Assessment of perfluoroalkyl substances concentration levels in wild bat guano samples.

Perfluoroalkyl substances (PFASs) are substances commonly used in the production of various everyday objects, including among others kitchen dishes, cosmetics, or clothes. They penetrate to the environment and living organisms causing disturbances in the functioning of many internal organs and systems. Due to environmental pollution, wildlife is also exposed to PFASs, but the knowledge about this issue is rather limited. The aim of this study was to evaluate the exposure of wild greater mouse-eared bats (Myotis myotis), living in Poland, to six selected PFASs: five perfluoroalkyl carboxylic acids (perfluorobutanoic acid-PFBuA, perfluoropentanoic acid-PFPeA, perfluorohexanoic acid-PFHxA, perfluoroheptanoic acid-PFHpA, perfluorooctanoic acid-PFOA) and perfluorooctane sulfonic acid (PFOS) through the analysis of guano samples with liquid chromatography with tandem mass spectrometry (LC-MS-MS) method. To our knowledge this is the first study concerning the PFASs levels in bats, as well as using guano samples to evaluate the exposure of wild mammals to these substances. A total of 40 guano samples were collected from 4 bats summer (nursery) colonies located in various parts of Poland. The presence of PFASs mentioned were detected in all colonies studied, and concentration levels of these substances were sampling dependent. The highest concentration levels were observed in the case of PFPeA [1.34 and 3060 ng/g dry weight (dw)] and PFHxA (8.30-314 ng/g dw). This study confirms the exposure of wild bats to PFASs.

Epidemiological characteristics and financial losses due to avian aspergillosis in households in the Almaty region, Republic of Kazakhstan.

Aspergillosis is a severe fungal disease that affects all species and ages of poultry and leads to significant economic losses within the poultry industry. The economic significance of aspergillosis is associated with direct losses due to poultry mortality, a decline in the production of meat and eggs, feed conversion, and poor growth of recovering poultry. Although a decrease in the production of poultry meat and eggs in Kazakhstan due to this fungal disease has been widely reported, studies on the consequent financial losses on affected farms (households) have not been carried out. This study aimed to estimate the financial losses and epidemiological parameters of avian aspergillosis among households affected by the disease in the Almaty region. To achieve the objectives of the research, a survey was conducted involving affected households from February 2018 to July 2019. The affected poultry were diagnosed based on clinical, macroscopical, and microscopical procedures, and once the infection was confirmed, household owners were interviewed. Data were collected from 183 household owners. The median incidence risk and fatality rates were 39 and 26% in chickens, 42 and 22% in turkeys, and 37 and 33% in geese, respectively, with young poultry having a higher incidence risk and fatality rate than adults. Approximately 92.4% of the household owners treated the affected poultry using natural folk methods and 7.6% of household owners used antifungal drugs and antibiotics, spending a median of US $35.20 (min US $0; max US $400) per household throughout the course of the infection. Egg production was reduced by a median of 58.3% when households were affected. The price of poultry fell by a median of 48.6% immediately after recovery due to weight loss. The median of the overall financial losses of households was US $198.50 (min US $11; max US $1,269). The majority of household owners (65%) did not replace their poultry, 9.8% of household owners replaced all their poultry, and the remaining 25.1% replaced only a proportion of the poultry lost at the time of the study. Newly acquired poultry were purchased from neighbors (10.9%), fellow villagers (50%), and state poultry farms (39.1%). This study demonstrates that aspergillosis has an immediate impact on subsistence household owners' livelihoods in the Almaty region of Kazakhstan.

Expression of TXNRD1, HSPA4L and ATP1B1 Genes Associated with the Freezability of Boar Sperm.

Cryopreservation is associated with increased oxidative stress, which is responsible for sperm damage. We analyzed the effect of cryopreservation on mRNA and protein expression of thioredoxin reductase 1 (TXNRD1), heat shock protein family A (HSP 70) member 4 like (HSPA4L) and sodium/potassium-transporting ATPase subunit beta-1 (ATP1B1) genes in boar sperm with different freezability. Boars were classified as having good and poor semen freezability (GSF and PSF, respectively), according to the assessment of post-thaw sperm motility. Total RNA was isolated from fresh pre-freeze (PF) and frozen-thawed (FT) sperm from five boars of the GSF and PSF groups, respectively. Quantification of TXNRD1, HSPA4L and ATP1B1 gene expression was performed by RT-qPCR analysis. Proteins extracted from sperm were subjected to Western blotting and SDS-PAGE analyses. Poor freezability ejaculates were characterized by significantly higher relative mRNA expression levels of TXNRD1 and HSPA4L in FT sperm compared with the fresh PF sperm. Furthermore, the relative mRNA expression level of ATP1B1 was significantly higher in the fresh PF sperm of the GSF group. Western blotting analysis revealed significantly higher relative expression of TXNRD1 protein in the fresh PF sperm of the GSF group, while HSPA4L protein expression was markedly increased in FT sperm of the PSF group. Electrophoretic and densitometric analyses revealed a higher number of proteins in the fresh PF and FT sperm of the PSF and GSF groups, respectively. The results of this study indicate that ATP1B1 mRNA expression in the fresh PF sperm is a promising cryotolerance marker, while the variations of TXNRD1 and HSPA4L protein expression in the fresh PF or FT sperm provide useful information that may help to elucidate their biological significance in cryo-damage.

Promoter polymorphisms in STK35 and IFT27 genes and their associations with boar sperm freezability.

We have shown that STK35 and IFT27 genes are differentially expressed in spermatozoa from boars with good and poor semen freezability (GSF and PSF, respectively). STK35 is a stress-related gene that is implicated in spermatogenesis, whereas IFT27 is a motility-related gene that is mainly involved in intracellular protein transport. In this study we hypothesized that polymorphic variants in the 5'-flanking regulatory regions of STK35 and IFT27 genes could contribute to differences in semen freezability. We also predicted the interactions of the polymorphic variants with transcription factors on the gene promoter activity, using bioinformatics. The 5'-flanking region sequences of the STK35 and IFT27 were PCR amplified and analyzed by Sanger sequencing method. Protein expression in STK35 and IFT27 was determined in pre-freeze (PF) and frozen-thawed (FT) spermatozoa, using western blotting analysis. Sanger sequencing revealed a single nucleotide polymorphism (SNP) rs327863835 (C > T) in STK35 promoter, while two SNPs (rs337563873, A > T; rs331520020, T > C) were detected in IFT27 promoter. STK35 and IFT27 promoter polymorphisms showed significant allele frequency differences between the GSF and PSF groups. Using bioinformatics approaches, we predicted that SNPs resulted in the generation of additional transcription factor binding sites for NFATC2, ELK1 and GR-ß, which appeared to enhance or repress the promoter activity of STK35 or IFT27 in either freezability group. Wide variations in STK35 and IFT27 protein expression were observed among the boars, however, significantly higher protein expression was detected in IFT27 in FT spermatozoa of the GSF group. We suggest that the upstream variants, detected in STK35 and IFT27 promoters, might regulate the transcriptional activity of the genes by affecting their potential binding of transcription factors. The results indicate that the allelic variants in STK35 and IFT27 could be considered as potential genetic markers for predicting boar sperm freezability.

Effect of Long-Acting Selenium Preparation on Health and Productivity of Sheep.

The aim of this study was to determine the effects of long-acting selenium (Se) preparation in sheep. The experimental material comprised Skudda ewes and their lambs. The animals were divided into two equal groups: C-control, and E-experimental. Between days 70 and 80 of pregnancy, group E ewes were injected with Se preparation (Barium Selenate Injection, BVP Animal Care, Ireland) at f 1 mL/50 kg body weight. Hematological, biochemical and immunological blood parameters as well as Se levels were analyzed in ewes. The growth rate of lambs, the dimensions of the musculus longissimus dorsi (MLD) cross-section, and fat thickness over the loin-eye area were also determined. It was found that barium selenate stimulated the mechanisms of humoral and cellular immunity. The injection was an effective form of Se supply, which was confirmed by its increased concentration in the blood serum of lactating ewes. The offspring of the experimental ewes were characterized by a faster growth rate, and they achieved significantly higher body weight (p ≤ 0.05) at 100 days of age. The lambs also had significantly higher parameters of MLD (p ≤ 0.05) at similar carcass fat content.

Molecular Characterisation of Uterine Endometrial Proteins during Early Stages of Pregnancy in Pigs by MALDI TOF/TOF.

The molecular mechanism underlying embryonic implantation is vital to understand the correct communications between endometrium and developing conceptus during early stages of pregnancy. This study's objective was to determine molecular changes in the uterine endometrial proteome during the preimplantation and peri-implantation between 9 days (9D), 12 days (12D), and 16 days (16D) of pregnant Polish Large White (PLW) gilts. 2DE-MALDI-TOF/TOF and ClueGOTM approaches were employed to analyse the biological networks and molecular changes in porcine endometrial proteome during maternal recognition of pregnancy. A total of sixteen differentially expressed proteins (DEPs) were identified using 2-DE gels and MALDI-TOF/TOF mass spectrometry. Comparison between 9D and 12D of pregnancy identified APOA1, CAPZB, LDHB, CCT5, ANXA4, CFB, TTR upregulated DEPs, and ANXA5, SMS downregulated DEPs. Comparison between 9D and 16D of pregnancy identified HP, APOA1, ACTB, CCT5, ANXA4, CFB upregulated DEPs and ANXA5, SMS, LDHB, ACTR3, HP, ENO3, OAT downregulated DEPs. However, a comparison between 12D and 16D of pregnancy identified HP, ACTB upregulated DEPs, and CRYM, ANXA4, ANXA5, CAPZB, LDHB, ACTR3, CCT5, ENO3, OAT, TTR down-regulated DEPs. Outcomes of this study revealed key proteins and their interactions with metabolic pathways involved in the recognition and establishment of early pregnancy in PLW gilts.

The Efficacy of a Long-Acting Injectable Selenium Preparation Administered to Pregnant Ewes and Lambs.

The aim of this study was to evaluate the effects of a long-acting selenium (Se) preparation administered to sheep. The experiment was conducted on 30 dams and 36 lambs divided into three equal groups of 10 dams and 12 lambs each: Control-C, and two experimental groups-E (Se administered to pregnant ewes) and EI (Se administered directly to lambs after the colostral period). The Se preparation (Barium Selenate Injection, BVP Animal Care, 50 mg/mL) was administered by injection at 1 mL/50 kg (1 mg Se/kg) body weight (BW) to group E ewes in the third month of pregnancy (between 70 and 90 days) and to group EI lambs between 4 and 7 days of age. The following parameters were determined: Se concentration in the blood of ewes, milk yield, milk composition, Se concentration in milk; hematological, biochemical, and immunological parameters and Se concentration in the blood of lambs; growth rate and in vivo measurements of lean meat and fat content in lambs. Barium selenate significantly improved the Se status of dams and lambs, regardless of whether it was administered to pregnant ewes or directly to lambs in the first week of their life. The milk of ewes receiving the Se preparation was characterized by higher concentrations of fat and dry matter. The Se preparation induced significant changes in immunological parameters, thus enhancing defense mechanisms in lambs. The Se preparation exerted more stimulatory effects on humoral and cellular immune responses when administered directly to lambs after the colostral period (group EI) than to pregnant ewes (group E). The results of this study indicate that the long-acting Se preparation delivers benefits to sheep by boosting their immunity and, therefore, improving performance.

Effect of a single, oral administration of selenitetriglycerides, at two dose rates, on blood selenium status and haematological and biochemical parameters in Holstein-Friesian calves.

BACKGROUND: Selenitetriglycerides are biologically active, organic forms of selenium formed as a result of the modification of selenic acid and sunflower oil. Studies in rats have shown that they are well absorbed and of low toxicity. There are no published studies on selenitetriglycerides supplementation in calves. RESULTS: In this study, selenitetriglycerides were administered once orally on the 2nd day of life at a dose of 0.5 or 1 mg Se/kg body weight to each of six Holstein-Friesian calves while six control calves were not supplemented. Blood for determination of selenium concentration, glutathione peroxidase activity, haematological parameters, aspartate aminotransferase, creatine kinase, and lactate dehydrogenase activities and glucose, total protein, albumin, triglycerides, cholesterol, urea, and creatinine concentration was collected before supplementation (day 0) and 1, 2, 5, 10 and 14 days after supplementation. Selenitetriglycerides administration increased (P < 0.01) serum selenium concentration in supplemented calves as early as day1, from a mean of 63.4 to 184.22 µg/l in calves receiving selenium at a dose of 0.5 mg/kg BW, and from 63.17 to 200.33 µg/l in calves receiving 1 mg/kg. Serum selenium concentrations remained significantly higher compared to the control group throughout the experiment. Glutathione peroxidase activity was higher in supplemented than control calves, significantly so in animals receiving the 1 mg/kg dose of Se on the 10th and 14th days (P < 0.05). There were no significant differences in the haematological and biochemical parameters between the groups. CONCLUSIONS: This experiment showed that supplementation with selenitetriglycerides could significantly improve blood selenium status in calves without adverse effects on haematological or biochemical parameters. These findings are essential prerequisites for future studies on selenitetriglycerides supplementation to manage clinical selenium deficiency in calves.

Differentiation of Subclinical Ketosis and Liver Function Test Indices in Adipose Tissues Associated With Hyperketonemia in Postpartum Dairy Cattle.

Past studies suggested that during early lactation and the transition period, higher plasma growth hormone (GH) levels in subclinical ketosis (SCK) might involve the initiation of body adipose tissues mobilization, resulting in metabolic disorders in ruminants particularly hyperketonemia. The upregulated GH mRNA expression in adipose tissue may take part in the adipolysis process in SCK-affected cows that paves a way for study further. This study aimed to characterize the plasma levels of GH, ß-hydroxybutyrate acid (BHBA) and non-esterified fatty acid (NEFA) and glucose (GLu) in ketotic cows and healthy control (CON) cows; to measure the liver function test (LFT) indices in ketotic and healthy CON cows, and finally the quantitative real-time PCR (qRT-PCR) assay of candidate genes expressed in adipose tissues of ketotic and healthy CON cows during 0 to 7 week postpartum. Three experiments were conducted. Experiment-1 involved 21 Holstein cows weighing 500-600 kg with 2-5 parities. Results showed that GH, BHBA, and NEFA levels in ketotic cows were significantly higher and the GLu level significantly lower. Pearson's correlation analysis revealed a significant positive correlation of GH with BHBA, NEFA, and GLu in ketotic and healthy CON cows. In experiment-2, dynamic monitoring of LFT indices namely, alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (GGT), total bilirubin (TBIL), direct bilirubin (DBIL), total protein (TP), albumin (ALB), globulin (GLOB) and albumin/globulin (A/G) were examined. The TBIL, DBIL, and GGT indices were significantly higher in ketotic cows and TP was significantly lower. In experiment-3, mRNA expression levels of GHR and peroxisome-proliferator-activated receptor alpha (PPARα) genes in adipose tissue were significantly upregulated in ketotic cows. However, the mRNA expression of insulin-like growth factor-I (IGF-1), insulin-like growth factor-I receptor (IGF-1R), and sterol regulatory element-binding protein-1c (SREBP-1c) genes in adipose tissue were downregulated in ketotic cows. Our study concluded that during postpartum, higher plasma GH levels in SCK cows might involve the initiation of body adipose tissue mobilization, resulting in hyperketonemia.

Influence of selenium administration to dry cows on selected biochemical and immune parameters of their offspring.

The study was performed on 16 Holstein-Friesian calves divided into two groups of eight animals each. The first group was composed of calves whose mothers did not receive selenium supplements (Se0). The second group consisted of calves whose mothers were administered intramuscular injections of a selenium and vitamin E supplement containing 0.5 of sodium selenite/ml and 50 mg of tocopherol acetate/ml in a single dose of 30 ml (Se30) ml, 10 days before the expected parturition date (10 ± 2 days). The calves were fed 2.5 L of the mother's colostrum administered by stomach tube 2 hr after birth and another 2 L 6 hr after birth. Blood from all calves was collected 7 times from external jugular vein (day 0-before colostrum administration and on the 2nd, 3rd, 4th, 7th, 14th and 21st days of life) for analyses of selenium, ceruloplasmin, transferrin, lactoferrin, immunoglobulin G (IgG) concentrations and gamma-glutamyl transferase (GGT) and lysozyme activity. Selenium concentration was significantly higher in calves whose mothers received selenium supplements than in the offspring of non-supplemented cows until 72 hr after birth (p ≤ .05). Lysozyme and GGTP activity and IgG concentration were significantly higher in the S30 group during the entire experiment (p ≤ .05). Supplementation of selenium to the mothers did not influence the ceruloplasmin, lactoferrin and transferrin levels in calves. A single injection of a selenium supplement administered to cows during late pregnancy increases selenium levels in calves and enhances passive transfer from the mother to the offspring.

Identification of neuropeptide y in superior cervical ganglion neurons that project to the oesophagus - A combined immunohistochemical labelling and retrograde tracing study in pigs.

Neuropeptide Y (NPY) is a neuronal active substance taking part in the regulation of gastrointestinal (GI) tract activity. This study used retrograde neuronal tracing and immunofluorescence methods to analyse NPY-positive neurons located in superior cervical ganglion and supplying the cervical oesophagus in the pig. The presence of NPY was observed in 30% of all neurons supplying the part of oesophagus studied. Probably the number of Fast Blue (FB) positive cells depends on the area of the wall injected with FB and the fragment of oesophagus studied. Therefore, the obtained results indicate that the described peptide is an important factor in the extrinsic innervation of this part of the GI tract.

Simvastatin-induced Changes in the Leukocytic System of Porcine Bone Marrow.

INTRODUCTION: Simvastatin is a substance which is commonly used as a medicine to reduce cholesterol level. Unfortunately, it shows numerous side effects. Simvastatin affects various internal organs, and among other detriments to health may cause persistent muscle weakness, osteolytic processes, headaches, and rashes. Until now knowledge of the influence of simvastatin on bone marrow cells has been rather scant and fragmentary. MATERIAL AND METHODS: During this experiment the numbers of all types of cells in the leukocytic system of porcine bone marrow were evaluated after 28 and 56 days of oral administration of simvastatin at a dose of 40 mg/day/animal. RESULTS: Simvastatin caused an increase in the number of all types of cells in the leukocytic system, and the most visible fluctuations concerned promyelocytes. CONCLUSION: Observations obtained during the present study indicated that the results of the action of simvastatin on porcine bone marrow differ from those observed in other mammal species, including human. This may be due to various metabolic pathways within the bone marrow in the particular species, but the exact mechanisms of these actions are unknown at the present time.

Applicability of the Protein-lipid Profile and Activity of Lactate Dehydrogenase Isoenzymes for Diagnosing Nutritional Muscular Dystrophy in Calves.

INTRODUCTION: In calves, hyposelenosis degenerates skeletal muscles in different parts of the body. The extent of damage to muscle cells can be diagnosed by determining the activity of creatine kinase (CK), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH). The aim of this study was to analyse variations in the serum levels of LDH isoenzymes in calves with nutritional muscular dystrophy (NMD), to determine the applicability of this parameter for diagnosing NMD, and to describe the influence of hyposelenosis on total protein (TP), triglyceride (TG), and cholesterol (CHOL) levels. MATERIAL AND METHODS: Two groups of calves (n = six animals per group) were used. After birth, control group calves (SC) were intramuscularly administered 10 ml of a preparation containing selenium (Se) and vitamin E, and experimental group animals (SE) that were not injected. Blood was collected after 5, 15, and 25 days, and the concentrations of Se, vitamin E, TP, TG, and CHOL and the activity of glutathione peroxidase (GSH-Px), CK, and LDH fractions were determined. RESULTS: Hypocholesterolaemia and elevated TG levels were found in SE group calves whose LDH fractions revealed a significant increase in LDH4 and LDH5 activity and a decrease in LDH1 activity when electrophoretically separated. CONCLUSION: Nutritional muscular dystrophy is accompanied by hypocholesterolaemia and elevated TG levels caused by muscle lipolysis. LDH4 and LDH5 activity parameters assist early diagnosis of NMD in calves.

Single nucleotide polymorphism discovery in bovine liver using RNA-seq technology.

BACKGROUND: RNA-seq is a useful next-generation sequencing (NGS) technology that has been widely used to understand mammalian transcriptome architecture and function. In this study, a breed-specific RNA-seq experiment was utilized to detect putative single nucleotide polymorphisms (SNPs) in liver tissue of young bulls of the Polish Red, Polish Holstein-Friesian (HF) and Hereford breeds, and to understand the genomic variation in the three cattle breeds that may reflect differences in production traits. RESULTS: The RNA-seq experiment on bovine liver produced 107,114,4072 raw paired-end reads, with an average of approximately 60 million paired-end reads per library. Breed-wise, a total of 345.06, 290.04 and 436.03 million paired-end reads were obtained from the Polish Red, Polish HF, and Hereford breeds, respectively. Burrows-Wheeler Aligner (BWA) read alignments showed that 81.35%, 82.81% and 84.21% of the mapped sequencing reads were properly paired to the Polish Red, Polish HF, and Hereford breeds, respectively. This study identified 5,641,401 SNPs and insertion and deletion (indel) positions expressed in the bovine liver with an average of 313,411 SNPs and indel per young bull. Following the removal of the indel mutations, a total of 195,3804, 152,7120 and 205,3184 raw SNPs expressed in bovine liver were identified for the Polish Red, Polish HF, and Hereford breeds, respectively. Breed-wise, three highly reliable breed-specific SNP-databases (SNP-dbs) with 31,562, 24,945 and 28,194 SNP records were constructed for the Polish Red, Polish HF, and Hereford breeds, respectively. Using a combination of stringent parameters of a minimum depth of ≥10 mapping reads that support the polymorphic nucleotide base and 100% SNP ratio, 4,368, 3,780 and 3,800 SNP records were detected in the Polish Red, Polish HF, and Hereford breeds, respectively. The SNP detections using RNA-seq data were successfully validated by kompetitive allele-specific PCR (KASPTM) SNP genotyping assay. The comprehensive QTL/CG analysis of 110 QTL/CG with RNA-seq data identified 20 monomorphic SNP hit loci (CARTPT, GAD1, GDF5, GHRH, GHRL, GRB10, IGFBPL1, IGFL1, LEP, LHX4, MC4R, MSTN, NKAIN1, PLAG1, POU1F1, SDR16C5, SH2B2, TOX, UCP3 and WNT10B) in all three cattle breeds. However, six SNP loci (CCSER1, GHR, KCNIP4, MTSS1, EGFR and NSMCE2) were identified as highly polymorphic among the cattle breeds. CONCLUSIONS: This study identified breed-specific SNPs with greater SNP ratio and excellent mapping coverage, as well as monomorphic and highly polymorphic putative SNP loci within QTL/CGs of bovine liver tissue. A breed-specific SNP-db constructed for bovine liver yielded nearly six million SNPs. In addition, a KASPTM SNP genotyping assay, as a reliable cost-effective method, successfully validated the breed-specific putative SNPs originating from the RNA-seq experiments.

Single Nucleotide Polymorphism Discovery in Bovine Pituitary Gland Using RNA-Seq Technology.

Examination of bovine pituitary gland transcriptome by strand-specific RNA-seq allows detection of putative single nucleotide polymorphisms (SNPs) within potential candidate genes (CGs) or QTLs regions as well as to understand the genomics variations that contribute to economic trait. Here we report a breed-specific model to successfully perform the detection of SNPs in the pituitary gland of young growing bulls representing Polish Holstein-Friesian (HF), Polish Red, and Hereford breeds at three developmental ages viz., six months, nine months, and twelve months. A total of 18 bovine pituitary gland polyA transcriptome libraries were prepared and sequenced using the Illumina NextSeq 500 platform. Sequenced FastQ databases of all 18 young bulls were submitted to NCBI-SRA database with NCBI-SRA accession numbers SRS1296732. For the investigated young bulls, a total of 113,882,3098 raw paired-end reads with a length of 156 bases were obtained, resulting in an approximately 63 million paired-end reads per library. Breed-wise, a total of 515.38, 215.39, and 408.04 million paired-end reads were obtained for Polish HF, Polish Red, and Hereford breeds, respectively. Burrows-Wheeler Aligner (BWA) read alignments showed 93.04%, 94.39%, and 83.46% of the mapped sequencing reads were properly paired to the Polish HF, Polish Red, and Hereford breeds, respectively. Constructed breed-specific SNP-db of three cattle breeds yielded at 13,775,885 SNPs. On an average 765,326 breed-specific SNPs per young bull were identified. Using two stringent filtering parameters, i.e., a minimum 10 SNP reads per base with an accuracy ≥ 90% and a minimum 10 SNP reads per base with an accuracy = 100%, SNP-db records were trimmed to construct a highly reliable SNP-db. This resulted in a reduction of 95,7% and 96,4% cut-off mark of constructed raw SNP-db. Finally, SNP discoveries using RNA-Seq data were validated by KASP™ SNP genotyping assay. The comprehensive QTLs/CGs analysis of 76 QTLs/CGs with RNA-seq data identified KCNIP4, CCSER1, DPP6, MAP3K5 and GHR CGs with highest SNPs hit loci in all three breeds and developmental ages. However, CAST CG with more than 100 SNPs hits were observed only in Polish HF and Hereford breeds.These findings are important for identification and construction of novel tissue specific SNP-db and breed specific SNP-db dataset by screening of putative SNPs according to QTL db and candidate genes for bovine growth and reproduction traits, one can develop genomic selection strategies for growth and reproductive traits.