A DNA Probe for the Detection and Identification of Bacillus sporothermodurans Using the 16S-23S rDNA Spacer Region and Phylogenetic Analysis of some Field Isolates of Bacillus which form Highly Heat Resistant Spores

Silva, Shyama de; Petterson, Bertil; Muro, Marilena Aquino de; Priest, Fergus G.

doi:10.1016/s0723-2020(98)80049-0

Cited by 27 publications

(30 citation statements)

References 34 publications

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“…All data discussed above deal only with the presence of spores in general and do not indicate possible sources of potentially highly heat-resistant spores at the dairy farm level. Although the isolation of B. sporothermodurans spores from various feeds after a heat treatment at 100°C has occasionally been reported (9,37,47), far less was known on sources and/or numbers of potentially highly heat-resistant spores. The present study shows that significant numbers of spores can indeed be recovered after a 30-min 100°C heat treatment.…”

Section: Discussionmentioning

confidence: 99%

“…Occasionally, B. sporothermodurans spores were reported in feed concentrate, silage, soy, and pulp (9,37,47). However, to prevent spoilage of milk and dairy products by aerobic spore-forming bacteria, it is important to minimize the initial contamination.…”

mentioning

confidence: 99%

“…In this study, at the dairy farm level, several spore-forming species, whose presence in an industrial setting has been previously reported, were found. More precisely, B. sporothermodurans (15,30), P. lactis (36), Brevibacillus borstelensis (9), B. sphaericus, B. licheniformis, and Brevibacillus brevis (4) spores were previously isolated from UHT milk. A. thermoaerophilus was previously isolated from an Austrian sugar beet factory (26), and representatives of the B. subtilis group, B. thermoamylovorans, Brevibacillus agri, and Brevibacillus borstelensis were isolated in a gelatin production process, where severe heat treatments are also used (6,8).…”

mentioning

confidence: 99%

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Incidence and Diversity of Potentially Highly Heat-Resistant Spores Isolated at Dairy Farms

Scheldeman

Pil

Herman

et al. 2005

Appl Environ Microbiol

121

105

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The presence of highly heat-resistant spores of Bacillus sporothermodurans in ultrahigh-temperature or sterilized consumer milk has emerged as an important item in the dairy industry. Their presence is considered undesirable since they hamper the achievement of commercial sterility requirements. By using a selective 30-min heat treatment at 100°C, 17 Belgian dairy farms were screened to evaluate the presence, sources, and nature of potentially highly heat-resistant spores in raw milk. High numbers of these spores were detected in the filter cloth of the milking equipment and in green crop and fodder samples. About 700 strains were isolated after the selective heating, of which 635 could be screened by fatty acid methyl ester analysis. Representative strains were subjected to amplified ribosomal DNA restriction analysis, 16S rRNA gene sequencing, percent G؉C content, and DNA-DNA reassociations for further identification. The strain collection showed a remarkable diversity, with representatives of seven aerobic spore-forming genera. Bacillus licheniformis and Bacillus pallidus were the most predominant species overall. Twenty-three percent of the 603 spore-forming isolates proved to belong to 18 separate novel species. These findings suggest that the selective heating revealed a pool of unknown organisms with a higher heat-resistant character. This study showed that high spore counts can occur at the dairy farm and that feed and milking equipment can act as reservoirs or entry points for potentially highly heat-resistant spores into raw milk. Lowering this spore load by good hygienic measures could probably further reduce the contamination level of raw milk, in this way minimizing the aerobic spore-forming bacteria that could lead to spoilage of milk and dairy products. Assessment and characterization of this particular flora are of great importance to allow the dairy or food industry to adequately deal with newly arising microbiological problems.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Incidence and Diversity of Potentially Highly Heat-Resistant Spores Isolated at Dairy Farms

Scheldeman

Pil

Herman

et al. 2005

Appl Environ Microbiol

121

105

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“…Typical colonies of B. sporothermodurans (20) and also spores were identified by phase-contrast microscopy and retained. All of the strains are listed in Table 1, including B. sporothermodurans type strain MB 581 (ϭ DSMZ 10599) and a raw milk isolate of Bacillus oleronius, the closest phylogenetic neighbor of B. sporothermodurans (4,10,22).…”

Section: Methodsmentioning

confidence: 99%

“…All these techniques showed that the different B. sporothermodurans strains isolated so far from European UHT-treated and sterilized milk are phylogenetically very closely related, forming the so-called HRS clone, named after the initial description of this highly heat-resistant spore-forming organism (8). Recently, B. sporothermodurans strains have also been isolated from raw milk and from animal feed (4,22,24). The majority of these farm isolates reacted negatively in the HRS-PCR of Herman et al (13) but could be assigned to B. sporothermodurans by a polyphasic approach and/or a new 16S rDNAbased specific PCR identification test (22).…”

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confidence: 99%

Genetic Heterogeneity in Bacillus sporothermodurans as Demonstrated by Ribotyping and Repetitive Extragenic Palindromic-PCR Fingerprinting

Guillaume-Gentil

Scheldeman²,

Marugg

et al. 2002

Appl Environ Microbiol

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Thirty-eight strains of Bacillus sporothermodurans isolated from ultra-high-temperature (UHT)-treated milk or sterilized milk (UHT isolates) and from animal feed or raw milk (farm isolates) were characterized by automated ribotyping and by repetitive extragenic palindromic (REP)-PCR fingerprinting. By investigating the genetic relationships among isolates from these various sources, the relative importance of different contamination sources could be evaluated. The results of the separate clustering analyses of the PvuII and EcoRI ribopatterns and the REP-PCR patterns were largely consistent with each other and revealed the existence of two main clusters; there was one homogeneous group containing all (REP-PCR) or most (ribotyping) of the UHT isolates, and there was a second more diverse group comprising the farm isolates. A combined threedimensional analysis of all data showed that three German UHT isolates did not belong to the compact group containing the majority of the UHT isolates. These results demonstrate that B. sporothermodurans is more heterogeneous than previously assumed and that most of the UHT isolates form a genetically distinct subgroup and are capable of producing highly heat-resistant spores. The close genetic relationship of these UHT isolates suggests a clonal origin of a few predominant strains of B. sporothermodurans that can be found in UHT-treated or sterilized milk products.

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Bacilli Associated with Spoilage in Dairy Products and Other Food

Heyndrickx¹,

Scheldeman²

2002

Applications and Systematics of Bacillus and Relatives

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A DNA Probe for the Detection and Identification of Bacillus sporothermodurans Using the 16S-23S rDNA Spacer Region and Phylogenetic Analysis of some Field Isolates of Bacillus which form Highly Heat Resistant Spores

Cited by 27 publications

References 34 publications

Incidence and Diversity of Potentially Highly Heat-Resistant Spores Isolated at Dairy Farms

Incidence and Diversity of Potentially Highly Heat-Resistant Spores Isolated at Dairy Farms

Genetic Heterogeneity in Bacillus sporothermodurans as Demonstrated by Ribotyping and Repetitive Extragenic Palindromic-PCR Fingerprinting

Bacilli Associated with Spoilage in Dairy Products and Other Food

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