scholarly journals Mycobacterium avium subsp. paratuberculosis ELISA Responses in Milk Samples from Vaccinated and Nonvaccinated Dairy Goat Herds in The Netherlands

2019 ◽  
Vol 6 (2) ◽  
pp. 58 ◽  
Author(s):  
Saskia Luttikholt ◽  
Karianne Lievaart-Peterson ◽  
Maaike Gonggrijp ◽  
Marian Aalberts ◽  
Gerdien van Schaik ◽  
...  
Keyword(s):  
The Netherlands ◽  
Mycobacterium Avium ◽  
Relative Sensitivity ◽  
Herd Size ◽  
Mycobacterium Avium Subsp ◽  
Dairy Goat ◽  
Dairy Goats ◽  
Milk Samples ◽  
Goat Population ◽  
Mycobacterium Avium Subsp Paratuberculosis

The aims of our study were to calculate the most appropriate cut-off value for milk samples in a serum-validated Mycobacterium avium subsp. paratuberculosis (MAP) ELISA and to analyze MAP ELISA responses in milk samples from vaccinated and nonvaccinated dairy goats in the Netherlands. Analyzed herds were representative for location and herd size of dairy goat herds in the Netherlands. A significantly higher proportion of the analyzed 49 herds were organic as compared with the total Dutch dairy goat population. First, the MAP ELISA was optimized using 992 paired serum and milk samples. At a cut-off of 25 S/P%, the relative sensitivity (Se) was 58.4% (n = 992, 95% CI: 48.8%−67.6%) and relative specificity (Sp) was 98.5% (n = 992, 95% CI: 97.5%−99.2%), as compared to serum ELISA results. The percentage of positively tested herds was 78.2% (n = 49, 95% CI: 63.4%−88.1%). The percentage of positive milk samples per herd (n = 22) was on average 4.6% (median, min, and max of 4.7%, 0.0%, and 10.7%, respectively). Average age of ELISA-positive (3.2 years) and -negative goats (3.2 years) was not different. Significantly more vaccinated goats tested positive (6.7%) as compared with nonvaccinated goats (1.1%). This study shows that a high number of vaccinated and nonvaccinated commercial dairy goat herds in the Netherlands have MAP-ELISA-positive goats.

scholarly journals Mycobacterium avium Subspecies paratuberculosis DNA and Antibodies in Dairy Goat Colostrum and Milk

2019 ◽  
Vol 6 (4) ◽  
pp. 96 ◽  
Author(s):  
Karianne Lievaart-Peterson ◽  
Saskia Luttikholt ◽  
Maaike Gonggrijp ◽  
Robin Ruuls ◽  
Lars Ravesloot ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Mycobacterium Avium Subspecies Paratuberculosis ◽  
Clinical Signs ◽  
Dairy Goat ◽  
Economic Losses ◽  
Dairy Goats ◽  
Convenience Sample ◽  
Milk Samples ◽  
Zoonotic Risk ◽  
Goat Population

Mycobacterium avium subspecies paratuberculosis (MAP) is endemic in the Dutch dairy goat population causing economic loss, and negatively influencing welfare. Moreover, there are concerns about a potential zoonotic risk. Therefore the industry’s objectives are to decrease MAP prevalence, limit economic losses as well as reduce the concentration of MAP in (bulk) milk. To diminish within-farm spread of infection, vaccination, age dependent group housing with separation of newborns from adults, as well as rearing on artificial or treated colostrum and milk replacers are implemented. However, the importance of MAP contaminated colostrum and milk as a route of infection in dairy goat herds is unknown. Therefore the aim of this study was to detect the presence of MAP DNA in colostrum and milk from dairy goats in infected herds. A convenience sample of 120 colostrum samples and 202 milk samples from MAP infected dairy goat herds were tested by IS900 real-time Polymerase Chain Reaction (PCR) for MAP DNA. Furthermore, 22 colostrum samples and 27 post mortem milk samples of goats with clinical signs consistent with paratuberculosis from known infected herds were tested. The majority of samples were from goats vaccinated against MAP. Positive or doubtful PCR results were obtained in none of the 120 and two of the 22 colostrum samples, and in eight of the 202 and four of the 27 milk samples Negative PCR results were obtained in the remaining 140 (99%) colostrum samples and 217 (95%) milk samples.

scholarly journals Molecular typing of Mycobacterium avium subsp. paratuberculosis (MAP) isolated from dairy goats in Brazil

2016 ◽  
Vol 140 ◽  
pp. 18-21 ◽  
Author(s):  
Marina de Castro Campos de Souza ◽  
Magna Coroa Lima ◽  
Isis de Freitas Espeschit Braga ◽  
David Germano Gonçalves Schwarz ◽  
Ana Paula de Souza Rodrigues ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Molecular Typing ◽  
Mycobacterium Avium Subsp ◽  
Dairy Goats ◽  
Mycobacterium Avium Subsp Paratuberculosis

Detection of Mycobacterium avium subsp. paratuberculosis (MAP) from Subclinical Caprine Paratuberculosis Cases of Odisha

2020 ◽  
Author(s):  
Sangram Biswal ◽  
Adya Prakash Rath ◽  
Shoor Vir Singh ◽  
Niranjana Sahoo ◽  
Saurabh Gupta ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Animal Health ◽  
Restriction Endonuclease Analysis ◽  
Mycobacterium Avium Subsp ◽  
Serum Samples ◽  
Blood Samples ◽  
Blind Review ◽  
Faecal Samples ◽  
Goat Population ◽  
Mycobacterium Avium Subsp Paratuberculosis

Paratuberculosis is caused by Mycobacterium avium subsp. Paratuberculosis (MAP) and is a chronic, intestinal tract infection in the ruminant sector globally. A total of 122 EDTA mixed blood samples, 121 serum and 16 pooled faecal samples were collected from farms of 4 different districts i.e. Nayagarh, Cuttack, Khordha and Angul and a blind review was conducted at the Animal Health Division, CIRG, Mathura. Microscopic examination of 16 pooled faecal samples revealed +2 reactivity to Acid-Fast Bacilli. All the serum samples were subjected to indirect ELISA. Out of them, 23 (19.01%), 85 (70.25%), shows strongly positive, positive, antibody titre respectively. EDTA blood samples of 23 ELISA-strongly positive were subjected to 413 bp IS900 PCR and 11 (9%) of them were found positive for Mycobacterium avium subsp. Paratuberculosis (MAP). MAP isolates were further subjected to genotyping using 608 bpIS1311 PCR and restriction endonuclease analysis (IS1311 PCR-REA) and 2 (1.64%) of them matched with “Indian Bison Type”. Genotyping of the isolates using IS1311 PCR-REA revealed that goat population of Odisha are primarily infected with “Indian Bison Type” strains.

scholarly journals Effects of freezing on ability to detect Mycobacterium avium subsp. paratuberculosis from bovine tissues following culture

2018 ◽  
Vol 30 (5) ◽  
pp. 743-746
Author(s):  
Caroline S. Corbett ◽  
Jeroen De Buck ◽  
Herman W. Barkema
Keyword(s):  
Mycobacterium Avium ◽  
Gold Standard ◽  
Frozen Storage ◽  
Mycobacterium Avium Subsp ◽  
Freezing And Thawing ◽  
Tissue Samples ◽  
Milk Samples ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Autopsy Tissues ◽  
Numerical Change

Mycobacterium avium subsp. paratuberculosis (MAP) is the bacterium that causes Johne’s disease in cattle. Although infected cattle can be identified by examining fecal, blood, or milk samples, the gold standard is identification of MAP in tissue samples postmortem. Although tissue samples are commonly frozen, the ability to detect MAP in frozen–thawed tissue samples has apparently not been reported. We therefore determined the ability to detect MAP in tissue samples following freezing. Tissue samples were collected from calves that were either inoculated (IN) 3 mo prior, or contact-exposed (CE) for 3 mo. Following autopsy, tissues were immediately processed for culture, followed by DNA extraction and detection by qPCR. Samples were categorized as positive or negative based on the cycle threshold (Ct) value. The remaining unprocessed tissue samples were frozen at −80°C. After 18 mo, 50 tissue samples designated MAP-positive were thawed and processed for detection of MAP. Four (8%) samples were qPCR-negative, and Ct values of the remaining 46 samples were higher after freezing. Given the small numerical change in Ct values for MAP-positive samples after 18 mo of frozen storage, freezing and thawing may have had some deleterious effects on MAP detection in tissues. Although the decrease in ability to detect MAP-positive samples was minor for IN calves, there may be a greater effect for CE calves that should be considered when freezing tissue samples.

scholarly journals A Comparative Study on the Efficiency of Two Mycobacterium avium subsp. paratuberculosis (MAP)-Derived Lipopeptides of L3P and L5P as Capture Antigens in an In-House Milk ELISA Test

Vaccines ◽  
2021 ◽  
Vol 9 (9) ◽  
pp. 997
Author(s):  
Sepideh Hosseiniporgham ◽  
Franck Biet ◽  
Christelle Ganneau ◽  
John P. Bannantine ◽  
Sylvie Bay ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Elisa Test ◽  
Cross Reactivity ◽  
Mycobacterium Avium Subsp ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Individual Level ◽  
Milk Samples ◽  
Milk Elisa ◽  
Mycobacterium Avium Subsp Paratuberculosis

Mycobacterium avium subsp. paratuberculosis (MAP) surface-exposed lipopeptides could be specific capture-antigen molecules targeting antibodies against MAP, in milk, through ELISA. Previous studies have revealed that MAP strains, isolated from sheep (S) or cow (C), could produce specific lipopeptides, L3P or L5P, respectively. In this study, we used L3P and L5P as capture antigens in an in-house milk ELISA (H-MELISA) to assess how these antigens perform, in comparison with other ELISA tests, on well-defined milk samples from MAP-infected sheep. The overall positivity rates of H-MELISA via L3P and L5P varied by the source of milk samples, in which, at bulk-tank-milk (BTM) level, the majority of positive cases (63.83%) reacted more against L5P, whereas a predominant number (69.14%) of milk samples were more responsive against L3P at the individual level. To clarify whether the positivity status of milk samples in H-MELISA L3P/L5P were predictive of MAP strain-types (S/C), strain-typing was carried out using PCR IS1311-restriction enzyme analysis. Although the presence of three MAP strains (S/C/bison types) was detected among the milk samples, the C-type (46.67%) and S-type (75%) MAP strains were detected with higher incidence among BTMs and individual milk samples, respectively. However, further examination on the H-MELISA L3P/L5P-positivity pattern of each C/S-type-MAP sample revealed that some samples had a reverse reactivity against both L3P and L5P. These results could be the consequence of either cross-reactivity between L3P and L5P (due to the similarity in the structures of the two epitopes) or simply a within-herd mixed infection with MAP strains of C and S types. These findings suggest that lipopeptide antigens could contribute a diagnostic test with optimal performance, considering the diversity of MAP strains.

scholarly journals A rapid phage assay for detection of viable Mycobacterium avium subsp. paratuberculosis in milk

2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Sepideh Hosseiniporgham ◽  
Lucio Rebechesu ◽  
Pierangela Pintore ◽  
Stefano Lollai ◽  
Maria Dattena ◽  
...  
Keyword(s):  
Magnetic Separation ◽  
Mycobacterium Avium ◽  
Limit Of Detection ◽  
Goat Milk ◽  
Mycobacterium Avium Subsp ◽  
Milk Samples ◽  
Sheep Milk ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Higher Sensitivity

AbstractParatuberculosis is an incurable gastroenteritis among ruminants that is promoted by Mycobacterium avium subsp. paratuberculosis (MAP), an acid-fast mycobacterium. To accelerate the detection of viable pathogen, a conventional (peptide mediated magnetic separation: PMS) and novel (phage-bead qPCR: PBQ) phage based assay was optimized. A superior limit of detection (LOD) of 10 MAP per 10 mL milk was suggested for PBQ compared to 100 cells/10 mL for PMS-phage assay. Via PBQ, viable MAP was found in 48.78% out 41 unpasteurized sheep and goat milk samples. Sheep milk samples (n = 29) that were tested by PMS-phage assay contained no viable MAP. The absence of viable MAP in milk collected from 21 of the recent sheep animals was also confirmed by PBQ after a 2-week gap. Although, the two phage assays comparably detected no viable MAP in the milk samples, MAP DNA and antibodies against MAP were recognized in milk and sera of some of these animals within two instances of sampling representing that some sheep animals were MAP shedders. In conclusion, PBQ and PMS-phage could be promising methods for the assessment of MAP viability in milk samples. However, PBQ was privileged over the PMS-phage assay due to the lower LOD, rapidity, higher sensitivity, lack of need to M. smegmatis and consequent virucidal treatment that are essential in PMS-phage assay for making lawn and inactivation of exogenous mycobacteriophages respectively.

scholarly journals Effects of Age and Environment on Adaptive Immune Responses to Mycobacterium avium subsp. paratuberculosis (MAP) Vaccination in Dairy Goats in Relation to Paratuberculosis Control Strategies

2019 ◽  
Vol 6 (3) ◽  
pp. 62 ◽  
Author(s):  
Ad Koets ◽  
Lars Ravesloot ◽  
Robin Ruuls ◽  
Annemieke Dinkla ◽  
Susanne Eisenberg ◽  
...  
Keyword(s):  
Immune Responses ◽  
Mycobacterium Avium ◽  
Diagnostic Tests ◽  
Control Strategies ◽  
Mycobacterium Avium Subsp ◽  
Dairy Goats ◽  
Adaptive Immune Responses ◽  
Adaptive Immune ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Free Environment

Paratuberculosis infection is caused by Mycobacterium avium subsp. paratuberculosis (MAP). In the Netherlands, 75% herd level prevalence of caprine paratuberculosis has been estimated, and vaccination is the principal control strategy applied. Most goat dairy farms with endemic paratuberculosis systematically vaccinate goat kids in the first months of life with a commercially available whole cell MAP vaccine. We hypothesized that the development of adaptive immune responses in goats vaccinated at young age depends on the environment they are raised in, and this has implications for the application of immune diagnostic tests in vaccinated dairy goats. We evaluated the early immune response to vaccination in young goat kids sourced from a MAP unsuspected non-vaccinated herd and raised in a MAP-free environment. Subsequently we compared these with responses observed in birth year and vaccination matched adult goats raised on farms with endemic paratuberculosis. Results indicated that initial adaptive immune responses to vaccination are limited in a MAP-free environment. In addition, adult antibody positive vaccinated goats raised in a MAP endemic environment are less likely to be IS900 PCR-positive as compared to antibody negative herd mates. We conclude that test-and-cull strategies in a vaccinated herd are currently not feasible using available immune diagnostic tests.

Modeling the Occurrence of Mycobacterium avium subsp. paratuberculosis in Bulk Raw Milk and the Impact of Management Options for Exposure Mitigation

2011 ◽  
Vol 74 (7) ◽  
pp. 1126-1136 ◽  
Author(s):  
CHRISTOPHE BOULAIS ◽  
RON WACKER ◽  
JEAN-CHRISTOPHE AUGUSTIN ◽  
MOHAMED HEDI BEN CHEIKH ◽  
FABRICE PELADAN
Keyword(s):  
Mycobacterium Avium ◽  
Raw Milk ◽  
Mycobacterium Avium Subsp ◽  
Dairy Herds ◽  
Management Options ◽  
Scientific Debate ◽  
Milk Samples ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Exposure Mitigation ◽  
The Impact

Mycobacterium avium subsp. paratuberculosis (MAP) is the causal agent of paratuberculosis (Johne's disease) in cattle and other farm ruminants. The potential role of MAP in Crohn's disease in humans and the contribution of dairy products to human exposure to MAP continue to be the subject of scientific debate. The occurrence of MAP in bulk raw milk from dairy herds was assessed using a stochastic modeling approach. Raw milk samples were collected from bulk tanks in dairy plants and tested for the presence of MAP. Results from this analytical screening were used in a Bayesian network to update the model prediction. Of the 83 raw milk samples tested, 4 were positive for MAP by culture and PCR. We estimated that the level of MAP in bulk tanks ranged from 0 CFU/ml for the 2.5th percentile to 65 CFU/ml for the 97.5th percentile, with 95% credibility intervals of [0, 0] and [16, 326], respectively. The model was used to evaluate the effect of measures aimed at reducing the occurrence of MAP in raw milk. Reducing the prevalence of paratuberculosis has less of an effect on the occurrence of MAP in bulk raw milk than does managing clinically infected animals through good farming practices.

scholarly journals Pooled Fecal Culture Sampling for Mycobacterium Avium Subsp. Paratuberculosis at Different Herd Sizes and Prevalence

2003 ◽  
Vol 15 (3) ◽  
pp. 233-241 ◽  
Author(s):  
G. van Schaik ◽  
S. M. Stehman ◽  
Y. H. Schukken ◽  
C. R. Rossiter ◽  
S. J. Shin
Keyword(s):  
Mycobacterium Avium ◽  
Disease Status ◽  
Herd Size ◽  
Pool Size ◽  
Mycobacterium Avium Subsp ◽  
Fecal Culture ◽  
Spreadsheet Model ◽  
High Prevalence ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Low Prevalence

A stochastic spreadsheet model was developed to obtain estimates of the costs of whole herd testing on dairy farms for Mycobacterium avium subsp. paratuberculosis ( Map) with pooled fecal samples. The optimal pool size was investigated for 2 scenarios, prevalence (a low-prevalence herd [≤5%] and a high-prevalence herd [≤5%]) and for different herd sizes (100-, 250-, 500- and 1,000-cow herds). All adult animals in the herd were sampled, and the samples of the individuals were divided into equal sized pools. When a pool tested positive, the manure samples of the animals in the pool were tested individually. The individual samples from a negative pool were assumed negative and not tested individually. Distributions were used to model the uncertainty about the sensitivity of the fecal culture at farm level and Map prevalence. The model randomly allocated a disease status to the cows (not shedding, low Map shedder, moderate Map shedder, and heavy Map shedder) on the basis of the expected prevalence in the herd. Pooling was not efficient in 100-cow and 250-cow herds with low prevalence because the probability to detect a map infection in these herds became poor (53% and 88%) when samples were pooled. When samples were pooled in larger herds, the probability to detect at least 1 (moderate to heavy) shedder was ≤90%. The cost reduction as a result of pooling varied from 43% in a 100-cow herd with a high prevalence to 71% in a 1,000-cow herd with a low prevalence. The optimal pool size increased with increasing herd size and varied from 3 for a 500-cow herd with a low prevalence to 5 for a 1,000-cow herd with a high prevalence.

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