Przemysław Sobiech

Distribution and chemical coding patterns of cocaine- and amphetamine-regulated transcript-like immunoreactive (CART-LI) neurons in the enteric nervous system of the porcine stomach cardia

The aim of this study was to determine the presence of cocaine- and amphetamine-regulated transcript-like immunoreactive (CART-LI) neurons and co-localisation of CART with vesicular acetylcholine transporter (VAChT), neuronal nitric oxide synthase (n-NOS), vasoactive intestinal polypeptide (VIP), substance P (SP) and leu-enkephalin (LENK) in the enteric nervous system of the porcine gastric cardia by using a double-labelling immunofluorescence technique. CART-LI neurons were observed in the myenteric plexus (18.2±2.6%). A dense network of CART-LI nerve fibers was mainly observed in the muscular layer. CART showed co-localization mainly with VAChT, n-NOS, VIP and to a lesser degree with LENK and SP. Distribution of CART and its co-localization with other neurotransmitters suggest that this peptide plays an important role in gastric motility in the pig.

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.

Influence of invasion of intracellular parasites on platelet response in dogs based on clinical cases

Abstract Blood morphology in dogs infected with intracellular parasites of the genera Babesia and Ehrlichia was examined. The parasites were detected in peripheral blood smears stained by the May-Grünwald-Giemsa method. During the parasitic invasion, a decrease in thrombocyte counts was observed. This was connected with forming platelet aggregates. It was noted that the number of giant platelets simultaneously increased.

MODULATION OF SYSTEMIC INNATE AND ADAPTIVE IMMUNE PARAMETERS IN WEANED PIGS BY SINGLE ORAL APPLICATION OF IMMUNOBIOTICS

The aims of this study were to evaluate the effects of peroral treatment of 4-week old pigs at weaning (Day 0) with a single dose of levamisole (LEVA) or polyoxyethyle-polyoxypropylene (POE-POP), well known immunobiotics (IBC) and promising alternatives to dietary antibiotic growth promoters (AGP), on systemic innate and adaptive immunity by determining phagocytosis (PHC) and microbicidity (MBC) efficiency of their monocytes (MO) and granulocytes (GR), changes of serum C-reactive protein (CRP) and haptoglobin (HpG) profiles and kinetics of CD4+ CD8+ T cells expression during 5 weeks following the treatments. Levels of CRP were decreased by either POE-POP or LEVA (P<0.05) at Day 7 and 21, respectively. LEVA-treated pigs had increased levels of HpG (P<0.05) at Day 14 and 21, whereas POE-POP-treated pigs had decreased and increased levels (P<0.05) at Day 7 and 21, respectively. Both IBC stimulated in vitro PHC of GR (from Day 7 to 35) and MBC of MO (at Day 35) from treated pigs (P<0.05). MO from POE-POP-treated pigs exhibited increased PHC (P<0.05) at Day 35, whereas GR from LEVA- or POE-POP-treated pigs showed increased MBC (P<0.05) at Day 7 and 35, respectively. The pigs treated by LEVA or POE-POP had higher proportions of CD4+ CD8+ T cells (P<0.05) from Day 14 to 35 or Day 28 to 35, respectively. Tested IBC showed capability of stimulating particularly cellular components of nonspecific and specific immunity during early postweaning period before pigs reach adult immunocompetence values, and thus could be considered as potent immunostimulators in swine production. Key words: synthetic immunobiotics; innate/adaptive immunity; weaned pig VPLIV NA SISTEMSKO PRIROJENO IN PRIDOBLJENO IMUNOST PRI ODSTAVLJENIH PUJSKIH PO ENKRATNEM DODATKU IMUNOBIOTIKOV PREKO PREBAVNEGA TRAKTA Namen raziskave je bil ugotoviti vpliv lavamisola (LEVA) in polyoxyethylen-polyoxypropylena (POE-POP), znanih imunobiotikov, ki bi jih lahko uporabljali v prasicereji za boljsi prirast namesto antibiotikov, na sistemsko prirojeno in pridobljeno imunost pri odstavljenih, 4 tedne starih pujskih. Vsem živalim smo LEVA in POE-POP dajali preko ust (peroralno) in tedensko v odobju petih tednov dodajanja imunobiotikov ugotavljali stopnjo fagocitoze in protimikrobno aktivnost monocitov in granulocitov, raven serumskega C-reaktivnega proteina (CRP), profil haptoglobina (HpG) ter raven prisotnosti limfocitov T, ki izražajo gena CD4 in CD8 v krvi. Rezultati so pokazali, da je bila raven serumskega CRP znižana pri pujskih, ki so prejemali tako LEVA kot POE-POP 7. in 21. dan raziskave (p < 0,05). Pujski, ki so prejemali LEVA, so imeli zvisan HpG 14. in 21. dan raziskave (p < 0,05), medtem ko so imeli pujski, ki so prejemali POE-POP, 7. dan raziskave HpG znižan, 21. dan pa zvisan (p < 0,05). Oba imunobiotika sta v pogojih in vitro dvignila stopnjo fagocitoze pri granulocitih in protimikrobno aktivnost monocitov na 35. dan raziskave (p < 0,05). Pujski, ki so prejemali en ali drugi imunobiotik, so imeli 35. dan raziskave visje vrednosti limfocitov T, ki so izražali CD4 in CD8, od vrednosti na 14. ali 28. dan raziskave (p < 0,05). Rezultati raziskave torej kažejo, da sta oba imunobiotika pozitivno vplivala na izbrane kazalce delovanja imunskega sistema in sta potencialno zanimiva za uporabo v prasicereji za spodbujanje delovanja imunskega sistema in s tem boljsi prirast prasicev. Kljucne besede: odstavljeni pujski; umetni imunobiotiki; prirojena imunost; pridobljena imunost;

Dietary supplementation with mannan oligosaccharide and clinoptilolite modulates innate and adaptive immune parameters of weaned pigs

The aim of this study was to assess modulatory effects of dietary supplements mannan oligosaccharide (MOS) and clinoptilolite (CPL) as potential alternatives to antibiotic growth promoters (AGP) given to 4-week old pigs at weaning (Day 0) on their innate/adaptive immunity by determining: alterations in C-reactive protein (CRP) and haptoglobin (HpG) serum levels, efficiency of blood monocytes (MO) and neutrophilic granulocytes (GR) for in vitro phagocytosis (PHC)/microbicidity (MBC) and proportion of extrathymic double positive CD4 CD8 (CD4+CD8+) T cells throughout 35 days of the study. Neither MOS nor CPL changed the serum concentrations of CRP, whereas that of HpG was significantly increased in the CPL supplemented pigs (p<0.05) at Day 35. Activity of PHA of GR was significantly increased by both dietary supplements (p<0.05) from Day 7 to Day 35. Also, the GR from pigs fed with both supplements had significantly increased MBC at Day 7 (p<0.05), but at Day 35 such an increase was observed only for CPL. The in vitro PHC/MBC of MO did not change in either group of supplemented pigs. The pigs supplemented with MOS had a significantly higher proportion of CD4+CD8+ T lymphocytes at Day 28 (p<0.05). Although both supplements showed a promising ability to stimulate rather innate than adaptive cellular immunity, it does not appear that any solely applied natural substance such as MOS or CPL in the current study could be a competitive alternative to conventional AGP for improving health and promoting growth in weaned pigs.

The effect of micronutrient deficiencies on the health status of transition cows

Minerals, in particular micronutrients such as copper, manganese, zinc, selenium and chromium, deliver health benefits for high-yielding dairy cows. The availability of nutrients is particularly important during the transition period (between 3 weeks prepartum to 3-4 weeks postpartum), which is a highly vulnerable time in the production cycle that determines the health status of lactating cows, their milk yield and reproductive performance. In cows, prolonged mineral deficiency can produce subclinical symptoms of nutrient deficiency and, if untreated, can lead to a clinical presentation of the disorder. Less severe deficiencies, in particular micronutrient deficiencies, are more difficult to identify because they induce only minor functional and structural changes in the body, but together with energy metabolism disorders, they can lead to oxidative stress and immunosuppression, in particular in the transition period. Micronutrient supplements reduce the harmful effects of free radicals because the majority of micronutrients form the active centers of antioxidant enzymes. Dietary supplementation promotes reproductive performance, healthy fetal development and immunity. Micronutrients alleviate bacterial infections associated with mastitis, lower somatic cell counts in milk and minimize the risk of metabolic diseases caused by a negative energy balance. This study discusses the role of the major micronutrients in cattle nutrition and their effect on the health status of transition cows.

Effect of Yeast Supplements On Selected Health-Promoting Properties of Lamb Meat

Abstract The objective of this study was to determine the effect of yeast preparations on selected properties of meat from suckling lambs. The experiment was conducted on 30 Kamieniec rams divided into three groups: I - control, II and III - experimental. Throughout the 100-day rearing period, the experimental animals were fed diets supplemented with Saccharomyces cerevisiae prebiotics: Inter Yeast S® brewers yeast in group II, and Biolex®-Beta S in group III. Fat content was determined by Soxhlet extraction. The fatty acid composition of intramuscular fat was determined by esterification followed by gas chromatography analysis. The content of vitamins A and E was determined by liquid chromatography. A significant increase in the content of fat, cholesterol, and vitamin E was noted in the meat of lambs of group II in comparison with control. The fat of animals in group II contained more fatty acids: C14:1, C18:2, C22:6 (P≤0.05), and C18:2 n-6 cis9trans11 (P≤0.01), and less C20:4 (P≤0.01). The fat of animals of group III contained more C18:0 and C22:6 fatty acids (P≤0.05), and less C18:0 (P≤0.05). The fat in this group had also lower concentrations of MUFA (P≤0.05), UFA, and, consequently, a lower UFA/SFA ratio (P≤0.05) in comparison with controls. The yeast-based supplements significantly affected selected health-promoting properties of lamb meat. The applied diet supplements had no significant effect on AI and TI values.