Microsatellite markers were employed to determine the genetic diversity and structure of eight goat breeds kept in the Czech Republic (White Shorthaired goat, Brown Shorthaired goat, Alpine, Anglo-Nubian) and Poland (Polish White Improved, Polish Fawn Improved goat, Sandomierska, Toggenburg). Across 398 individuals analyzed using 15 microsatellite markers, a total of 191 alleles were detected, with a mean of 7.108 alleles per locus. The average of observed and expected heterozygosity, fixation index, and Shannon’s information index were 0.684 ± 0.014, 0.696 ± 0.013, 0.014 ± 0.012, and 1.476 ± 0.037, respectively. The mean of overall genetic differentiation (FST) was 0.110, heterozygote deficiency among populations (FIT) was 0.125, and the inbreeding coefficient within the goat population (FIS) was 0.018. Analysis of molecular variance showed that 10% of the total genetic variation was attributed to differences among populations, 3% among individuals within populations, 86% within individuals, and 1% was observed among regions. A phylogenetic dendrogram and principal coordinate analysis (PCoA) revealed three main clusters, while STRUCTURE analysis identified two main gene pools. These findings provide critical insights for future genetic improvement, breeding program optimization, and conservation strategies for local landrace goat breeds, whose genetic resources are pivotal for maintaining livestock biodiversity.

In the mouse, spermatozoa are highly resistant to DNA damage, even when frozen without cryoprotectants, and can produce offspring when subsequently used for ICSI (intracytoplasmic sperm injection). It is not known whether the same applies to other mammals as well. For example, in the horse, even conventional sperm freezing is still very problematic and frequently leads to sperm immobility. It has, however, never been tested whether sperm immobility also mirrors sperm head DNA damage, and if so, to what extent. In our study, we evaluated the damage to DNA in horse frozen and thawed motile and immotile spermatozoa after their injection into ovulated mouse oocytes. In both groups, injected horse spermatozoa activated the mouse oocytes. This was followed by the extrusion of the second polar body (2 PB) and the formation of maternal pronuclei (Mo-fPN-mouse female pronucleus); in parallel, the horse sperm heads rapidly decondensed in the murine cytoplasm and formed paternal pronuclei (Ho-mPN-horse male pronucleus), which were larger than the female ones. With the exception of one stallion tested, DNA damage has been detected in almost all Ho-mPNs originating from immotile spermatozoa. DNA in motile (even sporadically) spermatozoa was mostly undamaged. Moreover, when the xenogeneic zygotes cleave to the two-cell stage, the incidence of micronuclei in blastomeres mirrors the extent of DNA damage in paternal pronuclei. In conclusion, and contrary to the mouse, where sperm DNA is very resistant to damage, we do not recommend the use of immotile horse spermatozoa for ICSI. On the other hand, even the sporadically motile mouse spermatozoa have no damaged DNA and can thus be used for intragenic ICSI.

There are circumstances in which mammalian spermatozoa are collected and/or long-term stored under suboptimal conditions, such as during posthumous sperm retrieval. As a consequence, the sperm can exhibit lower motility, be completely immotile, or have membranes that are more or less severely damaged. With the advent of ICSI (intracytoplasmic sperm injection), either the entire sperm or only the sperm head (e.g., in the mouse) can be directly injected into a mature oocyte, thereby making motility a less critical parameter. However, if sperm were collected and stored under suboptimal conditions, blind sperm injection into oocytes seems risky, especially when oocytes are also scarce, as in the case of endangered animals. The same applies to humans, where safety and ethical considerations are primary concerns. If such sperm are intended for reproductive purposes, DNA quality and integrity should be routinely evaluated, as they are among the most important factors for embryo viability. Although there are several standard tests to assess sperm quality, they may not be sufficient when working with to a certain extent compromised samples. Here, we discuss whether, in these specialized cases, interspecific ICSI could help expand the range of tests available to assess sperm normality.

A necessary step towards the development of genetic diversity is the protection of the valuable genetic resources of farm animals that are at risk of extinction. We analyzed 375 individuals of nine local sheep breeds bred in Central Europe (Carpathia area) from Czech Republic, Slovakia, Poland, Ukraine, and Romania using a panel of 13 microsatellite markers to investigate genetic differences and evaluate the genetic structure among and within breeds, thereby improving future breeding and conservation strategies. The mean number of alleles was 8.84, the mean number of effective alleles was 4.76, and the polymorphism information content (PIC) was 0.79. Diversity was measured using principal coordinate analysis (PCoA) as well as genetic structure, which revealed two main clusters. The first cluster was the Czech Wallachian sheep (CVA) and the Świniarka (SWI). The second cluster consisted the Improved Wallachian sheep (IVA), the Šumava sheep (SUM), the Slovak Wallachian sheep (SVA), the Polish Mountain sheep (POG), the Uhruska sheep (UHR), the Ukrainian sheep (UKR) and the Tsurcana sheep (TUR). The values of genetic distance and the fixation coefficient indicate sufficient differences between the analyzed breeds (Gst = 0.052 and Fst = 0.063). Negative values of the inbreeding coefficient also confirmed the predominance of outbreeding (Fis = −0.015). The results obtained may be helpful in breeding programs and conservation plans for local sheep breeds, as their genetic resources must be preserved to maintain an adequate level of biodiversity in animal husbandry.

The sperm heads of frozen/thawed horse spermatozoa were injected into ovulated mouse oocytes where they decondensed and formed paternal pronuclei in which the DNA damage has been evaluated. The motile spermatozoa exhibit none or minor level of DNA damage and the xenogenic zygotes cleave to the two-cell embryonic stage with regular nuclei and no micronuclei (MNs). On the other hand, the immotile spermatozoa exhibit typically high levels of DNA damage and blastomeres in two-cell stage embryos contain multiple MNs. Moreover, extremely high sperm DNA damage prevents even the cleavage to the two-cell stage.Keywords: keyword 1; keyword 2; keyword 3 (List three to ten pertinent keywords specific to the article yet reasonably common within the subject discipline.)

Although dairy goat production, characterized by traditional production on small farms, is an important source of income in the Czech Republic and Slovakia, locally adapted breeds have not been fully consolidated over the last 100 yr due to large fluctuations in population size and inconsistent breeding programs that allowed for different crossbreeding strategies. Our main objective in this study was therefore to assess the conservation status of 4 Czech (Alpine Goat, White Shorthair, Brown Shorthair, and Czech Landrace) and 1 Slovak (Slovak White Shorthair) local goat breeds, to analyze their population structure and admixture, and to estimate their relatedness to several neighboring breeds. Our analyses included 142 goats belonging to 5 local breeds genotyped with the Illumina 50K BeadChip, and 618 previously genotyped animals representing 15 goat breeds from Austria and Switzerland (all analyses based on 46,862 autosomal SNPs and 760 animals). In general, the conservation status of the Czech and Slovak local goat breeds was satisfactory, with the exception of the Brown Shorthair goat, as the analyzed parameters (heterozygosity, haplotype richness, runs of homozygosity-based inbreeding, and effective population size) were mostly above the median of 20 breeds. However, for all 5 Czech and Slovakian breeds, an examination of historical effective population size indicated a substantial decline about 8 to 22 generations ago. In addition, our study revealed that the Czech and Slovakian breeds are not fully consolidated; for instance, White Shorthair and Brown Shorthair were not clearly distinguishable. Considerable admixture, especially in Czech Landrace (effective number of parental clusters = 4.2), and low but numerous migration rates from other Austrian and Swiss breeds were found. These results provide valuable insights for future breeding programs and genetic diversity management of local Czech and Slovak goat breeds.

Mammary gland anatomy in small ruminants is very similar to that of cows; however, milk synthesis throughout lactation exhibits many functional particularities in small ruminants compared with that of cows. Goat’s milk is beneficial for human nutrition owing to the fatty acid composition, fat globule size, and conjugated linoleic acid content. As a raw material for dairy products, goat’s milk must be safe for human consumption. The number of mesophilic microorganisms, somatic cells, and selected mastitis pathogens should be limited. A prerequisite for the production of milk of high hygienic quality is the health of the mammary gland. Goat’s milk processing into cheese and other products is in the Czech Republic mostly performed on farms, partly for direct sales to consumers and partly for supplying selected stores. Revenues from dairy commodities represent the most important source of income for dairy goat farms. Mammary gland health has an important effect on the economics of dairy goat farms. Profitability can fall by up to 1/3 owing to indirect effects of udder health problems.

In contrast to the livestock industry, sperm cryopreservation has not yet been successfully established in the poultry industry. This is because poultry sperm cells have a unique shape and membrane fluidity, differing from those of livestock sperm. The objective of this review is to discuss the cellular and molecular characteristics of rooster spermatozoa as a cause for their generally low freezability. Furthermore, here, we discuss novel developments in the field of semen extenders, cryoprotectants, and freezing processes, all with the purpose of increasing the potential of rooster sperm cryopreservation. Currently, it is very important to improve cryopreservation of rooster sperm on a global scale for the protection of gene resources due to the incidence of epidemics such as avian influenza.

Wallachian and Sumava sheep are autochthonous breeds that have undergone a significant bottleneck effect and subsequent restoration efforts. The first objective of this study was to evaluate the degree of genetic variability of both breeds and, therefore, the current management of the breeding. The second was to determine whether these two breeds still retain their genetic uniqueness in relation to each other and other breeds, despite regenerative interventions. Our data consisted of 48 individuals of Sumava and 37 individuals of Wallachian sheep. The comparison data contained 25 other breeds (primarily European) from the HapMap dataset generated by the International Sheep Genomics Consortium. When comparing all 27 breeds, the Czech breeds clustered with 15 other breeds and formed a single branch with them according to Nei's distances. At the same time, however, the clusters of both breeds were integral and easily distinguishable from the others when displayed with principal component analysis (PCA). Population substructure analysis did not show any common genetic ancestry of the Czech national breeds and breeds used for regeneration or, eventually, breeds whose ancestral population was used for regeneration. The average values of FST were higher in Wallachian sheep (FST = 0.14) than in Sumava sheep (FST = 0.08). The linkage disequilibrium (LD) extension per autosome was higher in Wallachian than in Sumava sheep. Consequently, the Ne estimates five generations ago were 68 for Sumava versus 34 for Wallachian sheep. Both native Czech breeds exhibit a wide range of inbreeding based on the excess of homozygosity (FHOM) among individuals, from -0.04 to 0.16 in Sumava and from -0.13 to 0.12 in Wallachian. Average inbreeding based on runs of homozygosity was 0.21 in Sumava and 0.27 in Wallachian. Most detected runs of homozygosity (ROH) were less than 5 Mb long for both breeds. ROH segments longer than 15 Mb were absent in Wallachian sheep. Concerning putative selection signatures, a total of 471 candidate genes in Wallachian sheep within 11 hotspots and 653 genes within 13 hotspots in Sumava sheep were identified. Czech breeds appear to be well differentiated from each other and other European breeds. Their genetic diversity is low, especially in the case of the Wallachian breed. Sumava is not so threatened by low diversity but has a larger share of the non-native gene pool.

Due to that Czech indigenous goat breeds, together with Slovak indigenous goat breeds, are closed population, face this population loss of genetic diversity, because these populations are closed. The aim of this study was to perform a genome-wide diversity characterization using Illumina GoatSNP50 BeadChip of indigenous Czech Braun Shorthair and White Shorthair goat breeds together with international Alpine goat from Czech Republic and indigenous White Shorthair goat breed from Slovak Republic. For comparison was in this analysis included indigenous alpine goat breed from Austria and Switzerland. To determine key conservation risk parameters, we estimated inbreeding (FROH) and current and historical effective population size (Ne). The observed estimates for Czech and Slovak indigenous breed show similar values as other alpine goat breeds. In addition, by comparing genetic differences by Neighbour-net, no differences were recorded between the Czech-Slovak WSG. The study provides data and information utilizable in the management of conservation programs planned to reduce inbreeding and to minimize loss of genetic variability.

Xenogenic mammalian sperm heads injected into mouse ovulated oocytes decondense and form pronuclei in which sperm DNA parameters can be evaluated. We suggest that this approach can be used for the assessment of sperm DNA damage level and the evaluation of how certain sperm treatments (freezing, lyophilization, etc.) influence the quality of spermatozoa.

The COVID-19 crisis has had a negative impact on macroeconomic development in the Czech Republic, with the second quarter of 2020 seeing the most severe downturn since the formation of the Czech Republic in 1993. The Czech Statistical Office recorded a year-on-year decrease in GDP of 11% (CZSO, 2019). The analysis presented here focuses on the impact of the COVID-19 crisis on selected aspects of animal breeding and summarizes the situation up until 1 October 2020.

Sheep milk and products made from it are a source of valuable nutritional substances for human nutrition. In comparison to cow milk, sheep milk contains a higher content of milk fat and it is composited of a higher proportion of polyunsaturated fatty acids (PUFA), especially essential α-linolenic acid and conjugated linoleic acid (CLA). Also, a lower saturated fatty acid (SFA) content is present in sheep milk than in cow and goat milk (Moioli et al., 2012; Markiewicz-Kęszycka et al., 2013). Modification of the fatty acid (FA) profile in milk fat with a possibility of increasing the content of nutritionally essential FAs, such as n-3 FA especially CLA, has been an object of many types of research (Reynolds et al., 2006; Sánchez et al., 2010; Moioli et al., 2012; Buccioni et al., 2015). Crucial factors influencing the FA profile in sheep milk are feed composition, lactation stage, breed, and last but not least, genetic factors. Fatty acids essential for milk fat triacylglycerols (TAG) synthesis in the mammary gland are obtained in two ways. The one is de novo synthesis in the secretory cells of the ABSTRACT. The aim of the study was to evaluate the influence of single nucleotide polymorphism (SNP) in lipoprotein lipase gene (LPL) on the basic sheep milk parameters and the fatty acid profile in milk and yoghurt drinks made from it. The Primer Extension Analysis was used for genotyping 3 SNPs at positions: 74 (T/C), 130 (T/C) and 133 (T/A) in the 3 ́UTR region of the ovine LPL gene in 139 sheep of the East Friesian breed. According to the LPL genetic marker (CCCCAA and CTCTAT) 40 sheep were selected and divided into groups. Pooled milk was collected from May to June and used to make yoghurt drinks with probiotics and prebiotics according to the genetic combination. In the pooled samples used for yoghurt production, the contents of fat, protein, lactose and total solids have been determined. The profile of fatty acids in fat from milk pooled samples as well as from yoghurt drinks has been analysed. Differences in the composition of milk and fatty acid profile were shown. The content of fat and total solids was significantly higher in the milk with the CTCTAT genotype combination in comparison to that with CCCCAA genotype. Moreover, these samples of milk and yoghurt products had a statistically lower content of palmitic acid and a significantly lower content of hypocholesterolemic acids. Targeted production of sheep milk from animals with different genotypes of the LPL may positively influence the fatty acid profile in milk and products made from it. Received: 18 March 2020 Revised: 25 February 2021 Accepted: 1 March 2021

Milk production is influenced by many factors, including genetic and environmental factors and their interactions. Animal health, especially udder health, is usually evaluated by the number of somatic cells. The present study described the effect of polymorphisms in the ACACA, BTN1A1, LPL, and SCD genes on the daily milk yield, fat, and protein percentages and somatic cell count. In this study, 590 White Shorthaired (WSH) and Brown Shorthaired (BSH) goats were included. SNP genotyping was performed by PCR-RFLP and multiplex PCR followed by SNaPshot minisequencing analysis. The linear mixed model with repeated measurement was used to identify the genetic associations between the studied genes/SNPs and chosen traits. All selected genes were polymorphic in the tested goat populations and showed significant associations with milk traits. Only BTN1A1 (SNP g.599 A > G) showed a significant association with the somatic cell score. After Bonferroni correction, a significant effect of LPL g.300G > A on daily milk yield and fat percentage, LPL g.185G > T on protein percentage, and LPL G50C, SCD EX3_15G > A, and SCD EX3_68A > G on fat percentage was found. The importance of environmental factors, such as the herd-year effect, month of milking, and lactation order on all milk performance indicators was confirmed.

Czech local poultry breeds face high risks of extinction. Because these populations are closed, they are more likely to lose genetic diversity. The aim of this analysis was to determine the loss of genetic diversity in three Czech autochthonous poultry breeds. Pedigree data from a total of 1 932 Czech Gold Speckled Hens, 325 Czech White Geese and 111 Czech Crested Geese registered in studbooks between 2000 and 2018 were evaluated. Data were analysed to determine the major factors that affect the genetic variability of these breeds. The average num- bers of equivalent complete generations ranged from 2.53 to 4.82. The effective numbers of founders were from 29 to 59, representing from 43% to 62% of the total number of founders. The effective number of ancestors was estimated in the range of 21 to 41. The average inbreeding coefficient and relatedness coefficient (in parentheses) for the reference populations were 2.0% (6.5%), 1.9% (4.9% ) and 2.1% (9.3%), respectively. The results showed that the effective population size derived from the rate of inbreeding ranged from 46 to 108 and if derived from the rate of coancestry it ranged from 35 to 74. With regard to these results, the analysed breeds showed a high probability of allele loss and consequent loss of genetic diversity.

For the development of an appropriate program for the conservation of animal genetic resources, careful evaluation of the current genetic diversity in endangered breeds was performed. Pedigree data from a total of 38,288 animals registered in studbooks between 1970 and 2017 from two autochthonous Czech goat breeds, White and Brown Shorthair goats, were analysed to identify factors that may have affected the genetic variability of the breeds. Because the populations of both breeds are closed, there is concern about the loss of genetic variation. The average numbers of equivalent complete generations were 10 and 11 for White Shorthair goat and Bowen Shorthair goat, respectively. The effective numbers of founders contributing to the current genetic pool were 323 and 45, and the effective numbers of ancestors were 90 and 24, respectively. The average inbreeding and relatedness coefficients for the reference populations of the two breeds were 2% and 5.3%, respectively, for White Shorthair goats and 3.7 and 11.5% for Brown Shorthair goats. The corresponding estimates of the effective population size based on an increase in inbreeding or coancestry were 244 and 311, respectively, for White Shorthair goats and 82 and 87 for Brown Shorthair goats. Three kinds of inbreeding coefficients, the classical inbreeding coefficient, the new inbreeding coefficient and the ancestral inbreeding coefficient, were used to quantify the effects of inbreeding and purging on milk production in both analysed breeds. All tested inbreeding coefficients (FX, Fnew) showed a significantly negative influence on milk performance in the White Shorthair goat breed. The purging effect was confirmed based on the difference between FX and Fnew in White Shorthair goats and on the suitability of the statistical model in Brown Shorthair goats. These statistics suggest that genetic variability has decreased and inbreeding depression has occurred and that without changes in the breeding strategy, genetic variability might continue to decline.

Lipoprotein Lipase (LPL), a key enzyme in lipid metabolism, catalyses the hydrolysis of triglycerides (TG) from TG-rich lipoproteins, and serves a bridging function that enhances the cellular uptake of lipoproteins. By sequencing, we identified a four novel SNPs; one SNP involved a Gly -> Arg AA substitution at position 23 of the signal peptide, and three SNPs in the intron I (KP261023) of the LPL locus. A protocol for rapid efficient simultaneous genotyping of LPL variants using the primer extension method (PEA) was described, and a total of 230 animals belonging to two goat breeds: White Shorthaired goat (n = 177), and Brown Shorthaired goat (n = 103) were genotyped. A total 15 genotype combinations were detected in tested goat populations.

In nearly all somatic cells, the ribosome biosynthesis is a key activity. The same is true also for mammalian oocytes and early embryos. This activity is intimately linked to the most prominent nuclear organelles — the nucleoli. Interestingly, during a short period around fertilization, the nucleoli in oocytes and embryos transform into ribosome-biosynthesis-inactive structures termed nucleolus-like or nucleolus precursor bodies (NPBs). For decades, researchers considered these structures to be passive repositories of nucleolar proteins used by the developing embryo to rebuild fully functional, ribosome-synthesis competent nucleoli when required. Recent evidence, however, indicates that while these structures are unquestionably essential for development, the material is largely dispensable for the formation of active embryonic nucleoli. In this mini-review, we will describe some unique features of oocytes and embryos with respect to ribosome biogenesis and the changes in the structure of oocyte and embryonic nucleoli that reflect this. We will also describe some of the different approaches that can be used to study nucleoli and NPBs in embryos and discuss the different results that might be expected. Finally, we ask whether the main function of nucleolar precursor bodies might lie in the genome organization and remodelling and what the involved components might be.

In nearly all somatic cells, the ribosome biosynthesis is a key activity. The same is true also for mammalian oocytes and early embryos. This activity is intimately linked to the most prominent nuclear organelles-the nucleoli. Interestingly, during a short period around fertilization, the nucleoli in oocytes and embryos transform into ribosome-biosynthesis-inactive structures termed nucleolus-like or nucleolus precursor bodies (NPBs). For decades, researchers considered these structures to be passive repositories of nucleolar proteins used by the developing embryo to rebuild fully functional, ribosome-synthesis competent nucleoli when required. Recent evidence, however, indicates that while these structures are unquestionably essential for development, the material is largely dispensable for the formation of active embryonic nucleoli. In this mini-review, we will describe some unique features of oocytes and embryos with respect to ribosome biogenesis and the changes in the structure of oocyte and embryonic nucleoli that reflect this. We will also describe some of the different approaches that can be used to study nucleoli and NPBs in embryos and discuss the different results that might be expected. Finally, we ask whether the main function of nucleolar precursor bodies might lie in the genome organization and remodelling and what the involved components might be.

Sequence differences are considered to be the basic cause of developmental failure in interspecies embryos when more distant species are combined. However, other phenomena, such as insufficient or excessive quantity of specific cellular factors, might also influence the outcome. These effects are usually not considered. One of the organelles shown to contain different amount of proteins is the oocyte nucleolus-like body. Here we show that upon interspecies transfer, a single porcine nucleolus-like body is unable to support the development of a mouse parthenogenetic embryo derived from an enucleolated oocyte. However, when the amount of the porcine nucleolar material is increased to equalize the amount of mouse nucleolar material by transferring two nucleolus-like bodies, mouse embryos are able to pass the developmental block elicited by enucleolation. These embryos progress to the blastocyst stage at rates comparable to controls. Thus, using the model of an interspecies nucleolus-like body transplantation between mouse and pig oocytes, we show that an inadequate amount of nucleolar factors, rather than the species origin, affects the development. In a wider context of interspecies nuclear transfer schemes, the observed incompatibility between more distant species might not stem simply from sequence differences but also from improper dosage of key cellular factors.