scholarly journals Paratuberculosis.

1994 ◽  
Vol 7 (3) ◽  
pp. 328-345 ◽  
Author(s):  
C Cocito ◽  
P Gilot ◽  
M Coene ◽  
M de Kesel ◽  
P Poupart ◽  
...  
Keyword(s):  
Cell Wall ◽  
Pcr Amplification ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Polymorphism Analysis ◽  
Mycobacterial Antigen ◽  
Immune Reactions ◽  
Species Specific ◽  
Cellular Immune

Paratuberculosis (Johne's disease) is a chronic, wasting, widespread mycobacteriosis of ruminants. It involves extensive mycobacterial shedding, which accounts for the high contagiousness, and ends with a fatal enteritis. Decreases in weight, milk production, and fertility produce severe economic loss. The DNA of the etiological agent (Mycobacterium paratuberculosis) has a base composition (66 to 67% G+C) within the range of that of mycobacteria (62 to 70% G+C), a size (4.4 x 10(6) to 4.7 x 10(6) bp) larger than that of most pathogenic mycobacteria (2.0 x 10(6) to 4.2 x 10(6) bp), and a high relatedness (> 90%) to Mycobacterium avium DNA. However, the DNAs of the two organisms can be distinguished by restriction fragment length polymorphism analysis. M. paratuberculosis genes coding for a transposase, a cell wall-associated protein (P34), and two heat shock proteins have been cloned and sequenced. Nucleic acid probes (two of which are species specific) are used, after PCR amplification, for M. paratuberculosis identification in stools and milk. As in leprosy, with disease progression, cellular immune reactions decrease and humoral immune reactions increase. Cutaneous testing with sensitins, lymphocyte proliferation assays, and cytokine tests are used to monitor cellular immune reactions in paratuberculosis, but these tests lack specificity. Complement fixation, immunodiffusion, and enzymometric tests based on antibodies to M. paratuberculosis extracts, to mycobacterial antigen complex A36, to glycolipids, and to proteins help identify affected cattle but are not species specific. The carboxyl-terminal portion of the 34-kDa cell wall-associated A36 protein (P34) carries species-specific B-cell epitopes and is the basis for an enzyme-linked immunosorbent assay. Diagnostic tests for paratuberculosis are also used in Crohn's disease, a chronic human ileitis mimicking Johne's disease, in which isolates identified as M. paratuberculosis have been found.

scholarly journals Acute Plasmodium chabaudi chabaudi Malaria Infection Induces Antibodies Which Bind to the Surfaces of Parasitized Erythrocytes and Promote Their Phagocytosis by Macrophages In Vitro

1998 ◽  
Vol 66 (9) ◽  
pp. 4080-4086 ◽  
Author(s):  
Maria M. Mota ◽  
K. Neil Brown ◽  
Anthony A. Holder ◽  
William Jarra
Keyword(s):  
Parasite Clearance ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibody Activity ◽  
Parasite Line ◽  
Primary Role ◽  
Plasmodium Chabaudi ◽  
Species Specific ◽  
Cellular Immune ◽  
Variant Line

ABSTRACT CBA/Ca mice infected with 5 × 104 Plasmodium chabaudi chabaudi AS-parasitized erythrocytes experience acute but self-limiting infections of relatively short duration. Parasitemia peaks (∼40% infected erythrocytes) on day 10 or 11 and is then partially resolved over the ensuing 5 to 6 days, a period referred to as crisis. How humoral and cellular immune mechanisms contribute to parasite killing and/or clearance during crisis is controversial. Humoral immunity might be parasite variant, line, or species specific, while cellular immune responses would be relatively less specific. For P. c. chabaudi AS, parasite clearance is largely species and line specific during this time, which suggests a primary role for antibody activity. Accordingly, acute-phase plasma (APP; taken fromP. c. chabaudi AS-infected mice at day 11 or 12 postinfection) was examined for the presence of parasite-specific antibody activity by enzyme-linked immunosorbent assay. Antibody binding to the surface of intact, live parasitized erythrocytes, particularly those containing mature (trophozoite and schizont) parasites, was demonstrated by immunofluorescence in APP and the immunoglobulin G (IgG)-containing fraction thereof. Unfractionated APP (from P. c. chabaudi AS-infected mice), as well as its IgG fraction, specifically mediated the opsonization and internalization of P. c. chabaudi AS-parasitized erythrocytes by macrophages in vitro. APP from another parasite line (P. c. chabaudi CB) did not mediate the same effect against P. c. chabaudi AS-parasitized erythrocytes. These results, which may represent one mechanism of parasite removal during crisis, are discussed in relation to the parasite variant, line, and species specificity of parasite clearance during this time.

scholarly journals The collection of lymphatic fluid from the bovine udder and its use for the detection of Mycobacterium avium subsp. paratuberculosis in the cow

2011 ◽  
Vol 24 (1) ◽  
pp. 23-31 ◽  
Author(s):  
Johannes L. Khol ◽  
Pablo J. Pinedo ◽  
Claus D. Buergelt ◽  
Laura M. Neumann ◽  
Walter Baumgartner ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Fluid Collection ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Mycobacterium Avium Subsp ◽  
Enzyme Linked Immunosorbent Assay ◽  
Lymph Fluid ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Lymphatic Fluid ◽  
Elisa Result

The objective of the current study was to evaluate the feasibility of lymph collection from the bovine udder and to investigate if the lymphatic fluid might be of diagnostic value in cows infected with Mycobacterium avium subsp. paratuberculosis (MAP), the etiologic agent of paratuberculosis. Lymph fluid collection was attempted from 58 cows, and the reactions of the cows as well as the level of difficulty of the procedure were recorded in 56 animals. Lymph samples (51 in total) were tested for the presence of MAP by nested polymerase chain reaction. Collection of the lymphatic fluid caused no or mild signs of discomfort in 94.6% of the cows; in 51.8% of cows, lymphatic fluid was attained on the first attempt, while sample collection was unsuccessful in 12.1%. Mycobacterium avium subsp. paratuberculosis was detected in 43.1% of all lymph samples. The bacterium was present in 66.7% of cows with clinical Johne’s disease, in 42.8% of asymptomatic cows with a positive or suspicious enzyme-linked immunosorbent assay (ELISA) result in blood, and in 38.7% of cows with a negative ELISA result in blood. The present study shows that the procedure was well tolerated by most cows and can easily be performed on farm. The current report of the isolation of MAP from lymph fluid suggests that the present approach could be used for the early detection of Johne’s disease in cattle.

scholarly journals Immunoglobulin G1 Enzyme-Linked Immunosorbent Assay for Diagnosis of Johne's Disease in Red Deer (Cervus elaphus)

2005 ◽  
Vol 12 (12) ◽  
pp. 1401-1409 ◽  
Author(s):  
J. Frank T. Griffin ◽  
Evelyn Spittle ◽  
Christie R. Rodgers ◽  
Simon Liggett ◽  
Marc Cooper ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Cervus Elaphus ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Marker ◽  
Protein Antigens ◽  
Characteristic Analysis ◽  
Purified Protein Derivative ◽  
Subclinical Disease

ABSTRACT This study was designed to develop a customized enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of Johne's disease (JD) in farmed deer. Two antigens were selected on the basis of their superior diagnostic readouts: denatured purified protein derivative (PPDj) and undenatured protoplasmic antigen (PpAg). ELISA development was based on the antigen reactivity of the immunoglobulin G1 (IgG1) isotype, which is a highly specific marker for mycobacterial disease seroreactivity in deer. Sensitivity estimates and test parameters were established using 102 Mycobacterium paratuberculosis-infected animals from more than 10 deer herds, and specificity estimates were determined using 508 uninfected animals from 5 known disease-free herds. A receiver-operated characteristic analysis determined that at a cut point of 50 ELISA units, there was a specificity of 99.5% and sensitivities of 84.0% with PPDj antigen, 88.0% with PpAg, and 91.0% when the antigens were used serially in a composite test. Estimated sensitivity was further improved using recombinant protein antigens unique for M. paratuberculosis, which identified infected animals that were unreactive to PPDj or PpAg. While 80% of animals that were seropositive in the IgG1 ELISA had detectable histopathology, the assay could also detect animals with subclinical disease. The test was significantly less sensitive (75%) for animals that were culture positive for M. paratuberculosis but with no detectable pathology than for those with pathological evidence of JD (>90%). When the IgG1 ELISA was used annually over a 4-year period in a deer herd with high levels of clinical JD, it eliminated clinical disease, increased production levels, and reduced JD-related mortality.

scholarly journals A Novel Real-Time PCR-Based Screening Test with Pooled Fecal Samples for Bovine Johne’s Disease

2020 ◽  
Vol 58 (12) ◽  
Author(s):  
Satoko Kawaji ◽  
Reiko Nagata ◽  
Yasutaka Minegishi ◽  
Yumi Saruyama ◽  
Akiko Mita ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Pcr Assay ◽  
Serological Tests ◽  
Fecal Samples ◽  
Content Type ◽  
Cattle Herds

ABSTRACT Johne’s disease (JD) is an economically important infectious disease in livestock farming caused by Mycobacterium avium subsp. paratuberculosis. As an alternative to serological tests, which are used mainly for the screening of whole herds, we developed a novel ResoLight-based real-time PCR (RL-PCR) assay with pooled fecal samples for the detection of fecal shedders in cattle herds. The RL-PCR assay included an internal amplification control (IC) which was amplified using the same primer pair as the target molecule M. avium subsp. paratuberculosis IS900 and differentiated based on melting temperatures. Individual fecal suspensions were pooled and concentrated by centrifugation to avoid a loss of sensitivity by the dilution effect. Combined with a DNA extraction kit (Johne-PureSpin; FASMAC), no inhibition of PCR amplification was observed with up to 15 fecal samples in a pool. The detection limit of RL-PCR at a pool size of 10 was 10 M. avium subsp. paratuberculosis organisms per gram of feces, which was comparable to that of individual testing. A total of 2,654 animals in 12 infected herds were screened by individual antibody-enzyme-linked immunosorbent assay (ELISA) and the RL-PCR assay using pooled feces. Fifty animals were diagnosed with JD through the screening by RL-PCR, compared with only 5 by ELISA (which were also positive in RL-PCR). In 7 JD-free herds, the results of 4 out of 327 pools (1.2%) were invalid due to the lack of IC amplification, and then animals were confirmed negative individually. Our results suggest that implementation of herd screening by pooled RL-PCR would advance the monitoring and control of JD in cattle herds.

scholarly journals Control of Johne's Disease in Four Commercial Dairy Herds in Iowa

1989 ◽  
Vol 1 (3) ◽  
pp. 223-226 ◽  
Author(s):  
Charles O. Thoen ◽  
Larry A. Moore
Keyword(s):  
Potassium Chloride ◽  
Milk Production ◽  
Market Value ◽  
Control Program ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Economic Losses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dairy Herds ◽  
Serum Samples

A 6-year study was conducted in 4 dairy herds in Iowa in which Johne's disease was diagnosed previously. Fecal specimens were collected at 6-month intervals from animals 2 years of age and over for mycobacteriologic examination. Serum samples were obtained at 3-month intervals and tested by enzyme-linked immunosorbent assay (ELISA). The antigen used in the ELISA was a potassium chloride extract of a field strain of Mycobacterium paratuberculosis. The ELISA reactions were observed in 87% of the cows from which M. paratuberculosis was isolated. Dairy producers that participated in the Johne's control program reported reduced economic losses. Increased income was attributed to improved milk production, increased value of vaccinated animals sold as replacements to other dairy herds in which Johne's disease had been diagnosed, and the increased market value of slaughter animals removed from the herd.

scholarly journals Identification of Novel Seroreactive Antigens in Johne's Disease Cattle by Using the Mycobacterium tuberculosis Protein Array

2017 ◽  
Vol 24 (7) ◽  
Author(s):  
John P. Bannantine ◽  
Joseph J. Campo ◽  
Lingling Li ◽  
Arlo Randall ◽  
Jozelyn Pablo ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
The United States ◽  
Johne's Disease ◽  
Protein Array ◽  
Johne’S Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Content Type ◽  
A Genome ◽  
Elisa Testing ◽  
Close Phylogenetic Relationship

ABSTRACT Johne's disease, a chronic gastrointestinal inflammatory disease caused by Mycobacterium avium subspecies paratuberculosis, is endemic in dairy cattle and other ruminants worldwide and remains a challenge to diagnose using traditional serological methods. Given the close phylogenetic relationship between M. avium subsp. paratuberculosis and the human pathogen Mycobacterium tuberculosis, here, we applied a whole-proteome M. tuberculosis protein array to identify seroreactive and diagnostic M. avium subsp. paratuberculosis antigens. A genome-scale pairwise analysis of amino acid identity levels between orthologous proteins in M. avium subsp. paratuberculosis and M. tuberculosis showed an average of 62% identity, with more than half the orthologous proteins sharing >75% identity. Analysis of the M. tuberculosis protein array probed with sera from M. avium subsp. paratuberculosis-infected cattle showed antibody binding to 729 M. tuberculosis proteins, with 58% of them having ≥70% identity to M. avium subsp. paratuberculosis orthologs. The results showed that only 4 of the top 40 seroreactive M. tuberculosis antigens were orthologs of previously reported M. avium subsp. paratuberculosis antigens, revealing the existence of a large number of previously unrecognized candidate diagnostic antigens. Enzyme-linked immunosorbent assay (ELISA) testing of 20 M. avium subsp. paratuberculosis recombinant proteins, representing reactive and nonreactive M. tuberculosis orthologs, further confirmed that the M. tuberculosis array has utility as a screening tool for identifying candidate antigens for Johne's disease diagnostics. Additional ELISA testing of field serum samples collected from dairy herds around the United States revealed that MAP2942c had the strongest seroreactivity with Johne's disease-positive samples. Collectively, our studies have considerably expanded the number of candidate M. avium subsp. paratuberculosis proteins with potential utility in the next generation of rationally designed Johne's disease diagnostic assays.

scholarly journals Evaluation of Result Variability with a Commercial Johne's Disease Enzyme-Linked Immunosorbent Assay Kit and Repeat Testing of Samples

2002 ◽  
Vol 14 (5) ◽  
pp. 423-426 ◽  
Author(s):  
John M. Adaska ◽  
Claudia A. Muñoz-Zanzi ◽  
Sharon K. Hietala
Keyword(s):  
Immunosorbent Assay ◽  
Coefficient Of Variation ◽  
Mycobacterium Avium Subspecies Paratuberculosis ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Large Animal ◽  
Test Results ◽  
Elisa Kit ◽  
Individual Test

Two hundred one serum samples from individual dairy cows with a range of results on initial testing with a commercial Johne's disease enzyme-linked immunosorbent assay (ELISA) kit were repeat tested 5 times in each of 2 laboratories with kits produced by the same manufacturer. The results for the samples with all 10 replicates showed that the values for individual samples often had a coefficient of variation greater than 20%. As expected, the standard deviation for the results increased as the average value increased and the coefficient of variation was greater in samples with low mean values. The different lots of the commercial ELISA kit used in this study had a significant effect on both the optical density and the calculated sample to positive (S/P) ratio for test replicates. Based on the variability detected in S/P ratios of replicate samples, application of a single cutoff point to interpret individual test results as positive or negative for antibodies to Mycobacterium avium subspecies paratuberculosis could result in inconsistent classification of animals as positive or negative for Johne's disease. Such inconsistency in test interpretation leads to frustration in large animal veterinarians or producers trying to make management decisions based on individual test results. Instead of dichotomizing the test results as positive or negative based on a single cutoff value, reporting numerical values and supplying a classification scheme that includes a suspect category reflecting the uncertainty inherent in the test is recommended to provide more reliable result interpretation.

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