A semi-quantitative enzyme-linked immunosorbent assay for Rathayibacter toxicus, the bacterium involved in annual ryegrass toxicity, to assist in risk assessment of fodder for domestic use

2014 ◽  
Vol 65 (12) ◽  
pp. 1329 ◽  
Author(s):  
A. M. Masters ◽  
B. Samarasinghe ◽  
M. Kalkhoven ◽  
L. den Hollander ◽  
D. G. Palmer
Keyword(s):  
Immunosorbent Assay ◽  
Animal Health ◽  
Clinical Signs ◽  
Risk Level ◽  
Enzyme Linked Immunosorbent Assay ◽  
Annual Ryegrass ◽  
Department Of Agriculture ◽  
Domestic Livestock ◽  
Annual Ryegrass Toxicity ◽  
Risk Categories

A semi-quantitative enzyme-linked immunosorbent assay (ELISA) for the detection of Rathayibacter toxicus in hay or pasture was used to estimate the degree of R. toxicus bacterial gall contamination in hay or pasture that was unsuitable for export but that may be suitable for feeding to domestic livestock. Based on experience of testing of fodder samples from pastures where livestock showed clinical signs, or outbreaks of annual ryegrass toxicity (ARGT), four relevant levels of bacterial gall contamination were selected. Several 1-kg samples of hay with no contamination by R. toxicus were spiked with these four different amounts of bacterial galls to provide five different risk categories considered relevant to the occurrence of ARGT. Extracts of the spiked samples were assayed to determine the range of ELISA results to be expected in each category. To validate these risk categories, all cases of ARGT diagnosed between 2000 and 2012 by the Animal Health Laboratories, Department of Agriculture and Food Western Australia, associated with the submission of suspected toxic fodder were allocated to the risk categories on the basis of the recorded results for fodder. In ~15% and 79% of all cases, the fodder associated with outbreaks of ARGT fell into the ‘moderate risk’ and ‘high risk’ categories, respectively. The selected categories were considered to provide realistic estimations of the risk that fodder within them might cause ARGT if fed to livestock. This risk-level reporting has been adopted to enable informed decision making as to whether feeding a particular batch of hay or fodder to animals represents an acceptable risk.

An enzyme-linked immunosorbent assay for the detection of Rathayibacter toxicus, the bacterium involved in annual ryegrass toxicity, in hay

2006 ◽  
Vol 57 (7) ◽  
pp. 731 ◽  
Author(s):  
A. M. Masters ◽  
A. R. Gregory ◽  
R. J. Evans ◽  
J. E. Speijers ◽  
S. S. Sutherland
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Specific Antigen ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Annual Ryegrass ◽  
Capture Assay ◽  
Large Numbers ◽  
Soluble Polysaccharide ◽  
Annual Ryegrass Toxicity

An enzyme-linked immunosorbent assay (ELISA) for Rathayibacter toxicus is described. The development of a monoclonal antibody for a specific antigen from R. toxicus and a polyclonal antibody raised against the same R. toxicus preparation enabled a capture assay format. The assay is specific for a soluble polysaccharide produced by the bacterium and was found to be sensitive enough to detect antigen equivalent to less than one gall per kilogram of hay. The applicability of the assay to samples of pasture or hay is demonstrated. Cost-effective testing of large numbers of samples for the presence of R. toxicus is possible with the ELISA. This will assist stockowners, hay producers, and hay exporters in the management of the risk of annual ryegrass toxicity.

Development and application of polymerase chain reaction-based assays for Rathayibacter toxicus and a bacteriophage associated with annual ryegrass (Lolium rigidum) toxicity

2007 ◽  
Vol 47 (2) ◽  
pp. 177 ◽  
Author(s):  
M. C. Kowalski ◽  
D. Cahill ◽  
T. J. Doran ◽  
S. M. Colegate
Keyword(s):  
Polymerase Chain Reaction ◽  
Immunosorbent Assay ◽  
Potential Application ◽  
South Australia ◽  
Enzyme Linked Immunosorbent Assay ◽  
Annual Ryegrass ◽  
Lolium Rigidum ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Annual Ryegrass Toxicity

Annual ryegrass toxicity (ARGT) is responsible for significant stock losses in South Australia and Western Australia. The toxicity is caused by corynetoxins produced by the bacterium Rathayibacter toxicus (with the possible involvement of a bacteriophage), which infects annual ryegrass (Lolium rigidum). Polymerase chain reaction (PCR)-based assays, compatible with an existing enzyme-linked immunosorbent assay for the corynetoxins, have been developed and used to screen L. rigidum for both the presence of R. toxicus and for the bacteriophage isolate NCPPB 3778. The results from analysing bacterially infected galls from toxic grain screenings showed a positive correlation between the presence of the bacterium and corynetoxins but not with the bacteriophage. Analysis of pasture-derived samples of annual ryegrass showed about a 50% correlation of corynetoxins with bacterial presence and about a 5% correlation of phage with the presence of the bacterium. These observations support the potential application of the PCR-based assays in providing a useful, complementary tool in the assessment of the likelihood of pasture and feed to cause ARGT and to enable a better understanding of the complex aetiology of ARGT.

scholarly journals Interpretations of Antibody Responses toSalmonella enterica Serotype Enteritidis gm Flagellin in Poultry Flocks Are Enhanced by a Kinetics-Based Enzyme-Linked Immunosorbent Assay

1998 ◽  
Vol 5 (4) ◽  
pp. 550-555 ◽  
Author(s):  
Patrick L. McDonough ◽  
Richard H. Jacobson ◽  
John F. Timoney ◽  
Ahmed Mutalib ◽  
David C. Kradel ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
High Sensitivity ◽  
High Specificity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Sensitivity ◽  
Department Of Agriculture ◽  
Trace Back ◽  
Elisa Format ◽  
Commercial Poultry ◽  
Computer Controlled

ABSTRACT Many regulatory and diagnostic programs for the detection ofSalmonella enterica serotype Enteritidis infection in commercial poultry flocks have relied on rapid Pullorum agglutination tests to screen birds because of the shared antigens of S. enterica Enteritidis and S. enterica Pullorum and Gallinarum; however, the use of the enzyme-linked immunosorbent assay (ELISA) format affords better analytical sensitivity than crude agglutination tests. In this study, we adapted our earlier conventional indirect ELISA, using gm flagellin as the antigen, to a kinetics-based, computer-controlled ELISA (KELA). The KELA was used to screen for flagellin antibody from three commercial flocks: (i) a large flock involved in a U.S. Department of Agriculture trace back from a humanS. enterica Enteritidis foodborne outbreak (n = 3,209), (ii) a flock infected with the endemicS. enterica Enteritidis serotype but which also had multiple other salmonella serotypes (n = 65), and (iii) an S. enterica Pullorum-infected flock (n = 12). The first flock (S. entericaEnteritidis prevalence of 2.45% based on culture) provided a field test of the KELA and allowed the calculation of diagnostic sensitivity (D-Sn) and diagnostic specificity (D-Sp). With a cutoff of 10 (used for screening flocks [i.e., high sensitivity]), the KELA has a D-Sn of 95.2% and a D-Sp of 18.5%; with a cutoff of 140 (used in confirmatory flock testing [i.e., high specificity]), the KELA has a D-Sn of 28.0% and a D-Sp of 99.1%. We found that with a cutoff of 60 (D-Sn = 63.1%; D-Sp = 91.6%), we could eliminate reactions in the KELA caused by other non-S. enterica Enteritidis salmonellae. The KELA was also compared to two commercial rapid Pullorum tests, the Solvay (D-Sn = 94.9%; D-Sp = 55.5%) and the Vineland (D-Sn = 62.0%; D-Sp = 75.3%).

scholarly journals Detection of Antigenemia by Enzyme-Linked Immunosorbent Assay in Horses with Experimental Ehrlichia Risticii Infection

1993 ◽  
Vol 5 (1) ◽  
pp. 33-36 ◽  
Author(s):  
Richard E. Corstvet ◽  
Stephen D. Gaunt ◽  
Phillip A. Karns ◽  
David Burgermeister ◽  
Jere W. McBride ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Detection Method ◽  
Serum Antibody ◽  
Clinical Signs ◽  
Clinical Findings ◽  
Antigen Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infected Horse ◽  
Ehrlichia Risticii

Four horses were inoculated with Ehrlichia risticii contained in either infected murine P388 D1 cells or heparinized blood from an infected horse. All 4 horses produced serum antibody, plasma antigen, and clinical signs of the disease. An enzyme-linked immunosorbent assay was used to detect antibody in the serum and was also used in conjunction with an anti- E. risticii monoclonal antibody to detect antigenemia. These laboratory and clinical findings were correlated to determine the efficiency of the antigen detection method for discerning E. risticii infection.

scholarly journals A serosurvey of bluetongue and epizootic haemorrhagic disease in a convenience sample of sheep and cattle herds in Zimbabwe

2017 ◽  
Vol 84 (1) ◽  
Author(s):  
Stuart J.G. Gordon ◽  
Charlotte Bolwell ◽  
Chris W. Rogers ◽  
Godfrey Musuka ◽  
Patrick Kelly ◽  
...  
Keyword(s):  
Health Workers ◽  
Animal Health ◽  
Enzyme Linked Immunosorbent Assay ◽  
Convenience Sample ◽  
Domestic Livestock ◽  
Cattle Herds ◽  
Haemorrhagic Disease ◽  
First Time ◽  
Culicoides Vectors ◽  
Cattle And Sheep

A convenience sample of sheep and cattle herds around the cities of Harare, Kwekwe and Bulawayo, located in the Highveld region of Zimbabwe, was used to estimate the seroprevalence and sero-incidence of bluetongue virus (BTV) and epizootic haemorrhagic disease virus (EHDV) antibodies. A competitive enzyme-linked immunosorbent assay was used to identify serum antibodies against BTV and EHDV across three rainy seasons. The median sero-prevalence of BTV and EHDV antibodies in cattle was 62% (interquartile range [IQR]: 30–89) and 56% (IQR: 5–77), respectively. In sheep, the median sero-prevalence of BTV and EHDV was 41% (IQR: 19–63) and 0% (IQR: 0–21), respectively. Median sero-incidences of BTV and EHDV antibodies in cattle of 43% (IQR: 22–67) and 27% (IQR: 9–57) respectively were recorded. The median sero-incidence of BTV in sheep was 14% (IQR: 6–23). Based on these preliminary findings, animal health workers in Zimbabwe should continue to monitor the exposure rates of cattle and sheep to BTV and consider the possibility of strains emerging with increased pathogenicity. There are no previous published reports of antibodies against EHDV in Zimbabwe so the possibility of epizootic haemorrhagic disease existing in domestic livestock should now be considered by Zimbabwean animal health officials. Seroconversions to BTV and EHDV occurred predominantly at the end of each rainy season (March and April), which generally corresponds to high numbers of the Culicoides vectors. BTV isolations were made from three individual cows in two of the sentinel herds and all three were identified as serotype 3. This is the first time BTV serotype 3 has been recorded in Zimbabwe, although its presence in neighbouring South Africa is well documented.

scholarly journals Comparative examination of the serological response to bluetongue virus in diseased ruminants by competitive and double recognition enzime-linked immunosorbent assays

2018 ◽  
Vol 69 (3) ◽  
pp. 1088
Author(s):  
A. GAVRILOVIĆ ◽  
P. GAVRILOVIĆ ◽  
S. RADOJIČIĆ ◽  
D. KRNJAIĆ
Keyword(s):  
Blood Serum ◽  
Immunosorbent Assay ◽  
Clinical Signs ◽  
High Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Response ◽  
Rapid Diagnostics ◽  
Serum Samples ◽  
Perfect Agreement ◽  
First Time

Bluetongue (BT) is a viral non-contagious disease of ruminants which is transmitted by insects of the genus Culicoides. In recent years, BT has been a serious threat to livestock and to the economies of European countries. In Serbia the disease appeared for the first time in 2001, and after a 12 year period of freedom, it broke out again in 2014. Considering the actuality of this infectious disease, especially the need for prompt and rapid diagnostics, the aim of this paper was to determine the possibility of detecting the serological response in sheep and cattle with manifested clinical signs of the disease using two different methods: double recognition enzyme-linked immunosorbent assay (sELISA) and competitive enzyme-linked immunosorbent assay (cELISA). A total of 105 blood serum samples of cattle and sheep, which had exhibited clinical signs of BT during 2014, were taken for examination from a serum bank. Out of 74 blood serum samples of sheep and 31 blood serum samples of cattle, 52 samples of sheep and 18 samples of cattle tested positive using sELISA, while 50 samples of sheep and 18 samples of cattle gave positive reactions with cELISA. The results confirm the high sensitivity of sELISA which detected 4% more seropositive sheep in comparison with cELISA. Using Cohen’s kappa statistical analysis, almost perfect agreement was determined between the results (k>0,81) obtained by cELISA and sELISA.

Adaptation and evaluation of an ELISA for Trypanosoma evansi infection (surra) in elephants and its application to a serological survey in Thailand

Parasitology ◽  
2017 ◽  
Vol 145 (3) ◽  
pp. 371-377 ◽  
Author(s):  
MARGOT CAMOIN ◽  
ARTHUR KOCHER ◽  
PIANGJAI CHALERMWONG ◽  
SARAWUT YANGTARRA ◽  
NIKORN THONGTIP ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Trypanosoma Evansi ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Survey ◽  
Diminazene Aceturate ◽  
Survey Period ◽  
Frequent Relapses ◽  
Close Proximity ◽  
Positive Threshold ◽  
Domestic Livestock

SUMMARYTrypanosoma evansi, the causative agent of surra, is widespread in domestic livestock and wildlife in South East Asia. Surra can affect cattle, buffaloes, horses and also Asian elephants (Elephas maximus). Despite the ‘threatened to extinction’ CITES status of elephant, surra's impact has not been thoroughly assessed yet in this species. This work offers to adapt an antibody enzyme-linked immunosorbent assay (ELISA) protocol, to detect Trypanosoma evansi antibodies in elephant serum. The test was validated with 365 negative-reference samples, which allowed the determination of a 16% positive threshold. The test was applied to a serological survey including 375 individuals. The estimated global seroprevalence was 2·1% (95% CI 1·1–4·2%). Therefore, surra does not appear to be endemic in Thai domestic elephants, but occasional outbreaks were reported to our laboratory during the survey period. These outbreaks seemed to be linked to close proximity to cattle or buffaloes, and led to severe clinical signs in elephants. Frequent relapses were observed after treatment with diminazene aceturate, the only trypanocide drug currently available in Thailand. Therefore, care should be taken to keep elephants away from bovine reservoirs, and to monitor the disease in this endangered species. ELISA proved to be reliable for screening purposes as well as for post-treatment monitoring.

scholarly journals Detection of Anaplasma antibodies in wildlife and domestic species in wildlife-livestock interface areas of Kenya by major surface protein 5 competitive inhibition enzyme-linked immunosorbent assay

2008 ◽  
Vol 75 (3) ◽  
Author(s):  
J.J.N. Ngeranwa ◽  
S.P. Shompole ◽  
E.H. Venter ◽  
A. Wambugu ◽  
J.E. Crafford ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Competitive Inhibition ◽  
Surface Membrane ◽  
Clinical Signs ◽  
Surface Protein ◽  
Domestic Animal ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Domestic Species ◽  
Major Surface

The seroprevalence of Anaplasma antibodies in wildlife (eland, blue wildebeest, kongoni, impala, Thomson's gazelle, Grant's gazelle, giraffe and plains zebra) and domestic animal (cattle, sheep and goat) populations was studied in wildlife / livestock interface areas of Kenya. Serum samples were analyzed by competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA), using a recombinant antigen (MSP-5) from Anaplasma marginale surface membrane. A monoclonal antibody, FC-16, was used as the primary antibody, while anti-mouse conjugated to horseradish peroxidase was used as the secondary antibody. The results indicate a high seroprevalence in both wildlife and livestock populations, in contrast to earlier reports from Kenya, which indicated a low seroprevalence. The differences are attributed to the accurate analytical method used (CI-ELISA), as compared with agglutination techniques, clinical signs and microscopy employed by the earlier workers.

Lactate Dehydrogenase as Safe Endpoint Cooking Indicator in Poultry Breast Rolls: Development of Monoclonal Antibodies and Application to Sandwich Enzyme-Linked Immunosorbent Assay (ELISA)

1993 ◽  
Vol 56 (2) ◽  
pp. 120-124 ◽  
Author(s):  
MOHAMED M. ABOUZIED ◽  
CHENG HSING WANG ◽  
JAMES J. PESTKA ◽  
DENISE M. SMITH
Keyword(s):  
Monoclonal Antibodies ◽  
Lactate Dehydrogenase ◽  
Immunosorbent Assay ◽  
Polyclonal Antibodies ◽  
Sandwich Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Department Of Agriculture ◽  
Poultry Products ◽  
Chicken Muscle ◽  
Assay Detection

A sandwich enzyme-linked immunosorbent assay (ELISA) was developed to detect lactate dehydrogenase (LDH) as a marker protein for verifying endpoint cooking of uncured poultry products. Monoclonal antibodies were prepared against chicken muscle LDH and used with rabbit polyclonal antibodies developed against turkey or chicken muscle LDH for capture and detection in the assay, respectively. Minimum assay detection limits for turkey and chicken muscle LDH were 1 ng/ml. Turkey and chicken muscle LDH, but not LDH from other species cross reacted in the ELISA. The ELISA was further verified using extracts of turkey breast rolls processed to internal temperatures between 68.3 and 72.1°C. The LDH content of extracts diluted 3- to 6-fold was below 15 ng/ml for turkey rolls processed to 70.9 and 72.1°C. At a 6-fold dilution, LDH content of extracts from rolls processed to 69.7°C was approximately 10 times greater than those processed to 70.9°C. A survey of market precooked poultry products indicated assay validity with precooked turkey roast, but not turkey hams with maximum internal temperature requirements of 68.3°C. Results suggested the sandwich ELISA should be applicable for determining whether turkey breast rolls are processed to the required U.S. Department of Agriculture endpoint temperature of 71.1°C.

Sign in / Sign up

Export Citation Format

Share Document