The technical execution of single-cell RNA sequencing (scRNA-seq) in liver tissue poses notable challenges due to the inherent fragility of hepatocytes, the distinct metabolic zonation, and the rapid activation of stress-response mechanisms during tissue dissociation. These complications are further exacerbated in porcine liver, where the large size of hepatocytes and elevated endogenous RNase activity resemble human hepatic tissue, yet are particularly vulnerable to mechanical and enzymatic stresses. Conventional scRNA-seq methodologies frequently yield diminished cell viability, RNA degradation, and skewed representation of hepatic subpopulations when applied to porcine models. We introduce a reproducible and species-optimized protocol for conducting scRNA-seq on porcine liver. This methodology incorporates cold-active enzymatic dissociation, low-shear mechanical manipulation, RNase suppression, and debris-minimized density separation to maintain cellular integrity and transcriptomic accuracy. A brief in vitro stabilization phase further improves cell viability and mitigates ambient RNA contamination, yielding single-cell suspensions amenable to droplet-based systems, such as the 10× Genomics v3.1 and v4 chemistries. Benchmarking outcomes reveal high viability rates (>80%), substantial transcript complexity, and consistent recovery of both parenchymal and non-parenchymal populations with diminished stress-response profiles. This workflow establishes a standardized approach for high-fidelity, cross-institutional scRNA-seq investigations and bolsters translational research applications in toxicology, pharmacokinetics, and hepatoprotective therapies. Acknowledgement This research is financed and supported by the scientific project NCN-OPUSLAP (UMO-2021/43/I/NZ9/02612) entitled: Multilevel molecular analysis of the hepatoprotective effect of medicinal herbs extracts in prevention of liver dysfunction caused by aflatoxin B1 in pig as an animal model (in-vivo), and hepatocyte cell culture analysis in human and pig (in-vitro).
Background: Newcastle disease (ND) caused by virulent avian avulavirus 1 (AAvV-1) within the family paramyxoviridae, affects variety of avian species around the globe as well as responsible for significant economic losses in the poultry industry. DOI: http://dx.doi.org/10.12775/TRVS.2020.001
Background: Bovine chemokine C-X-C motif legend 8 (CXCL8) also known as interleukin 8 (IL8) is a chemotactic factor that attracts neutrophils, basophils, and T-cells, in re-
Background: Ever since the development of first next-generation genome sequencer (NGS) in 2005, there are rapid developments of high throughput next-generation genome
The aim of the study was to evaluate the pregnancy rate after application of the OvSynch (OVS) programme depending on the reliability of the programme implementation and the number of cows treated at the same time. Taking into account the reliability of the programme, cows were divided into five groups. Females in group I were inseminated after the initial dose of GnRH, i.e. 1-6 days after the start of the programme, in group II – on days 7-9 of the OVS protocol, after administration of prostaglandin, in group III – on day 10, after completion of the full OVS protocol and timely insemination, and in group IV – on days 11-12, after completion of the full protocol and insemination according to estrous symptoms. Cows in group V were inseminated at a later period. Additionally, cows were divided into VI groups depending on the number of animals treated simultaneously during the periodic veterinary visit. Groups A, B, C, D, E and F consisted respectively of 1, 2, 3, 4, 5 and 6 cows treated on the same day. Examination for pregnancy was performed twice – on the 28th and 56th day after insemination using an iSkan ultrasound device (Draminski) equipped with a 7.5 MHz linear probe. Large differences in mating rates were shown between farms. Taking into account the timely execution of the OvSynch programme, the highest – 42.3% – mating rate was recorded in cows in which the OVS programme was executed correctly. In the other groups it was significantly (P<0.05) lower. In the case of simultaneous OvSynch in 6 cows, the percentage of correctly executed programs amounted to 83.3% and was significantly higher than in the case of treatment at smaller groups. Using the OvSynch program, the highest pregnancy rates are guaranteed by timely administration of the prescribed injections and insemination at the appointed time. Deviation from the protocol results in a worsening of the pregnancy rate. A higher number of treated cows guaranteed a higher percentage of correctly executed OvSynch programmes. Reasons for this situation were discussed.
Background: Studies have shown the influence of the Red and White Belted meat breeding boars’ usage regimes on the quality of sperm products under the effect of “Humilid” biologically active feed supplement. The data obtained in the control group of breeding boars are explained by their intensive use and significant consumption of body resources to maintain sexual activity. Due to the adaptogenic properties of the drug, which had a positive effect on the quality of sperm production in the experimental group of animals on the 30th day of the experiment, the body’s resources were mobilized. It is established that consumption of biologically and ecologically safe feed supplements of humin nature permits to obtain high-quality and complete sperm products under different regimes of using breeding boars. Methods: For the experiment, 10 animals of similar age and live weight were selected, 5 of which were the control group that did not receive the 'Humilid’ drug and 5 animals that received a biologically active feed supplement with food. During the experiment, different regimes of boar usage were applied (once, twice and three times a week) for 60 days. The experiment was divided into three stages (preparatory, main, and final), during which sperm quality indices were monitored: ejaculate volume, sperm concentration, total number, and number of live sperm, their motility, and survival during incubation. Results: In the previous period, the following dynamics in the effect of the effect of the regimens on sperm quality was observed: the twofold regime showed a decrease in ejaculate volume in the control group by 18.7 % and in the experimental group by 7.2 % compared to a single semen collection regimen. In the main period, the decrease in ejaculate volume was 17.6 % (p<0.001) and 5.4 % (p<0.001), respectively, and at the final stage – by 15.8 % (p<0.001) and 9.1 % (p<0.001). At that time, sperm motility and survival rates were the highest in animals that received “Humilid” biologically active feed supplement and had a sexual load twice a week. Sperm motility in the preparatory period in the control group was 90.4 % and in the experimental group – 95.0 %, survival – 84.0 % and 90.8 %, respectively. During the main phase of the experiment, sperm motility was within the range of 85.9 % and 98.8 % and survival – 78.5 % and 95.3 %, respectively. In contrast, in the final stage, sperm motility was 89.6 % and 92.0 % and survival was 82.0 % and 88.9 %, respectively.
Tuberculosis (TB) is a chronic pulmonary disease caused by different representatives of Mycobacterium. The fight against bovine TB has been implemented within the framework of the 'One Health’ concept. ‘One Health’ is based on cooperation between broad medical professions, including medicine and veterinary medicine to prevent prevalence of zoonotic diseases. Analysing the development of these disciplines throughout history, we can say that they common origin. Up to the second half of the 18th century, it was often the same people treating both humans and animals. Veterinary medicine diverged from medicine in the second half of the 18th century, after the first veterinary schools were established. However, it occurred that some diseases affecting humans and animals required close cooperation between medicine and veterinary medicine. At first, the link between human and cattle TB has not been made. Due to the chronic nature of TB, it usually took quite long between contact with the sick animal or its products and the onset of the disease. But, after Robert Koch attributed the origin of the disease to mycobacteria, the progress has been made in identification of the pathogenicity of human and bovine TB. Effective measures of limiting spread of TB had to combine treatment of infected people with eliminating the reservoirs of the germ, which were and are animals. Due to coordinated efforts within ‘One Health’, about a hundred years later, many countries were announced as TB-free.
Background: The puberty period in pigs depends on the functional activity of the endocrine system, which has an effect on the blood through the connective tissue. It has been established that during puberty period, the steroidal and thyroid hormones regulate the processes of metabolism and proliferation, differentiation and apoptosis of cells. The aim of the research was to find out the effects of the morphological composition of blood under the influence of steroidal and thyroid hormones during the process of puberty in different directions of productivity in the pigs.Methods: Experiments were performed on two groups of clinically healthy piglets from Pietrain and Large White (LW) breed. To assess the steroidal and thyroid hormone levels, the morphological blood was collected from the anterior hollow vein at the puberty ages of 120, 150, 180 and 210 days, when their live weights achieved to 100 kg.Results: The predominance of hemoglobin content and the amount of erythrocytes in the blood were observed higher (p <0,05) in Pietrain piglets than LW piglets on the 120 and 150 days of puberty development, in contrast to the general decrease in leukocytes in piglets of both breeds at the age of 150 days with the next gradual increase. The age-related changes in the leukocyte formula were directly related to the direction of animal productivity. In the LW breed, from the beginning of the experiment, there was an increase in the number of rodenuclear neutrophils at 210 days of puberty, followed by a sharp decreasing on 36.4%. In Pietrain breed, the level of the rodenuclear neutrophils decreased on 28.6% (p <0.01) at the age of 150 days, followed by a sharp increasing over the coming months. From the puberty ages of 150 days to the 210 days, the concentration of estradiol decreased in the first genotype: two times (p <0.05); in the second genotype: 1,9 times (p <0,01). The content of thyroxine and triiodothyronine increased in the blood serum of Pietrain (p <0.01) and LW (p <0.001) piglets at the puberty ages of 120 to 180 days. Study found that the amount of progesterone in LW piglets was higher relatively to Pietrain piglets in the all investigated periods. The content of estradiol in piglets of both breeds from the 120 and 150 days of puberty tended to decrease by 2.8 times (p <0.001) in LW piglets, and 1.4 times (p <0.05) in Pietrain piglets, and the testosterone level increases, 2.1 (p <0,05) and 1,9 (p <0,05) times respectively. The significant influence of progesterone on the number of lymphocytes and common leukocytes in Pietrain piglets was established. The level of estradiol significantly influenced the number of granulocytes in Pietrain piglets at the age of 150 days (r = 0.92 and 0.99), and in LW piglets, the maximum correlation level was observed at the puberty age of 150 days (r = 0, 61) and 180 days (r = 0.94).
Background: Due to the growing amount of biological data, it is often necessary to select the most optimal estimation method for DNA sequence alignment across livestock species. One of the most important benches of genomics is to modelling homology between considered DNA sequences. A multiple sequence alignment is a potent tool for molecular and evolutionary biology, and there are several programs and algorithms applicable for this purpose. The purpose of this paper was to study the most commonly used DNA alignment algorithms to select the optimal tool dedicated for short sequences.Methods: Four steps of bioinformatics pipelines were considered to benchmark the algorithms for multiple DNA sequence alignment across livestock species: 1) selection of reference genome sequences of ARS1.2 for cattle, EquCab3.0 for horse and vicPac2 for alpaca with a low E-value using TBLASTn 2) removing gaps for these sequences 3) alignment of obtained sequences using examined algorithms 4) matching the quality of aligned sequences with sequences of reference genomes by more software. The time of computation was archived for the whole analysis. The seven programs were utilized, each based on different alignment algorithms, namely: ClustalO, ClustalW, Kalign, MAFFT, MUSCLE, Probcons and T-Coffee.Results: The result obtained in this study showed that the fastest is progressive algorithms such as Kalign or MUSCLE-FAST. Moreover, the iterative algorithms like MAFFT and MUSCLE revealed a higher quality of the alignment. The T-Coffee and Probcons programs were computational cost-effective; simultaneously, they were generating a medium-quality calculation in a relatively long time. The best quality of alignment was shown by iterative variants of the MAFFT program; however, the speed of the calculations was relatively low. The fastest algorithm was Kalign, making alignment much faster than the competitors, but achieving average results in the quality of the alignment. The average speed ratio concerning the quality of the analyzed algorithms was obtained by the progressive version of MAFFT, NS1.Conclusions: We conclude that the results of this study can be used to re-alignment of variant primers in new livestock genome releases.
This study investigated the effects of dexmedetomidine on bodily functions in animals by prolonged use. The studies were carried out on laboratory mice, which were divided into 2 groups: the first (test) one was administered administered orally by the Dexmedetomidine drug at a 0.01 ml dose, the second (control) one did not introduce any drugs. During the experiment, control over both groups’ mice’s coordination of movements, metabolic parameters were carried out, and the bodyweight was measured. Vital signs monitoring was carried out in 3 stages: 1) before drug administration; 2) 15 minutes after administration; and 3) after awakening, respectively. Results revealed that in 2-3 days, the frequency of respiratory movements began to increase by 19,1%, the awakening began to increase, and the general condition (refusal to eat, inactivity) changed for the worse. There was a 11% decrease in the test group on the last day after the next body weight measurement, while in the control group, on the contrary, the weight increased by almost 5%, and also at autopsy, changes were found in the tissues of the lungs and liver. The research aimed to identify the toxic effects of drugs in the category of α2-adrenergic receptor agonists, and at the moment it is possible to talk about some toxic effects of this group, but research needs to continue.
Diseases of the muscular-skeletal system in horses concerning the tendons, especially the suspensory ligament (interosseous muscle), have a significant impact on the possibility of subsequent use of sporting horses in recreation and sport. Therefore, accurate diagnosis and proper treatment is a key to solve this issue. The article describes the causes, clinical symptoms, treatment and diagnostic possibilities of interosseous muscle injuries in horses.
Animal production is of major economic importance in a number of European countries, including Poland. In terms of swine production, there are over 1.3 billions of swine worldwide. The EU is the second largest pork producer with over 186 millions of swine. In Poland, according to the data from EUROSTAT, in June 2020, the number of pigs exceeded 11.43 million, showing an increase of 6.0% compared to the same period in the previous year. The observed international fluctuation in the produced pigs’ number is mainly related to the lack of profit in production due to the occurrence of infectious diseases, with African Swine Fever (ASF) amongst them. ASF in Poland has been a major threat for pig production for the eight years. The main problems in ASF eradication in Poland are due to the high density of wild boar. Other factors, responsible for long-distance ASF spread are related to human-mediated activity and lack of awareness of pig producers. The observations from the eight years of ASF epizootic in Poland indicate that the disease could not have been effectively controlled in wild boar population and could only be restricted in domestic pig population following stringent biosecurity measures. The only solution for future sustainable pig production in Poland seems to be strict collaboration between pig producers, veterinary inspection, and hunters.
Background: The aim of this study was to investigate the relationship between CXCL8 (CXCL8c.105A>G and CXCL8c.210C>T) SNP polymorphism and the clinical mastitis and production trait in Polish HF cattle.Methods: The trait-associated CXCL8 SNPs polymorphism study was carried out in the Polish HF bulls (n=151) by PCR-RFLP methods as described in the previous issue of this journal. The phenotypic records (assessment year 2017) of clinical mastitis, functional and fertility trait were collected from the Research Institute of Animal Production, Balice, Poland (http://www.izookrakow.pl).Results: Overall, the trait-associated study revealed no relationship between the CXCL8c.210C>T SNP polymorphism and the breeding value of selected clinical mastitis and fertility trait of breeding bulls. However, TT genotypes showed low levels of significance of differences for the breeding values of somatic cells count (p = 0.27) and stillborn calving of heifers (p=0.27). Similarly, the trait-associated study showed no correlation between the CXCL8c.105A>G polymorphism and the breeding value of selected clinical mastitis and fertility trait of breeding bulls. However, AA genotypes showed low levels of significance of differences for the somatic cell count (p = 0.12), ease calving of heifers (p = 0.14), the difficulty calving of heifers (p = 0.17), live-born calves of heifers (p = 0.21), and AG genotypes showed the low levels of significance of differences for the stillborn calving in heifers (p = 0.21).Conclusions: Study concludes that trait-associated studies of CXCL8 polymorphism did not identify highly significant effects on clinical mastitis and fertility trait in the investigated breeding bulls of Polish HF cattle.
Background: With modern requirements for housing agricultural animals particularly pigs, the manifestation of normal physiological functions significantly changes, especially reproductive one, which is manifested in an insufficiently expressed sexual cycle, reduced resistance due to technological stress. There is a need to develop effective reproductive biotechnologies in pig using organic stimulants of entomological origin, among which a significant place is given to the homogenate of drone larvae (HDL).The aim of the study was to determine the dietary effects of HDL on the formation of prooxidant-antioxidant homeostasis in Large White (LW) gilts,as well as to optimize reproductive function during experimental feedingof HDL.Methods: In the study, gilts representing Large White breed were investigated. A total of 20 gilts representing two groups. The first feeding group (n=10) of LW gilts was fed with the control normal diet, and the second feeding group (n=10) of LW gilts wasfed with the supplement diet i.e., the biological additive of homogenate drone larvae in quantity of 0.5g daily. To assess prooxidant-antioxidant homeostasis in pigs, blood samples were taken from the anterior vena cava into the diestrus phase i.e., 11 days from the beginning of the second cycle after the determination of the immobility reflex and the estrus phase i.e., 24 days after the start of estrus after the determination of the immobility reflex. Intracervical artificial insemination of gilts was performed during 24 and 36 hours after the determination of the immobility reflex, and depending on the manifestation of signs of estrus.Results:Study found an intensification of peroxidation processes during the period of puberty in LW gilts , which wasmanifested in an increase in the content of diene conjugates on the 180th and 210th days of development by 63.6% and 44.1%, respectively.This is confirmed by a significant increase in the number of secondary products of peroxidation - TBA-active complexes in these periods of their development. Studyrevealed that during puberty of gilts the antioxidant defense system wasthe most labile from 6 to 8 months of age, where the variability of PAG components wassignificant in terms of increased activity of superoxide dismutase (p <0.01) and catalase (p <0.05), as well as the amount of reduced glutathione. Moreover, additional feeding of HDLcontributedto the probable slowing down of peroxidation processes in blood, which was confirmed by the lower content of diene conjugates (p <0.05-0.01) and TBA-active complexes (p <0.01-0.001).Conclusions: Study concluded that experimental supplementary feeding ofHDLdiet to LWgiltsstimulatedthe reproductive function during their puberty by reducing the time to the first estrous cycle for artificial insemination, thus improved the fertility in LW gilts.
This case report describes an episode of acute encephalopathy in a cat treated with metronidazole due to chronic gastrointestinal disease (giardiasis). Metronidazole had been administered at a dose of 28 mg/ kg/day for 7 days. Cat showed signs of head pressing, reduced consciousness, ataxia, seizures, lack of menace response and progressive weakness. Complete blood cell count revealed haemolytic anaemia, serum biochemistry, coagulation profile, blood pressure measurement, urinalysis, were unremarkable. Metronidazole administration was immediately discontinued; supportive care consisted of fluid therapy, blood transfusion was also performed. The neurological status of the patient improved rapidly within 72 h. Anaemia improved in next 7 days, however transfusion and steroid treatment was necessary. Two weeks post intoxication abdominal ultrasound revealed abnormal hepatic pattern, enlarged mesenteric lymph nodes and lack of normal intestinal layering. A biopsy had been taken during explorative laparotomy and histopathologic examination revealed gastrointestinal and hepatic lymphoma. The aim of this case report is to present that cats suffering from multiple conditions may show metronidazole intoxication even when recommended dosage and treatment length is used. We cannot exclude that metronidazole was responsible for autoimmune haemolytic anaemia.
Transcriptome data presented in this article is associated with the research article entitled “Single nucleotide polymorphism discovery in bovine pituitary gland using RNA-seq technology” published in PLOS One [1]. Herein, we provide raw and analysed RNA-seq data of pituitary gland tissues from three cattle breeds, viz., Polish-HF, Polish Red and Hereford
Transcriptome analysis using high-throughput next-generation sequencing (HT-NGS) technology provides the capability to understand global gene expression variations through a wide range of tissue samples in domesticated animals. We provide raw and analysed data for transcriptomic analysis of liver tissues from Polish-HF, Polish Red and Hereford cattle breeds, obtained by RNA-seq. High-quality sequencing data have been analysed using our bioinformatics pipeline which consists of FastQC for quality controls, Trimmomatic for trimming, and BWA version 0.7.5-r404 for alignment to the Bos taurus reference genome, SAMtools for SNPs identifications, and differentially expressed genes (DEGs) identification using DEseq and edgeR pipelines after adjustment for false-discovery rate (FDR) with adjusted two-sided p values <0.01 and the trimmed mean of M values (TMM) normalisation method. The data accompanying the published manuscript describing the SNPs and DEGs identification in the bovine liver transcriptome of cattle breeds. Raw FASTq files for the RNA-seq libraries are deposited in the NCBI Sequence Read Archive (SRA) and have been assigned BioProject accession PRJNA312148. Raw and processed RNA-seq data were deposited and made publicly available on the Gene Expression Omnibus (GEO; GSE114233).
Background: The mastitis is one of the most important diseases affecting losses of milk yield and functional longevity resulting in economic losses in dairy farms. The cow's inflammation of the mammary gland is a consequence of complex processes in the innate immune system, activation of several cells and accompanying factors that lead to the elimination of microorganism invaders. The study aimed to investigate genetic background by the genome-wide association study (GWAS) for identified copy number variations (CNVs) in one of the most important economically disorders – the mastitis in dairy cattle.Methods: We investigated 682 Polish-HF cows previously genotyped and analyzed using the following steps: i) identification of CNVs across all the cows based on ARS-UCD1.2 genome map, ii) to perform the GWAS with the utilization of logistic regression models, iii) functional analysis for identified CNVs to indicate putative candidate genes. The cow's genotypes were deposited in the Polish cattle SNP genotypes database (cSNP-db). The phenotype database information covered the veterinary history of mastitis was collected from four experimental farms belonging to the National Institute of Animal Production, Balice, Poland.Results: Study identified 326 CNVs of which only 28 CNVs were significant with p-value <0.05. The most significant CNVs were identified for BTA17, BTA22, and BTA25, respectively. However, the most interesting from the functional point of view of previously reported CNVs overlapped with genes associated with mastitis were loaded on BTA03 (MSH4), BTA18 (FOXC2, FOXL1, MTHFSD) and BTA27 (TLR3), respectively.Conclusions: The study clearly concluded that the identification of candidate genes associated with mastitis using of CNVs is feasible. Therefore, one can apply this GWAS approach between the common CNVs and mastitis as a case-control experiment. Our study identified few genes like MSH4, MTHFSD, FOXC2, FOXL1, and TLR3 which were overlapped with tree CNVs to functionally assess and might be considered as the candidate genes for mastitis in Polish HF cattle population. Moreover, obtained results supported by the different genome and transcriptome analysis could lead to a better understood of the genetic background of mastitis.
Introduction and objective. With regard on its’ proprieties, the food of animal origin is subject to a special supervision of the Polish Veterinary Inspection. Acquisition of such products requires strict observation of legal norms. The paper focuses on the sanitary state of slaughterhouses and butcheries, with regard to the supervision held over these institutions by the Veterinary Inspection. Material and methods The numerical data being subject to the present research reveal its activity in the years 2005 – 2016, divided into 2 sub-terms, each covering the 6-years period (report No. RRW-5). Statistical analysis was performed, comparing statistical trends (number of given administrative decisions, both warning and forbidding) to the number of supervised and controlled establishments. Results and conclusions. The Veterinary Inspection applies administrative tools, adequate to the dangers affecting consumers. As a result of this, the Inspection extorts the improvement of sanitary state or it eliminates incorrect abattoirs from the market. In the light of conducted analysis, the sanitary state of institutions processing foodstuffs of animal origin can be rated positively, thanks to the proper supervision. Results and observations were described in scope of worldwide tendencies in veterinary supervision of food safety. It is advisable to compare the results obtained in this work with the assessment of final product offered for consumption to the end consumer.
The article provides data on the morphometric changes in the kidneys of pigeons with colibacteriosis. Patho-morphological examination of the kidneys revealed focal hyperemia of the kidney cortical area and edema of the loose fibrous tissue. The convoluted renal tubule epitheliocytes were enlarged in size. In this case, the tubules get narrow and become slit-shaped. The convoluted tubules epithelium showed signs of granular dystrophy. In the chronic course of colibacteriosis, granuloma formation and signs of chronic interstitial nephritis were observed, in which the loose connective tissue of the cortical area is replaced by fibrous tissue. Morphometric analysis revealed that in the experimental group there was a decrease in the nephrocyte diameter (p= <0.05) to 98.06±0.46µm and the nephrocyte area to 6319.38±9.62µm. This results in a significant reduction (p= <0.05) of the sub-capsular space to 11.14±0.16µm compared to the control group. The distal and proximal convoluted tubules diameter also decreased to 127.43±20.78μm and 39.50±1.63μm, respectively. Accordingly, the area of the distal and proximal convoluted tubules decreased significantly (p= <0.05) to 10164.10±0.43µm and 2974.43±4.01µm, respectively. The patho-genetic effect of E. coli toxins revealed the renal tubular obstruction and slowing of the reabsorption and urinary excretion processes resulting in a reduction of glomerular filtration capacity of the kidney. Due to the prolonged course of salmonellosis occurring during the latent course, the area of renal filtration was also reduced. Accumulation of nitrogen metabolism products that enter the bloodstream caused impaired water-salt metabolism and acid-base equilibrium.
