Membrane Filtration of Milk for Counting Spores of Clostridium tyrobutyricum

Bourgeois, Claude; Parc, Odile Le; Abgrall, Brigitte; Cleret, J. J.

doi:10.3168/jds.s0022-0302(84)81605-7

Cited by 20 publications

(8 citation statements)

References 11 publications

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“…Nevertheless, filtration of milk has been reported in several studies for isolation of bacteria (Bourgeois et al, 1984;Christiansson et al, 1997). These studies focused on using a combination of detergents, enzymes, and other treatments to simplify the composition and structure of milk.…”

Section: Filtration With Maldi Analysismentioning

confidence: 99%

Analysis of Bacillus anthracis Spores in Milk Using Mass Spectrometry

Whiteaker

Karns²,

Fenselau

et al. 2004

Foodborne Pathogens and Disease

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New approaches for identifying biological threat agents in raw milk using spectroscopy were tested using Bacillus anthracis (BA) Sterne strain spores seeded into unpasteurized bulk tank milk. Direct filtration onto Tyvek membranes provided the optimal filtration approach from raw milk, but detection limits were not ideal. When beads coated with anti-BA antibodies were mixed with spores in raw milk, the beads were capable of concentrating the spores that could be later detected and characterized by MALDI spectroscopy based on presence of previously characterized small acid-soluble proteins (SASP's). This approach could provide a very rapid assessment of whether milk or milk products have been purposefully contaminated with BA spores. This work was fundamentally a proof-of-concept study demonstrating feasibility of the approach in milk. Other parameters could be changed to potentially lower detection limits, and additional studies are currently underway to improve the approach.

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Section: Filtration With Maldi Analysismentioning

confidence: 99%

Analysis of Bacillus anthracis Spores in Milk Using Mass Spectrometry

Whiteaker

Karns²,

Fenselau

et al. 2004

Foodborne Pathogens and Disease

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“…Naturally contaminated milk from the Institut Technique du Gruyere (ITG Rennes, France) was also used. These samples were firstly submitted to a thermal treatment followed by filtration according to the procedure described by Bourgeois et al (1984). The thermal treatment, associated with the action of Triton (Sigma, France) and trypsin (Difco Laboratories, France), leads to a better milk filtration by breaking up greasy globules and vegetative cells (bacterial and somatic).…”

Section: Membrane Treatment and Milk Filtrationmentioning

confidence: 99%

“…tyrobutyrirum. Another method which has been developed (Bourgeois et al 1984) involves high pressure membrane filtration of milk followed by growing the bacteria on the membrane. This technique gives results within 48 h but is not selective enough and cannot distinguish Cl.…”

Section: Introductionmentioning

confidence: 99%

Monoclonal antibody detection of Clostridium microcolonies directly on membrane used for milk filtration

et al. 1997

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1997. The normal procedure for bacterial colony detection requires a nitrocellulose transfer step after membrane filtration and culture to prevent the development of a high background during the immunodetection. In this paper, we describe a modification of the basic protocol that omits the transfer step and reduces the risk of background. Previous observations indicated that interactions between milk components (principally cream) and membrane are responsible for the high non-specific staining observed. Experiments were performed to remove lipid components or to block the membrane binding sites before milk filtration. Samples of milks of different origin (collected at different times of the year) and different membranes were tested. The results obtained showed that removing lipids did not significantly improve the test but, on the contrary, led to an antigen diffusion. Incubation of the membrane in 0.1% (w/v) of Tween 20 in phosphate-buffered saline before milk filtration prevented non-specific binding, and allowed performance of the detection without any noticeable background.

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“…Methods to detect C. tyrobutyricum are lengthy and not specific. Among the variables reported in literature are agars, incubation temperatures, and incubation times (Bester and Claasans 1970;Bester and Lombard 1990;Bourgeois et al 1984;Kleter et al 1984;Kruyswick 1996;and Thualt et al 1991). One consistent factor in these previous studies has been the use of an anaerobic environment for growth of C. tyrobutyricum.…”

Section: Introductionmentioning

confidence: 99%

APPLICATION OF A DOUBLE TUBE SYSTEM FOR ENUMERATION OF CLOSTRIDIUM TYROBUTYRICUM

Schmidt

Thakur

Jiang

et al. 2000

Rapid Methods Automation Mic

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Clostridium tyrobutyricum, a spore‐forming, gram‐positive, anaerobic bacterium, is considered to be the main organisms responsible for the late spoilage of cheese by gas formation. Most methods for detecting C. tyrobutyricum are based on spore germination and vegetative growth and take 4–7 days plus an identification step for confirmation. The purpose of this study was to develop a faster detection method using a Double Tube System. Because no selective medium is available for detection of C. tyrobutyricum, three media (Reinforced Clostridial, AC, and Tomato Juice) were compared using two strains of C. tyrobutyricum and one strain of C. sporogenes. Each 4 day‐old test strain was inoculated on duplicated plates of each agar that were then placed in anaerobic jars or in the double‐tube systems for 2–4 days at 30 or 37C. All three agars consistently supported growth of the test strains. Counts did not differ with incubation at 30 or 37C and were comparable using the conventional anaerobic jar or a Double Tube System. However, in the Double Tube System, colonies could be counted accurately at least 6 h earlier than on the plates in anaerobic jars.

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Membrane Filtration of Milk for Counting Spores of Clostridium tyrobutyricum

Cited by 20 publications

References 11 publications

Analysis of Bacillus anthracis Spores in Milk Using Mass Spectrometry

Analysis of Bacillus anthracis Spores in Milk Using Mass Spectrometry

Monoclonal antibody detection of Clostridium microcolonies directly on membrane used for milk filtration

APPLICATION OF A DOUBLE TUBE SYSTEM FOR ENUMERATION OF CLOSTRIDIUM TYROBUTYRICUM

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