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Open access

R. Urban-Chmiel, R. Pyz-Łukasik, A. Dudzic and A. Wernicki

Abstract

In view of the significant role of Hsp70 in protecting the organism against the destructive effects of stress, and the possibility of using this protein as a marker of the infarction process in the heart, the aim of this study was to conduct an evaluation of the expression of 70kDa heat shock proteins (Hsp70) and the concentration of TBARS (thiobarbituric acid reactive substances) and nitric oxide ions (NO), determined as nitrite ions, as markers of oxidative stress in hearts obtained from healthy pigs following slaughter and pigs which had died during or immediately after transport with symptoms of sudden cardiac death.

The material consisted of hearts obtained from 90 pigs following slaughter and from pigs which had died. Oxidative stress was determined in heart lysates based on the concentration of TBARS and nitrite ions. Expression and concentration of Hsp70 were determined using SDS-PAGE, Western blotting, and ELISA.

Expression of Hsp70 was observed in hearts lysates obtained from slaughtered pigs and from those which had died with symptoms of sudden death. The strongest reaction in the Western Blotting was noted in hearts lysates from pigs with no pathological changes. The highest TBARS concentration was observed in lysates from hearts in pigs which had died during or immediately after transport. The highest concentration of NO ions, determined as nitrite ions, was noted in hearts from pigs with myocardial infarction lesions. The significant decrease observed in Hsp70 concentration in heart tissue obtained from the pigs which had died in comparison to the hearts from healthy pigs indicates the important role of this protein in protecting the heart muscle against the destructive effects of stress, which limits the occurrence of post-stress cardiomyopathy in pigs following transport.

Open access

Andrzej Wernicki, Renata Urban-Chmiel, Andrzej Puchalski and Marta Dec

Abstract

Immune and oxidative parameters were evaluated as indicators of the influence of stress on the occurrence of respiratory syndrome in feedlot calves. The study was carried out on 60 2 and a half-month-old Simmentaler calves transported at feedlot. Mean daily feed consumption, daily weight gain, and behavioural activity were evaluated. Blood was collected on days 1, 3, 7, 14, 21, and 28 of feedlot. Serum NO ions, lipid peroxidation, acute phase proteins, IgG, and IgM were determined. The calves showed a decrease in feed consumption during the first 7 d of the feedlot. Sporadic stereotypies were noted. NO concentration showed a significant (P < 0.05) increase on days 7 and 14. Concentration of thiobarbituric acid reactive substances (TBARS) increased (P < 0.05) on days 1 and 3. IgM concentration was found to be considerably lower on days 14 and 28. Serum haptoglobin level showed a significant increase in stressed calves on days 1, 3, 7, and 14 of the feedlot. Based on these results, it can be suggested that stressors associated with transport and adaptation to the feedlot induce a stress reaction in calves, resulting in behavioural disorders, reduced weight gain, suppression of the humoral immunity and increased morbidity during the first weeks. These parameters seem to be crucial in evaluating the animals' health and welfare.

Open access

A. Puchalski, R. Urban-Chmiel, M. Dec and A. Wernicki

Abstract

Iron-regulated outer membrane proteins (IROMPs) in Mannheimia haemolytica A1, which function as a receptor for complexes containing iron ions, are induced by iron deficiency in the growth environment of the bacteria. Densitometric analysis of SDS-PAGE separation showed expression of IROMPs of 71, 77, and 100 kDa in the case of bacteria grown in a medium with 2,2-dipyridyl. The electrophoregrams obtained in 2-DE separations confirmed the presence of protein fractions with these molecular weights and isoelectric points ranging from 5.4 to 6.4. The results of the study also confirmed the ability of M. haemolytica A1 proteins involved in iron uptake to induce a protective immune response. In Western blot with serum from convalescent calves naturally infected with M. haemolytica A1, distinct reactions were obtained for IROMPs of 71, 77, and 100 kDa.

Open access

Urszula Kosikowska, Dagmara Stepien-Pysniak, Dorota Ozga, Andrzej Wernicki and Anna Malm

ABSTRACT

Bacillus spp. can be regarded as a rare component of the nasal mucosa microflora. The aim of this study was to identify Bacillus spp. from the nasal mucosa of healthy adults living in the suburban area near Lublin using the matrix-assisted laser desorptionionization time-of-flight mass spectrometry (MALDI-TOF MS) system.

A total of 11 bacterial isolates from the nasal specimens were cultured. The following species were identified using the routine microbiological methods: Staphylococcus aureus (3 isolates), S. epidermidis (1 isolate), S. intermedius (1 isolate) and Staphylococcus spp. (1 isolate). Moreover, 2 strains of Escherichia coli were isolated. Besides, 3 isolates of Bacillus spp. were found. These isolates were characterized by means of MALDI-TOF MS, resulting in highly specific mass spectral fingerprints and these were identified as B. pumilus, B. safenis and B. licheniformis. It was observed that all studied Bacillus spp. isolates only had the masses in common at 3864 ± 2, 7727 ± 2, and 14301 ± 4. The spectra of B. safensis and B. pumilus showed peaks at m/z 4914 ± 3, 6621 ± 3 and 14291 ± 2, which were absent in the spectrum of B. licheniformis. For B. safenis and B. pumilus, other potential biomarkers could be found at m/z 12620 and 16668, respectively.

Open access

Renata Urban-Chmiel, Rafał Stachura, Piotr Hola, Andrzej Puchalski, Marta Dec and Andrzej Wernicki

Abstract

The aim of the study was to evaluate the effect of flunixin and florfenicol administered in combination with vitamin E or C on selected leukocyte immune mechanisms and on the inflammatory process during the first few weeks in the feedlot. Fifty calves divided into 5 groups (n = 10) received florfenicol and flunixin with vitamin E or C. Blood was collected on the 1st, 3rd, 7th, 14th, 21st, and 28th d of the experiment. Intracellular metabolism (NBT), apoptosis, chemotaxis, susceptibility to M. haemolytica leukotoxin, and expression of β2-integrins were determined in leukocytes. The symptoms of respiratory tract infection were observed in 40% of calves in control group, while in the other groups the morbidity rate ranged from 10% to 20%. Leukocytes showed decreased NBT, and the mean values for apoptosis ranged from 14% to 24%. The lowest percentage of apoptotic cells was observed in the calves that received florfenicol with flunixin and vitamins E and C. The chemotactic activity confirmed the significant inhibitory effect of the preparations on migration of the cells. A significant decrease (P ≤ 0.05) in the susceptibility of leukocytes to leukotoxin was noted in the group that received florfenicol and flunixin with vitamin E. Expression of β2-integrin receptors was the lowest in calves receiving florfenicol with flunixin and vitamin E or C. The application of an antibiotic and a non-steroidal anti-inflammatory drug with antioxidants protected the leukocytes involved in defence against M. haemolytica virulence factors and effectively limited oxidative stress in the calves.

Open access

Andrzej Wernicki, Renata Urban-Chmiel, Jerzy Rola, Wojciech Socha, Diana Stęgierska, Marta Dec and Andrzej Puchalski

Abstract

The study was performed on nasal swabs, tracheal samples, and sera obtained from young beef heifers aged between 6 and 12 months, from farms in eastern and south-eastern Poland. The samples were evaluated using bovine herpesvirus 1 (BHV-1) ELISA kits (ELISA BHV1 antibody and ELISA BHV1 antigen) and PCR. Among all the animals examined, 37 (32.2%) were positive in the ELISA BHV1 antigen test. The presence of BHV-1 was confirmed by PCR in 42 (36.5%) animals. In the ELISA BHV1 antibody test, 39 (33.9%) seropositive animals were identified. The presence of BHV-1 positive samples was observed in all the examined breeds of young cattle. There were no significant differences (P ≤ 0.05) in BHV-1 positive samples. The results indicate that the incidence of BHV-1 infections in feedlot cattle herds studied was 32.2%-36.5%, which suggests that preventive measures should be implemented in order to limit transmission of the virus.