor could introduce organism in stress reaction. Adrenal glands of broilers exposed to sound stress characterized by marked hyperemia, hypertrophy of interrenal cells, smaller affinity for staining of chromaffin cells compared with control groups. According to our results, degree of histological changes of adrenal gland under the influence of sound stress depends on the length of exposure and age of chickens. Cilj ovog rada je da ispita uticaj različite dužine trajanja zvučnog stresa na histološku građu nadbubrežnih žlezda brojlerskih pilića u različitim uzrastima. Ogled je izveden na 90 HYBRO brojlerskih pilića, podeljenih u 9 grupa u skladu sa uzrastom i dužinom izlaganja stresu. Pilići su zvučnom stresu bili izlagani u zvučno izolovanim objektima uz korišćenje požarnog alarmnog zvona (95dB) u trajanju od 120 min svakog dana. Histološka građa nadbubrežnih žlezda je analizirana svetlosnim mikroskopom. Rezultati ukazuju da je nadbubrežna žlezda osetljiva na ovu vrstu stresa i da zvuk, kao stresor, može da uvede organizam u stresnu reakciju. Nadbubrežne žlezde pilića izlaganih zvučnom stresu karakteriše značajna hiperemija, hipertrofija interrenalnih ćelija, manji afinitet prema bojenju hromafinih ćelija u odnosu na kontrolnu grupu. U skladu sa ovim rezultatima, stepen histoloških promena nadbubrežne žlezde pod uticajem zvučnog stresa zavisi i od dužine izlaganja i uzrasta pilića.

Eggs of broiler (Ross 308) and layer (Hy-Line Brown) chicken hybrids were incubated and tissue samples from the leg muscle (m. biceps femoris) and breast muscle (m. pectoralis profundus) were taken on embryonic days 9, 12, 15, 18 and 20, as well as on the first day post hatching. Histological preparations were made from the samples and the diameter of muscle cells, the volume density of connective tissue of muscles, and the nucleocytoplasmic ratio of muscle cells were recorded. Comparison of the diameter of muscle cells of leg muscle and the breast muscle between broiler and layer chicken during the period of late incubation and day 1 post hatching showed the diameter of the examined muscle cells was greater in layers, while the nucleocytoplasmic ratio of muscle cells was higher in broilers. No differences were established between these two strains in the volume density of connective tissue in both muscles examined. Such results are explained by the prolonged proliferation of myoblasts and delayed differentiation of muscle cells in broilers, which leads to a greater development of the skeletal musculature during the postnatal period. These processes in the embryonic period of development are the cause of differences in the morphodynamics of embryonic development of the skeletal musculature between broilers and layers presented in this paper.

The aim of the present study was to investigate the possibility of increasing boar's reproductive exploitation by using AI doses of doubly reduced volume and sperm count in the intrauterine AI procedure. The experiment was conducted at a commercial pig farm in Serbia in 2014. Classic intracervical insemination (ICI) was performed by using 50 mL or 100 mL volume doses containing 4 × 109 or 2 × 109 progressively motile spermatozoa. The same volumes and sperm numbers per dose were used with intrauterine insemination (IUI). Each dose combination was used to inseminate 30 sows. Intrauterine insemination with AI doses of reduced volume (50 mL) and sperm count (2 × 109) did not produce a statistically significant difference (P < 0.05) in the farrowing rate (76.7%) as compared with 4 × 109 spermatozoa in the same volume (83.3%) or with insemination by doses of 100 mL with a 2 × 109 (83.3%) or a 4 × 109 sperm count (86.7%). The number of live-born piglets (10.82) was larger following IUI using a 50 mL volume dose with a 2 × 109 sperm count as compared with ICI with the same AI dose volume and sperm count (9.85). The results show that the use of reduced AI dosages provides an opportunity for the swine industry to considerably exploit the reproductive potential of genetically superior boars.

42nd European-Society-for-Medical-Oncology Congress (ESMO), Sep 08-12, 2017, Madrid, Spain

It is proved that the protein expression of IGF-1 is detected in several tissues including liver, brain, eye, lung, pancreas and muscle. This growth factor exerts autocrine and paracrine effect during embryonic and postnatal development and is involved in muscle development. The aim of this study was to determine the level of protein expression of IGF-1 in breast muscle during embryonic and postnatal development of broiler chickens. For the experiment were used fertilized eggs and broiler chickens of heavy hybrid "Ross 308", originating from parent flock at the age of 54 weeks. Samples of breast muscle were taken from embryos during the 15th and 19th day of embryonic development and 3rd and 21st day of postnatal development from 5 birds in each age category. From all of the embryos and chickens the right side of the brest muscles were individually sampled. The muscle tissue was macerated and then homogenized in buffer with protease inhibitors. After that immunoprecipitation, electrophoresis and transfer the Western blot analysis were performed on all samples. For visualization of proteins the membranes were incubated in chemiluminescent substrate. The results indicate statistically significant increase in protein expression of IGF-1 in breast muscle on the 19th day of embryonic development and 3rd of postnatal development in compared to 15th day of embryonic development (p

Embryonic stage of development, in conditions of intensive broiler production, is a period in which the dynamics of postnatal development and the productive performance can be affected significantly. Green monochromatic light and thermal conditioning are the most commonly used treatments that have given satisfactory results in postnatal development. The aim of this study was to examine the effect of these treatments on meat quality of broiler chickens. Our study is based on the four experimental groups (control and three different treatments). Each group was incubated in separate incubator with 80 eggs, with controlled incubation parameters (temperature, humidity, ventilation). The control group was incubated under standard conditions (37.8 ?C, without light). The first treatment consisted of thermal conditioning (raising the temperature to 39,8 ?C for 3 h on 16th, 17th and 18th day of incubation). Another treatment consisted of lighting a green monochrome light (diffused LED lighting intensity of 0.1 W/m2) from 5th until 14th day of incubation, in the intermittent mode (15 min. light, 15 min. darkness), and from 15th day of incubation under constant light regime. The third treatment was a combination of first two treatments, with the temperature and lighting changed by these regimes. Our results indicate a significantly higher average body weight, protein content in meat, and less progressive loss of moisture (drip loss) in all treated groups. The applied treatments had no effect on mortality, conversion, or the content of fat and ash in meat. The results also indicate a significant synergistic effects achieved by the simultaneous application of both treatments.

Heat stress (HS) is one of the greatest problems in contemporary chicken production and it entails significant economic losses. The purpose of this paper is to examine the influence of early-age thermal conditioning (ETC), vitamin C (Vit C) supplementation, and their combination on the production characteristics and specific quality parameters of the meat of broilers which were exposed to chronic HS in the last two weeks of breeding. Four hundred broilers (Cobb 500) of both sexes were divided into 4 experimental groups. Group C was given Vit C (2 g/L) dissolved in water from day 22 until the end of production. Group T was exposed to ETC for a period of 24 h at the temperature of 38 +/- 1 degrees C and 40%-60% relative humidity on the fifth day of breeding. Group TC was the combination of the groups T and C, while group K was the control group. The results indicate that ETC, independently or in combination with Vit C, improves the production characteristics in terms of feed conversion reduction (P < 0.05). The examined treatments increase the volume (P < 0.05) of certain parts of the body (carcass, legs and thighs, and back) with regards to total body weight, especially in group TC. Regarding meat quality determined according to pH value and meat color criteria (CIEL*), the best results were also observed in group TC, followed by groups T and C. These results justify the use of Vit C and ETC in summer conditions, when heat stress is expected; however, the best results can be achieved by combining these two methods, thus producing a synergistic effect.