Leader investigator of the task: prof. dr hab. Wojciech Niżański
Investigators: dr Wiesław Bielas, dr Agnieszka Partyka, dr Sylwia Prochowska
Summary: The aim of the project is post-mortem collection of epididymal wisent semen and its cryopreservation for further use for artificial insemination and in vitro fertilization. Collected spermatozoa (both fresh and frozen thawed) are evaluated with modern techniques such as Computer Assssted Sperm Analysis (CASA) and flow cytometry. This approach allows for deeper understanding of gametes’ biology and cryobiology in this species. Semen cryopreservation led to the creation of wisent semen bank. Fertilizing ability of samples stored in this bank was confirmed by in vitro fertilisation and obtaining the first wisent blastocyst.
Keywords: wisent, spermatozoa, semen bank
Project NCN 2015/17/B/NZ8/02411.
2016-2019
Leader investigator of the task: prof. dr hab. Wojciech Niżański
Investigators: dr hab. Michał Dzięcioł, lek. wet. Martyna Woszczyło, lek. wet. Barbara Pieczewska
Summary: The aim of the project is the investigation of the semiochemical communication in the relationship to reproductive behaviour using dog (Canis familiaris) as a model animal. The main target is the description of the sex pheromones produced in bitches during heat that modify the typical males’ behaviour. There is still a lack of knowledge concerning its chemical composition, which if discovered would allow to produce its commercial equivalents, useful in regulation of reproduction. The results would be used not only in dogs, but also in other canines including endangered species. The capacity of pheromones in modifying the behaviour and physiology of the receiving individual usually are easy to observed and interpret. Hence the relationships there can be considered as a model and the methods developed to describe their action and identification would be a background for further investigations on other groups of pheromones (social or repelling pheromones).
Keywords: canine, pheromone, semiochemistry, sexual behaviour
Leader investigator of the task: prof. dr hab. Wojciech Niżański
Investigators: dr Agnieszka Partyka, dr Sylwia Prochowska, Joanna Kochan, Wiesława Młodawska, Agnieszka Nowak, Anna Migdał, Józef Skotnicki, Teresa Grega, Marcin Pałys
Summary: The aim of the project is to implement the assisted reproductive techniques into gene preservation programmes of wild felids. The cooperation with University of Agriculture in Kraków and the Zoologiacal Garden in Kracow allows to collect skin samples of wild and domestic cats for the fibroblast culture. Obtained cells will be later cryopreserved and stored in the bank of fibroblast. The in vitro fertilisation will be attempted using domestic cat’s gametes as a model animal, as well as the somatic cloning will be performed using domestic cat’s oocytes and the previously obtained and preserved fibroblasts. The expected result is to increase the novelty and effectiveness of the genetic resources conservation programmes especially for the local population of Eurasian lynx and European wildcat. The development of the cloning technique using somatic cells of wild cats enables for state of the art of conservation programmes
Keywords: cat, wild felids, genetic resources protection, ICSI, cloning
2015- 2016
Leader investigator: prof. dr hab. Wojciech Niżański
Investigator: lek. wet. Barbara Pieczewska
Summary: The aim of the project was to assess the safety of the commonly used deworming medication for dogs during pregnancy and for new born puppies up to weaning period. The results showed no negative effects of the investigated substances included in MilPro preparation (milbemycin oxime and praziquantel) on the pregnancy and puppies’ development. There were not noticed higher risk of pregnancy pathology, malformations or puppies deaths in animals receiving the drug every 2 weeks starting and the beginning of the pregnancy and up to weaning. The investigations were carried out in the agreement with Virbac company, Deparment R&D.
Keywords: antihelmintics, praziquantel, milbemycin oxime, dog, pregnancy, pup
Project NCN N308 576240
2011-2014
Leader investigator: prof. dr hab. Wojciech Niżański
Investigators: dr Sylwia Prochowska, dr hab. Małgorzata Ochota, prof. uczelni, dr hab. Agnieszka Partyka, prof. uczelni
Summary: The aim of the research was to describe the properties, fertilising ability and the suitability for assisted reproductive techniques of frozen-thawed cat’s (Felis domesticus) spermatozoa, obtained from urethra and compared to the epididymal spermatozoa. The research would show whether the semen obtained from the urethra, after the administration of an alpha-mimetic agent, which cause contractions of the vas deferent is useful in obtaining the good quality semen suitable for in vitro fertilization, artificial insemination and obtaining embryos good for transfer. Additionally, the research should provide the basis for establishment of the genetic reserve of male felids (semen bank).
Keywords: cat, semen, sperm, cryopreservation, in-vitro fertilization, apoptosis
Leader investigator: prof. dr hab. Wojciech Niżański
Investigator: dr Joanna Pasikowska
Summary: The study assessed the effectiveness of treatment of benign prostatic hyperplasia (BPH) using two drugs with different way of action – osaterone acetate (OA, the androgen receptor blocker) and deslorelin acetate (DA, used for pharmacological castration in males). It was demonstrated that the DA affected the prostate for 12 months, while chemical castration lasted for 6 months. This gives the opportunity to use a stud dog when the chemical castration effect wears off. It was also demonstrated that the DA is extremely effective in treatment of BPH causing prostate involution comparable to the size observed in surgically castrated males. Whereas OA was shown to be effective in inducing temporary gland involution without losing the fertility. There were also designed algorithms and ways to diagnose and monitor the therapy with the use of Color Doppler.
2013-2014
Leader investigator: prof. dr hab. Wojciech Niżański
Investigator: lek. wet. Honorata Ziętek
Summary: the aim of the study was to check the levels of serum progesterone in bitches using the RIA (reference method) and semi-quantitative ELISA test easy to use in field conditions. The studies shown that the similarity in the obtained results between both tests ranged around 80%. Therefore, the semi-quantitative test seems to be very useful for field and every day practical use.
Keywords: progesterone, semi-quantitative ELISA test, females
Project NCN N308 573339
2010-2013
Leader investigator: prof. dr hab. Wojciech Niżański
Investigators: dr Marta Gotowiecka, dr hab. Agnieszka Partyka, prof. uczelni
Summary: The aim of the project was to determine the impact of reactive oxygen species (ROS) on dogs’ spermatozoa subjected to preservation in low temperatures and in liquid state, and to determine the influence of those procedures as well as the presence of seminal plasma on the self-antioxidant defence mechanisms. Studies showed the relationship between the oxidative stress in semen cryopreserved and stored in liquid state and sperm parameters. Additionally, the role of diluents supplementation with enzymatic and low-molecular compounds was investigated. The investigations helped in improving the earlier developed preservation methods for dogs’ semen.
Keywords: dog, semen, seed preservation, reactive oxygen species, antioxidants, lipid peroxidation, apoptosis
Project NCN N308 576240.
2011-2013
Leader investigator: prof. dr hab. Wojciech Niżański
Investigators: dr Agnieszka Antończyk, dr hab. Agnieszka Partyka, prof. uczelni
Summary: The aim of the research was to introduce computer assisted semen analysis (CASA) for veterinary practice, in particular to describe kinetics of spermatozoa movement. The research made it possible to design the initial settings for CASA system to asses spermatozoa in fresh and cryopreserved semen. Moreover, the comparative parameters for sperm motility in fresh and frozen-thawed semen were described, which extended the current knowledge on sperm cryobiological features and helped to understand the nature of cryo-injuries of male gametes. Also, automated sperm head morphology analysis (ASMA) was performed for the first time in dogs’ spermatozoa. The morphometric changes in dogs’ spermatozoa occurring as a result of the cryopreservation procedures were described. The research helped also to develop freezeability indicators with the use of computer assisted methods. CASA and ASMA algorithms were designed to help in identifying spermatozoa as good or bad freezers.
Keywords: semen, dog, CASA, ASMA, cryopreservation, suitability for cryopreservation markers
Project NCN UMO- 2011/03/B/NZ9/03412.
2012–2015
Leader investigator: dr hab. Małgorzata Ochota, prof. uczelni
Investigators: mgr Barbara Smalec, mgr Anna Pasieka
Summary: The project was designed to develop suitable methods for cats’ embryo vitrification. The research were the foundation for cryopreservation of wild cats’ embryos. The investigations brought valuable data on molecular changes occured in cryopreserved embryos. Hence found the answer which cryopreservation technique is the most suitable for feline embryos as well as how far the apoptotic changes were induced by low temperature storage.
Keywords: cat, embryo, vitrification, apoptosis, in vitro
Project NCN 49/14-W/2011/G.
2011-2014
Leader investigator: prof. dr hab. Jan Twardoń
Investigators: dr Marta Gotowiecka, dr hab. Agnieszka Partyka, prod. uczelni
Summary: The aim of the investigation was to design a biocybernetic model of a dairy cow. The model of cow’s body used in reproduction may than be used in designing cybernetic system aiding the productivity, health and reproduction in dairy herds. Such a system is based on data obtained in dairy herds and will be a centre of analytical model for monitoring the cows’ condition and in herd management. Studying the correlation of various reproductive data with the biochemical and physical parameters was necessary to develop mathematic formulas which help to investigate the efficacy in herd management.
Keywords: biocybernetic model, cow, physiological functions
Project NCN UMO-2011/01/B/NZ5/04173
2011-2014
Leader investigator: prof. dr hab. Roland Kozdrowski
Investigators: dr Justyna Buczkowska, dr Monika Sikora, lek. wet. Katarzyna Wojtysiak
Summary: The aim of the project was to compare different diagnostic methods used for diagnosis of the subclinical endometritis in mares. There were compared the results of bacteriological and cytological examination of samples taken with the use of uterine cytobrushes, microbiological cultures and cytological examination of endometrial biopsy in relation to histopathology results used as the reference method. The project evaluated the expression of genes involved in the metabolism of arachidonic acid (PTGS2, PGES, PGFS) the products of arachidonic acid metabolism (prostaglandins, praostacyclins, leukotrienes) and Toll-like receptors 2 and 4 (Toll-like receptor 2 and 4). Moreover, the proinflammatory cytokines (TNF alpha, IL-1 beta, IL-6) and changes in gene expression for these cytokines and their receptors were investigated. The projects aimed also to determine the optimal procedure to diagnose the subclinical uterine inflammation in mares.
Keywords: mare, endometritis, diagnosis, inflammatory cytokines
Project NCN N N311 5300 40
2011-2014
Leader investigator: dr hab. Agnieszka Partyka, prof. uczelni
Investigators: prof. dr hab. Wojciech Niżański, dr inż. Joanna Bajzert, prof. dr hab. Ewa Łukaszewicz
Summary: this project investigated the influence of the cysteine and superoxide dismutase addition into the diluent used for rooster semen cryopreservation and the biological value of the semen in vitro (sperm characteristics, antioxidant enzyme activity, gene expression: pro-apoptotic Bac and Boc, anti-apoptotic Bcl-2 and Mcl-2, and the expression of heat shock proteins HSP60 and 70) and in vivo (artificial insemination of hens). The research allowed to develop the composition of the diluent with added antioxidants used during cryopreservation of rooster semen, which in future may help to optimize semen management to limit the gamete damage.
Keywords: cryopreservation, antioxidants, apoptosis, rooster semen
Project NCN UMO- 2012/05/B/NZ9/00907
2013-2016
Leader investigator: prof. dr hab. Marek Świtoński
Investigator: prof. dr hab. Wojciech Niżański
Summary: Studies will help to look for the causes of inherited reproduction disorders. The project is a part of the cooperation with the University of Life Sciences in Poznań (Prof. Marek Świtoński) using patients of the Department of Reproduction in Wroclaw. The project includes in-depth clinical and endocrinological examinations of patients with abnormalities of genital organs, combined with morphological, ultrasound (USG, color Doppler, CT) and histopathological investigation of gonads, as well as genetic and molecular analysis. The studies are to determine the genetic causes of sexual developmental disorders and estimate the degree of inheritance in case of intersexualism.
Keywords: genetic disorders, developmental abnormalities, reproductive organs, intersex, kariotype
Project MNiSW Nr N N308 008 31/3067
2007-2009
Leader investigator: prof. dr hab. Wojciech Niżański
Investigator: dr hb. Agnieszka Partyka, prof. uczelni
Summary: The aim of the study was to describe the quality of the insemination doses used in the national insemination centres for sows. The research was performed on the boar semen preserved in liquid state using long- and short-lasting diluents. The study used computer-assisted sperm analysis (CASA) and flow cytometry. It was shown using SYBR-14 / PI fluorochrome that the boar sperm membrane is extremely stable with unchanged integrity for almost 10 days incubation. Whereas, the spermatozoa motility decreased significantly within the first 2 – 3 days of incubation, regardless the diluent used. This project allowed for development of the gold standards for boar semen assessment using CASA and flow cytometry devices.
Key words: boar, semen, preservation in liquid state, CASA, flow cytometry
Projekt MNiSW Iuventus Plus 0401/IP1/2011/ 71
2012-2014
Leader investigator: dr Agnieszka Partyka
Investigators: prof. dr hab. Wojciech Niżański, mgr Maciej Strojecki
Summary: The aim of this project was to determine the optimal concentration of a chosen cyclodextrin or cyclodextrin-cholesterol complex added to the rooster semen before freezing, that would improve the viability and motility of spermatozoa after thawing. Moreover, there was investigated the effect of modifying cholesterol content (removal/addition) in cell membranes of rooster sperms on: 1) viability, motility and acrosome status of gametes before and after cryopreservation; 2) cholesterol content in sperm cell membranes before and after cryopreservation, and 3) changes in fluidity and the degree of lipid ordering in the sperm cell membranes subjected to this procedure. The obtained results will be used to design the most suitable treatment of semen before cryopreservation to limit the effect of physicochamiacal and biological factors affecting male gametes.
Keywords: cholesterol, cyclodextrin, cholesterol, membrane fluidity, cryopreservation and rooster semen
