B and T lymphocytes in rabbits change according to the sex and throughout the year.

The European rabbit (Oryctolagus cuniculus) is a good model in biomedicine used in research on several human diseases. The reference values of B and T cells and their subpopu- lations are very important to understand how the adaptive immune system is responding to infectious agents. The aim of this study was to determine values of B and T cells and their subpopulations in Polish mixed-breed rabbits, considering seasons of the year and sex. The study was performed on 200 Polish mixed-breed rabbits and the percentage of B and T lymphocytes was measured cytometrically using mouse anti-rabbit antibodies. The study revealed that the season of the year and sex of the animals affected the percentage of B- and T-cells and their subpopulations in peripheral blood. Statistically significant values of CD19+ B-cells in spring and autumn, of T CD5+ cells in spring and winter, of T CD4+ in spring, summer, autumn and winter, of T CD8+ in winter and of T CD25+ in spring were noted. Generally the highest values were found mainly in warm part of the year, while the lowest in colder months. A statistical significance was also observed between males and females - changes were found in T CD4+ and T CD25+ lymphocytes in spring, T CD8+ cells in winter and higher percentage was generally obtained in females than in males. The only exception was the T CD5+ subpopulation in which no differences were observed between the sexes and throughout the year. This is the first paper on adaptive immune system cell values in the European rabbit of domestic breeds.

The first detection of the sequence of bacteria from the Simkaniaceae family in surface waters in Poland.

One of the most common reasons for horse lameness is subchondral bone cysts (SBCs), which are especially evident in young horse athletes. It is believed that SBC development is strongly associated with an individual's bone growth and/or bone microstructure impairment. Current methods of SBC treatment include pharmacological treatment or surgical procedures which may allow the bone within the cyst to rebuild and be restored to properly developed bone tissue. Thus, we propose filling the SBCs with a 3D complex of alginate hydrogel and autologous adipose derived mesenchymal stem cells (ASCs). We have observed at the in vitro level, that this hydrogel complex induces osteogenic and chondrogenic differentiation potential through the upregulation of bone morphogenetic protein, osteopontin, collagen type I and aggrecan mRNA levels. Moreover, we detected the creation of a 3D extracellular matrix (EM). To investigate the complex in vivo, we chose 8 horses of varying age suffering from SBC, which resulted in lameness, to undergo experimental surgery. We documented the horses' clinical appearance, lameness and radiographic appearance, to determine that there was clinical improvement in 87.75% of the patients (n=7, out of 8 horses) 6 months postoperatively and 100% (n=8, out of 8 horses) a year after surgery. These results are promising for the potential of this procedure to become the standard in SBC treatment.

Values of apoptosis of lymphocytes and granulocytes in peripheral blood of Polish mixed-breed rabbits in the annual cycle.

The objective of the study was to determine values of apoptosis for peripheral blood lymphocytes and granulocytes, including dependency on the season of the year, in Polish mixed-breed rabbits and in mixed-breed rabbits with the addition of blood of meat-breed rabbits. The study was carried out in four seasons of the year (spring, summer, autumn, winter) involving 120 Polish mixed-breed rabbits - group I, and 120 Polish mixed-breed rabbits with addition of meat-breed rabbit blood - group II. Assessment of apoptosis of lymphocytes and granulocytes was performed using as FACScan cytometer by Becton Dickinson with FACSDiva software (USA), using as ApoFluor® Green Caspase reagent kit (MP Biomedicals, USA) to detect the activity of the total caspase pool 1, 3, 4, 5, 6, 7, 8, 9 in granulocytes and lymphocytes of rabbit peripheral blood. The results for apoptosis of lymphocytes and granulocytes in peripheral blood in the animals investigated (group I and II) were subjected to statistical analysis with the t'Student test at p=0.05. It was noted that, in rabbits from group I, the values for apoptosis of lymphocytes were the highest in winter and autumn (36.02% and 31.24%, respectively), and the lowest in spring and summer (26.73% and 22.72%, respectively), whereas in the case of granulocytes the highest values were in summer and spring (14.69% and 12.95%, respectively), and the lowest in winter and autumn (8.16% and 8.57%, respectively). In mixed-breed rabbits with the addition of meat-breed blood (group II), the values for apoptosis of lympocytes were the highest in spring (29.13%), and the lowest in summer (25.43%); whereas in the case of granulocytes the highest values were in summer and spring (14.0% and 11.15%, respectively), and the lowest in autumn and winter (7.46% and 7.64%, respectively).

White and red blood cell picture in rabbits experimentally infected with strains of the rabbit haemorrhagic disease (RHD) virus without or with variable haemagglutination capacity.

The aim of the study was to establish if haemagglutination of rabbit haemorrhagic disease virus (RHDV) affects haematological picture of peripheral blood in rabbits and the pathogenicity of the virus. The study analyzed white and red blood cell picture in rabbits experimentally infected with two non-haemagglutinating (HA-) RHDV strains (Frankfurt and Asturias) and one strain with variable haemagglutination capacity (HA+/-) (Hagenow). Studies with HA- and HA +/- are rare and relate only to 4 HA- strains (2 RHDV: BLA and Rainham; 2 RHDVa: Pv97 and 9905) and 1 HA+/- RHDV strain: ZD, where less changes in haematological indices and less pathogenicity were observed. We found that changes caused by HA- Frankfurt strain were related to the number of neutrophils and thrombocytes, while in HA- strain Asturias, in thrombocytes and leukocytes. Changes evoked by HA+/- Hagenow strain pertained to the number of eosinophils, thrombocytes, leukocytes, monocytes, and concentration of hemoglobin. Mortality caused by the Frankfurt strain was 100% between 36 and 48 h post infection (p.i.), while that caused by Asturias strain was 100% between 24 and 36 h p.i., and that observed in case of Hagenow strain was 90% between 36 and 48 h p.i. The changes in haematological picture caused by the HA- and HA+/- RHDV strains were less intensive than those found in case of the HA+ RHDV strains, which cannot be confirmed for pathogenicity, and is not in line with the existing hypothesis suggesting higher pathogenicity in HA+ viruses.

Natural immunity factors in Polish mixed breed rabbits.

Mixed-breed rabbits in Poland are widely used for diagnostic and scientific research and as utility animals, therefore there is a need to know their immunological status, as well as their haematological status. In this study natural immunity factors were analyzed in Polish mixed-breed rabbits and Polish mixed-breed rabbits with addition of blood of meet-breed, considering the impact of sex and season of the year (spring, summer, autumn, winter) using measurement of non-specific cellular and humoral immunity parameters in peripheral blood. The study has revealed that there is a variety between the two commonly used mixed-breed types of rabbits, especially when sex and season is concerned, which is crucial for using these animals in experiments.

Hematological parameters in Polish mixed breed rabbits with addition of meat breed blood in the annual cycle.

In the paper we studied haematologic values, such as haemoglobin concentration, haematocrit value, thrombocytes, leucocytes: lymphocytes, neutrophils, basophils, eosinophils and monocytes in the pheral blood in Polish mixed-breeds with addition of meat breed blood in order to obtain the reference values which are until now not available for this animals. In studying this indices we took into consideration the impact of the season (spring, summer, autumn, winter), and sex of the animals. The studies have shown a high impact of the season of the year on those rabbits, but only in spring and summer. Moreover we observed that the sex has mean impact on the studied values of haematological parameters in those rabbits. According to our knowledge, this is the first paper on haematologic values in this widely used group of rabbits, so they may serve as reference values.

B-cell and T-cell values in peripheral blood in Polish mixed-breed rabbits with addition of blood of meet breeds.

In Poland, rabbit is a highly valued animal, due to dietetic and flavour values of its meat, but above all, rabbits tend to be commonly used laboratory animals. The aim of the study was developing standards for counts of B-cells with CD19+ receptor, T-cells with CD5+ receptor, and their subpopulations, namely T-cells with CD4+, CD8+ and CD25+ receptor in the peripheral blood of mixed-breed Polish rabbits with addition of blood of meet breeds, including the assessment of the impact of four seasons of the year and animal sex on the values of the immunological parameters determined. The results showed that the counts of B- and T-cells and their subpopulations in peripheral blood remain within the following ranges: for CD19+ B-cells: 1.05 - 3.05%, for CD5+ T-cells: 34.00 - 43.07%, CD4+ T-cells: 23.52 - 33.23%, CD8+ T-cells: 12.55 - 17.30%, whereas for CD25+ T-cells: 0.72 - 2.81%. As it comes to the season of the year, it was observed that it principally affects the values of CD25+ T-cells, while in the case of rabbit sex, more changes were found in females.

Values of white and red blood cell parameters in Polish mixed breed rabbits in the annual cycle.

The aim of the study was to develop reference values for such haematologic values as haemoglobin concentration, haematocrit value, volume of erythrocytes, thrombocytes, leucocytes, lymphocytes, neutrophils, basophils, eosinophils and monocytes in peripheral blood in Polish mixed-breed rabbits, considering the impact of the season (spring, summer, autumn, winter), and sex of the animals. The results reveal, that the season of the year has a small impact in Polish mixed-breed rabbits, and it principally referred to spring and summer. Finally it was shown, that sex has average impact on the studied values of haematological parameters.

Lymphocytes T and B in rabbits infected with RHD virus.

The aim of this study was to determine the differences in immunological response of animals infected with different antigenic variants of the virus--three haemagglutinating (Vt97, Triptis, Hartmannsdorf) and two non-haemagglutinating (Pv97, 9905 RHDVa). The specific immunological response was measured by the dynamics of changes in the amount of lymphocytes T (with CD5+, CD4+, CD8+, CD25+ receptor) and B (with CD19+ receptor). The study showed differences in immunogenicity of the analysed RHDV antigenic variants, which allowed them to be divided into groups of: more immunogenic strains, including non-haemagglutinating 9905 RHDVa and haemagglutinating Vt97 and Triptis variants; and less immunogenic strains, including the haemagglutinating Hartmannsdorf variant and the non-haemagglutinating Pv97 variant. Such a result may indicate that the agglutination capacity of red blood cells might not be a factor impacting the number of T and B lymphocytes.

Apoptosis of granulocytes and lymphocytes in peripheral blood in rabbits infected with haemagglutinating and non-haemagglutinating antigenic variants of the RHD (rabbit haemorrhagic disease) virus.

This paper attempts to study the dynamics of apoptosis of granulocytes and lymphocytes in peripheral blood in rabbits infected with haemagglutinating (Vt97, Triptis, Hartmannsdorf) and non-haemagglutinating (Pv97, 9905 RHDVa) antigenic variants of the RHD virus. The pathogenicity of those antigenic variants was also assessed by recording the mortality of the infected animals. The animals were infected with antigenic variants and blood was sampled at hour 0, 4, 8, 12, 24, 36 p.i. and the percentage of apoptotic granulocytes and lymphocytes was measured with the use of flow cytometry. The results of the study showed that apoptosis is included during RHDV infection, as the number of apoptotic granulocytes and lymphocytes increases throughout the experiment; depending on the antigenic variant, apoptosis joins in at 4-8-12 h p.i. and lasts until 24-36 h p.i. Furthermore, the mortality of rabbits infected with the examined strains of RHD virus varied from 30% to 100%. This study performed for the first time in this manner, indicates the importance of apoptosis during infection with the RHD virus.

Platelets--an important element of the immune system.

Platelets are anucleate cells derived from the megakaryocyte series, and have long been considered only as cells responsible for coagulation and the fibrinolysis process. However, recently more data shows that they are also effector cells in the inflammatory response and important elements of the immunological response. Platelets store and release many biologically active substances, including growth factors, cytokines and chemokines (tab. 1), which actively affect i.a. elements of the immune system, and thus become regulators of immunity and mediators of inflammatory response. Their impact on the immune system cells is also associated with the induction of leucocytes and progenitor cells to the site of pathogen permeation or vascular injury inflow, as well as endothelial cells. Interacting with neutrophils, monocytes and lymphocytes, they not only activate them, but also form platelet-leukocyte aggregates that immobilise pathogens and prevent their spreading. Furthermore, platelets are capable of absorbing pathogens, affecting anti-infection immunity of the system. It is also assumed that the presence of receptors on their surface, such as Toll-like receptors (TLRs), affects their initiation and activity of the immunological response.

Real time PCR detection of rabbit haemorrhagic disease virus in rabbits infected with different European strains of RHDV.

The paper concerns the use of a novel, very effective diagnostic method, a real-time PCR for diagnosis of a viral agent causing viral haemorrhagic disease in rabbits - RHDV. Until now, the method was widely used for detecting many different viruses, both DNA, and RNA, but as far as RHDV is concerned, there are not many records of such use. This study aimed at the detection of 17 different strains from different European regions, differing in biological features and mortality. The study confirmed that real-time PCR is an applicable and effective method for diagnosis of RHDV, irrespective of the stains' features.

Autophagy in physiological and pathological processes--selected aspects.

This paper describes a model of cell death, called autophagy, one among other typical and atypical processes of cell death. This phenomenon is present in the organism, from conception until death, and is conditioned by many genes of ATG family, or mTOR kinase and specific proteins, like BNIP3. This process plays a very important role not only in physiological functions of the organism but also in pathological, such as Alzheimer or Huntington disease, as well as diseases caused by viruses.

Lymphocyte subpopulations and apoptosis of immune cells in rabbits experimentally infected with a strain of the RHD virus having a variable haemagglutination capacity.

The paper describes the immunological response in the matter of percentage of T cells (receptor CD5+) and subpopulations (Th with receptor CD4+, Tc/Ts with receptor CD8+, T with receptor CD25+) and B cells with receptor CD19+, as well as the percentage of apoptotic granulocytes and lymphocytes, in rabbits experimentally infected with the Hagenow strain of the RHD virus. The material chosen for the experiment is special, as among all strains of RHD virus, there are only two strains which carry the variable haemagglutination capacity of red cells. The results of the study show that the Hagenow strain gives an untypical picture of T and B lymphocytes, whereas the results in inducing apoptosis seems to corespond with previous data, confirming the inclusion of apoptosis from 4 h p.i. and the intensity of the phenomenon being higher in granulocytes.

Phagocytosis of neutrophils in rabbits infected with antigenic variants of RHD (rabbit haemorrhagic disease) virus.

The present study was aimed at determining changes in chosen elements of phagocytosis in rabbits infected with 3 antigenic variants of RHD - Hartmannsdorf, Pv97 and 9905, which differed in haemagglutination ability. The animals were tested for phagocytosis parameters, and the results revealed that the examined strains showed the differences. These variations regarded mainly Pv97 strain, as the intensity of the changes were 5 times stronger in comparison to strain Hartmannsdorf and 9905. As all of the strains examined are signified as antigenic variants, we have stated that this feature does not determine their immunological picture. The results suggest the existence of immunological dissimilarities among strains of the RHD virus, which was revealed for the first time in antigenic variants.

White and red blood cells picture in rabbits experimentally infected with RHD virus.

Four strains of RHDV assigned as haemagglutinating (Vt97 and Hartmannsdorf) and non-haemagglutinating (Pv97 and 9905) antigenic variants were examined for dynamic changes in the values of white and red blood cells indexes. The study showed differences among strains examined that were not depending on haemagglutination property.

Cytometric analysis of lymphocytes T and B in rabbits infected with non-haemagglutinogenic strains of RHD virus (rabbit haemorrhagic disease)--Rainham, Frankfurt and Asturias.

The present paper refers to the cytometric analysis of lymphocytes T (with receptor CD5+), Th (with receptor CD4+), Tc/Ts (with receptor CD8+), lymphocytes CD25+ and lymphocytes B with receptor CD19+ in rabbits experimentally infected with strains of RHD virus--Rainham, Frankfurt and Asturias, not having haemagglutinogenic capacities, which makes them unique, as haemagglutinogenic capacity is a classic and typical property of most strains of this virus. The study was performed in the dynamic system, drawing blood samples from animals at hour 0, namely before the administration of the viral antigen, and then at 4, 8, 12, 24 and 36 h after the infection. The study indicated that Rainham and Asturias strains of RHD virus cause a similar amount of changes as the most immunogenic haemagglutinogenic strains CAMP V-561 and CAMP V-562 of the RHD virus do. In contrast, the Frankfurt strain of the RHD virus is characterised with 5-6-fold lower reactivity in this respect and is most similar to the least immunogenic haemagglutinogenic strain CAMP V-558 of the RHD virus.

Immune response in farm animals infected or immunised with bacteria of Chlamydia sp. and Chlamydophila sp. genus.

Bacteria from genera Chlamydia (Ch.) and Chlamydophila (Chl.) are very pathogenic and may infect humans and animals. They also may cause latent infection, especially in animals. In this paper we discuss the non-specific and specific cellular and humoral immunity in farm animals, after infection or immunisation with Chlamydia sp. and Chlamydophila sp. bacteria. It has been shown, that the infection or immunisation with the microorganisms influenced the activity of polimorphonuclear cells (PMN) and mononuclear cells (MN) in the process of phagocytosis. It has also been shown that the bacteria influenced the amount and activity of lysozyme, activities of myeloperoxidase and lysosomal enzymes. Infection or immunisation with the microorganisms was demonstrated to affect numbers of lymphocytes T and B and those of their subpopulation as well as the activity of cytokines and levels of serum and secreted immunoglobulins. The changes were detected just a few hours after infection or immunisation and persisted for a few days to a few decades.

Values of selected immune and haematological parameters in healthy rabbits.

In the paper, values of selected indices of non-specific immunity as well as white and redblood cell patterns were presented in healthy rabbits at the age of 3 to 4,5 months. The purpose of the study was to provide reference values in healthy rabbits being frequent research model in the studies of several diseases in humans and animals.

Selected indices of non-specific humoral immunity in rabbits experimentally infected with non-hemagglutinogenic strain of VHD (viral hemorrhagic disease) virus.

This study aimed at an evaluation of selected indices of non-specific humoral immunity, myeloperoxidase and lysozyme, in rabbits following administration of a non-hemagglutinogenic strain of VHD virus. The results obtained indicate participation of the mechanisms in response to the administered antigen.