Svaki kirurški zahvat narušava homeostazu organizma, koja se očituje u imunološkim, neuroendokrinim i metaboličkim promjenama u organizmu. Pretpostavlja se da jačina odgovora organizma ovisi i o jačini traume tijekom operativnih zahvata. Cilj istraživanja je bio utvrditi antioksidacijski status kuja prije i poslije ovarijektomije. U istraživanje je bilo uključeno petnaest kuja pacijenata Klinike za porodništvo i reprodukciju Veterinarskog fakulteta u Zagrebu u kojih je učinjena elektivna ovarijektomija u svrhu kastracije. Kujama je krv vađena prije operacije te poslije prvog, trećeg i sedmog postoperativnog dana kako bi se izmjerio ukupni antioksidacijski status. Svim kujama je učinjena i kompletna krvna slika, biokemija i C-reaktivni protein (CRP). Ukupan antioksidacijski status određivan je pomoću TAS test kitova. Proizvođač kitova preporučuje svakom laboratoriju standardiziranje referentnih vrijednosti. Zbog toga je učinjena analiza 30 seruma zdravih kuja koje nisu bile operirane te je kao referentna vrijednost korištena njihova srednja vrijednost +/- dvije standardne devijacije. Prosječna referentna vrijednost iznosila je 1,55 mmol/L. Vrijednosti ukupnog antioksidacijskog statusa nakon operacije u svim promatranim razdobljima nisu se statistički znatnije razlikovale. Uspoređujući minimalne i maksimalne vrijednosti u svim promatranim razdobljima, zaključili smo da nije bilo velikih razlika u vrijednostima ukupnog antiosksidacijskog statusa u operiranih kuja. Na temelju naših rezultata možemo zaključiti da ovarijektomija nije prouzročila oksidacijski stres.

AimTo investigate the impact of synthetic electrospun polyurethane (PU) and polycaprolactone (PCL) nanoscaffolds, before and after hydrolytic surface modification, on viability and differentiation of cultured human eye epithelial cells, in comparison with natural scaffolds: fibrin and human amniotic membrane.MethodsHuman placenta was taken at elective cesarean delivery. Fibrin scaffolds were prepared from commercial fibrin glue kits. Nanoscaffolds were fabricated by electrospinning. Limbal cells were isolated from surpluses of human cadaveric cornea and seeded on feeder 3T3 cells. The scaffolds used for viability testing and immunofluorescence analysis were amniotic membrane, fibrin, PU, and PCL nanoscaffolds, with or without prior NaOH treatment.ResultsScanning electron microscope photographs of all tested scaffolds showed good colony spreading of seeded limbal cells. There was a significant difference in viability performance between cells with highest viability cultured on tissue culture plastic and cells cultured on all other scaffolds. On the other hand, electrospun PU, PCL, and electrospun PCL treated with NaOH had more than 80% of limbal cells positive for stem cell marker p63 compared to only 27%of p63 positive cells on fibrin.ConclusionNatural scaffolds, fibrin and amniotic membrane, showed better cell viability than electrospun scaffolds. On the contrary, high percentages of p63 positive cells obtained on these scaffolds still makes them good candidates for efficient delivery systems for therapeutic purposes.

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.

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The aim of this study was to investigate the effects of exogenous melatonin on libido and semen quality parameters in bucks during the non-breeding season. Twelve bucks of the French alpine breed from 1.5 to 4 years of age were assigned into melatonin (MG) and control (CG) groups, with 6 bucks in each group. The experimental period was 3 months (March-May), divided into six periods of 15 days each. The bucks in the MG group received four melatonin implants at the end of March. Two semen samples were taken from the bucks by artificial vagina once per week and their libido estimated. Volume and spermatozoa concentration, their mass motility and motility, proportion of live and total abnormal and forms with abnormal head and tail were determined in the obtained ejaculate samples. The total number of spermatozoa and functional spermatozoa fraction in the ejaculate was also calculated. The MG bucks had significantly higher mass motility and motility of spermatozoa in the first half of April, and a higher proportion of live spermatozoa in the first and second half of April (p < .05). Differences in libido intensity were not significant. The results indicated that the application of melatonin significantly improved the qualitative parameters of semen in bucks, as seen in increased mass motility, motility of spermatozoa and proportion of live spermatozoa shortly following melatonin insertion. Therefore, the results of the current study are novel regarding the use of melatonin treatment during the non-breeding season to improve the qualitative parameters of ejaculates in bucks.

The aim of this research was to investigate the ovarian influence on the success of two superovulatory protocols in goats. Fifteen goats were used twice in the study in a crossover design: once within the control group where standard protocol was applied, and the other time in a modified, day 0 group, where no dominant follicle was present and the follicular cohort was homogenous ("Day 0 protocol"). All the animals were synchronized with vaginal pessaries and superovulated with 6 decreasing doses of pFSH. Although the day 0 protocol significantly decreased the presence of the dominant follicle before the start of the superovulation treatment; it did not improve the superovulatory success. Fifty-four percent of goats in the control group had a dominant follicle which exerted a negative influence on the number of transferable embryos. The other 46% of goats from the same group had a very good superovulatory response, which improved the overall response of the control group in comparison with the day 0 group. Another reason for the lower superovulatory success of the day 0 protocol was the smaller number of second and third category follicles (3.5 -5.5 mm in size) at the onset of the superovulation protocol, that positively correlated with the number of the transferable embryos (r=0.67; P<0.05). In Boer goats, for the success of superovulatory treatment, the number of follicles of a certain size before the start of the superovulatory treatment has an effect as important as the effect of dominance. The application of the FSH hormone should start at the time when a suitable number of follicles belonging to the second category are present on the ovaries when performing the day 0 protocol.

Simple Summary: Haematological tests are an important diagnostic tool for animal diseases. However, little is known beyond the standardly used haematological methods in sheep, especially those in which the detection of sheep red blood cells (RBCs) shape changes is crucial. Our goal is to obtain sheep RBC morphometric parameters as well as RBC subpopulations based on their morphometric parameters. Morphometric parameters of RBC size and shape were determined from stained blood smears using SFORM, a computer-assisted program for automated cell morphometric measurements. Based on their morphometric parameters, three RBC subpopulations were obtained using principal component and cluster analysis: the smallest and most elongated RBCs, the biggest and most rounded ones, and RBCs of average size and shape. When the values of RBC haematological parameters were higher or above the physiological range, a significantly higher proportion of both average size and shape RBCs, as well as the biggest and most rounded RBCs, was obtained. Since they were obtained from healthy animals, these results indicated the importance of determining morphometric parameters of RBCs and the proportion of each RBC subpopulation, which could serve as a basis for future possibilities in the diagnostic interpretation of haematological disorders in sheep, especially those for which the detection of shape changes in ovine RBCs is crucial.Abstract: Data concerning the morphometric parameters of sheep red blood cells (RBCs) obtained using computer-assisted image analysis have not yet been investigated, and there are no data on any analyses of ovine RBC subpopulations based on their morphometric parameters. The aims of this study are to determine the values of RBC haematological and morphometric size and shape parameters, to form groups according to the obtained values of haematological parameters; to determine the differences in RBC morphometric parameters between the formed groups, and to determine RBC subpopulations and their respective proportions in the formed groups. Thirty-six blood samples were collected from the jugular vein of clinically healthy Lika pramenka sheep, aged between 2 and 5 years. Haematological parameters including haemoglobin (HGB), haematocrit (HTC), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular Animals 2019, 9, 1130 2 of 15 haemoglobin concentration (MCHC), and RBC distribution width were analysed using a haematology analyser. Haematological parameters were categorized into two groups: those with lower values or values below the physiological range (Groups 1) and groups with higher values or values above the physiological range (Groups 2). Morphometric parameters of RBCs were determined from stained blood smears using SFORM, a computer-assisted program. Significantly higher values of RBC area, outline, convex, minimal and maximal radius, as well as length and breadth were established in Groups 2 compared to Groups 1 of HGB, HCT, MCV, MCH, and MCHC, respectively. Based on the morphometric p...

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