Application of a Fluorescence Microscopy Technique for Detecting Viable Mycobacterium avium ssp. paratuberculosis Cells in Milk

被引：8

作者：

Akineden, Oe. ^{[1
]}

Weirich, S. ^{[1
]}

Abdulmawjood, A. ^{[2
]}

Failing, K. ^{[3
]}

Buelte, M. ^{[1
]}

机构：

[1] Univ Giessen, Inst Vet Food Sci, D-35392 Giessen, Germany

[2] Univ Vet Med, Inst Food Qual & Food Safety, D-30173 Hannover, Germany

[3] Univ Giessen, Unit Biomath & Data Proc, D-35392 Giessen, Germany

来源：

FOOD ANALYTICAL METHODS | 2015年 / 8卷 / 02期

关键词：

Mycobacterium avium ssp Paratuberculosis; Viable count; Auramine orange; Fluorescence staining; Real-time PCR; Rapid detection; TIME PCR ASSAY; SUBSP-PARATUBERCULOSIS; INACTIVATION; ENUMERATION; BACTERIA; DECONTAMINATION; VIABILITY; INFECTION; PRODUCTS;

D O I：

10.1007/s12161-014-9918-3

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

The aim of the present study was to develop a fast and reliable fluorescence staining technique in order to accurately determine the number of viable Mycobacterium avium ssp. paratuberculosis (MAP) cells in milk. Milk was artificially inoculated with known amounts of four different MAP strains. Different dilutions were tested by the combination of 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) and auramine orange (AO) staining. To validate this combined fluorescence staining technique, two additional methods were applied: culture and the FASTPlaqueTB (TM) assay. The detection limit of the combined CTC and AO staining was 10(2) colony-forming unit (CFU) mL(-1) for spiked ultra-high-temperature (UHT) milk and 10(3) CFU mL(-1)for spiked raw milk (probability of detection > 95 %). Combined CTC and AO staining provides the opportunity to determine the total cell count of acid fast bacteria as well as the number of respiratory active cells in milk within 8 h.

引用

页码：499 / 506

页数：8

共 35 条

[1]

Akineden O, 2011, J FOOD SAF FOOD QUAL, V62, P5

[2] UV light inactivation of Mycobacterium avium subsp paratuberculosis in milk as assessed by FASTPlaqueTB phage assay and culturev [J].

Altic, Leslie C. ;

Rowe, Michael T. ;

Grant, Irene R. .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2007, 73 (11) :3728-3733

[3] LIVE/DEAD® BacLight™:: application of a new rapid staining method for direct enumeration of viable and total bacteria in drinking water [J].

Boulos, L ;

Prévost, M ;

Barbeau, B ;

Coallier, J ;

Desjardins, R .

JOURNAL OF MICROBIOLOGICAL METHODS, 1999, 37 (01) :77-86

[4] Auramine orange stain with fluorescence microscopy is a rapid and sensitive technique for the detection of cervical lymphadenitis due to mycobacterial infection using fine needle aspiration cytology: a case series [J].

Cheng, AG ;

Chang, A ;

Farwell, DG ;

Agoff, SN .

OTOLARYNGOLOGY-HEAD AND NECK SURGERY, 2005, 133 (03) :381-385

[5]

Dixon W. J, 1993, BMDP STAT SOFTWARE M, V1

[6]

Dixon W. J, 1993, BMDP STAT SOFTWARE M, V2

[7]

Dundee L, 2001, LETT APPL MICROBIOL, V33, P173, DOI 10.1046/j.1472-765X.2001.00979.x

[8] Contamination of food products with Mycobacterium avium paratuberculosis: a systematic review [J].

Eltholth, M. M. ;

Marsh, V. R. ;

Van Winden, S. ;

Guitian, F. J. .

JOURNAL OF APPLIED MICROBIOLOGY, 2009, 107 (04) :1061-1071

[9] Rapid Assessment of the Viability of Mycobacterium avium subsp paratuberculosis Cells after Heat Treatment, Using an Optimized Phage Amplification Assay [J].

Foddai, Antonio ;

Elliott, Christopher T. ;

Grant, Irene R. .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2010, 76 (06) :1777-1782

[10] Optimization of a Phage Amplification Assay To Permit Accurate Enumeration of Viable Mycobacterium avium subsp paratuberculosis Cells [J].

Foddai, Antonio ;

Elliott, Christopher T. ;

Grant, Irene R. .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2009, 75 (12) :3896-3902

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