Anna Domosławska, Sławomir Zduńczyk, Wojciech Niżański, Andrzej Jurczak and Tomasz Janowski
Thirty clinically healthy dogs with poor semen quality were used in the study. Fifteen dogs were supplemented daily with selenium (0.6 mg/kg organic selenium from yeast) and vitamin E (5 mg/kg) per os for 60 d. The control group (15 dogs) was not supplemented. Semen was collected from all dogs by manual manipulation on days 0, 30, 60, and 90. The sperm concentration and motility parameters were evaluated with a Hamilton Thorne sperm analyser, version IVOS 12.3. For the assessment of sperm morphology, Diff-Quik stain was used. The percentage of live and dead spermatozoa was estimated on dried smears stained with eosin-nigrosin. The concentration of spermatozoa, most motility parameters determined (PMOT, VSL, VCL, ALH, BCF, RAPID, MEDIUM, SLOW, and STATIC), and the percentage of spermatozoa morphologically normal and live increased significantly (P < 0.05) after 60 d of supplementation. In the control group, there were no changes in motility parameters while the concentration and total sperm count decreased over the duration of the study. In conclusion, supplementation with selenium and vitamin E for 60 d can improve the quality of semen in dogs with lowered fertility.
Anna Domosławska, Sławomir Zduńczyk, Wojciech Niżański and Tomasz Janowski
Semen quality parameters of infertile and fertile dogs were compared. Sperm concentration and semen motility parameters were measured by the Hamilton-Thorne Semen Analyser IVOS 12.3. The spermatozoal morphology and the percentage of live spermatozoa were examined microscopically. Forty-six dogs of various breeds were examined. Twenty dogs had a conception failure within last year. These dogs had a history of prior normal fertility. Twenty six fertile dogs served as control. All animals underwent a clinical examination as well as ultrasonography. Sperm concentration was significantly lower in infertile dogs than in fertile dogs. For most determined motility parameters (MOT, PMOT, VAP, VSL, VCL, BCF, RAPID, STATIC) significant differences between infertile and fertile dogs were found. The percentage of spermatozoa with normal morphology also significantly differed between both groups. Ultrasonography of the prostate gland and testes revealed no pathological conditions. The testicular degeneration was assumed to be a possible cause of infertility in these dogs. The present study showed that the most sperm CASA motility parameters were significantly lower in infertile dogs in comparison to the fertile ones, and confirmed the usefulness of the Hamilton-Thorne Semen Analyser for a quick and objective analysis of sperm concentration and motility in dogs.
Michał Dzięcioł, Agnieszka Noszczyk-Nowak, Katarzyna Michlik, Wojciech Niżański and Grzegorz Dejneka
The aim of the study was to evaluate the influence of sildenafil citrate administrated intravaginaly on the blood flow in the bovine uterus during dioestrus. Uterine blood flow was examined in six healthy adult cows. Sildenafil was administrated intravaginaly to each co w between the 6th and 8th d of the ovarian cycle, in the form of vaginal suppositories containing 100 mg of active substance at a dose of 100, 200, or 300 mg per animal. Uterine perfusion was estimated by the colour Doppler examination, and obtained results were analysed with the Pixel Flux Software (Chameleon, Germany). Moreover, cardiovascular parameters were also evaluated. Animals were examined before and five times after drug application (two times at 15 min intervals, and three times at 2 h intervals). A placebo suppository was also given to the cows. The analysis of the intensity and velocity of blood flow in the uterus proved that sildenafil administrated intravaginaly significantly increased blood flow in the uterus and the effect of increased perfusion was observed for 4 h and 30 min after administration. The effect of increased uterine perfusion was observed after low as well as high doses of sildenafil. Significant changes in the cardio-vascular parameters were not detected. There were no changes in the uterine perfusion as well as in cardiovascular parameters after placebo administration.
Joanna Kochan, Wojciech Niżański, Nei Moreira, Zalmir Silvino Cubas, Agnieszka Nowak, Sylwia Prochowska, Agnieszka Partyka, Wiesława Młodawska and Józef Skotnicki
With the exception of the domestic cat, all felid species (Felidae) are currently threatened with extinction in their natural habitat. To develop effective and optimal wild cat conservation programmes with assisted reproductive technology (ART) it is necessary to combine advances from different disciplines of science, starting from the biology of the species, through research into the population and habitat, assisted reproductive technologies, establishment of gene banks, developing bioinformatic systems, and ending with biodiversity and endangered species management. In the last few years knowledge of felid reproduction has expanded considerably thanks to comparative studies utilising the domestic cat as a research model for endangered wild cats. Basic reproductive techniques utilised in both domestic cat breeding and rescuing wild felid populations that are threatened with extinction include semen collection and cryopreservation, artificial insemination, oocyte collection, in vitro maturation, in vitro fertilisation, somatic cloning, and embryo transfer. The main directions in which assisted reproductive technologies are being developed in wild cat conservation implementations and the contribution of Polish research centres in advancing these methods are presented.
Michał Dzięcioł, Thomas Scholbach, Ewa Stańczyk, Justyna Ostrowska, Wojciech Kinda, Magdalena Woźniak, Wojciech Atamaniuk, Piotr Skrzypczak, Wojciech Niżański, Andrzej Wieczorek, Jacob Scholbach and Zdzisław Kiełbowicz
The aim of this study was to investigate the usefulness of new software Pixel Flux (PXFX) for clinical evaluation of tissue perfusion in the field of reproduction in dogs. The experiment was performed on six adult Beagle dogs. Different organs and tissues of the animals were examined with the MyLab25 Gold ultrasound system. Blood flow in the ovary, testicle, prostate, the ramification of the penile artery, and the network of blood vessels of the pampiniform plexus were examined with the use of colour coded Doppler technique, and obtained data was evaluated with the PXFX software. The more objective digital evaluation of data obtained with colour Doppler sonography through the application of dynamic tissue perfusion measurements provides new opportunities for diagnosis, as well as continuous monitoring of the function of the examined tissues and organs. The use of PXFX software is strongly indicated as a tool in small animal practice as an additional method for evaluation of tissue perfusion, especially in the cases when other methods like pulsed wave Doppler techniques are difficult to be performed
Wiesława Młodawska, Patrycja Mrowiec, Beata Grabowska, Joanna Waliszewska, Joanna Kochan, Agnieszka Nowak, Anna Migdał, Wojciech Niżański, Sylwia Prochowska, Agnieszka Partyka, Marcin Pałys, Teresa Grega and Józef Skotnicki
Dermal fibroblasts are commonly used as donors of genetic material for somatic cell nuclear transfer in mammals. Basic fibroblast growth factor (bFGF) is a cytokine that regulates proliferation and differentiation of different cell types. The study was aimed at optimizing the cell culture protocol for cat dermal fibroblasts by assessing the influence of culture media and different doses of bFGF on proliferation of fibroblasts and their viability in terms of cell banking and somatic cloning of felids. In Experiment I, skin biopsies of domestic cats were cultured in DMEM (D) and/or DMEM/F12 (F), both supplemented with 5 ng bFGF/ml (D-5, F-5, respectively). After the primary culture reached ~80% of confluency, the cells were passaged (3–4 times) and cultured in media with (D-5, F-5) or without (D-0, F-0) bFGF. To determine the optimal doses of bFGF, in Experiment II, secondary fibroblasts were cultured in DMEM with 0 (D-0), 2.5 (D-2.5), 5 (D-5) or 10 (D-10) ng bFGF/ml. The results showed that in D-5 the cells proliferated faster than in D-0, F-5 and F-0. Due to their poor proliferation, passages IV were not performed for cells cultured in F-0. In experiment II, a dose-dependent effect of bFGF on proliferation of cat dermal fibroblasts was found. In D-5 and D-10, the cells exhibited higher (P<0.05) proliferation compared with D-0. In D-2.5 the cells showed a tendency to proliferate slower than in D-5 and D-10 and at the same faster than in D-0. In conclusion. DMEM supplemented with bFGF provides better proliferation of domestic cat dermal fibroblasts culture than DMEM/F12. Supplementation of culture medium with bFGF has a beneficial effect on cat dermal fibroblast proliferation and could be recommended for addition to culture media.