A direct PCR detection method for Clostridium tyrobutyricum spores in up to 100 milliliters of raw milk

Herman, Lieve; Block, John H.; Waes, G.

doi:10.1128/aem.61.12.4141-4146.1995

Cited by 48 publications

(16 citation statements)

References 14 publications

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“…The specificity of Ct1F and Ct1R primer sets used in this study to C. tyrobutyricum is confirmed by Herman et al(1995) using 13 species of Clostridium. However, recently Kobayashi et al (2017) proposed a novel species C. pabulibutyricum which is closely related to C. tyrobutyricum.…”

Section: Specific Identification Of C Tyrobutyricum By Pcr Analysismentioning

confidence: 57%

“…The main cause of late blowing is C. tyrobutyricum, and its detection rate is particularly important in cheese production (Klijn, Nieuwenhof, Hoolwerf, van der Waals, & Weerkamp, 1995;Sheehan, 2011). C. tyrobutyricum was identified by species-specific PCR by Herman, Block, and Waes(1995). The cells were harvested and resuspended in sterile water and centrifuged at 9,500× g for 1 min, then DNA was extracted by InstaGene matrix (Bio-Rad, Hercules, CA), then used as a template DNA.…”

Section: Specific Identification Of C Tyrobutyricum By Pcr Analysismentioning

confidence: 99%

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One year investigation of the prevalence and diversity of clostridial spores in raw milk from the Tokachi area of Hokkaido

Komori

Ohkubo

Katano

et al. 2018

Animal Science Journal

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We investigated the seasonal prevalence and diversity of clostridial spores in raw milk from the Tokachi area of Hokkaido. Samples of raw milk were collected quarterly from May 2013 through February 2014. The mean clostridial spore count for the raw milk from 336 milk tankers was 27.6 CFU/100 ml. The clostridial species isolated most frequently from raw milk samples was Clostridium tyrobutyricum. The dominant species was C. tyrobutyricum regardless of the season. The percentage of samples with low spore counts (<10 CFU/100 ml) was highest (60.9%) during winter (February) and lowest (34.5%) in autumn (November). In comparison, the percentage of samples with high spore counts (>100 CFU/100 ml) was highest (5.7%) in autumn (November) and lowest (1.1%) during spring (May).

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Section: Specific Identification Of C Tyrobutyricum By Pcr Analysismentioning

confidence: 57%

Section: Specific Identification Of C Tyrobutyricum By Pcr Analysismentioning

confidence: 99%

One year investigation of the prevalence and diversity of clostridial spores in raw milk from the Tokachi area of Hokkaido

Komori

Ohkubo

Katano

et al. 2018

Animal Science Journal

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“…Aside from the MPN method, only a few alternatives are available. A highly specific PCR method detects low numbers of C. tyrobutyn'curn; however, quantification based on that technology is not possible (Herman et al 1995;Kiljn ef al. 1995).…”

Section: Introductionmentioning

confidence: 99%

Rapid Enumeration of Clostridial Spores in Raw Milk Samples Using an Impedimetric Method

Fontana¹,

Busiello²,

Bisotti³

et al. 2002

Rapid Methods Automation Mic

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Late spoilage of cheese is due to gas formation during lactic acid fermentation by spore‐forming, gram‐positive, anaerobic clostridia of the species Ciostridium tyrobutyricum, Clostridium butyricum and Clostridium sporogenes. Since small numbers of such clostridial spores readily cause considerable losses in cheese production, spore numbers of fewer than 100 spores/liter must be determined reliably. Until recently, the only reliable method available was the time‐consuming (7 days) and cumbersome Most Probable Number Method (MPN). The objective of this study was to examine the feasibility of using impedance technology as an alternative method for the enumeration of clostridial spores. Three to fifteen replicates of 7.5–12.0 mL samples were tested using an impedimetric method with and without the addition of Oxyrase to generate anaerobic conditions within the impedance measurement cells. Results were obtained in less than 48 h. Data derived from the rapid impedance method were statistically comparable to those obtained using the reference method (MPN).

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“…One consistent factor in these previous studies has been the use of an anaerobic environment for growth of C. tyrobutyricum. Though recent research has focused on using polymerase chain reaction technology to identify this microorganism, it is still not commonly used for quality testing within cheese processing plants (Herman et al 1995;Klijn et al 1995). Ali and Fung (1991) reported that the Fung Double Tube (FDT) method, which utilizes a small tube inserted into a larger tube with selective agar, could be used to enumerate C. perfringens in ground beef.…”

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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A direct PCR detection method for Clostridium tyrobutyricum spores in up to 100 milliliters of raw milk

Cited by 48 publications

References 14 publications

One year investigation of the prevalence and diversity of clostridial spores in raw milk from the Tokachi area of Hokkaido

One year investigation of the prevalence and diversity of clostridial spores in raw milk from the Tokachi area of Hokkaido

Rapid Enumeration of Clostridial Spores in Raw Milk Samples Using an Impedimetric Method

APPLICATION OF A DOUBLE TUBE SYSTEM FOR ENUMERATION OF CLOSTRIDIUM TYROBUTYRICUM

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