Acute phase response in cattle infected with Anaplasma marginale

This study was undertaken to evaluate the acute phase responses via the assessment of the concentration of serum sialic acids (total, lipid bound and protein bound), inflammatory mediators (IFN-γ and TNF-α) and acute phase proteins (Hp and SAA) in 20 adult crossbred cattle naturally infected by Anaplasma marginale. The infected animals were divided into 2 subgroups on the basis of parasitemia rate (<20% and >20%). Also, as a control group, 10 clinically healthy cattle from the same farms were sampled. Our data revealed significant decreases in red blood cell count (RBC), hematocrite (PCV) and hemoglobine (Hb) in infected cattle compared to healthy ones. Conversely, the concentrations of Hp, SAA, ceruloplasmin, fibrinogen, serum sialic acids and the circulatory IFN-γ and TNF-α were increased in the diseased cattle (P<0.05). In addition, it was evident that the progression of parasitemia in infected cattle did not induce any significant alterations in the hematological indices (RBCs, PCV and Hb) and the concentrations of Hp, SAA, ceruloplasmin and fibrinogen. SAA was the most sensitive factor to change in the diseased cattle. Therefore, increase in SAA concentration may be a good indicator of inflammatory process in cattle naturally infected with Anaplasma marginale.     

Evaluation of some acute phase proteins in cattle naturally infected with Babesia bigemina

Bovine babesiosis due to Babesia bigemina infection induces systemic inflammation, evidenced by increased sialic acid (SA) levels and declined cholinesterase activity. The current study was undertaken to assess further indicators of the systemic inflammation in the naturally infected cattle. To this end, serum levels of some selected acute phase-proteins (APPs) including serum amyloid A (SAA), haptoglobin (Hp), ceruloplasmin (Cp), and fibrinogen were measured. Additionally, sensitivity and specificity of the APPs were calculated by receiver operating characteristic curve. The correlation among APPs, SA and cholinesterase activity were also assessed. Our previous blood specimens were used to measure APPs. Briefly, the diseased animals were divided into two groups according to the parasitemia: 12 mildly (20 % <) and 8 severely (20 %>) infected animals. Moreover, 10 healthy animals as the control were included. The levels of all APPs were measured to be significantly elevated in a parasitemia burden-dependent fashion as compared to the control. Furthermore, all the APPs showed 100 % specificity, but only SAA and Cp had 100 % sensitivity. A strong and positive correlation was calculated between the APPs and SA; however, cholinesterase activity was inversely correlated with AAPs and SA. In conclusion, inflammatory reactions play a pivotal role in the pathogenesis of bovine babesiosis and APPs can be considered as the potential indicators of inflammation.     

Serum haptoglobin concentration in cattle

To obtain a basal concentration of serum Haptoglobin (Hp) in cattle in Taiwan, Hp concentrations were measured from serum samples collected from 10 healthy heifers, every week for one year. The values were also compared with those collected from 15 cows diagnosed with postpartum metritis. The heifers were successfully impregnated by artificial insemination six months after the tests. Hp concentrations were also measured in the serum collected from 11 other cows within 3 weeks after parturition. The Hp assay developed in this study gave a good correlation (r=0.893)with Western blotting. The Hp concentration of 454 serum samples from the 10 heifers had a mean value of 83.6 +/- 34.1 mg/l, and there was no significant difference among individual heifers. The basal value of Hp in heifers was calculated as less than 73.6 mg/l. No significant difference in Hp concentration was observed among the 10 heifers during cold and warm seasons (19.8 +/- 2.2 degrees C vs 27.3 +/- 1.4 degrees C), or before and after pregnancy. The mean serum Hp concentration from cows suffering from postpartum reproductive disorders was 1133.5 +/- 627.1 mg/l, which was significantly greater than the serum of healthy heifers and postpartum cows (104.6 +/- 61.0 mg/l) (P<0.05). These results demonstrate that Hp concentration may be a useful indicator for cows with postpartum reproductive disorders.     

Bovine haptoglobin: single radial immunodiffusion assay of its polymeric forms and dramatic rise in acute-phase sera.

Using purified bovine haptoglobin (Hp) and specific antisera, a single radial immunodiffusion (SRID) assay method has been developed to measure the serum Hp level in cattle. Bovine Hp is a highly polymerized protein showing heterogeneous molecular forms in serum. After treatment with cysteine or glutathione, Hp was partially reduced to a homogeneous form, suitable for SRID assay. This method gives values comparable to those obtained by hemoglobin-binding capacity assay, and has the advantage of being simple and convenient. Although serum Hp was not detectable in healthy cattle, it was found more than 50-fold after invasive surgery, indicating that Hp is a characteristic acute-phase protein in cattle.     

Preliminary studies of serum acute-phase protein concentrations in hematologic and neoplastic diseases of the dog

Serum concentrations of acute-phase proteins (APPs): haptoglobin (Hp), ceruloplasmin (Cp), serum amyloid A (SAA), and C-reactive protein (CRP) were determined in healthy dogs (n = 15) and dogs with different diseases grouped as acute inflammation (I, n = 12), hematologic neoplasias (HT, including leukemia and lymphoma, n = 16), nonhematologic neoplasias (NHT, including epithelial, mesenchymal, and mixed, n = 20), and autoimmune hemolytic anemia (AIHA, n = 8). SAA and CRP were analyzed using commercially available enzyme-linked immunosorbent assay (ELISA) kits, and Hp and Cp were measured using colorimetric methods, all previously validated for use in dogs. Increased concentrations of all APPs were observed in all groups of diseased dogs, but statistical significance only was observed with Hp (I, P < .001; HT, P < .05), Cp (I, P < .05; AIHA, P < .01), and CRP (I, P < .001; HT, P < .001; AIHA, CRP P < .05). High variability in individual APPs within each group of diseases was found with no significant differences between leukemia and lymphoma as well as among different types of neoplasia. The AIHA group had smaller increases in Hp, SAA, and CRP but higher concentrations of Cp. When follow-up of individual cases was possible, a decrease in APPs generally was found in cases with favorable outcome. The results of this study suggest that neoplasia and hematologic diseases such as AIHA should be considered as possible causes of mild increases in APPs in dogs. Measurement of APPs may be helpful to assess clinical evolution and monitor treatment of these processes.     

Serum amyloid A and haptoglobin levels in bovine amyloidosis

Serum amyloid A (SAA) and haptoglobin (Hp) levels were determined in 25 cows suffering from amyloidosis. SAA levels in cows with amyloidosis ranged between < 0.3 and 225.8 microg/ml, with a median level of 105.1 microg/ml, and Hp levels ranged between < 20 and 1860 microg/ml, with a median level of 950 microg/ml. These levels were significantly higher than the levels observed in healthy cows (SAA levels ranged from < 0.3 to 13.5 microg/ml, with median of 1.4 microg/ml, and Hp levels were undetectable in all cases), but were not significantly different from the levels observed in control cows with chronic inflammation. There was a significant correlation between SAA and Hp levels in cows with chronic inflammation , but not in cows with amyloidosis. It was concluded that the serum SAA levels in cows with amyloidosis might be changed by some factor other than inflammation.     

Acute phase protein response in goats

Acute phase proteins (APPs) are important diagnostic indicators of inflammatory disturbances in animals. The objectives of the current study were to validate analytical methods for measuring haptoglobin (Hp), serum amyloid A (SAA), acid soluble glycoprotein (ASG), fibrinogen, and albumin concentrations in goats and to evaluate their response to an inflammatory stimulus in this species. Intra- and interassay coefficients of variation (CVs) were in the range 0.07-9.31% and 1.83-12.68%, respectively, for all APPs and showed good precision. All assays determined APPs in a linear manner at different sample dilutions with high correlation coefficients with the exception of fibrinogen, which was measured by the heat precipitation method. Subcutaneous injection of turpentine oil induced an increase in Hp, SAA, ASG, and fibrinogen serum concentrations and a decrease in albumin concentration.     

Serum Cardiac Troponin I Concentrations in Cattle with Cardiac and Noncardiac Disorders

Background: Making a clinical diagnosis of pericarditis in cattle is difficult and additional diagnostic tests are needed to evaluate cattle with suspected pericarditis. Serum cardiac troponin I (cTnI) concentrations are increased in cattle with pericarditis, but the utility of measuring serum cTnI concentrations in cattle with suspected pericarditis in cattle remains unclear.
Objectives: To determine if serum cTnI concentrations in cattle can be used to differentiate pericarditis from other cardiac disorders and noncardiac thoracic diseases.
Animals: Seventy-seven clinically diseased cattle and 19 healthy control cattle.
Methods: Serum cTnI concentrations were measured using an Immunlite Troponin I immunometric chemiluminescent assay in consecutive cases of postmortem-confirmed pericarditis (n = 18), endocarditis (n = 15), chronic suppurative pneumonia (n = 13), congenital heart disease (n = 10), reticulitis (n = 3), mediastinal abscess (n = 7), thymic lymphoma (n = 6), and caudal vena cava thrombosis (n = 5). Serum cTnI concentrations were measured in 19 healthy cattle.
Results: Although serum cTnI concentrations were significantly higher in cattle with pericarditis compared with healthy cattle, they were not significantly different from concentrations in cattle with endocarditis, congenital cardiac disease, mediastinal abscess, reticulitis, caudal vena cava thrombosis, or chronic suppurative pneumonia.
Conclusions: Serum cTnI cannot be used to distinguish cattle with pericarditis from cattle with other primary cardiac diseases. In addition, serum cTnI concentrations cannot distinguish between cattle with primary cardiac diseases and those with other noncardiac, intrathoracic disorders.     

Low levels of haptoglobin and putative amino acid sequence in Taiwanese Lanyu miniature pigs

Porcine haptoglobin (Hp) is an acute phase protein. Its plasma level increases significantly during inflammation and infection. One of the main functions of Hp is to bind free hemoglobin (Hb) and inhibit its oxidative activity. In the present report, we studied the Hp phenotype of Taiwanese Lanyu miniature pigs (TLY minipigs; n=43) and found their Hp structure to be a homodimer (beta-alpha-alpha-beta) similar to human Hp 1-1. Interestingly, Western blot and high performance liquid chromatographic (HPLC) analysis showed that 25% of the TLY minipigs possessed low or no plasma Hp level (<0.05 mg/ml). The Hp cDNA of these TLY minipigs was then cloned, and the translated amino acid sequence was analyzed. No sequences were found to be deficient; they showed a 99.7% identity with domestic pigs (NP_999165). The mean overall Hp level of the TLY minipigs (0.21 +/- 0.25 mg/ml; n=43) determined by enzyme-linked immunosorbent assay (ELISA) was markedly lower than that of domestic pigs (0.78 +/- 0.45 mg/ml; p<0.001), while 25% of the TLY minipigs had an Hp level that was extremely low (<0.05 mg/ml). In addition, the initial recovery rate (first 40 min) in the circulation of infused fluorescein isothiocyanate (FITC)-Hb was significantly higher in the TLY minipigs with extremely low Hp levels than those with high levels. This data suggests that the low concentration of Hp-Hb complex is responsible for the higher recovery rate of Hb in the circulation. TLY minipigs have been used as an experimental model for cardiovascular diseases; whether they can be used as a model for inflammatory diseases, with Hp as a marker, remains a topic of interest. However, since the Hp level varies significantly among individual TLY minipigs, it is necessary to prescreen the Hp levels of the animals to minimize variation in the experimental baseline. The present study may provide a reference value for future use of the TLY minipig as an animal model for inflammation-associated diseases.     

Natural Antibodies in Sera of Mink Before and After The Development of Aleutian Disease (Viral Plasmacytosis)

The effect of infection of mink with aleutian disease virus on the level of natural antibodies in the serum was investigated. The level of natural antibodies to chicken red blood cells was increased following infection but there was no correlation between the degree of hypergamma globulinemia in the diseased mink and the increase in titers. On the other hand, serum levels of natural hemolytic antibodies to sheep red blood cells in mink did not increase during the course of aleutian disease. These data indicate that the aleutian disease virus does not stimulate a broad spectrum of pre-existing antibody producing cells.     

Correlation of serum concentration of alpha 1-acid glycoprotein with lymphocyte blastogenesis and development of experimentally induced or naturally acquired hepatic abscesses in cattle.

Changes in serum alpha 1-acid glycoprotein (alpha 1AG) concentration in cattle with hepatic abscesses were observed, and function of alpha 1AG was evaluated, particularly its influence on cellular immune response. Test cattle (n = 4) were inoculated with Fusobacterium necrophorum, control cattle (n = 2) were inoculated with inactivated bacteria, and naturally affected cattle (n = 11) were found in a slaughterhouse. Determination of alpha 1AG was made by use of a single radial immunodiffusion method. The action on lymphocyte blastogenesis was determined by [3H]thymidine incorporation. Cultured lymphocytes from healthy cattle were treated with variable concentrations of alpha 1AG purified from serum obtained from cattle with hepatic abscesses and suppression of blastogenesis stimulated by each of 3 mitogens was measured. In cattle with experimentally induced abscesses, serum alpha 1AG concentration increased for 7 to 10 days after F necrophorum inoculation, its change being parallel to that of sialic acid. High concentration of alpha 1AG was found in naturally affected cattle and was highly correlated to sialic acid concentration. Suppression of lymphocyte blastogenesis in cattle with experimentally induced hepatic abscesses was highly correlated to serum alpha 1-AG concentration.     

Analysis by enzyme-linked immunosorbent assay and 2-dimensional electrophoresis of haptoglobin in the high-density lipoprotein fraction in cows

Haptoglobin (Hp) is a hemoglobin (Hb)-binding acute-phase protein. Besides its relevance in inflammation, Hp is involved in the regulation of lipid metabolism. In cattle, in addition to the lipoprotein-deficient fraction, Hp is distributed in high-density lipoprotein (HDL) and very high-density lipoprotein (VHDL) fractions. The purpose of this study was to determine Hp concentrations in the lipoprotein fractions using an enzyme-linked immunosorbent assay (ELISA) based on the affinity with Hb, and also to detect structural differences of HDL Hp from that in the lipoprotein-deficient fraction using 2-dimensional electrophoresis. When purified Hp was used as the antigen for the ELISA, the detection limit was 7.4 ng/ml and linearity was obtained from 14.8 to 475 ng/ml. The correlation coefficient between the ELISA and single radial immunodiffusion was 0.884. The ELISA was shown to be applicable to evaluate Hp concentrations in the lipoprotein fractions. Hp concentrations in the lipoprotein fractions were in the range of 0.94 to 8.77 microg of Hp/ml (n = 4), and concentration ratios were 0.2 to 0.3% of whole serum Hp. Of the lipoprotein fractions, Hp was most abundant in HDL, moderate in VHDL and faint in chylomicrons, the very low-density lipoprotein fraction and low-density lipoprotein fraction. By 2-dimensional electrophoresis, alpha- and beta-chains of serum Hp were each separated into 5 spots, and their isoelectric point (pI) values were from 5.05 to 6.28 in the alpha-chain and from 5.92 to 6.95 in the beta-chain. The pI values of HDL Hp were indistinguishable from those of serum Hp. These results indicate that the ELISA based on the affinity with Hb is useful for evaluating Hp concentrations in lipoprotein fractions, and also suggest that HDL Hp is structurally similar to that in the lipoprotein-deficient fraction.     

The correlations among serum tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and sialic acids with peripheral lymphocytes in bovine tropical theileriosis

The infection with protozoan parasite Theileria annulata induces changes triggering the activation and/or proliferation of the host lymphocytes. In order to find out the possible correlations among peripheral circulatory lymphocytes, cytokine activities and the level of sialic acids, 50 dairy Holstein cattle, naturally infected with T. annulata, were divided into 4 subgroups according to their parasitemia rates (<1%, 1–3%, 3–5% and >5%). Also, ten non-infected cattle were sampled as control group. Blood samples were taken from jugular vein into acid citrate dextrose-containing tubes for measuring hematological parameters and B and T (CD₄ and CD₈) cell populations and without anticoagulant for TNF-α, IFN-γ and sialic acid concentrations. Remarkable decreases observed in red blood cells (RBCs), white blood cells (WBCs) and packed cell volume (PCV) in infected cattle compared to healthy ones (P < 0.05). Also, with increase in parasitemia rate, total lymphocytes and monocytes alleviated in the diseased groups. By contrast, total neutrohpils and the concentrations of TNF-α, IFN-γ and total sialic acids were significantly elevated (P < 0.05) in infected animals. Accordingly, the circulatory populations of CD₄ and CD₈ T cells and B cells showed a substantial decrease, while a significant increase was observed in T (CD₄ and CD₈) cells in cattle infected with <1% parasitemia rates. Decreased circulatory T cell population shows the ineffective responses of T cells to the stimulatory cytokines such as IFN-γ or TNF-α. On the other hand, the elevation of cytokines (particularly IFN-γ) and sialic acids have presumably an inhibitory role on circulatory B cell population in infected cattle. In addition, a high level of sialic acid concentration indicates the probable role of sialic acid to regulate the parasite-host cell adhesion during sporozoites invasion.     

Development of an enzyme immuno assay for the determination of porcine haptoglobin in various body fluids: testing the significance of meat juice measurements for quality monitoring programs

Quantification of haptoglobin (Hp), an acute phase protein, in blood is presently discussed as being useful to monitor animal health. We developed an enzyme immuno assay (EIA) which is specific for porcine Hp, is not impaired by hemolytic samples and is sufficiently sensitive to be applied in meat juice. Hp was purified from porcine serum by affinity chromatography on hemoglobin Sepharose followed by gel filtration. A specific rabbit antiserum was obtained. In a competitive approach, biotinylated porcine Hp was used as tracer and incubated with Hp standard or sample in microtiter plates. The limit of detection was 0.02 mg/l, parallelism of sample dilutions was proven; recovery of Hp added to serum samples was 96.4 +/- 4.7%. The coefficients of intra and inter-assay variation were 3.3 (n=5) and 10.2% (n=16), respectively. Hp was reliably quantified in blood serum and plasma, whole blood, saliva and meat juice. For healthy pigs of different ages (4 weeks and 6 months), mean Hp concentrations of about 0.5-0.7 mg/ml were observed. To test the significance of Hp measurements in other matrices, samples were obtained from fattening pigs or from slaughter pigs. Blood serum or plasma was collected in parallel. In whole blood, Hp concentrations were about 40% lower than in plasma, but were closely related (n=24,r=0.85,P<0.001). Saliva Hp concentrations ranged between 0.3 and 3.0 microg/ml and were marginally related with blood plasma concentrations (n=93,r=0.35,P<0.001). From 106 hybrid slaughter pigs (100-110 kg) blood and muscle samples (diaphragmatic pillar, d.p.; m. brachiocephalicus, m.b.) were collected. Meat juice was obtained after freezing and thawing. Concentrations were 0.39+/-0.5 mg/ml in serum and 0.04+/-0.06 mg/ml in meat juice. Hp concentrations in blood were closely correlated with those in d.p. juice (P<0.001,r=0.750) and m.b. juice (P<0.001,r=0.776). In view of the many reports on Hp measurements being predictive for animal health even in the subclinical range, we conclude that Hp quantification in meat juice might be useful to assess meat quality at slaughter and further along the processing chain in terms of animal health.     

Cerebrospinal fluid S-100B concentrations in normal and diseased cattle

We measured the concentrations of S-100B, a marker protein used in humans to detect brain damage, in the cerebrospinal fluid (CSF) of clinically normal cattle (n=15, mean age +/- SD: 31.8 +/- 37.5 months) and of cattle with various inflammatory disorders (n=43, 70.6 +/- 31.9 months). The mean +/- SD CSF S-100B level was 2.9 +/- 1.6 ng/ml in the normal group and 7.0 +/- 7.4 ng/ml in the diseased group. Thirteen diseased cattle that had developed no obvious neurological signs showed abnormally high S-100B concentrations (> 8.0 ng/ml), whereas the two cattle with neurological disorders did not. No particular disease could be related to the S-100B rise. Therefore, it remains inconclusive whether measurement of CSF S-100B concentration is useful in veterinary neurological diagnosis.     

Increase of serum lipid-associated sialic acid concentration in dogs with neoplasms

Serum concentrations of lipid-associated sialic acid (LASA) were determined in sera obtained from 26 apparently healthy dogs (group 1), 56 dogs with nonneoplastic disorders (group 2), and 100 dogs with neoplasms (benign [group 3], malignant [group 4]), using a simple, reliable, rapid spectrophotometric procedure. The LASA concentrations were significantly increased (P less than 0.01) in group 4 dogs (malignant neoplasms) when compared with those in dogs from groups 1, 2, and 3. The sensitivity of the test was between 71.5% and 100%, and specificity was 92.3% for group 1, 67.8% for group 2, and 80.6% for group 3. A preliminary study concerning variations of serum LASA values as a marker after surgical treatment of tumor showed a good correlation between effectiveness of therapy and serum LASA concentrations.     

Serum concentration of sialic acids in naturally occurring ovine babesiosis

This study is designated to assess the effect of the severity of Babesia ovis infection on sialic acid concentration in blood sera in naturally infected sheep. Infected animals (diseased group) comprised 38 Iranian fat-tailed sheep, about 1–3 years old, naturally infected with B. ovis, divided into four subgroups with respect to parasitemia rates (low 0.1–0.3 %, moderate 0.4–0.9 %, high 1–2.5 %, and very high >2.5 %). The parasitological diagnosis was confirmed using PCR analysis. As a control group, ten clinically healthy sheep reared under the same management and environmental conditions were also sampled. Hematological parameters and the concentrations of total sialic acid (TSA), lipid-bound sialic acid (LBSA), and protein-bound sialic acid (PBSA) were measured in both groups. Compared to controls, sialic acid concentrations showed significant increase (p?Parasitemia rate was positively correlated with sialic acid concentrations. This study demonstrated that B. ovis infection induced marked and persistent elevations of serum sialic acid concentrations. It seems that increase of serum sialic acid concentrations during parasitemia alter receptor-ligand interactions, which are known to play important role in immune response. Furthermore, sialic acid would indirectly inhibit the action of leukocytes and consequently promote the evasion of the immune response and persistence of the parasite in the host. This factor could influence the parasite-host cell adhesion, but further detailed biochemical investigations are needed to precisely explain the exact role of sialic acid in invasion process of the parasite to the host cells.     

Plasma prolidase activity as a possible diagnostic index of chronic hepatic abscess in cattle

The usefulness of prolidase as a biochemical parameter to represent the chronic state of hepatic abscess was discussed in eight cattle experimentally inoculated with Fusobacterium necrophorum and 18 spontaneously affected cattle. Blood was daily collected to measure the plasma prolidase activity and sialic acid level from the experimental cattle for 90 days after inoculation. In three out of four cattle affected with hepatic abscess, prolidase activity began to rise about 40 days after inoculation, and maintained high activity till 90 days. In the same cattle the sialic acid concentration increased from 7 to 10 days after inoculation, and gradually returned to the normal value 50 days after it. Another cow showed a similar change in early stage of experiment, but prolidase activity decreased after 70 days and sialic acid concentration maintained high level till 90 days. In two cattle, which showed scars but no abscess on autopsy, the prolidase activity increased temporarily from 40 to 55 days after inoculation. In the control cattle inoculated with an inactivated bacterial suspension, neither the sialic acid level nor the prolidase activity showed any large variation in the experimental period. Among the spontaneously affected cattle, those with a high sialic acid level revealed normal prolidase activity and those with a normal sialic acid level had high prolidase activity.     

Comparative cardiac pathological changes of Atlantic salmon (Salmo salar L.) affected with heart and skeletal muscle inflammation (HSMI), cardiomyopathy syndrome (CMS) and pancreas disease (PD)

The heart is considered the powerhouse of the cardiovascular system. Heart and skeletal muscle inflammation (HSMI), cardiomyopathy syndrome (CMS) and pancreas disease (PD) are cardiac diseases of marine farmed Atlantic salmon (Salmo salar) which commonly affect the heart in addition to the skeletal muscle, liver and pancreas. The main findings of these diseases are necrosis and inflammatory cells infiltrates affecting different regions of the heart. In order to better characterize the cardiac pathology, study of the inflammatory cell characteristics and cell cycle protein expression was undertaken by immunohistochemistry. Immunohistochemistry was performed on paraffin embedded hearts from confirmed diseased cases applying specific antibodies. The inflammatory cells were predominantly CD3⁺ T lymphocytes. The PD diseased hearts exhibited moderate hypoxia inducible factor-1α (HIF1α) immuno-reaction that suggested tissue hypoxia while recombinant tumor necrosis factor-α (rTNFα) antibody identified putative macrophages and eosinophilic granulocytes (EGCs) in addition to endocardial cells around lesions. There were strong to low levels of major histocompatibility complex (MHC) class II immunostaining in the diseased hearts associated with macrophage-like and lymphocyte-like cells. The diseased hearts expressed strong to low levels of apoptotic cells identified by caspase 3 and terminal deoxynucleotidyl transferase nick-end labeling (TUNEL) staining. The strong signals for proliferative cell nuclear antigen (PCNA) and TUNEL, and moderate levels of caspase 3 immuno-reactivity suggested a high cell turnover where DNA damage/repair might be occurring in the diseased hearts. Interestingly, the apparently similar cardiac diseases exhibited differences in the immunopathological responses in Atlantic salmon.     

Mycobacterium avium subsp. paratuberculosis infection in cattle in Austria, diagnosis with culture, PCR and ELISA

Serum samples from healthy, infected (n=11) and diseased (n=2) cattle as well as positive (n=17) and negative (n=41) reference sera were tested for antibodies to Mycobacterium avium subsp. paratuberculosis with two ELISA-methods (A-ELISA, Allied Monitors, Fayette, USA; H-ELISA, Institute of Microbiology and Animal Diseases, Veterinary University Hannover). Fecal samples of these animals were examined by PCR and culture. Also field serum samples found to be positive (n=664) or inconclusive (n=1589) by A-ELISA during a survey done on 11028 cattle of 2757 farms at different districts in Austria were retested with H-ELISA (Gasteiner et al., 1999). In both ELISA-methods total agreement between antibody detection and shedding of M. avium subsp. paratuberculosis (PCR, culture) in cases of diseased animals during the testing period was found. In subclinically infected animals H-ELISA showed a better correlation with the results of PCR and fecal culture. Reference serum samples of culturally negative cattle were negative in 98% by H-ELISA and in 82% by A-ELISA, and those of positive animals were positive in 59% by H-ELISA and in 82% by A-ELISA. The 664 A-ELISA positive field serum samples were positive in 20.5%, inconclusive in 32.5% and negative in 47% by H-ELISA. A-ELISA inconclusive sera gave positive reactions by H-ELISA in 5.2%, negative in 74.8% and inconclusive results in 20%. The highest prevalence of antibodies (7.9% by A- and 2.2% by H-ELISA) against M. avium subsp. paratuberculosis were found in cattle at the age of six and seven years. Seropositive animals were found at all tested ages. The A-ELISA gave two to three times more positive reactors than the H-ELISA. Also both tests showed the highest prevalence of reagents among Holstein Friesian (6.2% by A-ELISA, 2.5% by H-ELISA) followed by other cattle breeds. Seropositive cattle were observed in all districts of Austria in 3. 3-7.1% and in 0.5-1.8% of herds according to A- and H-ELISA, respectively.