20 research outputs found
Pilot-scale crossflow-microfiltration and pasteurization to remove spores of Bacillus anthracis (Sterne) from milk
High-temperature, short-time pasteurization of milk
is ineffective against spore-forming bacteria such as
Bacillus anthracis (BA), but is lethal to its vegetative
cells. Crossflow microfiltration (MF) using ceramic
membranes with a pore size of 1.4 μm has been shown
to reject most microorganisms from skim milk; and,
in combination with pasteurization, has been shown to
extend its shelf life. The objectives of this study were
to evaluate MF for its efficiency in removing spores
of the attenuated Sterne strain of BA from milk; to
evaluate the combined efficiency of MF using a 0.8-μm
ceramic membrane, followed by pasteurization (72°C,
18.6 s); and to monitor any residual BA in the permeates
when stored at temperatures of 4, 10, and 25°C
for up to 28 d. In each trial, 95 L of raw skim milk
was inoculated with about 6.5 log10 BA spores/mL of
milk. It was then microfiltered in total recycle mode
at 50°C using ceramic membranes with pore sizes of
either 0.8 μm or 1.4 μm, at crossflow velocity of 6.2 m/s
and transmembrane pressure of 127.6 kPa, conditions
selected to exploit the selectivity of the membrane.
Microfiltration using the 0.8-μm membrane removed
5.91 ± 0.05 log10 BA spores/mL of milk and the 1.4-
μm membrane removed 4.50 ± 0.35 log10 BA spores/
mL of milk. The 0.8-μm membrane showed efficient
removal of the native microflora and both membranes
showed near complete transmission of the casein proteins.
Spore germination was evident in the permeates
obtained at 10, 30, and 120 min of MF time (0.8-μm
membrane) but when stored at 4 or 10°C, spore levels
were decreased to below detection levels (≤0.3 log10
spores/mL) by d 7 or 3 of storage, respectively. Permeates
stored at 25°C showed coagulation and were
not evaluated further. Pasteurization of the permeate
samples immediately after MF resulted in additional
spore germination that was related to the length of
MF time. Pasteurized permeates obtained at 10 min of
MF and stored at 4 or 10°C showed no growth of BA
by d 7 and 3, respectively. Pasteurization of permeates
obtained at 30 and 120 min of MF resulted in spore
germination of up to 2.42 log10 BA spores/mL. Spore
levels decreased over the length of the storage period
at 4 or 10°C for the samples obtained at 30 min of MF
but not for the samples obtained at 120 min of MF.
This study confirms that MF using a 0.8-μm membrane
before high-temperature, short-time pasteurization
may improve the safety and quality of the fluid milk
supply; however, the duration of MF should be limited
to prevent spore germination following pasteurization
Table_1_The quorum sensing peptide BlpC regulates the transcription of genes outside its associated gene cluster and impacts the growth of Streptococcus thermophilus.DOCX
Bacteriocin production in Streptococcus thermophilus is regulated by cell density-dependent signaling molecules, including BlpC, which regulates transcription from within the bacteriocin-like peptide (blp) gene cluster. In some strains, such as S. thermophilus ST106, this signaling system does not function properly, and BlpC must be supplied exogenously to induce bacteriocin production. In other strains, such as S. thermophilus B59671, bacteriocin (thermophilin 110 in strain B59671) production occurs naturally. Here, transcriptomic analyses were used to compare global gene expression within ST106 in the presence or absence of synthetic BlpC and within B59671 to determine if BlpC regulates the expression of genes outside the blp cluster. Real-time semi-quantitative PCR was used to find genes differentially expressed in the absence of chromosomal blpC in the B59671 background. Growth curve experiments and bacteriocin activity assays were performed with knockout mutants and BlpC supplementation to identify effects on growth and bacteriocin production. In addition to the genes involved in bacteriocin production, BlpC affected the expression of several transcription regulators outside the blp gene cluster, including a putative YtrA-subfamily transcriptional repressor. In strain B59671, BlpC not only regulated the expression of thermophilin 110 but also suppressed the production of another bacteriocin, thermophilin 13, and induced the same YtrA-subfamily transcriptional repressor identified in ST106. Additionally, it was shown that the broad-spectrum antimicrobial activity associated with strain B59671 was due to the production of thermophilin 110, while thermophilin 13 appears to be a redundant system for suppressing intraspecies growth. BlpC production or induction negatively affected the growth of strains B59671 and ST106, revealing selective pressure to not produce bacteriocins that may explain bacteriocin production phenotype differences between S. thermophilus strains. This study identifies additional genes regulated by BlpC and assists in defining conditions to optimize the production of bacteriocins for applications in agriculture or human and animal health.</p