PubMed Health⌕ Search

Biomedical subjects

S H Flint

Publications and source records attributed to S H Flint.

At least 19 recordsLinked to original sources
PubMed2004

Evaluation of the effect of cleaning regimes on biofilms of thermophilic bacilli on stainless steel.

AIMS: To determine the mechanism for both the removal and inactivation of 18-h biofilms of a thermophilic Bacillus species that optimally grows at 55 degrees C on stainless steel. METHODS AND RESULTS: The cleaning strategies tested were based on biofilm biochemistry and physiology, and focused on the chemistry of the cleaners, the duration and temperature of the cleaning process and a combination of various cleaners. The success of the cleaning regimes was determined based on the removal of cells and organic debris and the elimination of viable cells. The results confirmed that a caustic (75 degrees C for 30 min) and acid (75 degrees C for 30 min) wash, relied upon heavily in most food processing industries for cleaning-in-place systems, was successful in removing these biofilms. However, any changes in the concentrations of these cleaners or the temperature of cleaning drastically affected the overall outcome. Alternative cleaning agents based on enzymatic or nonenzymatic breakdown of cellular proteins or polysaccharides, surfactant action, use of oxidative attack and free radicals varied in degrees of their success. Combining proteolytic action with surfactants increased wetability and therefore enhanced the cleaning efficiency. CONCLUSIONS: Several procedures, including caustic/acid and enzyme based cleaners, will be satisfactory, provided that the correct process parameters are observed i.e. concentration, time, temperature and kinetic energy (flow). Confirmation of these results should be carried out in a pilot plant through several use/clean cycles. SIGNIFICANCE AND IMPACT OF THE STUDY: Confidence in standard and alternative cleaning procedures for food manufacturing plant to prevent contamination with thermophilic bacilli that threaten product quality.

Bacillus↗
PubMed2003

Physiology of biofilms of thermophilic bacilli-potential consequences for cleaning.

Thermophilic Bacillus species readily attached and grew on stainless steel surfaces, forming mature biofilms of >10(6.0) cells/cm2 in 6 h on a surface inoculated with the bacteria. Clean stainless steel exposed only to pasteurized skim milk at 55 degrees C developed a mature biofilm of >10(6.0) cells/cm2 within 18 h. When bacilli were inoculated onto the steel coupons, 18-h biofilms were 30 microm thick. Biofilm growth followed a repeatable pattern, with a reduction in the numbers of bacteria on the surface occurring after 30 h, followed by a recovery. This reduction in numbers was associated with the production of a substance that inhibited the growth of the bacteria. Variations in the environment, including pH and molarity, affected the viability of the cells. Chemicals that attack the polysaccharide matrix of the biofilm were particularly effective in killing and removing cells from the biofilm, demonstrating the importance of polysaccharides in the persistence of these biofilms. Treatment of either the biofilm or a clean stainless steel surface with lysozyme killed biofilm cells and prevented the attachment of any bacteria exposed to the surface. This suggests that lysozyme may have potential as an alternative control method for biofilms of these bacteria.

Animals↗
PubMed2002

Comparison between Streptococcus macedonicus and Streptococcus waius strains and reclassification of Streptococcus waius (Flint et at. 1999) as Streptococcus macedonicus (Tsakalidou et al. 1998).

Two species of dairy streptococci, Streptococcus waius and Streptococcus macedonicus, were originally characterized by 16S-23S intergenic spacer sequence analysis, random amplified polymorphic DNA fingerprinting, PFGE analysis and DNA-DNA reassociation experiments. All genetic data suggested that S. waius strains belong to the previously described species S. macedonicus. Likewise, the phenotypic characterization showed that strains of S. macedonicus and S. waius were highly related and easily differentiated from the closest phylogenetic neighbour, Streptococcus bovis, principally by their failure to produce a blackening reaction in medium containing aesculin. The utilization of maltose and cellobiose by S. macedonicus/S. waius strains allowed their differentiation from the most studied dairy species, Streptococcus thermophilus. On the basis of genetic and phenotypic data S. macedonicus and S. waius species should be considered synonyms and S. macedonicus has the priority.

Bacterial Typing Techniques↗
PubMed2001

Rapid detection of Bacillus stearothermophilus using impedance-splitting.

An impedance splitting method was used to detect Bacillus stearothermophilus in suspension and attached to stainless steel surfaces. The effects of bacterial metabolism on the impedance of the culture medium and the ionic layers of the measuring electrodes were recorded using the BacTrac 4000 microorganism growth analyser. Impedance changes were measured at 55 degrees C. Seven of the eight media produced changes in the electrode impedance (E-value) and all media produced negligible changes in the impedance of the culture medium (M-value). Good correlations were obtained between cell numbers and the E-value measured over 18 h (r > 0.9) for the two strains of B. stearothermophilus tested in trypticase soy broth. The E-value correlations were used to estimate the numbers of both vegetative and spore forms of B. stearothermophilus as either planktonic or adhered cells. For the detection of B. stearothermophilus using impedance, only methods where the E-value impedance is recorded, can be used.

Bacterial Adhesion↗
PubMed2001

Factors influencing attachment of thermophilic bacilli to stainless steel.

AIMS: This project aimed to investigate the mechanism of attachment of the vegetative cells and spores of thermophilic bacilli to stainless steel with a view to devising strategies to limit biofilm development and survival. METHODS AND RESULTS: Spores and vegetative cells of bacterial isolates were exposed to protein denaturing agents (sodium dodecyl sulphate (SDS) and trypsin) and polysaccharide removing agents (sodium metaperiodate, trichloroacetic acid (TCA) and lysozyme). Treatment with sodium metaperiodate, TCA and lysozyme increased the number of vegetative cells attaching in many of the strains studied, while SDS and trypsin decreased attachment. Spores attached to stainless steel in greater numbers than vegetative cells, and the various treatments had less effect on this attachment than for vegetative cells. Viability of the cells or spores was not an important factor in attachment, as cells and spores rendered non-viable also attached to stainless steel in similar numbers. Coating the stainless steel with skim milk proteins decreased the attachment of both vegetative cells and spores. There was no correlation between the degree of attachment and the amount of extracellular polysaccharide (EPS) produced by each strain, surface hydrophobicity or zeta potential of vegetative cells or spores, though spores were found to be more hydrophobic than vegetative cells. CONCLUSIONS: The results suggest that biofilm formation by these thermophilic bacilli is probably a multifactorial process, and that cell-surface proteins play a very important role in the initial process of attachment during the formation of biofilms by these bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: This information will provide direction for developing improved cleaning systems to control biofilms of thermophilic bacilli in dairy manufacturing plants.

Bacillus↗
PubMed2001

Functional grouping of thermophilic Bacillus strains using amplification profiles of the 16S-23S internal spacer region.

Molecular and biochemical assays were used to determine the identification of thermophilic bacilli isolated from New Zealand milk powder. One hundred and forty one isolates of thermophilic bacilli were classified into six species using biochemical profiles. Geobacillus stearothermophilus represented 56% of the isolates. All isolates were also analysed by randomly amplified polymorphic DNA (RAPD) analysis, with 45 types identified. Amplification of the 16S-23S rDNA internal spacer region produced two to eight amplification products per strain. The patterns from gel electrophoresis of the internal spacer region amplicons formed two major groupings suggesting the possibility of two distinct species. Partial sequences of 16S rDNA from representatives from each group were compared with sequences in GeneBank and were found to match the 16S rDNA sequences of B. flavothermus and G. thermoleovorans. Primers were designed for these species and used to screen an arbitrary selection of 59 of the dairy isolates. This enabled the identification of 28 isolates as B. flavothermus and 31 isolates as Geobacillus species and these appear to be the predominant isolates in the New Zealand milk powder samples examined. Comparison of the fragment pattern generated by amplification of the 16S-23S rDNA internal spacer region is a simple method to differentiate thermophilic Bacillus species associated with the dairy industry.

Animals↗
PubMed1999

Streptococcus waius sp. nov., a thermophilic Streptococcus from a biofilm.

Thermophilic streptococci were isolated from biofilms on stainless steel samples exposed to pasteurized skimmed milk and from dairy products from a dairy manufacturing plant. The phenotypic characters of these isolates were distinct from those of other thermophilic streptococci of dairy origin (Streptococcus thermophilus and Streptococcus bovis). Genotypic data [restriction endonuclease analysis, ribotyping, random amplified polymorphic DNA (RAPD) profiles, DNA-DNA hybridization and G + C contents] support the classification of these isolates as a new species. The sequence of the 16S rRNA was compared with that of 29 species of streptococci and shown to be significantly different. The sequence of the 16S-23S rRNA intergenic spacer region also differed from published sequences of closely related species. A fluorescent in situ hybridization probe prepared to a specific part of the 16S rRNA gene sequence was able to distinguish the unknown isolates from reference isolates of S. thermophilus and S. bovis. It is proposed that these thermophilic streptococcal isolates from a dairy environment be classified in the genus Streptococcus as a new species, Streptococcus waius (from waiu, the New Zealand Maori word for milk). The type strain is 3/1T (= NZRCC 20100T).

Animals↗
PubMed1997

Use of the Malthus conductance growth analyser to determine numbers of thermophilic streptococci on stainless steel.

The use of the Malthus conductance growth analyser for the detection of Streptococcus bovis attached to stainless steel surface was evaluated. A comparison between the results from acridine orange epifluorescence direct counts, swab recovery viable count and conductance estimates of attached cell concentrations, based on calibrations for planktonic cells, showed that the conductance results were up to 2 log10 greater than the epifluorescence results and the swab counts. The growth rates of planktonic and attached cells were similar over 16 h using the Malthus technique. This suggests that the Malthus technique detects more attached cells of Strep. bovis than epifluorescence microscopy or swab recovery.

Acridine Orange↗
PubMed1997

The influence of cell surface properties of thermophilic streptococci on attachment to stainless steel.

The quality of milk products is threatened by the formation of biofilms of thermophilic streptococci on the internal surfaces of plate heat exchangers used in milk processing. Although attachment to stainless steel surfaces is one of the first stages in the development of a biofilm, the mechanisms involved in attachment have not been reported. The cell surface properties of 12 strains of thermophilic streptococci were examined to determine their importance in attachment to stainless steel surfaces. Hydrophobicity, extracellular polysaccharide production and cell surface charge varied between the different strains but could not be related to numbers attaching. Treating the cells with sodium metaperiodate, lysozyme or trichloroacetic acid to disrupt cell surface polysaccharide had no effect on attachment. Treatment with trypsin or sodium dodecyl sulphate to remove cell surface proteins resulted in a 100-fold reduction in the number of bacteria attaching. This result suggests that the surface proteins of the thermophilic streptococci are important in their attachment to stainless steel.

Bacterial Adhesion↗
PubMed1996

The sub-typing of Listeria monocytogenes isolates from food, environments surrounding food manufacturing sites, and clinical samples in New Zealand using multilocus enzyme electrophoresis.

Listeria monocytogenes strains isolated from food (cheese, meat and fish), environments surrounding food manufacturing sites, and clinical samples from throughout New Zealand were analyzed by multilocus enzyme electrophoresis. One hundred and twenty four strains were divided into 56 electrophoretic types, containing primarily clinical isolates and environmental isolates.

Bacterial Typing Techniques↗
PubMed1996

A comparison between starch and polyacrylamide gels for the analysis of Listeria monocytogenes using multilocus enzyme electrophoresis.

Forty-seven Listeria monocytogenes isolates were analysed using multilocus enzyme electrophoresis in two laboratories. Both assayed for the same six enzymes, but one used a starch gel method and the other polyacrylamide gels. The starch gel method distinguished six electrophoretic types whereas the polyacrylamide gel method produced 17 different electrophoretic types. The polyacrylamide gel method was more discriminatory than the starch gel method.

Adenylate Kinase↗
PubMed1995

Evaluation of the TECRA Escherichia coli O157 visual immunoassay for tests on dairy products.

The TECRA visual immunoassay was compared with an enrichment plating procedure according to the United States Food and Drug Administration procedures, for the detection of Escherichia coli in dairy foods. Both methods were used to detect E. coli O157 which had been inoculated into cheeses, milk powders, caseins and whey products. A centrifugation step was needed to test whey products according to the TECRA method. The results from both test methods were in complete agreement.

Dairy Products↗
PubMed1986

Dual infections of red deer (Cervus elaphus) by Leptospira interrogans serovars copenhageni and hardjo.

On a property in the Nelson District, blood and urine samples were taken from red deer (Cervus elaphus) from which low (<1:100) antibody titres to serovar copenhageni and suspected leptospiral abortions had previously been reported. A total of 27 hinds were sampled. Microscopic agglutination test (MAT) titres in sera ranging from 1:32 to 1:128 were found in six animals. Thirteen leptospiral isolations were made from nine of the 27 urine samples. Four of these were typed as copenhageni and nine as hardjo. Two cultures were prepared from each urine sample and hardjo and copenhageni were both isolated from single urine samples from two animals. None of the 27 deer had serum MAT titres at 1:32 or above to copenhageni.

Journal Article↗