Rapid and Sensitive Method To Identify Mycobacterium avium subsp. paratuberculosis in Cow's Milk by DNA Methylase Genotyping

Mundo, Silvia Leonor; Gilardoni, Liliana Rosa; Hoffman, Federico José; Lopez, Osvaldo J.

doi:10.1128/aem.02719-12

Cited by 9 publications

(10 citation statements)

References 55 publications

(56 reference statements)

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“…An infected goat sheds up to 10 8 MAP per gram of feces (Windsor, 2015). Also colostrum and milk are considered infectious; however, in contrast to cattle, colostrum apparently plays a minor role in goats (Leonor Mundo et al, 2013). In a recent Dutch survey, MAP could not be detected in any of 121 colostrum samples, whereas 95% contained MAP antibodies (Lievaart-Peterson, 2017).…”

Section: Prevention and Controlmentioning

confidence: 99%

Paratuberculose bij melk- en dwerggeiten: een onderschat probleem?

Tuerlinckx¹,

Vicca²,

Deprez³

et al. 2018

Vlaams Diergeneeskundig Tijdschrift

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Paratuberculosis is a chronic intestinal disease affecting goats and other ruminants worldwide. The objective of this review was to summarize current knowledge on the prevalence, diagnostic possibilities and possible prevention and control measures for paratuberculosis in dairy and pygmy goats in Europe. The herd level prevalence of Mycobacterium avium subspecies paratuberculosis (MAP) in dairy goat farms is very high, namely 71%, 63% and 86% in Germany, France and the Netherlands, respectively. The prevalence in pygmy goats is undocumented. Antibody ELISA, fecal culture and PCR testing are readily available for diagnosis. A highly suggestive finding for paratuberculosis is the presence of enlarged intestinal lymph nodes on abdominal ultrasonography. To control the disease a combination of ‘test and cull’ and environmental hygienic measures are recommended. In contrast to cattle, vaccination is available and might be an additional tool in the control program for goats. Vaccine antibodies are no longer detectable after one year, and therefore serological monitoring can be continued on vaccinating herds. The worrisome prevalence data from neighboring countries indicate the need for a prevalence study on dairy goat farms in Belgium.

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Section: Prevention and Controlmentioning

confidence: 99%

Paratuberculose bij melk- en dwerggeiten: een onderschat probleem?

Tuerlinckx¹,

Vicca²,

Deprez³

et al. 2018

Vlaams Diergeneeskundig Tijdschrift

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“…paratuberculosis strains sequenced. Mundo et al 2013 M. avium subsp. paratuberculosis was specifically concentrated from milk samples by immunomagnetic separation prior to performing PCR.…”

Section: Molicotti Et Al 2013mentioning

confidence: 99%

Crohn's disease and related inflammatory diseases: from many single hypotheses to one "superhypothesis"

Hruška

Pavlík

2014

Vet. Med. - Czech

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ABSTRACT:The aetiology of Crohn's disease and paratuberculosis are the subjects of intensive study and also frequently, of dispute. However, a number of other nosological entities have a similar history, namely type 1 diabetes, multiple sclerosis, sarcoidosis, asthma, psoriasis, spondylarthritis, Blau syndrom etc. The zoonotic risk of Mycobacterium avium subsp. paratuberculosis (MAP) has been discussed for more than one hundred years. "The problem remains open, further research is needed", is the sentence which seems to be obligatory in the conclusions of many scientific articles. A number of hypotheses have been suggested, all with a grain of truth in them. The infection hypothesis has many supporters and opponents, but it does not fit to all Crohn's disease cases. The contribution of the genetic factor has been admitted a long time ago and has been experimentally confirmed by recent excellent studies. An environmental factor is expected and has been often mentioned, but has yet to be discovered. Muramyl dipeptide, derived from peptidoglycans of the bacterial cell wall is one of the triggers, mentioned in connection with chronic inflammatory diseases. The immunomodulatory ability of this compound has been recognised for decades and is exploited in Freund's adjuvant. A critical amount of muramyl dipeptide can affect immunity during some bacterial infections but the long latent period between infection and onset of the clinical form of the disease could explain why a causative relationship between the primary infection and chronic inflammation is not considered. Different species of mycobacteria can be found in the environment, in water, dust, soil and aerosol. Although severe infections with mycobacteria have been described, these species are not thought to be typical zoonotic pathogens. Muramyl dipeptide derived from mycobacteria obviously plays a starring role as a bacterial trigger in the aetiology of many autoimmune and autoinflammatory diseases. Paratuberculosis in cattle and other ruminants is a source of enormous contamination of the environment but also of milk and meat by MAP. Muramyl dipeptide from mycobacteria, namely MAP, and Crohn's disease as a representative of diseases often called civilization threats, are important pieces of the gigantic puzzle. Mycobacteria in the environment and foodstuffs have to be acknowledged as a public health risk, which can never be completely eliminated. There is no reason to push the panic button, but we must learn how to live together with this microorganism, how the pool of immunomodulator sources can be diminished, and how the pathogenic relationship between triggers and target tissues can be disrupted. The dissemination of knowledge, the availability of rapid and inexpensive tools for identification of mycobacteria in different matrices, and the establishment of a maximal allowed limit for mycobacteria in milk and meat should contribute to food safety and consumer protection.Keywords: aerosolisation; asthma; autoimmune diseases; autoinflammatory diseases; bacteri...

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“…,; Mundo et al . ). Since 2012, a real‐time PCR system has been approved for the direct detection of MAP in feces samples in Germany (FLI ).…”

Section: Introductionmentioning

confidence: 97%

“…In animals that excrete the pathogen in large quantities, the detection is also possible by means of direct PCR from fecal samples. It is possible to use PCR systems based on IS900, which has, however, been reported to show false-positive results (M€ obius et al 2008a,2008bMundo et al 2013). Since 2012, a real-time PCR system has been approved for the direct detection of MAP in feces samples in Germany (FLI 2012).…”

Section: Introductionmentioning

confidence: 99%

Development and validation of a loop‐mediated isothermal amplification assay—a rapid and sensitive detection tool forMycobacterium aviumsubsp.paratuberculosisin small ruminants

et al. 2019

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Aims The aim of this study was to design an assay for the identification of Mycobacterium avium subsp. paratuberculosis (MAP) to be used in faeces and milk samples of small ruminants with a loop‐mediated isothermal amplification (LAMP) system, as a time‐saving and user‐friendly method in contrast to real‐time PCR. Methods and Results For the detection of MAP in milk and faeces of small ruminants, we developed a set of primers, specific for the target gene ISMap02. The analytical sensitivity of LAMP, when targeting ISMap02, showed a DNA detection limit of 10 fg μl−1. After performing spiking experiments with two MAP reference strains, DSM 44133 and ATCC 19698T, the limit of detection, using the LAMP protocol described herein were 3·8 MAP CFU per ml milk and 12·5 MAP CFU per gram faeces. All LAMP results during the establishment of the assay were compared to those of the real‐time PCR results. An internal amplification control was incorporated into the assay to exclude false‐negative results produced and had no significant negative impact on the analytical sensitivity. Validation of the assay was confirmed by testing field samples of faeces and revising the results with real‐time PCR. Conclusion Our study conducted the first MAP detection system with a LAMP targeting ISMap02. Due to the positive results we encourage the use of LAMP in combination with ISMap02, when detecting MAP in faeces samples, as an alternative to targeting other genes as f57 or IS900. Further research on MAP detection in different matrices like raw milk, tissue or sperm with this system is recommended. Significance and Impact of the Study This study provides new achievements in MAP diagnostic. Especially small ruminants do not show signs of diarrhoea until the terminal stage of the illness. The greatest task in fighting MAP is to rule out animals, which shed MAP with faeces and milk before showing symptoms of Johne's disease. Worldwide there is a need to eradicate animals, which are low MAP shedders to stop the illness spreading in animal holdings. MAP detection with LAMP is time saving, easy to use, does not need expensive equipment, as, for example, PCR kits and can be used without access to laboratories. The target gene ISMap02 was shown to be a specific insertion element for MAP and is a reliable aim in future MAP detection studies.

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Rapid and Sensitive Method To Identify Mycobacterium avium subsp. paratuberculosis in Cow's Milk by DNA Methylase Genotyping

Cited by 9 publications

References 55 publications

Paratuberculose bij melk- en dwerggeiten: een onderschat probleem?

Paratuberculose bij melk- en dwerggeiten: een onderschat probleem?

Crohn's disease and related inflammatory diseases: from many single hypotheses to one "superhypothesis"

Development and validation of a loop‐mediated isothermal amplification assay—a rapid and sensitive detection tool forMycobacterium aviumsubsp.paratuberculosisin small ruminants

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