Introduction: The goal of this study was to assess the influence of vaccination against enteric redmouth disease on oxidative stress biomarkers and antioxidant defence in the muscle tissue of rainbow trout (Oncorhynchus mykiss Walbaum) vaccinated against Yersinia ruckeri in the first and second month after immunisation. Material and Methods: Healthy fish were vaccinated orally with inactivated whole cells of a virulent strain of Y. ruckeri. One and two months after immunisation the muscle samples were collected. Results: No significant difference was noted in lipid peroxidation level in either the first or second month after vaccination, while aldehydic and ketonic derivatives of oxidatively modified proteins (OMB) in the vaccinated group were significantly lower in the second month compared to those in the first month after vaccination (P < 0.05). The content of ketonic derivatives of OMB in muscles in the first month after immunisation was higher compared to untreated control. All these culminated in a depletion of glutathione peroxidase (GPx) activity and low level of total antioxidant capacity (TAC). Conclusion: Correlations between catalase activity and lipid peroxidation and TAC confirmed the pivotal role of catalase in antioxidant defence during immunisation. From a broader perspective, it is suggested that immunisation of fish with Yersinia vaccine is associated with induced free radical formation and oxidative stress. Free radicals would therefore be at least partially responsible for the induction of both humoral and cellular elements of the immunity and increased protective immunity against Y. ruckeri infection.

Introduction: The goal of this study was to assess the influence of vaccination against enteric redmouth disease on oxidative stress biomarkers and antioxidant defence in the muscle tissue of rainbow trout (Oncorhynchus mykiss Walbaum) vaccinated against Yersinia ruckeri in the first and second month after immunisation. Material and Methods: Healthy fish were vaccinated orally with inactivated whole cells of a virulent strain of Y. ruckeri. One and two months after immunisation the muscle samples were collected. Results: No significant difference was noted in lipid peroxidation level in either the first or second month after vaccination, while aldehydic and ketonic derivatives of oxidatively modified proteins (OMB) in the vaccinated group were significantly lower in the second month compared to those in the first month after vaccination (P < 0.05). The content of ketonic derivatives of OMB in muscles in the first month after immunisation was higher compared to untreated control. All these culminated in a depletion of glutathione peroxidase (GPx) activity and low level of total antioxidant capacity (TAC). Conclusion: Correlations between catalase activity and lipid peroxidation and TAC confirmed the pivotal role of catalase in antioxidant defence during immunisation. From a broader perspective, it is suggested that immunisation of fish with Yersinia vaccine is associated with induced free radical formation and oxidative stress. Free radicals would therefore be at least partially responsible for the induction of both humoral and cellular elements of the immunity and increased protective immunity against Y. ruckeri infection.

Introduction: Due to the high heterogeneity of canine lymphoma, the aim of the present study was to test in vitro the chemosensitivity of canine high-grade primary lymphoma cells to various cytostatic drugs commonly used to treat dogs: 4-HO-cyclophosphamide, doxorubicin, dexamethasone, prednisolone, vincristine, etoposide, chlorambucil, lomustine, and cytosine arabinoside. Material and Methods: To determine the cell viability and drug ability to induce apoptosis two different tests were used: an MTT assay and annexin V/propidium iodide staining. Results: Both in vitro tests were found to be useful tools. Significant differences in the sensitivity, depending on the drug type, between B-, T- and mixed/null-type lymphoma cells were found for the majority of the tested drugs. B-type cells were the most sensitive in vitro, whereas T-type cells seemed to be the most resistant. Doxorubicin, chlorambucil, etoposide, and vincristine most strongly reduced the cell viability and induced apoptosis. Conclusion: In vitro assays, such as the MTT test and especially the annexin V/PI assay, may be useful tools for predicting a response to the treatment of high-grade lymphoma in dogs or improving the treatment outcomes in individual animals.

Introduction: Due to the high heterogeneity of canine lymphoma, the aim of the present study was to test in vitro the chemosensitivity of canine high-grade primary lymphoma cells to various cytostatic drugs commonly used to treat dogs: 4-HO-cyclophosphamide, doxorubicin, dexamethasone, prednisolone, vincristine, etoposide, chlorambucil, lomustine, and cytosine arabinoside. Material and Methods: To determine the cell viability and drug ability to induce apoptosis two different tests were used: an MTT assay and annexin V/propidium iodide staining. Results: Both in vitro tests were found to be useful tools. Significant differences in the sensitivity, depending on the drug type, between B-, T- and mixed/null-type lymphoma cells were found for the majority of the tested drugs. B-type cells were the most sensitive in vitro, whereas T-type cells seemed to be the most resistant. Doxorubicin, chlorambucil, etoposide, and vincristine most strongly reduced the cell viability and induced apoptosis. Conclusion: In vitro assays, such as the MTT test and especially the annexin V/PI assay, may be useful tools for predicting a response to the treatment of high-grade lymphoma in dogs or improving the treatment outcomes in individual animals.

Introduction: The current study was designed to detect Mycoplasma agalactiae (Ma), Mycoplasma mycoides subsp. capri (Mmc), Mycoplasma capricolum subsp. capricolum (Mcc) and Mycoplasma putrefaciens (Mp) in sheep and goats with clinical signs consistent with contagious agalactia.Material and Methods: A total of 299 samples were collected from 55 mixed herds in Azarbaijan-e-Sharghi province, Iran. Samples were examined using PCR and culture methods.Results: The findings showed that in 40 herds at least one sample was positive by PCR or culture method. Moreover, out of 274 sheep samples, 101 were proved to be positive using the PCR technique and 76 were found positive using the culture method. Out of 25 goat samples, 10 were found positive using PCR and 9 were positive through the culture method. Less than 20% of isolated mycoplasmas were Ma. Ma was detected from almost all studied regions in the province while Mmc, Mcc, and Mp were detected only in a very limited area that was deemed by the research group the mixed infection zone.Conclusion: In vaccination or eradication projects, it would be more economical to focus on mixed infection zones. Further investigation on mixed infection zones could facilitate better understanding of contagious agalactia epidemiology.

Introduction: The current study was designed to detect Mycoplasma agalactiae (Ma), Mycoplasma mycoides subsp. capri (Mmc), Mycoplasma capricolum subsp. capricolum (Mcc) and Mycoplasma putrefaciens (Mp) in sheep and goats with clinical signs consistent with contagious agalactia.

Introduction: To identify novel pathways involved in the pathogenesis of ketosis, an isobaric tag for relative and absolute quantitation/mass spectrometry was used to define differences in protein expression profiles between healthy dairy cows and those with clinical or subclinical ketosis.Material and Methods: To define the novel pathways of ketosis in cattle, the differences in protein expression were analysed by bioinformatics. Go Ontology and Pathway analysis were carried out for enrich the role and pathway of the different expression proteins between healthy dairy cows and those with clinical or subclinical ketosis.Results: Differences were identified in 19 proteins, 16 of which were relatively up-regulated while the remaining 3 were relatively down-regulated. Sorbitol dehydrogenase (SORD) and glyceraldehyde-3-phosphate dehydrogenase (G3PD) were up-regulated in cattle with ketosis. SORD and G3PD promoted glycolysis. These mechanisms lead to pyruvic acid production increase and ketone body accumulation.Conclusion: The novel pathways of glycolysis provided new evidence for the research of ketosis.

Introduction: To identify novel pathways involved in the pathogenesis of ketosis, an isobaric tag for relative and absolute quantitation/mass spectrometry was used to define differences in protein expression profiles between healthy dairy cows and those with clinical or subclinical ketosis.

Introduction: Peroxiredoxin 6 (Prdx6) is a bifunctional protein and a unique 1-Cys Prdx of the peroxiredoxin family. The expression and regulation of Prdx6 are implicated in numerous physiological and pathological processes.Material and Methods: Eight stepwise truncated DNA fragments obtained from the 5′-flank region of the Prdx6 gene were prepared and subcloned into the pSEAP2-Enhancer vectors. To investigate the transcriptional activity of the truncated DNA fragments, the recombinant plasmids were transfected into the COS-1 cells and the transcriptional activity was measured via assaying the expression of the reporter gene of the secreted alkaline phosphatase.Results: A 3.4 kb 5′-upstream flank region of the Prdx6 gene was cloned and sequenced. The region from −108 nt to −36 nt of the 5′-flanking region of the Prdx6 gene contained basal transcriptional activity.Conclusion: This result provides the basis for further studies on the gene regulation of the Prdx6-mediated biological processes and on screening for the transacting factors that interact with cis-acting elements of the Prdx6 gene promoter.

Introduction: Peroxiredoxin 6 (Prdx6) is a bifunctional protein and a unique 1-Cys Prdx of the peroxiredoxin family. The expression and regulation of Prdx6 are implicated in numerous physiological and pathological processes.

Introduction: The article presents a rapid and simple analytical procedure for determination of four tetracyclines (oxytetracycline, tetracycline, chlortetracycline, and doxycycline) in animal medicated feedingstuffs. Material and Methods: Two-gramme samples were extracted by a Na₂EDTA-McIlvaine buffer (pH 4)/methanol mixtures (40/60, v/v). The determination was achieved by liquid chromatography using a Zorbax Eclipse XDB C18 analytical column with mass spectrometer detection (LC-MS). Results: Recoveries of the antibiotics from spiked feed samples ranged from 78.2% to 113.5%. The LOD and LOQ for tetracyclines in feeds ranged from 2.8 to 4.2 and from 4.3 to 5.7 mg/kg, respectively. Conclusion: The method was successfully validated and proved to be efficient, precise, and useful for quantification of tetracyclines in medicated feedingstuffs.

Introduction: The article presents a rapid and simple analytical procedure for determination of four tetracyclines (oxytetracycline, tetracycline, chlortetracycline, and doxycycline) in animal medicated feedingstuffs. Material and Methods: Two-gramme samples were extracted by a Na2EDTA-McIlvaine buffer (pH 4)/methanol mixtures (40/60, v/v). The determination was achieved by liquid chromatography using a Zorbax Eclipse XDB C18 analytical column with mass spectrometer detection (LC-MS). Results: Recoveries of the antibiotics from spiked feed samples ranged from 78.2% to 113.5%. The LOD and LOQ for tetracyclines in feeds ranged from 2.8 to 4.2 and from 4.3 to 5.7 mg/kg, respectively. Conclusion: The method was successfully validated and proved to be efficient, precise, and useful for quantification of tetracyclines in medicated feedingstuffs.

Introduction: The aim of the study was to investigate the presence of human papillomaviruses (HPV), mouse mammary tumour virus (MMTV), Epstein-Barr virus (EBV), and human polyomavirus BK in canine mammary tumours (CMTs) and to correlate the results of histopathological classification with the results of virological examination. Material and Methods: Eighty CMTs and ten normal canine mammary gland samples were evaluated using histopathological methods and TaqMan real-time PCR analysis. Results: The results indicated that all mammary tumours and normal mammary tissue samples were negative for HPV16 and other HPV, EBV, human polyomavirus, and human mammary tumour virus strains. Conclusion: Further studies should be performed to investigate the existence of other strains of HPV, EBV, and human polyomavirus in CMTs.

Introduction: The aim of the study was to investigate the presence of human papillomaviruses (HPV), mouse mammary tumour virus (MMTV), Epstein-Barr virus (EBV), and human polyomavirus BK in canine mammary tumours (CMTs) and to correlate the results of histopathological classification with the results of virological examination. Material and Methods: Eighty CMTs and ten normal canine mammary gland samples were evaluated using histopathological methods and TaqMan real-time PCR analysis. Results: The results indicated that all mammary tumours and normal mammary tissue samples were negative for HPV16 and other HPV, EBV, human polyomavirus, and human mammary tumour virus strains. Conclusion: Further studies should be performed to investigate the existence of other strains of HPV, EBV, and human polyomavirus in CMTs.

Introduction: In the present study apoptosis was investigated in the cornu ammonis and cerebellum of 10 dogs naturally infected with rabies virus. Diagnosis of rabies was based on the results of fluorescent antibody staining and experimental inoculation.Material and Methods: The paraffin tissue sections were stained with haematoxylin and eosin, avidin-biotin complex peroxidase (ABC-P), and terminal deoxynucleotidyl transferase biotin-dUTP nick end-labelling (TUNEL) methods.Results: Histopathological examination revealed encephalomyelitis of varying severity and the presence of Negri bodies. Dense rabies antigens were determined in the motor neurons with ABC-P method. On the other hand, Bcl-2 protein and Bax protein gave positive reaction in seven and five cases, respectively. TUNEL staining demonstrated very marked apoptotic changes in the nuclei of neurons localised deep in the substantia alba of the cerebellum. Similar changes were also determined in perivascular mononuclear cells and glia cells within the substantia alba. No apoptopic changes were found in the motor neurons of the cornu ammonis.Conclusion: The absence of apoptotic changes in the neurons was considered to be the consequence of the necrotic changes that developed in these neurons.

Introduction: In the present study apoptosis was investigated in the cornu ammonis and cerebellum of 10 dogs naturally infected with rabies virus. Diagnosis of rabies was based on the results of fluorescent antibody staining and experimental inoculation.

The aim of the study was to assess the changes of blood parameters in 12 three-week-old Polish Merino sheep subjected to experimental jaagsiekte sheep retrovirus (JSRV) infection.Material and Methods: Haematological (WBC with leukocyte subpopulations: GRA, LYM, MID, and RBC, MCV, MCH, MCHC, HGB, HCT, PLT, and MPV) and biochemical blood parameters (acid/base balance, cation/anion content, and gasometry) were determined in blood samples collected one month after JSRV infection, then at four-week intervals for five consecutive months.Results: A decrease in RBC, HCT, MCV, PLT, MPV, and LYM values in comparison with controls was found in the last month of observation. On the other hand, at the same time, an increase in HGB, MCH, MCHC, WBC, MID, and GRA indices was observed. Moreover, at the end of experiment blood gasometric indices such as pCO₂, HCO₃, and tCO₂, and Na and K ion concentrations were higher in the affected lambs than in the healthy animals. The pH values of the challenged animals exhibited less alkaline character than in the case of controls, which was associated with a decrease in O₂% saturation. However, the majority of differences between JSRV inoculated and control groups was not statistically significant.Conclusion: The observed changes in the examined blood parameters can be considered as prodromal symptoms in the preclinical phase of adenocarcinoma development associated with JSRV infection.

The aim of the study was to assess the changes of blood parameters in 12 three-week-old Polish Merino sheep subjected to experimental jaagsiekte sheep retrovirus (JSRV) infection.

Introduction: The objective of this study was to describe a pharmacokinetic–pharmacodynamic (PK/PD) approach for determination of a rational dosage of ampicillin (AMP) and depletion of the antibiotic residues in milk after intramammary administration to cows.Material and Methods: The cows came from different farms from the Lublin Province area. They (n = 9) received 5 g of the drug, containing 75 mg of AMP sodium in physiological solution, through a syringe tube by intramammary administration. Following single intramammary administration, the milk samples (5 mL) were collected after 2, 4, 6, 8, 10, 24, 36, 48, and 60 h. The liquid chromatography-mass spectrometry analysis was performed on the Agilent 1200 system connected to an AB Sciex API 4000™ mass spectrometer. The pharmacokinetic analysis of the concentrations of the antibiotic in milk was performed using software Phoenix® WinNonlin® 6.4. Calculations were made in non-compartmental (slopes, highest, amounts, and moments) and compartmental analysis.Results: The pharmacokinetic characteristics of AMP after intramammary administration indicate rapid elimination of the drug from milk. The mean residence time had a several-fold lower value than the designated elimination half-life and amounts to only 3.4 h. The concentration of the drug in the milk dropped relatively quickly and the process was very dynamic.Conclusion: The conducted research confirms the rationale of using the PK/PD model in order to verify the dosing regimen for other antibiotic groups and various indicators of the applied PK/PD model.

Introduction: The objective of this study was to describe a pharmacokinetic–pharmacodynamic (PK/PD) approach for determination of a rational dosage of ampicillin (AMP) and depletion of the antibiotic residues in milk after intramammary administration to cows.