Outcome of Three Commercial Serum ELISAs and Faecal Detection of Mycobacterium avium subsp. paratuberculosis in Consecutive Samples from a Cattle Herd with Low Prevalence of Paratuberculosis (Johne’s Disease)

Khol, J. L.; Geisbauer, E.; Wassertheurer, M.; Revilla‐Fernández, Sandra; Damoser, J.; Österreicher, E.; Dünser, Michael; Kleb, Ulrike; Baumgartner, Walter

doi:10.1111/j.1865-1682.2011.01256.x

Cited by 8 publications

(4 citation statements)

References 23 publications

(36 reference statements)

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“…When blood samples with serological borderline ELISA per cent values were regarded negative, the agreement between ELISA and faecal snPCR was poor ( r = 0.26, P < 0.05). These results agree with a recent study testing 33 cows suspected to be infected with MAP and 30 cows classified as MAP‐free by three commercial serum ELISAs (Khol et al., 2012). The reason for more frequent positive PCR than serological test results is probably due to paratuberculosis being mainly a local disease in the subclinical phase remaining restricted over years to the gut tissues.…”

Section: Discussionsupporting

confidence: 91%

A Longitudinal Study to Characterize the Distribution Patterns ofMycobacterium aviumssp.paratuberculosisin Semen, Blood and Faeces of a Naturally Infected Bull by IS 900 Semi-Nested and Quantitative Real-Time PCR

Munster

Völkel

Wemheuer

et al. 2012

Transboundary and Emerging Diseases

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Johne's disease is caused by Mycobacterium avium ssp. paratuberculosis (MAP) and has been recognized as an important bacterial infection in ruminants. Although MAP has been detected in semen and within the reproductive organs of bulls, the bacterial distribution and shedding patterns are currently not well characterized. Our investigation was performed to detect and quantify MAP in faeces, semen and blood samples repeatedly drawn from a naturally infected but asymptomatic 18-month-old German Simmental breeding bull candidate over a period of 3 years (June 2007-November 2010). Qualitative and quantitative polymerase chain reaction (PCR) techniques were used to correlate the presence and matrix-specific amounts of MAP. In total, 65 sampling dates were selected. Mycobacterium avium ssp. paratuberculosis was detected intermittently in all matrices with MAP-free intervals of up to 18 weeks by an IS900 semi-nested PCR. The number of MAP-positive results from semen and blood samples was higher than from faecal samples. A quantitative polymerase chain reaction detected the highest MAP contents in faeces (10(3) -10(6) MAP/g), while lower amounts were found in semen and blood samples (10(2) -10(5) MAP/ml). Although no significant agreement was calculated between the presence of MAP in faeces and blood, a statistically significant positive correlation between its occurrence in semen and blood was determined (r = 0.38, P < 0.05, n = 29). The present study contributes to a more detailed understanding of MAP distribution patterns in faeces, semen and blood of a subclinically infected breeding bull candidate. It highlights the possible role of breeding bulls as a source of MAP transmission and indicates the need for further monitoring and hygienic measures to prevent the spread of the infection via semen.

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Section: Discussionsupporting

confidence: 91%

A Longitudinal Study to Characterize the Distribution Patterns ofMycobacterium aviumssp.paratuberculosisin Semen, Blood and Faeces of a Naturally Infected Bull by IS 900 Semi-Nested and Quantitative Real-Time PCR

Munster

Völkel

Wemheuer

et al. 2012

Transboundary and Emerging Diseases

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“…Furthermore, the commercial ELISA used did not show any antibody response throughout the whole investigation period of the bull. These results agree with a recent study, in which 30 of 33 cows were classified as MAP‐free by three commercial serum ELISAs, although their MAP infection was previously confirmed by culture and PCR (Khol et al., ). Such negative ELISA results could be explained by the fact that ELISAs often used in the routine diagnostic of paratuberculosis are highly specific but lack in sensitivity for MAP‐infected animals, especially in the subclinical stage of the disease.…”

Section: Discussionsupporting

confidence: 73%

Distribution ofMycobacterium aviumsubsp.paratuberculosisin a Subclinical Naturally Infected German Fleckvieh Bull

Fechner

Schafer

Wiegel

et al. 2015

Transbound Emerg Dis

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Although it has been known for years that Mycobacterium avium subsp. paratuberculosis (MAP) is detectable in the reproductive organs and semen of infected bulls, only few studies have been conducted on this topic worldwide. This study surveyed the MAP status of a bull, naturally infected due to close contact with its subclinically infected parents over a period of 4 years. From the age of 7 weeks to necropsy, faecal, blood and, after sexual maturity, semen samples were drawn repeatedly. Already at the first sampling day, MAP-DNA was detected in faeces by semi-nested PCR. True infection was confirmed by the detection of MAP-DNA in blood at the age of 40 weeks. In total, MAP-DNA was present in 25% faecal (34/139), 16% blood (23/140) and 5% semen (4/89) samples, including MAP-free intervals of up to 9 weeks. MAP genome equivalents (MAP-GE) of up to 6.3 × 10 /g faeces and 1.8 × 10 /ml blood were determined. Cultivation of MAP occurred only in three of 137 faecal and two of 109 blood, but never in semen samples. Over the whole period, the bull was a serological negative MAP shedder. During necropsy, 42 tissue samples were collected. Neither macroscopic nor histological lesions characteristic of a MAP infection were observed. Cultivation of MAP in tissue sections failed. However, MAP-DNA was spread widely in the host, including in tissues of the lymphatic system (7/15), digestive tract (5/14) and the urogenital tract (5/9) with concentrations of up to 3.9 × 10 MAP-GE/g tissue. The study highlighted the detection of MAP in male reproductive organs and semen. It supports the hypothesis that bulls may probably transmit MAP, at least under natural mating conditions. In artificial insemination, this might not be relevant, due to antibiotics included currently in semen extenders. However, the survivability of MAP in this microenvironment should be investigated in detail.

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“…A limitation of past longitudinal studies is the relative low sampling frequency, which may miss rapid fluctuations that occur in the intervals (typically 6 mo to 1 yr) between sampling events in control plans. Khol et al (2012) performed a monthly follow-up of MAP fecal shedding and antibodies for almost 1 yr to assess the outcome of commercial serology tests. However, the herd under study had a low prevalence (9% by real-time PCR), and more than 80% of the tested cows were MAP-negative in feces.…”

Section: Introductionmentioning

confidence: 99%

Longitudinal study of Mycobacterium avium ssp. paratuberculosis fecal shedding patterns and concurrent serological patterns in naturally infected dairy cattle

Navarro-González

Fourichon

Blanquefort³

et al. 2019

Journal of Dairy Science

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Mycobacterium avium ssp. paratuberculosis (MAP) is the etiological agent of paratuberculosis, a disease that affects ruminants worldwide. Despite global interest in the control of this disease, gaps exist in our knowledge of fecal shedding patterns and concurrent serological patterns. This longitudinal study in dairy cattle herds with high MAP seroprevalence in France aimed at accurately describing fecal shedding patterns over 1 year; relating those shedding patterns to individual animal characteristics (age, breed, parity); and exploring the association between fecal shedding patterns and serological patterns. To describe temporal fecal shedding patterns and continuity of shedding, along with the standard quantitative PCR (qPCR) threshold cycle we used a cutoff value that related to low or nonculturable fecal shedding. We also defined a threshold cycle indicative of shedding in high quantities to describe infection progression patterns. Twenty-one herds completed the study, and 782 cows were tested 4 times each. We obtained 4 sets of paired fecal qPCR and serum ELISA results from 757 cows. Although we targeted highly likely infectious animals, we found a large diversity of shedding patterns, as well as high variability between herds in the proportion of animals showing a given pattern. The fecal qPCR results of almost 20% of the final study sample were positioned at least once in the range that indicated low or nonculturable fecal shedding (between the adjusted and the standard cutoff value). Although these animals would typically be classified as non-shedders, they could be important to infection dynamics on the farm. Animals that shed at least twice consecutively and animals that shed in high quantities rarely reverted to negativity. Repeated fecal qPCR can be used to detect temporal fecal shedding traits, and the decision to cull an animal could practically be based on temporal, semiquantitative results. Overall, we found a mismatch between fecal shedding and ELISA seropositivity (637 animals were ELISAnegative 4 times, but only 13% of those animals were qPCR-negative 4 times). We found that having more than 2 ELISA-positive samples was strongly related to persistent and continuous shedding. We suggest that although serological testing is much less sensitive than qPCR, it can also be used, particularly over the course of multiple testing events, to identify animals that are most likely to contribute to the contamination of the farm environment.

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Outcome of Three Commercial Serum ELISAs and Faecal Detection of Mycobacterium avium subsp. paratuberculosis in Consecutive Samples from a Cattle Herd with Low Prevalence of Paratuberculosis (Johne’s Disease)

Cited by 8 publications

References 23 publications

A Longitudinal Study to Characterize the Distribution Patterns ofMycobacterium aviumssp.paratuberculosisin Semen, Blood and Faeces of a Naturally Infected Bull by IS 900 Semi-Nested and Quantitative Real-Time PCR

A Longitudinal Study to Characterize the Distribution Patterns ofMycobacterium aviumssp.paratuberculosisin Semen, Blood and Faeces of a Naturally Infected Bull by IS 900 Semi-Nested and Quantitative Real-Time PCR

Distribution ofMycobacterium aviumsubsp.paratuberculosisin a Subclinical Naturally Infected German Fleckvieh Bull

Longitudinal study of Mycobacterium avium ssp. paratuberculosis fecal shedding patterns and concurrent serological patterns in naturally infected dairy cattle

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