Research

Leader: prof. dr hab. Wojciech Niżański

Investigators: dr hab. Agnieszka Partyka,prof. uczelni, dr Sylwia Prochowska, dr Wiesław Bielas

Keywords: semen, chilling, cryopreservation, CASA, flow cytometry

Research concerning various species of companion animals (dogs, cats), farm animals (boars, roosters, minks, chinchillas) and wild (mainly wild ungulates, including European bison). Spermatozoa are evaluated using computer assisted sperm analysis (CASA) and a flow cytometer, to assess such parameters as mitochondrial potential, DNA structure, intensification of oxidative stress or apoptotic changes.

In the studies on the optimization of semen preservation, diluents addition of various substances (including antioxidants, polyphenols and nanoparticles) were tested. The influence of the semen plasma on sperm quality subjected to the cryopreservation was also investigated. The results of the research will help to optimize pre-treating procedures of semen subjected for cryostorage, that should reduce the impact of many sperm-damaging factors, and thus improve the survival and motility of frozen-thawed gametes.

Based on the obtained results, cryopreservation methods suitable for different species of companion, farm and wild animals were developed. In 1997 there was established The Companion and Wild Animal Semen Bank in the Reproduction Department in Wrocław. Since 2013, the team has participated in the European bison protection program, establishing and operating the bison semen bank, collecting semen post mortem from cauda epididymis of male bison.

Investigations carried out as a part of this project:

Project no.271. 3.10.2017, 07.08.2017 „A complex project of the protection of wisent by National Forests”, financed by Forests Funds
No N N308 576540: „The quality, level of apoptosis and in vitro fertilizing ability of frozen-thawed cat’s spermatozoa collected form the urethra and epididymis”, leader W. Niżański
0401/IP1/2011/ 71: „The modification of cholesterol content in cell membranes of rooster spermatozoa for the improvement of viability and motility of gametes after cryopreservation.”, leader. A. Partyka
5300/B/P01/2011/40: „The effect of antioxidants addition during cryopreservation of rooster semen on sperm characteristics in vitro and in vivo and the expression of genes related to apoptosis”, leader A. Partyka
N N308 573339: „The antioxidant status and the level of lipid peroxidation in canine semen subjected to biotechnical procedures”, leader W. Niżański

Leader: prof. dr hab. Wojciech Niżański

Keywords: ultrasound scanning, elastography, Colour Doppler, biopsy, infertility, new therapeutic methods

Studies carried out in the Department of Reproduction include reproduction of companion animals and both the diagnosis and therapy of reproductive system diseases.

In dogs, the main points of interest concern the endometrial biopsy, rarely used in dogs, as well as the latest imaging techniques, including ultrasound examination using the Colour Doppler function to study blood flow parameters in ovarian vessels and the uterus. For the first time the reference values of vascular flows in pregnant bitches were established. Moreover, various biometric formulas for pregnancy assessment and delivery date were investigated. Additionally, the usefulness of mammary gland tumors treatment using the cold atmospheric plasma and cryotherapy in dogs and cats are investigated.

The team carries out also research requested by private companies including clinical trials of drugs (Suprelorin, Yposane, Milbemax) and diagnostic tests (Ovulation test). In cooperation with prof. dr hab. Marek Świtoński (Poznań University of Environmental and Life Sciences) the genetic basis of disorders of sex development is also investigated.

Investigations carried out as a part of this project:

1/2015. “Prospective study in the tolerance of a combination of Milbemycine Oxime and Praziquantel in Breeding, Lactating Bitches and in newborn puppies.” Leader W. Niżański, company Virbac
UMO-2012/05/B/NZ9/00907: „The investigation of genetic base of the reversed sex syndrome in dogs with 78, XX karyotype”. Leader Dr hab. M. Świtoński, investigator W. Niżański
1/2013: „“Assessment of performances of the test Ovulation test for the semi-quantitive dosage of progesterone in bitches”. Leader W. Niżański, company BTV
1/2012: “Comparison study on the effectiveness of osaterone acetate and deslorelin acetate on prostate gland in dogs”. Leader W. Niżański, company Virbac

Leader: prof. dr hab. Wojciech Niżański

Investigator: lek. wet. Jacek Mrowiec, dr Wiesław Bielas

Keywords: artificial insemination, intrauterine insemination, follicular insemination

Our team for many years have been carrying out studies on ways to improve the conditions of intrauterine insemination and support the development of early pregnancy by means of hormonal preparations, antibiotics and non-steroidal anti-inflammatory drugs. The methodology of intrauterine insemination with the use of a rigid endoscope was developed, which lead to noticeable improvement in the effectiveness of insemination in relation to routine methods.

Additionally, some research concerns the alternative methods of cattle insemination, i.e. follicular insemination were performed.

Investigations carried out as a part of this project:

KBN 3 PO6K 004 23: „The efficacy of artificial insemination of bitches in relations to in vitro characteristics of dog spermatozoa”. Leader W. Niżański
B030/0044/16 ”The analysis of efficacy of intrafollicular artificial insemination in dairy cows”. Leader J. Mrowiec
B030/0060/17 ”The influence of follicular liquid on survival, morphological parameters and motility characteristics of bull spermatozoa”. Leader J. Mrowiec

Leader: prof. dr hab. Wojciech Niżański

Investigators: dr Małgorzata Ochota, dr Agnieszka Partyka, dr Sylwia Prochowska, lek. wet. Olga Rodak

Keywords: assisted reproductive techniques, felidae, in vitro, embryos

In 2011, within the collaboration with University of Agriculture in Kraków and Foundation Municipal Park and the Zoological Garden in Cracow, the team has started the research concerning the use of assisted reproduction techniques (ART) for saving the populations of wild felids. The tests are carried out on a domestic cat (Felis catus) used as a model animal. There have been developed biotechniques and culture media for female and male gametes to optimize maturation, fertilization (IVF and ICSI) and in vitro culture. Effective protocols for semen freezing and vitrification of oocytes and embryos have been developed – allowing cells and tissues to be stored for an unlimited period of time, thereby contributing to the conservation of genetic resources of wild felines threatened with extinction. Additionally, the tested gametes and embryos are subjected to detailed evaluation using modern techniques (flow cytometry, fluorescence microscopy).

Currently, research concentrates to develop the ovarian cortex tissue cryopreservation using vitrification and programmed freezing, as well as somatic cloning using the nuclear transfer method (NT).

The protocols developed for the domestic cat have already been successfully used in wild cats which died in zoological gardens. As the result, the gene bank of wild felids was created in 2015, in which fibroblasts, semen and oocytes from ocelot, tiger, lion, jaguarundi, lynx, serval, manul, caracal and some domestic cat breeds were stored. It is the first one in Poland and unique in Europe somatic cell bank of a domestic cat and wild felids (similar centre exists only in Uppsala), courtesy of the cooperation with the ZOO Kraków.

Investigations carried out as a part of this project:

No PBS3/B8/16/2015: „Increasing the innovativeness and effectiveness of programs for the conservation of genetic resources of wild felids by creating a bank of cells and implementing methods of in vitro embryo production into practice." Leader W. Niżański
No 2011/03/B/NZ9/03412: „The research on the influence of low temperatures on the developmental competence of vitrified feline embryos, as a one of strategies for saving genetic resources of Felidae family.” Leader M. Ochota

Leader: dr hab. Michał Dzięcioł

Investigators: lek. wet. Martyna Woszczyło, lek. wet. Barbara Pieczewska

Keywords: dog, semiochemicals, pheromones

The aim of the project is the identification and verification of the semiochemical activity of the components of vaginal secretions in bitches in heat carried out on the dog’s model (Canis familiaris).

The research carried out so far concerned the suitability of synthetic animal pheromones to stimulate sexual behaviour of male dogs. The new software was used for the objective assessment of blood flow relative to the active substances and pheromones way of action, measured using the Color Coded Doppler (PixelFlux Software). The influence of antibiotic therapy on the attractiveness of vaginal discharge in bitches and the possibility to use male dogs to identify bitches in optimal mating time were also examined. The ongoing research concerns the separation of semiochemical substances responsible for the attractiveness of bitches in heat.

Investigations carried out as a part of this project:

No UMO-2015/17/B/NZ8/02411: „The mechanisms of semiochemical communication in canidae in the relationship to the reproductive behaviour: the research on the domestic dog (Canis familiaris) as a model species.” Leader W. Niżański

Leader: prof. dr hab. Roland Kozdrowski
Investigators: dr Justyna Buczkowska, lek. wet. Monika Sikora, lek. wet. Katarzyna Wojtysiak

Keywords: mare, metritis, endometrial biopsy, endometrial cytology

The research focused on improving the diagnosis of chronic uterine inflammation in mares. Special attention was paid to:
-performing and evaluating of the uterine flush for the assessment of the inflammation level.
-the usefulness of a uterine biopsy in assessing changes of the endometrium. There was developed the reliable protocol utilising uterine biopsy in regards of cycle phase and specimens assessment protocol.
-immunohistochemistry tests as an additional diagnostic tool for metritis.
-acute phase proteins: alpha-amyloid and haptoglobin for the diagnosis of subclinical uterine inflammation.

Currently, the research focuses to determine the role of cytokines in the pathogenesis of after-mating metritis, as well as their potential use in diagnostics. Differences in the concentration of cytokines in uterine flushes from healthy and ill mares will allow early diagnosis and, consequently, the implementation of faster and more effective therapy.

Investigations carried out as a part of this project:

UMO-2011/01/B/NZ5/04173: „The assesment of endometrium of mares showing subclinical endometritis based on microbiological, cytological, histological and molecular analysis.” Leader R. Kozdrowski

prof. dr hab. Jan Twardoń, lek. wet. Jacek Mrowiec, lek. wet. Agnieszka Bartoszewicz

Keywords: dairy cows, udder, mastitis

Our team has been carrying out studies on reproduction of dairy cattle. Their focus is on the udder health and the milk quality in dairy cattle farms. The identification of atypical mammary gland pathogens: Streptococcus canis and Staphylococus microti was performed in order to determine their pathogenic importance and methods of its effective control. Currently conducted research concerns the possibility of using itraconazole in the therapy of fungal mastitis in cows. Research also focuses on improving the reproduction indices on farms utilising a new, fast method to determine the progesterone level and evaluation of energy potential (energy balance) in the perinatal period in high yielding cows.

Investigations carried out as a part of this project:

No PBS2/B8/15/2014: „Immunological tests for detection and measurements of progesterone in cows’ milk.” Leader J. Twardoń
No 49/14-W/2011/G: „Analysis of correlation of chosen biochemical and clinical parameters of dairy cows for the establishment of biocybernetic model of cow’s organism.” Leader. J. Twardoń

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