The European eel Anguilla anguilla is a highly important market species which is also increasingly in danger of extinction in Europe. One of the ways of protecting the species in the natural environment, while maintaining its market supply, is its aquaculture, e.g. due to much higher survival rate under controlled conditions than in the wild. However, this can be done only when an effective artificial reproduction biotechnique is developed. The aim of this study was to use an ultrasound device to determine the sex and to perform a biopsy of the gonads, which is a part of complete protocol of eel artificial reproduction. The findings indicate that ultrasonography is highly useful in determining the sex in migrating eel (100% of sureness) and in performing high-precision biopsy of the gonads. The present method gives the possibility to quick determine the maturity of the female gonads. The application of ultrasonography (USG) is totally safe for fish and a portable ultrasound device can be used in both the laboratory and in the field.

Common parasites of the European bison include gastro-intestinal and pulmonary nematodes, liver flukes (Fasciola hepatica), tapeworms, and protozoa of the genus Coccidia. This study compared the extensiveness and intensities of European bison parasitic invasions in three north-eastern Polish forests in different seasons and queried the role of parasitological monitoring in sanitary and hygienic control of feeding places. Faecal samples were collected in the Białowieża, Knyszyńska, and Borecka Forests between 2014 and 2016, as were some from an area neighbouring the Białowieża Forest outside the Natura 2000 protected area. Parasites were detected in individual samples with the flotation, decanting and Baermann methods. The eggs of Trichostrongylidae, Aonchotheca sp., Nematodirus sp., Strongyloides spp., Trichuris sp., Moniezia spp., and Fasciola hepatica; the larvae of Dictyocaulus viviparus; and the oocytes of Eimeria spp. were identified. Significant variation in invasion intensity and diversity was seen by origin and season. The relationships were assessed first by univariable tests and next multivariately, when origin and season emerged as the major risk factors for exposure to most of the parasites. The differences in the level of parasitic infection between the forests did not have implications for its sufficiency to cause clinical symptoms. However, the associations and risk factors found enable the necessary preventive measures to be taken to protect the E. bison from exposure or decrease the risks. Additionally, parasitological monitoring is appropriate as the method of sanitary and hygienic control of European bison winter feeding places. Threats to public health through adventitious invasions by zoonotic factors such as F. hepatica have been identified.

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.

Introduction The addition of low-molecular-weight antioxidants during the freezing process improves post-thaw sperm quality. The high antioxidant potential of cryopreserved semen could have a positive effect on the motility, viability, and energy status of sperm cells and their ability to bind to the zona pellucida of oocytes. The aim of the study was to determine the effects of different concentrations and combinations of vitamins E and C in a semen extender on selected quality parameters of frozen-thawed canine spermatozoa. Material and Methods The experimental material was the semen of four mixed-breed dogs. Sperm viability (motility, plasma membrane integrity, and mitochondrial function) was examined at 0, 60, and 120 min in semen samples supplemented with the extender and in the controls. Results Combined supplementation with vitamins C + E at a concentration of 200 + 200 μM /1 × 10⁹ spermatozoa had the most profound effect on total sperm motility, linear motility, and the percentage of spermatozoa with intact plasma membrane and active mitochondria. Conclusion The synergistic activity of vitamins E and C had a more beneficial influence on the quality of frozen–thawed sperm than these non-enzymatic antioxidants applied separately.

Repeat breeding is a serious reproductive disorder in dairy cattle. The causes of repeat breeding are multifactorial and there are two main mechanisms: failure of fertilisation or early embryo death, mainly due to poor quality of oocytes and an inadequate uterine environment. Many methods have been used to increase the pregnancy rate for repeat breeder cows, such as intrauterine infusion of antibacterial agents or antibiotics, hormonal treatments for oestrus synchronisation and induction of ovulation, and progesterone supplementation or induction of accessory corpus luteum; however, the results were inconsistent between studies. Embryo transfer (ET) has the capability to minimalise the effects of poor oocyte quality and unfavourable uterine environments on early embryo development during the first seven days after ovulation in repeat breeder cows, and several studies showed that ET significantly improved the pregnancy rate in this group of animals. Thus, ET can be considered an option to increase the conception rate in repeat breeder dairy cows.

The left ovary of chicken embryos was removed and incubated in culture medium with a thymidine analogue, bromodeoxyuridine (BrdU), in vitro. In addition, fertile chicken eggs were injected with BrdU via the extraembryonic vessels and incubated for 24 hours. The ovaries were then processed for immunohistochemical localization of calbindin-D28K (a 28-kd vitamin D-dependent calcium-binding protein) and BrdU. Calbindin-D28K was detected in the germinal epithelium and in cells surrounding the oogonia and oocytes (future granulosa cells) of the embryonic chicken ovary. However, Brdu was observed in the nucleus of the oogonia and oocytes of the chicken embryonic ovaries. Comparison of the 2 adjacent sections, immunostained for calbindin-D28K and BrdU consecutively, indicated that BrdU, the marker for cell proliferation was not detected in calbindin-D28K-containing cells, namely, germinal epithelium and future granulosa cells, in the ovary of chicken embryos. These results suggested that calbindin-D28K-containing cells in the ovary were not in the process of cell division during the 24-hour incubation of chicken embryos.

To optimize the most efficient method for porcine in vitro maturation (IVM), we compared the effects of supplementing extracellular vesicles (EVs) derived from porcine follicular fluid (pFF). The cumulus oocyte complexes were grouped into 4 groups with different supplementations as following: pFF (G1), pFF-depleted EVs (G2), EVs (G3) and control (G4) groups. After IVM with different supplementations, maturation rates and the developmental competences of porcine oocytes and blastocyst development were investigated. Additionally, glutathione (GSH) and reactive oxygen species (ROS) levels were measured in mature oocytes. The EVs were isolated and characterized with cryo-TEM and nanoparticle tracking analysis. The pFF significantly affected the maturation rate, whereas the presence of EVs did not show notable difference in the maturation rates. Although there were numerical increases in the measured parameters in EV and pFF-depleted EVs groups, no significant differences were observed between them. The EV group showed similar oocyte maturation rate for both positive and negative control groups. The GSH was not different among the groups, but ROS levels were significantly lower in pFF-supplemented group when compared with other groups with the highest level in the control group. G2 group wasn’t significantly different G1 and G3 group. G3 group wasn’t significantly different from G2 and G4 group. This suggests that EVs in IVM medium which probably effected partially to protect against oxidative stress and potentially enhance the quality of oocytes. This study indicates that the EVs in pFF play a significant role in improving the efficiency of oocyte maturation in porcine.

Infertility is a major concern affecting a huge population worldwide. Aneuploidy is one of the leading causes of the infertility, usually originated from errors that happens in meiosis I as chromosomal misalignment and abnormal chromosomal segregation. Recent research showed that cathepsins family play an important role in affecting the oocytes developmental competence in many species. Cathepsin B inhibition improved bovine developmental competence of the oocytes, especially in the low-quality oocytes. Here, we are the first to introduce the significance of another member of this family, cathepsin L (CTSL) in the process of oocyte maturation using mouse oocyte model.  In this study we evaluated the expression of CTSL during oocyte meiotic maturation and the effect of CTSL inhibition using specific inhibitor “SCP110” on the subsequent maturation process. We found, CTSL is expressed in all stages of Meiosis I. In addition, inhibition of CTSL resulted in delaying the maturation time by elongation the time required for extrusion of the polar body (PBE). Moreover, CTSL inhibition reduced the maturation rate compared to the control group as we found a significant decrease in PBE percent. Trying to understand the reason of the lower maturation rates, we stained the chromosomes and spindles at metaphase I stage, we found a significant increase in chromosomal misalignment at metaphase plate. Finally, we evaluated matured oocytes quality after CTSL inhibition, importantly, aneuploidy incidence was increased significantly after CTSL inhibition. In summary, CTSL is required for normal meiosis completion in oocytes and production of healthy euploid egg.

Although the developmental potential of oocytes is related to oocyte quality, whether the expression of specific genes is altered in oocytes of different quality and in resulting embryos is not known. Semi-quantitative reverse transcription-polymerase chain reaction was used to compare the relative abundance of 2 transcripts for housekeeping proteins (β-actin and ribosomal protein L30) and 3 transcripts for growth factor ligand or receptors (platelet derived growth factor receptor α (PDGFRα), basic fibroblast growth factor (bFGF)), in mature bovine oocytes of high versus low developmental potential. The transcripts for L30, PDGFRα, and bFGF in 16-cell embryos originating from these oocytes were also examined. No significant effect of oocyte quality was detected for any of the transcripts examined from oocytes or 16-cell embryos. In conclusion, a lower developmental potential of oocytes with advanced signs of atresia, was not associated with a lower level of abundance of the transcripts examined.

Immunoreactivity for 28 kd vitamin D-dependent calcium-binding protein (calbindin-D28k) has been localized in the germinal epithelium and cells surrounding oogonia and oocytes (future granulosa cells) of developing and growing ovaries of chicken embryos. The protein first appeared prominently in the germinal epithelium of the developing left ovary in 8-day embryos. At the twelfth day of incubation, cells surrounding oogonia and oocytes reacted intensely for calbindin-D28k. The number and intensity of calbindin-D28k-containing cells increased in both types of cells as the embryos further developed. Calbindin-D28k remained in the germinal epithelium throughout the study period observed (up to 10 weeks). However, the protein was present transiently in the future granulosa cells. It gradually decreased after hatching, and was virtually absent from granulosa cells in a 10-week old chicken. Compared with the known process of onset of sexual development, these results indicated possible involvement of calbindin-D28k in the early phases of oogenesis in chicken ovaries.