Concentrations of the acute phase proteins haptoglobin (Hp) and C-reactive protein (CRP) were measured in the serum, saliva and meat juice of pigs experimentally infected with a porcine reproductive ...and respiratory syndrome virus (PRRSv) field isolate. Sixteen PRRSv-free pigs were inoculated IM, killed in groups of four at 7, 14, 21 and 24
days post-inoculation (dpi), and samples of blood, saliva and diaphragmatic muscle were collected. Four non-infected controls were killed at 24
dpi. Significant differences in lung lesions were found between PRRSv-inoculated animals and controls. Changes in the concentrations of Hp and CRP in serum, saliva and meat juice samples were similar, peaking at 21
dpi. The correlations found suggest that the measurement of Hp and CRP in saliva and meat juice could serve as complementary, or possibly alternative, biomarkers of pig herd-health.
Transforming growth factor β (TGFβ) is an immunomodulatory cytokine which is able to modulate the host immune response eliciting an inefficient response against pathogens. In this sense, the role of ...this cytokine in porcine reproductive and respiratory syndrome (PRRS) has been poorly studied and the reported results are contradictory. Thus, in the present study, the expression of TGFβ was analysed both at tissue (lymphoid organs and lung) and serum level to study its correlation with the expression of PRRS virus (PRRSV). To carry out this study, 32 pigs were inoculated with the European PRRSV field isolate 2982 and sequentially killed from 0dpi to the end of the study (24dpi). Blood and tissue samples were collected to determine the expression of PRRSV and TGFβ. PRRSV was detected in inoculated animals from 3dpi until the end of the study, however TGFβ was not detected in sera from inoculated animals. Contrary, an increase of TGFβ antigen was observed both in the lymphoid organs and in the lung of PRRSV-inoculated pigs when compared with control group. Since TGFβ play a role as an immunomodulatory cytokine of the immune response and also in the differentiation of regulatory T cells (Tregs), the upregulation of the TGFβ at tissue level may play a role in the impairment of the host immune response observed during PRRS, being observed a significant correlation between PRRSV and TGFβ expression at lung level.
The changes in peripheral blood mononuclear cells (PBMCs) have been studied in several reports in an attempt to determine the immune response against porcine reproductive and respiratory syndrome ...virus (PRRSV) infection. However, how these changes are evoked after PRRSV infection has not yet been clarified. The aim of this study was to analyze the changes seen in lymphocyte subsets and immunomodulatory cytokine expression in pigs after an acute experimental infection with a European PRRSV field isolate. Pigs were inoculated intramuscularly with PRRSV field isolate 2982. Samples from blood, medial retropharyngeal and tracheobronchial lymph nodes, and spleen were collected at different time points for flow cytometry studies and for cytokine expression by ELISA. CD21(+) cell counts increased in PBMCs and tracheobronchial lymph node cells from 17 to 24 dpi, coinciding with an increase in PRRSV-specific antibody titer in blood. CD3(+) T-cell counts increased mainly due to an enhancement of CD4(-)CD8(high) and CD4(+)CD8(+) T cells. CD4(-)CD8(low) T cells were decreased in all organs studied, whereas CD4(+)CD8(-) T cells decreased only in the spleen. The drop in viremia correlated with an enhancement of CD4(-)CD8(high) T cells, and with a higher expression of interleukin-10 (IL-10) and interleukin-12 p40 (IL-12 p40). No efficient interferon-gamma (IFN-gamma) response was detected during the acute phase of the infection, and the expression of interferon-alpha (IFN-alpha) was late and reached its maximum expression once the viremia decreased. These results point to IL-10 and IL-12 as cytokines that might play a significant role in the PRRSV immune response, as may CD4(-)CD8(high) T cells.
A cat was presented with right head tilt and circling. The lack of expression of virus antigens did not support the postmortem diagnosis of encephalomyelitis pointing to a diffuse primary central ...nervous system T-cell lymphoma on the basis of CD3 and CD45R co-expression with absence of CD79α staining.
Correspondence: 1 Corresponding Author: Jaime Gómez-Laguna, Departamento de Anatomía y Anatomía Patológica Comparadas, Facultad de Veterinaria, Universidad de Córdoba, Campus de Rabanales, Edificio ...de Sanidad Animal, 14014 Córdoba, Spain, e-mail: v92golaj{at}uco.es
The current study was carried out to set up a fast and specific technique for porcine tuberculosis diagnosis in formalin-fixed, paraffin-embedded tissues. A retrospective study was carried out using 54 samples fixed in 10% neutral buffered formalin from 29 slaughtered Iberian pigs. Most of the pigs showed tissue samples positive to immunohistochemical staining (70.4%), and mycobacteria were detected within or near the necrotic cores of the lesions. However, diagnosis by this technique was time-consuming and tedious because of the paucibacillar nature of porcine tuberculous lesions. Classic polymerase chain reaction (PCR) was unsuccessful in mycobacteria genome amplification in all of the examined samples; however, real-time PCR amplified the mycobacteria genome in 23 of 29 examined pigs, identifying the Mycobacterium tuberculosis complex in all but one, which amplified Mycobacterium avium complex. Moreover, when reamplification of the DNA was performed, classic PCR amplified the mycobacteria genome in all the examined pigs (29/29), identifying the M. tuberculosis complex in 28 of 29 studied pigs and M. avium complex in only 1 pig. Results of the current study point out that both real-time and classic PCR assays, with genome reamplification, represent sensitive, fast, and specific diagnostic tools for porcine tuberculosis in formalin-fixed, paraffin-embedded tissues.
Key Words: Formalin-fixed granulomatous polymerase chain reaction tuberculosis
RT-qPCR is the method of choice for the accurate detection of low quantities of mRNA due to its higher sensitivity and specificity. Most of the previously published reports about swine cytokine gene ...expression lack information regarding the validation of the technique, which impedes the potential implementation by new users. This study was focussed on the technical validation of already published RT-qPCR assays for swine proinflammatory (IFN-α, IFN-γ, IL-12p35, IL-12p40 and TNF-α) and immunomodulatory (IL-10 and TGF-β) cytokines, and on defining the best qPCR amplification conditions for simultaneous amplification of the selected cytokines from several porcine tissue samples. The tested RT-qPCR assays here are sensitive (Efficient close to 2, Correlation coefficient higher than 0.95 and a Limit Of Detection below 305-100 mRNA copies), robust (Coefficint of Variation and Factor of Discrimination means were lower than 5 and 3%, respectively) and highly useful for the study of immune swine responses.
Tuberculosis-like lesions (TBL) in pigs have been associated with microorganisms other than mycobacteria. In this work a histopathological and microbiological evaluation of TBL in pigs is shown. A ...total of 352 samples belonging to 171 pigs totally condemned at slaughterhouse due to generalized TBL were sampled and selected for analysis. Pyogranulomatous (56.2%) and granulomatous lesions (20.2%) were observed in all analysed organs. Most of the granulomas observed in both lymph nodes and lungs belonged to more advanced stages of development (stages III and IV) whereas in the liver and the spleen most of lesions belonged to intermediate stages (stages II and III). Different microorganisms were simultaneously detected from TBL in the 42.7% of the animals. Mycobacterium tuberculosis complex (MTC) (38%), coryneform bacteria (40.3%) and streptococci (28.1%) were the main groups of microorganisms detected after bacteriological analysis, with Trueperella pyogenes and Streptococcus suis as the most frequently isolated species. Mycobacteria belonging to MTC were the most frequently detected pathogens in granulomatous and pyogranulomatous lesions in submandibular lymph nodes (32.7%) and coryneform bacteria were the microorganisms more frequently isolated from lungs (25.9%) and spleen samples (37.2%). These results may provide new insights into the pathogenesis and diagnosis of this pathology. The importance of coryneform bacteria and streptococci in such processes must be evaluated in future studies.
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Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Apoptosis is a strictly regulated mechanism of cell death that involves a complex network of biochemical pathways. Whether a cell undergoes apoptosis or not depends on a delicate balance of anti- and ...pro-apoptotic stimuli. This phenomenon can be induced by two different pathways: intrinsic and extrinsic pathways. The main aim of this study was to determine the ideal fixative and antigen retrieval method in porcine paraffin embedded tissues for the immunohistochemical detection of apoptosis mediators, from both extrinsic and intrinsic pathways. Tonsil, retropharyngeal lymph node and lung tissue samples were fixed in 10% neutral buffered formalin, Bouin solution and zinc salts fixative (ZSF) and different unmasking methods were carried out. Both 10% neutral buffered formalin and ZSF resulted as the fixatives of election to study apoptosis phenomena. Tween 20 (0.01% in PBS), citrate buffer (microwave, pH 6.0) and/or protease type XIV were the antigen retrieval methods which displayed better labelling. Our results allow to deep in the knowledge of apoptosis and its role in the pathogenesis of porcine diseases.
The aim of this work was to perform a complete study of maedi-visna virus (MVV) infected mammary glands of naturally-infected sheep, and to determine if cells other than macrophages undergo a ...productive viral infection in this organ. Fifteen seropositive and two seronegative ewes were selected from MVV-infected flocks on the basis of clinical indurative mastitis and three sheep from an MVV-free flock. Within the mammary gland, MVV-positive cells were located by immunohistochemistry in the stroma and the epithelial alveolar barrier, most likely the ovine mammary epithelial cells (OMEC) of the acini. In situ hybridization confirmed these findings. Ultrastructural studies showed the presence of lentivirus-like particles budding off the cell surface in the alveolar barrier and also free in the acinar lumen. The presence of mammary histopathological lesions and MVV together with clear indications of productive infection (demonstration of a cytopathic effect in OMEC cultures and infection of co-cultures) were observed in the 15 seropositive and one of the seronegative sheep from the infected flock. These findings demonstrate that the OMEC were infected in vivo and probably underwent productive infection when studied ex-vivo. The OMEC of MVV-free sheep, which had subsequently been infected in vitro with MVV, also showed productive infection when challenged in vitro, confirming the replication of MVV in OMEC in vitro. The presence of MVV-infected OMEC in the mammary gland from infected animals, the productive infection in these OMEC and the release of lentiviral particles to the acinar lumen may have relevance in the pathogenesis and transmission of MVV infection.
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