This study investigated the effect of sex-sorting and cryopreservation on post-thaw characteristics and fertility of red deer (Cervus elaphus) sperm for the first time. Semen was collected by electroejaculation from 10 mature stags during the breeding season, and each ejaculate split into four experimental groups: Bulk sorted spermatozoa, sorted but not sexed (BSS); sorted high purity X-spermatozoa (XSS); sorted high purity Y-spermatozoa (YSS); and, control non-sorted spermatozoa (NS). Following, all samples were frozen over liquid nitrogen. Two straws per stag and sample type were analyzed immediately post-thaw and following a 2-h incubation period at 37 °C. Post-thaw total motility (TM) as assessed by CASA was not different (P < 0.05) among NS, BSS and YSS sperm. For XSS, post-thaw TM was lower (39%, P < 0.05) than that for NS (54%) or BSS (50%), but similar (P > 0.05) to that of YSS (47%) sperm. The percentage of apoptotic spermatozoa as assessed by PI/YO-PRO-1 and flow cytometry analysis, was higher (17%, P ≤ 0.05) for XSS sperm than NS (12%), BSS (13%) and YSS (14%) sperm. Following incubation there were no differences (P > 0.05) in TM or percent apoptosis among treatments. Post-thaw chromatin stability calculated as the DNA fragmentation index (%DFI) was similar among treatments; following incubation %DFI increased in all except YSS, which displayed the lowest value (P < 0.05). Artificial insemination of synchronized hinds yielded 44, 52 and 62% delivery rates for YSS, NS and standard frozen-thawed sperm, respectively (P < 0.05). Notably, 93 and 55% of fawns born were males for the YSS and NS spermatozoa, respectively (P < 0.05). In summary, Y-sorted sperm displayed acceptable post-thaw sperm evaluation parameters and the expected offspring sex ratio. More studies are needed to understand the source of sperm damage that may compromise the fertility of Y-sorted red deer sperm.

• MeSH
• fragmentace DNA MeSH
• kryoprezervace metody   veterinární   MeSH
• motilita spermií MeSH
• poměr pohlaví MeSH
• předvýběr pohlaví veterinární   MeSH
• průtoková cytometrie veterinární   MeSH
• spermie fyziologie   MeSH
• uchování spermatu metody   veterinární   MeSH
• umělá inseminace veterinární   MeSH
• vysoká zvěř * MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

In this study we compared the proteomes of macrophages and heterophils isolated from the spleen 4 days after intravenous infection of chickens with Salmonella Enteritidis. Heterophils were characterized by expression of MMP9, MRP126, LECT2, CATHL1, CATHL2, CATHL3, LYG2, LYZ and RSFR. Macrophages specifically expressed receptor proteins, e.g. MRC1L, LRP1, LGALS1, LRPAP1 and a DMBT1L. Following infection, heterophils decreased ALB and FN1, and released MMP9 to enable their translocation to the site of infection. In addition, the endoplasmic reticulum proteins increased in heterophils which resulted in the release of granular proteins. Since transcription of genes encoding granular proteins did not decrease, these genes remained continuously transcribed and translated even after initial degranulation. Macrophages increased amounts of fatty acid elongation pathway proteins, lysosomal and phagosomal proteins. Macrophages were less responsive to acute infection than heterophils and an increase in proteins like CATHL1, CATHL2, RSFR, LECT2 and GAL1 in the absence of any change in their expression at RNA level could even be explained by capturing these proteins from the external environment into which these could have been released by heterophils.

• MeSH
• kur domácí metabolismus   mikrobiologie   MeSH
• kvantitativní polymerázová řetězová reakce veterinární   MeSH
• makrofágy metabolismus   MeSH
• nemoci drůbeže metabolismus   mikrobiologie   MeSH
• proteom MeSH
• protilátky heterofilní metabolismus   MeSH
• průtoková cytometrie veterinární   MeSH
• regulace genové exprese MeSH
• Salmonella enteritidis * MeSH
• salmonelová infekce u zvířat metabolismus   mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

While haematological variation is well known in birds, variation in avian breeds (distinct morphotypes of the same species) remains unexplored. Poultry breeds, in particular, may show interesting evolutionary patterns and economically-relevant physiological differences. We performed a comparative examination of blood cellular composition in five chicken (Gallus gallus domesticus) breeds: Araucana, Booted bantam, Czech, Minorca and Rosecomb bantam. In standard-environment-reared hens whole-blood flow cytometry revealed remarkable differences in most erythrocyte- and leukocyte-related parameters. We identified two extremes: Czech, a European breed, with a low heterophil/lymphocyte (H/L) ratio and high CD4+ levels, and Araucana, a South-American breed, with a high H/L ratio and high relative monocyte count. Such variation may reflect a combination of artificial and natural selection acting on health- and stress-related traits in domestic populations. Different breeds have evolved different immunological adaptations reflecting their original need to fight pathogens and physiological constraint resulting from dissimilar physiological trade-offs.

• MeSH
• CD4-pozitivní T-lymfocyty MeSH
• druhová specificita MeSH
• kur domácí krev   imunologie   MeSH
• počet leukocytů veterinární   MeSH
• počet lymfocytů veterinární   MeSH
• protilátky imunologie   MeSH
• průtoková cytometrie veterinární   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

In pigs, the epitheliochorial placenta does not allow transfer of maternally derived antibodies or immune cells to the fetus. Thus, piglets are dependent on intake of colostrum for acquisition of passive immunity during the neonatal period. As well as immunoglobulin G (IgG), cellular components of colostrum, mainly lymphocytes, can enter the systemic circulation and secondary lymphoid organs of the neonate. In order to understand the function and immunological role of these cells, a flow cytometric study was undertaken to characterise the cellular profile and phenotype of T cells and NK cells present in porcine colostrum. The results indicated that the greatest numbers of lymphocytes were found on the first day of lactation. The predominant cell types in colostrum were CD8(+) single positive T cells (53.6%), followed by CD4(+)CD8(+) double positive T cells (21.1%), CD2(+)CD8(+) γδ T cells (15.0%) and NK cells (13.5%). CD4(+) single positive T cells (4.4%) and other γδ T cell subpopulations (1.8% CD2(-)CD8(-) and 0.4% CD2(+)CD8(-)) were present in colostrum at low levels. Although the profile of the T cell subpopulations during the first 3 days of lactation remained constant, the absolute numbers of T and NK cells decreased significantly in the first few hours of lactation. Expression of CCR7, CD11b, CD25, CD45RA and MHC class II was used to assess the activation status of T and NK cells in colostrum. T cell subpopulations expressed markers consistent with an effector memory phenotype, indicating that these were antigen-experienced cells. The phenotype of colostral T and NK cells suggests a role in mucosal immunity and potentially in transfer of passive immunity from sow to piglet.

• MeSH
• antigeny imunologie   MeSH
• buňky NK imunologie   MeSH
• fenotyp MeSH
• imunita získaná od matky * MeSH
• kolostrum imunologie   MeSH
• poporodní období imunologie   MeSH
• průtoková cytometrie veterinární   MeSH
• Sus scrofa imunologie   MeSH
• T-lymfocyty imunologie   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Monocytes play an essential role in the defense against bacterial pathogens. Bone marrow (BM) and peripheral blood (PB) monocytes in pigs consist of the main "steady-state" subpopulations: CD14 hi/CD163-/SLA-DR- and CD14 low/CD163+/SLA-DR+. During inflammation, the subpopulation of "inflammatory" monocytes expressing very high levels of CD163, but lacking the SLA-DR molecule (being CD14 low/CD163+/SLA-DR-) appears in the BM and PB and replaces the CD14 low/CD163+/SLA-DR+ subpopulation. However, current knowledge of monocyte migration into inflamed tissues in pigs is limited. The aim of the present study was to evaluate the distribution of "inflammatory" CD14 low/CD163+/SLA-DR- monocytes during experimental inflammation induced by Actinobacillus pleuropneumoniae (APP) and a possible role for chemokines in attracting "inflammatory" CD14 low/CD163+/SLA-DR- monocytes into the tissues. Monocyte subpopulations were detected by flow cytometry. Chemokines and chemokine receptors were detected by RT-qPCR. The "steady-state" monocytes were found in the BM, PB, spleen and lungs of control pigs. After APP-infection, "inflammatory" monocytes replaced the "steady-state" subpopulation in BM, PB, spleen and moreover, they appeared in an unaffected area, demarcation zone and necrotic area of the lungs and in tracheobronchial lymph nodes. They did not appear in mesenteric lymph nodes. Levels of mRNA for various chemokines with their appropriate receptors were found to be elevated in BM (CCL3-CCR1/CCR5, CCL8-CCR2/CCR5, CCL19-CCR7), necrotic area of the lungs (CCL3-CCR1, CCL5-CCR1/CCR3, CCL11-CCR3, CCL22/CCR4) and tracheobronchial lymph nodes (CCL3-CCR1) and therefore they could play a role in attracting monocytes into inflamed tissues. In conclusion, "inflammatory" monocytes appear in different lymphoid tissues and the lungs after APP infection in pigs. Various chemokines could drive this process.

• MeSH
• Actinobacillus pleuropneumoniae fyziologie   MeSH
• antigeny diferenciační myelomonocytární metabolismus   MeSH
• CD antigeny metabolismus   MeSH
• chemokiny genetika   metabolismus   MeSH
• infekce bakteriemi rodu Actinobacillus imunologie   mikrobiologie   veterinární   MeSH
• lymfoidní tkáň metabolismus   MeSH
• messenger RNA genetika   MeSH
• monocyty cytologie   metabolismus   MeSH
• nemoci prasat imunologie   mikrobiologie   MeSH
• plíce metabolismus   MeSH
• prasata MeSH
• průtoková cytometrie veterinární   MeSH
• receptory buněčného povrchu metabolismus   MeSH
• receptory chemokinů genetika   metabolismus   MeSH
• zánět mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The present study describes the distinct bone marrow (BM) and peripheral blood (PB) monocyte subpopulations detected by seven-colour flow cytometry. Mononuclear phagocytes were identified as viable CD172a(+) SWC8(-) CD203a(-) mononuclear leukocytes. After that, monocyte subpopulations were differentiated by using CD14, CD163 and SLA-DR markers. Four distinct monocyte subpopulations were found in the BM and PB. Based on the discovered populations two possible maturation pathways have been proposed. The first pathway was characterised by release of CD14(hi) CD163(-) SLA-DR(-) BM monocytes into the PB where they matured into CD14(low) CD163(+) SLA-DR(+) monocytes. In the alternative pathway the monocytes finalised their phenotypical maturation in the BM and then they were released into the PB as CD14(low) CD163(+) SLA-DR(+) cells. In Salmonella-infected piglets, the population of CD14(low) CD163(+) SLA-DR(+) monocytes was elevated in the BM and mesenteric lymph nodes (MLN), suggesting the role of this population in pathogenesis of Salmonella infection in pigs.

• MeSH
• buňky kostní dřeně klasifikace   cytologie   MeSH
• membránové proteiny MeSH
• monocyty klasifikace   cytologie   MeSH
• nemoci prasat krev   mikrobiologie   MeSH
• prasata MeSH
• průtoková cytometrie metody   veterinární   MeSH
• Salmonella enteritidis MeSH
• salmonelová infekce u zvířat krev   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Adenosine is a well described anti-inflammatory modulator of immune responses. The aim of the present study was to describe the role of common adenosine agonist 5'-N-ethylcarboxamidoadenosine (NECA) in cytokine production by main porcine T cell subpopulations. TNF-α, IFN-γ, IL-2 and IL-10 were detected by multicolor flow cytometry together with cell surface markers CD3, CD4 and CD8. It was found that NECA inhibits (in a dose-dependent manner) production of pro-inflammatory TNF-α and Th1-associated cytokines IFN-γ, IL-2 in all concanavalin A-stimulated T cell subpopulations. Moreover, production of IL-10 was potentiated in all T cell subpopulations tested. These corresponded well with the fact that all T cell subsets expressed mRNA for adenosine receptor (AR) subtypes to comparable extents. Contrary to concanavalin A-stimulated cells, NECA had a moderate effect on PMA-stimulated T cells, suggesting that AR in pigs acts via signaling pathways not associated with protein-kinase C. Non-selective antagonist CGS15943 as well as allosteric modulator SCH202676 failed to reverse the effect of NECA in pigs. In conclusion, NECA has an anti-inflammatory effect on porcine T cell subpopulations.

• MeSH
• adenosin-5'-(N-ethylkarboxamid) farmakologie   MeSH
• adenosin agonisté   fyziologie   MeSH
• antigeny CD3 fyziologie   MeSH
• CD4-pozitivní T-lymfocyty účinky léků   fyziologie   MeSH
• CD8-pozitivní T-lymfocyty účinky léků   fyziologie   MeSH
• chinazoliny farmakologie   MeSH
• interferon gama biosyntéza   fyziologie   MeSH
• interleukin-10 biosyntéza   fyziologie   MeSH
• interleukin-2 biosyntéza   fyziologie   MeSH
• konkanavalin A farmakologie   MeSH
• prasata imunologie   MeSH
• průtoková cytometrie veterinární   MeSH
• T-lymfocyty - podskupiny účinky léků   fyziologie   MeSH
• T-lymfocyty účinky léků   fyziologie   MeSH
• TNF-alfa biosyntéza   fyziologie   MeSH
• triazoly farmakologie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zánět patofyziologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Very little is known about the occurrence of immune system cells in the canine uterus. The aim of this study was to generate information about lymphocyte subsets that are present in the healthy canine uterus and that are recruited under inflammatory conditions caused by pyometra. Using immunohistochemistry and flow cytometry, a significant influx of γδ T lymphocytes was found in pyometra samples mainly due to recruitment of γδ(+)/CD8(-) T lymphocytes. The relative expression of genes encoding selected cytokines/chemokines was evaluated in samples from healthy and pyometra-affected uteri. Expression of pro-inflammatory cytokines (including IL-1β, TNF-α, IL-8, IL-17 and IFN-γ) and chemokines (including CXCL10, CCL4 and CCL5) was upregulated in pyometra samples confirming the presence of inflammation. In contrast, the expression of the homeostatic chemokine CCL25 and of the anti-inflammatory cytokine IL-10 was downregulated and unchanged, respectively.

• MeSH
• chemokiny genetika   imunologie   metabolismus   MeSH
• cytokiny genetika   imunologie   metabolismus   MeSH
• imunohistochemie veterinární   MeSH
• kvantitativní polymerázová řetězová reakce veterinární   MeSH
• nemoci psů imunologie   metabolismus   MeSH
• průtoková cytometrie veterinární   MeSH
• psi genetika   imunologie   metabolismus   MeSH
• pyometra imunologie   metabolismus   veterinární   MeSH
• regulace genové exprese MeSH
• T-lymfocyty - podskupiny imunologie   metabolismus   MeSH
• uterus imunologie   metabolismus   patofyziologie   MeSH
• zvířata MeSH
• Check Tag
• psi genetika   imunologie   metabolismus   MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The zoonotic characteristic of Mycobacterium avium subsp. avium (MAA) represents a veterinary and economic problem in infected pigs. In this study, we analysed cell-mediated immunity six months after experimental infection by measuring interferon-γ (IFN-γ) production and by performing lymphocyte transformation tests after in vitro re-stimulation with the MAA-derived antigen. At the same time, IFN-γ-producing cells were characterised by flow cytometry. In MAA-infected animals, the production of IFN-γ increased in response to the MAA antigen in the blood, spleen and mesenteric lymph nodes. Similarly, a positive antigen-driven response was detected by the proliferation assay. In contrast, IFN-γ production and proliferation was undetectable after stimulation with the MAA antigen in uninfected control animals. These results indicate that both methods can be used for the identification of individual MAA-infected pigs. Using flow cytometry, we found that double-positive CD4(+)CD8(+) lymphocytes were the major T lymphocyte subset producing IFN-γ after in vitro re-stimulation.

• MeSH
• aktivace lymfocytů imunologie   MeSH
• buněčná imunita imunologie   MeSH
• interferon gama fyziologie   MeSH
• Mycobacterium avium imunologie   MeSH
• nemoci prasat imunologie   mikrobiologie   MeSH
• polymerázová řetězová reakce s reverzní transkripcí veterinární   MeSH
• prasata imunologie   MeSH
• průtoková cytometrie veterinární   MeSH
• T-lymfocyty - podskupiny imunologie   MeSH
• tuberkulóza imunologie   mikrobiologie   veterinární   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Five days after birth of a viable Fell pony filly, yellow watery diarrhoea appeared without any signs of systemic disease. Four days later the diarrhoea ceased. On 11th day, the animal showed apathy, and a few days later, the foal was very lethargic, suffered from muscular weakness and severe watery diarrhoea that reappeared. The illness did not respond to therapy. At the age of 21 days the filly spontaneously died under symptoms of intestinal colic and pneumonia. Haematological examinations revealed lower numbers of erythrocytes as well as non-selective lymphopenia. Phagocytic activity was slightly increased, lymphocyte activity was inhibited. Histopathology showed severe alteration of the lymphatic organs. T and B lymphocytes and antigen-presenting macrophages were not arranged in characteristic areas, and the quantity of these cells was lower than would be expected. Histopathological changes in lymphatic organs resembled those described in the literature as severe combined immunodeficiency.

• MeSH
• fatální výsledek MeSH
• financování organizované MeSH
• koně MeSH
• lymfopenie diagnóza   imunologie   veterinární   vrozené   MeSH
• nemoci koní diagnóza   imunologie   vrozené   MeSH
• novorozená zvířata MeSH
• počet lymfocytů veterinární   MeSH
• průtoková cytometrie veterinární   MeSH
• syndromy imunologické nedostatečnosti diagnóza   imunologie   veterinární   vrozené   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• kazuistiky MeSH