Detection of mycobacterium avium subsp. paratuberculosis in cheeses from small ruminants in Tuscany

Galiero, Alessia; Fratini, Filippo; Ματαράγκα, Αντωνία; Turchi, Barbara; Nuvoloni, Roberta; Ikonomopoulos, John; Cerri, Domenico

doi:10.1016/j.ijfoodmicro.2015.10.029

Cited by 27 publications

(16 citation statements)

References 24 publications

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“…The ability of MAP to exist in a variety of phenotypes, some with a high resistance to killing (Grant and Rowe, 2004), has increased the importance of providing accurate quantitative estimates of viable counts when testing for the presence of this pathogen in food (Botsaris et al, 2016; Galiero et al, 2016; Ricchi et al, 2016), animal and human samples (Timms et al, 2016). MAP is widely accepted as a difficult organism to culture reproducibly and accurately, particularly at low loads (Hines et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Improved Culture Medium (TiKa) for Mycobacterium avium Subspecies Paratuberculosis (MAP) Matches qPCR Sensitivity and Reveals Significant Proportions of Non-viable MAP in Lymphoid Tissue of Vaccinated MAP Challenged Animals

et al. 2017

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The quantitative detection of viable pathogen load is an important tool in determining the degree of infection in animals and contamination of foodstuffs. Current conventional culture methods are limited in their ability to determine these levels in Mycobacterium avium subspecies paratuberculosis (MAP) due to slow growth, clumping and low recoverability issues. The principle goal of this study was to evaluate a novel culturing process (TiKa) with unique ability to stimulate MAP growth from low sample loads and dilutions. We demonstrate it was able to stimulate a mean 29-fold increase in recoverability and an improved sensitivity of up to three logs when compared with conventional culture. Using TiKa culture, MAP clumping was minimal and produced visible colonies in half the time required by standard culture methods. Parallel quantitative evaluation of the TiKa culture approach and qPCR on MAP loads in tissue and gut mucosal samples from a MAP vaccine-challenge study, showed good correlations between colony counts (cfu) and qPCR derived genome equivalents (Geq) over a large range of loads with a 30% greater sensitivity for TiKa culture approach at low loads (two logs). Furthermore, the relative fold changes in Geq and cfu from the TiKa culture approach suggests that non-mucosal tissue loads from MAP infected animals contained a reduced proportion of non-viable MAP (mean 19-fold) which was reduced significantly further (mean 190-fold) in vaccinated “reactor” calves. This study shows TiKa culture equates well with qPCR and provides important evidence that accuracy in estimating viable MAP load using DNA tests alone may vary significantly between samples of mucosal and lymphatic origin.

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Section: Introductionmentioning

confidence: 99%

Improved Culture Medium (TiKa) for Mycobacterium avium Subspecies Paratuberculosis (MAP) Matches qPCR Sensitivity and Reveals Significant Proportions of Non-viable MAP in Lymphoid Tissue of Vaccinated MAP Challenged Animals

et al. 2017

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“…The discrepant results could be attributed to the fact that AGID is less sensitive than ELISA (OIE, 2104). On the contrary, paratuberculosis has been already assessed in this region not only in goats (Cerri et al, 2002) and in sheep (Galiero et al, 2015), but Map has been identified in small ruminant dairy products (Galiero et al, 2016). Based on the evidences recorded by previous studies concerning the presence of Map in food chain (Savi et al, 2015;Ricchi et al, 2016) and its potential role in the development of human diseases (Sechi and Dow, 2015;Koriem, 2016), it is advisable to pasteurize bovine milk before sale or cheese making.…”

Section: Discussionmentioning

confidence: 99%

“…With reference to this, in Italy, as suggested by previous studies, not only bovine raw milk, but also pasteurized milk is a significant source of Map exposure for consumers (Giacometti et al, 2012;Serraino et al, 2014a;Serraino et al, 2014b). Unfortunately, while the disease has been highlighted in the Tuscan caprine and ovine population (Galiero et al, 2015;Galiero et al, 2016), no official data have been published on the occurrence of paratuberculosis in cattle in Tuscany.…”

Section: Introductionmentioning

confidence: 97%

Serological survey of paratuberculosis in dairy cattle in Garfagnana district (Tuscany)

Galiero¹,

Fratini²,

Bravi³

et al. 2018

J Hellenic Vet Med Soc

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Mycobacterium avium subsp. paratuberculosis (Map) is the agent of a chronic, progressive granulomatous enteritis in ruminants known as Johne’s disease or paratuberculosis. Nowadays, the interest regarding this pathogen is increasing not only because Map causes economic losses, but it has also been suggested as a potential risk factor for the development of human diseases such as Crohn disease, autoimmune diseases like type-1 diabetes, sarcoidosis, multiple sclerosis and Hashimoto’s thyroiditis. The aim of our study was to determine the presence of paratuberculosis in Garfagnana (Tuscany, Italy) where raw milk is not submitted to pasteurization, but it is directly sold to consumers or it is destined for cheese making without the employment of heat treatment. The survey was conducted in Garfagnana district where there are 17 herds which produce and market bovine milk for direct human consumption. Serum samples (n=162) were obtained form 16 herds and were analyzed performing ELISA ID screen® Paratuberculosis Indirect screening test (ID.VET, Montpellier, France) and positive samples were tested with ELISA ID screen® Paratuberculosis Indirect confirmation test (ID.VET, Montpellier, France), according to the instructions provided by the manufacturer. The analysis performed by ELISA revealed that the true seroprevalence was 29.1% at the herd level and 4.6% at the animal level. In our opinion, although ELISA is a very useful tool to screen herds for paratuberculosis, more studies should be carried out in our territory combining serological data with cultural and PCR analysis.

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“…Pillai ve Jayarao (37) Süt IS900 F(5 ' -GATGGCCGAAGGAGATTG-3 ' ) R(5 ' -CACAACCACCTCCGTAACC-3 ' ) Slana ve ark (21) Süt IS900 F(5 ' -ACCCGCTGCGAGAGCAATCGCTGC-3 ' ) R(5 ' -ACGTCGGCGTGGTCGTCTGCTGGG-3 ' ) Gilardoni ve ark (38) Peynir IS900 F(5 ' -AATGACGGTTACGGAGGTGGT-3 ' ) R(5 ' -GCAGTAATGGTC GGCCTTACC-3 ' ) Galiero ve ark (39) Süt bazlı bebek maması IS900 F(5 ' -GAAGGGTGTTCGG GGCCGTCGCTTAGG-3 ' ) R(5 ' -GGCGTTGAGGTCGATCGCCCACGT GAC-3 ' ) Botsaris ve ark (40) Süt f57 F(5 ' -GCCCATTTCATCGATACCC-3 ' ) R(5 ' -GTACCGAATGTTGTTGTCAC-3 ' ) Slana ve ark (21) Süt f57 F(5 ' -GGTCGCGTCATTCAGAATC-3 ' ) R(5 ' -TCTCAGACAGTGGCAGGTG-3 ' ) Botsaris ve ark (41) Süt, süt tozu, peynir f57 F(5 ' -CCG CGA TCC CAA AAG TTG-3 ' ) R(5 ' -CTC GTA GCT GCC GAT TCA TG-3 ' ) Donaghy ve ark (33) 7. Grant IR., Rowe MT., Dundee L., Hitchings E.,…”

Section: Crohn Hastalığı Ve Mycobacterium Avium Subsp Paratuberculosismentioning

confidence: 99%

Mycobacterium avium subsp. paratuberculosis’in Süt ve Süt Ürünlerinde Tespit Yöntemleri

Telli¹,

Güner²

2018

Atatürk Üniversitesi Veteriner Bilimleri Dergisi

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Öz: Mycobacterium avium kompleksi üyelerinden olan Mycobacterium avium subsp. paratuberculosis (MAP) çok çeşitli hayvan türlerinde Johne hastalığının (paratüberkülozis) etiyolojik etkenidir. Paratüberkülozun klinik belirtilerini gösteren ve asemptomatik taşıyıcı hayvanlar etkeni süt, dışkı ve spermleriyle yaymaktadırlar. Süt direkt olarak etkeni taşıyabildiği gibi özellikle dışkı kaynaklı indirekt bulaşmalar da gerçekleşebilmektedir. Sütün yanı sıra peynir, yoğurt, krema, tereyağı, dondurma vb. süt ürünleri de etkeni taşıyabilmektedir. Süt sığırlarındaki Johne hastalığı ile insanlarda görülen Crohn hastalığı insidenslerinde dünya genelinde artış görülmektedir. Yüksek klinik ve patolojik benzerlikler bulunan bu hastalıklar arasındaki olası ilişkide gıdaların rolünün yorumlanmasında ve güvenilir sonuçlar alınmasında MAP'ın süt ve süt ürünlerinde tespit yöntemleri önem arz etmektedir. Bu amaçla süt ve süt ürünlerinde Ziehl-Neelsen boyama, floresan mikroskop, katı faz sitometri ve biyolüminesans gibi direkt tanı ve kültür yöntemleri ile MAP etkenleri araştırılabilmektedir. Bu yöntemlerin yanı sıra radyoimmün testler (RIA), enzim bağlı immün testler (ELISA), agar jel immünodifüzyon testi (AGID) ve komplement fiksasyon testi (CF test) gibi serolojik teknikler ve polimeraz zincir reaksiyonu (PCR), nükleik asit hibridizasyonu, immunomagnetic separation PCR, Nested PCR, Real-Time PCR gibi moleküler teknikler veya bu tekniklerin kombinasyonlarıyla hızlı tanı yöntemleri geliştirilmiştir.

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Detection of mycobacterium avium subsp. paratuberculosis in cheeses from small ruminants in Tuscany

Cited by 27 publications

References 24 publications

Improved Culture Medium (TiKa) for Mycobacterium avium Subspecies Paratuberculosis (MAP) Matches qPCR Sensitivity and Reveals Significant Proportions of Non-viable MAP in Lymphoid Tissue of Vaccinated MAP Challenged Animals

Improved Culture Medium (TiKa) for Mycobacterium avium Subspecies Paratuberculosis (MAP) Matches qPCR Sensitivity and Reveals Significant Proportions of Non-viable MAP in Lymphoid Tissue of Vaccinated MAP Challenged Animals

Serological survey of paratuberculosis in dairy cattle in Garfagnana district (Tuscany)

Mycobacterium avium subsp. paratuberculosis’in Süt ve Süt Ürünlerinde Tespit Yöntemleri

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